Journal of Toxicology and Environmental Health-Part B-Critical Reviews最新文献

{"title":"Atrazine's effects on mammalian physiology.","authors":"Anna G Holliman, Laci Mackay, Vinicia C Biancardi, Ya-Xiong Tao, Chad D Foradori","doi":"10.1080/10937404.2025.2468212","DOIUrl":"https://doi.org/10.1080/10937404.2025.2468212","url":null,"abstract":"<p><p>Atrazine is a chlorotriazine herbicide that is one of the most widely used herbicides in the USA and the world. For over 60 years atrazine has been used on major crops including corn, sorghum, and sugarcane to control broadleaf and grassy weed emergence and growth. Atrazine has exerted a major economic and environmental impact over that time, resulting in reduced production costs and increased conservation tillage practices. However, widespread use and a long half-life led to a high prevalence of atrazine in the environment. Indeed, atrazine is the most frequent herbicide contaminant detected in water sources in the USA. Due to its almost ubiquitous presence and questions regarding its safety, atrazine has been well-studied. First reported to affect reproduction with potential disruptive effects which were later linked to the immune system, cancer, stress response, neurological disorders, and cardiovascular ailments in experimental models. Atrazine impact on multiple interwoven systems broadens the significance of atrazine exposure. The endeavor to uncover the mechanisms underlying atrazine-induced dysfunction in mammals is ongoing, with new genetic and pharmacological targets being reported. This review aims to summarize the prominent effects of atrazine on mammalian physiology, primarily focusing on empirical studies conducted in lab animal models and establish correlations with epidemiological human studies when relevant. In addition, current common patterns of toxicity and potential underlying mechanisms of atrazine action will be examined.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"1-40"},"PeriodicalIF":6.4,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143525187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Uncovering layer by layer the risk of nanoplastics to the environment and human health.","authors":"Joana Cepeda da Silva Antunes, Paula Sobral, Vasco Branco, Marta Martins","doi":"10.1080/10937404.2024.2424156","DOIUrl":"10.1080/10937404.2024.2424156","url":null,"abstract":"<p><p>Nanoplastics (NPs), defined as plastic particles with dimensions less than 100 nm, have emerged as a persistent environmental contaminant with potential risk to both environment and human health. Nanoplastics might translocate across biological barriers and accumulate in vital organs, leading to inflammatory responses, oxidative stress, and genotoxicity, already reported in several organisms. Disruptions to cellular functions, hormonal balance, and immune responses were also linked to NPs exposure in <i>in vitro</i> assays. Further, NPs have been found to adsorb other pollutants, such as persistent organic pollutants (POPs), and leach additives potentially amplifying their advere impacts, increasing the threat to organisms greater than NPs alone. However, NPs toxic effects remain largely unexplored, requiring further research to elucidate potential risks to human health, especially their accumulation, degradation, migration, interactions with the biological systems and long-term consequences of chronic exposure to these compounds. This review provides an overview of the current state-of-art regarding NPs interactions with environmental pollutants and with biological mechanisms and toxicity within cells.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"63-121"},"PeriodicalIF":6.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142820022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Review of non-invasive biomarkers as a tool for exposure characterization in human health risk assessments.","authors":"N Shilnikova, F Momoli, N Karyakina, D Krewski","doi":"10.1080/10937404.2024.2428206","DOIUrl":"10.1080/10937404.2024.2428206","url":null,"abstract":"<p><p>Blood and urine are historically the most frequent matrices used for measuring chemical levels in human biomonitoring studies. As biomonitoring programs are refreshed, consideration of specific priority substances and specific population targets provide opportunities for inclusion of alternative non- or minimally invasive matrices. This review describes methods used in health risk assessment to characterize exposure and risk based upon biomarkers from noninvasive matrices other than urine or blood, including human milk, hair, fingernails, toenails, exhaled breath, deciduous teeth, sweat, semen, meconium, and feces. Illustrative examples of these methods relevant to chemical management are provided. This review suggests that, although these alternative noninvasive biomarkers are not frequently used in human health risk assessment at present, these biomarkers may prove useful in (1) characterizing exposure and health risk in vulnerable populations, (2) cumulative risk assessments, and (3) community-based risk assessments, depending upon the substance of concern. To incorporate alternative noninvasive biomarkers into human health risk assessments with confidence, more research is needed to improve our knowledge of the relationships between external dose, internal dose, and biologic consequent effects in matrices other than blood and urine.