Medicine in Omics最新文献

{"title":"Identification of effect of gene mutations related to telomere maintenance in gastric cancer patients","authors":"Anju R. Nath,&nbsp;Jeyakumar Natarajan","doi":"10.1016/j.meomic.2023.100025","DOIUrl":"10.1016/j.meomic.2023.100025","url":null,"abstract":"<div><h3>Background</h3><p>Mutations in gastric cancer may remain harmless despite mutations that can turn oncogenic. Although omics technologies have supplied a clear view of mutations and driver genes in cancers, the pathology of gastric cancer still is unknown.</p></div><div><h3>Objective</h3><p>This study aims to understand mutations of