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"122-150"},"PeriodicalIF":6.4,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142741121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ecotoxicological bioassays with terrestrial plants: a holistic view of standards, guidelines, and protocols.","authors":"Leonardo Mendes da Silva, Larissa Fonseca Andrade-Vieira","doi":"10.1080/10937404.2024.2440876","DOIUrl":"https://doi.org/10.1080/10937404.2024.2440876","url":null,"abstract":"<p><p>Terrestrial and aquatic ecosystems face various chemicals that might induce acute and/or long-term harm. To assess these impacts, ecotoxicological bioassays are essential. However, bioassays using animals, particularly mammals, are costly, time-consuming, and raise ethical concerns. In this context, terrestrial plants emerge as a viable alternative to conventional assays. Thus, the aim of this review was to address the history and evolution of plant bioassays, highlighting the main regulations, guidelines, and protocols governing the use of terrestrial plants in ecotoxicological tests. Initially, plant bioassays were employed to assess the cytogenotoxic effects of chemicals, gaining prominence with the GENE-TOX program in the 80s. Subsequently, plants were used in allelopathy bioassays and in studies aimed to examine the ecotoxicity of pesticides in soil. Currently, ecotoxicological bioassays with plants are regulated by specific standards, such as ASTM E1963-22, EPA 600/3-88/029, EPS 1/RM/45, ISO 11269-1, ISO 11269-2, ISO 17126, ISO 18763, ISO 29200, ISO 22030, OECD-208, OECD-227, OCSPP 850.4100, OCSPP 850.4230, OCSPP 850.4800 and OPPTS 850.4200. The existing protocols standardize bioassays in greenhouse and lab environments, and the duration of the tests varies from hours to months. The main ecotoxicological parameters to be analyzed after exposure include germination percentage, survival rate, root length, aerial part length, fresh mass of exposed plants, and phytotoxicity symptoms. In addition, the absorption rate of substances and genotoxic and mutagenic effects might also be assessed. Therefore, data in this review demonstrate that terrestrial plants represent an important tool in the analysis of environmental risks associated with chemicals and might serve as crucial allies in modern ecotoxicology.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"1-39"},"PeriodicalIF":6.4,"publicationDate":"2025-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142933298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Incorporating new approach methods (NAMs) data in dose-response assessments: The future is now!","authors":"En-Hsuan Lu, Ivan Rusyn, Weihsueh A Chiu","doi":"10.1080/10937404.2024.2412571","DOIUrl":"10.1080/10937404.2024.2412571","url":null,"abstract":"<p><p>Regulatory dose-response assessments traditionally rely on <i>in vivo</i> data and default assumptions. New Approach Methods (NAMs) present considerable opportunities to both augment traditional dose-response assessments and accelerate the evaluation of new/data-poor chemicals. This review aimed to determine the potential utilization of NAMs through a unified conceptual framework that compartmentalizes derivation of toxicity values into five sequential Key Dose-response Modules (KDMs): (1) point-of-departure (POD) determination, (2) test system-to-human (e.g. inter-species) toxicokinetics and (3) toxicodynamics, (4) human population (intra-species) variability in toxicodynamics, and (5) toxicokinetics. After using several \"traditional\" dose-response assessments to illustrate this framework, a review is presented where existing NAMs, including <i>in silico</i>, <i>in vitro</i>, and <i>in vivo</i> approaches, might be applied across KDMs. Further, the false dichotomy between \"traditional\" and NAMs-derived data sources is broken down by organizing dose-response assessments into a matrix where each KDM has Tiers of increasing precision and confidence: Tier 0: Default/generic values, Tier 1: Computational predictions, Tier 2: Surrogate measurements, and Tier 3: Direct measurements. These findings demonstrated that although many publications promote the use of NAMs in KDMs (1) for POD determination and (5) for human population toxicokinetics, the proposed matrix of KDMs and Tiers reveals additional immediate opportunities for NAMs to be integrated across other KDMs. Further, critical needs were identified for developing NAMs to improve <i>in vitro</i> dosimetry and quantify test system and human population toxicodynamics. Overall, broadening the integration of NAMs across the steps of dose-response assessment promises to yield higher throughput, less animal-dependent, and more science-based toxicity values for protecting human health.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"28-62"},"PeriodicalIF":6.4,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11614695/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142401770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>In vitro</i> models to evaluate multidrug resistance in cancer cells: Biochemical and morphological techniques and pharmacological strategies.","authors":"Maria Fernanda Madrid, Eleicy Nathaly Mendoza, Ana Lizeth Padilla, Celia Choquenaira-Quispe, Celina de Jesus Guimarães, João Victor de Melo Pereira, Francisco Washington Araújo Barros-Nepomuceno, Ingredy Lopes Dos Santos, Claudia Pessoa, Manoel Odorico de Moraes Filho, Danilo Damasceno Rocha, Paulo Michel Pinheiro Ferreira","doi":"10.1080/10937404.2024.2407452","DOIUrl":"10.1080/10937404.2024.2407452","url":null,"abstract":"<p><p>The overexpression of ATP-binding cassette (ABC) transporters contributes to the failure of chemotherapies and symbolizes a great challenge in oncology, associated with the adaptation of tumor cells to anticancer drugs such that these transporters become less effective, a mechanism known as multidrug resistance (MDR). The aim of this review is to present the most widely used methodologies for induction and comprehension of <i>in vitro</i> models for detection of multidrug-resistant (MDR) modulators or inhibitors, including biochemical and morphological techniques for chemosensitivity studies. The overexpression of MDR proteins, predominantly, the subfamily glycoprotein-1 (P-gp or ABCB1) multidrug resistance, multidrug resistance-associated protein 1 (MRP1 or ABCCC1), multidrug resistance-associated protein 2 (MRP2 or ABCC2) and cancer resistance protein (ABCG2), in chemotherapy-exposed cancer lines have been established/investigated by several techniques. Amongst these techniques, the most used are (i) colorimetric/fluorescent indirect bioassays, (ii) rhodamine and efflux analysis, (iii) release of 3,30-diethyloxacarbocyanine iodide by fluorescence microscopy and flow cytometry to measure P-gp function and other ABC transporters, (iv) exclusion of calcein-acetoxymethylester, (v) ATPase assays to distinguish types of interaction with ABC transporters, (vi) morphology to detail phenotypic characteristics in transformed cells, (vii) molecular testing of resistance-related proteins (RT-qPCR) and (viii) 2D and 3D models, (ix) organoids, and (x) microfluidic technology. Then, <i>in vitro</i> models for detecting chemotherapy MDR cells to assess innovative therapies to modulate or inhibit tumor cell growth and overcome clinical resistance. It is noteworthy that different therapies including anti-miRNAs, antibody-drug conjugates (to natural products), and epigenetic modifications were also considered as promising alternatives, since currently no anti-MDR therapies are able to improve patient quality of life. Therefore, there is also urgency for new clinical markers of resistance to more reliably reflect <i>in vivo</i> effectiveness of novel antitumor drugs.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"1-27"},"PeriodicalIF":6.4,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142373390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neutrophils in toxicology: a forgotten field.","authors":"Pablo Scharf, Michael Aschner, Sandra Farsky","doi":"10.1080/10937404.2024.2431692","DOIUrl":"https://doi.org/10.1080/10937404.2024.2431692","url":null,"abstract":"<p><p>Neutrophils are the most abundant leukocytes in humans and essential for innate immune responses despite a short lifespan in the bloodstream. A complex and tightly regulated production of neutrophils is required to maintain host defense. This process involves intricate signaling between the bone marrow, blood, and tissue clearance. Deficiency or excessive neutrophil infiltration impairs host defenses. Historically, neutrophils were viewed as initial effectors in innate immune responses. Recent discoveries have expanded our understanding of neutrophil biology, identifying multiple activation states and subsets. These subsets may switch phenotypes based on the composition of the microenvironment and might exhibit reverse migratory behavior, moving from inflamed tissues back into the bloodstream. This versatility poses neutrophils as key players in (1) signaling for host defenses, (2) trained immunity, (3) tissue repair, and (4) cancer biology. Disturbances in neutrophil production, responsiveness, apoptosis, and cell removal significantly affect inflammatory diseases and cancer progression. Environmental factors may directly affect the immune system and trigger the onset of many diseases; however, the precise mechanisms underlying the impact of xenobiotics on neutrophil production and functions remain unclear. This review aimed to summarize the current knowledge on neutrophil ontogeny, plasticity, and roles in inflammation, tissue repair, and cancer, emphasizing their susceptibility to different sources of xenobiotic exposures.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"1-32"},"PeriodicalIF":6.4,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142689367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neuroendocrine contribution to sex-related variations in adverse air pollution health effects.","authors":"Devin I Alewel, Urmila P Kodavanti","doi":"10.1080/10937404.2024.2383637","DOIUrl":"10.1080/10937404.2024.2383637","url":null,"abstract":"<p><p>Air pollution exposure is ranked as a leading environmental risk factor for not only cardiopulmonary diseases but also for systemic health ailments including diabetes, reproductive abnormalities, and neuropsychiatric disorders, likely mediated by central neural stress mechanisms. Current experimental evidence links many air pollution health outcomes with activation of neuroendocrine sympathetic-adrenal-medullary and hypothalamic-pituitary-adrenal (HPA) stress axes associated with resultant increases in adrenal-derived hormone levels acting as circulating mediators of multi-organ stress reactions. Epidemiological and experimental investigations also demonstrated sex-specific responses to air pollutant inhalation, which may be attributed to hormonal interactions within the stress and reproductive axes. Sex hormones (androgens and estrogens) interact with neuroendocrine functions to influence hypothalamic responses, subsequently augmenting stress-mediated metabolic and immune changes. These neurohormonal interactions may contribute to innate sex-specific responses to inhaled irritants, inducing differing individual susceptibility. The aim of this review was to: (1) examine neuroendocrine co-regulation of the HPA axis by gonadal hormones, (2) provide experimental evidence demonstrating sex-specific respiratory and systemic effects attributed to air pollutant inhalation exposure, and (3) postulate proposed mechanisms of stress and sex hormone interactions during air pollution-related stress.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"287-314"},"PeriodicalIF":6.4,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141793934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Local and systemic effects of microplastic particles through cell damage, release of chemicals and drugs, dysbiosis, and interference with the absorption of nutrients.","authors":"Eleonore Fröhlich","doi":"10.1080/10937404.2024.2406192","DOIUrl":"https://doi.org/10.1080/10937404.2024.2406192","url":null,"abstract":"<p><p>Microplastic particles (MPs) have been detected in a variety of environmental samples, including soil, water, food, and air. Cellular studies and animal exposures reported that exposure to MPs composed of different polymers might result in adverse effects at the portal of entry (local) or throughout the body (systemic). The most relevant routes of particle uptake into the body are oral and respiratory exposure. This review describes the various processes that may contribute to the adverse effects of MPs. Only MPs up to 5 µm were found to cross epithelial barriers to a significant extent. However, MPs may also exert a detrimental impact on human health by acting at the epithelial barrier and within the lumen of the orogastrointestinal and respiratory tract. The potential for adverse effects on human health resulting from the leaching, sorption, and desorption of chemicals, as well as the impact of MPs on nutritional status and dysbiosis, are reviewed. <i>In vitro</i> models are suggested as a means of (1) assessing permeation, (2) determining adverse effects on cells of the epithelial barrier, (3) examining influence of digestive fluids on leaching, desorption, and particle properties, and (4) role of microbiota-epithelial cell interactions. The contribution of these mechanisms to human health depends upon exposure levels, which unfortunately have been estimated very differently.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":"27 8","pages":"315-344"},"PeriodicalIF":6.4,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142331320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An integrative exploration of environmental stressors on the microbiome-gut-brain axis and immune mechanisms promoting neurological disorders.","authors":"Hajar Heidari, David A Lawrence","doi":"10.1080/10937404.2024.2378406","DOIUrl":"10.1080/10937404.2024.2378406","url":null,"abstract":"<p><p>The microbiome-gut-brain axis is altered by environmental stressors such as heat, diet, and pollutants as well as microbes in the air, water, and soil. These stressors might alter the host's microbiome and symbiotic relationship by modifying the microbial composition or location. Compartmentalized mutualistic microbes promote the beneficial interactions in the host leading to circulating metabolites and hormones such as insulin and leptin that affect inter-organ functions. Inflammation and oxidative stress induced by environmental stressors may alter the composition, distribution, and activities of the microbes in the microbiomes such that the resultant metabolite and hormone changes are no longer beneficial. The microbiome-gut-brain axis and immune adverse changes that may accompany environmental stressors are reviewed for effects on innate and adaptive immune cells, which may make host immunity less responsive to pathogens and more reactive to self-antigens. Cardiovascular and fluid exchanges to organs might adversely alter organ functionality. Organs, especially the brain, need a consistent supply of nutrients and clearance of debris; disruption of these exchanges by stressors, and involvement of gut microbiome are discussed regarding neural dysfunctions with Alzheimer's disease, autistic spectrum disorders, viral infections, and autoimmune diseases. The focus of this review includes the manner in which environmental stressors may disrupt gut microbiota leading to adverse immune and hormonal influences on development of neuropathology related to hyperhomocysteinemia, inflammation, and oxidative stress, and how certain therapeutics may be beneficial. Strategies are explored to lessen detrimental effects of environmental stressors on central and peripheral health navigated toward (1) understanding neurological disorders and (2) promoting environmental and public health and well-being.</p>","PeriodicalId":49971,"journal":{"name":"Journal of Toxicology and Environmental Health-Part B-Critical Reviews","volume":" ","pages":"233-263"},"PeriodicalIF":6.4,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141591890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}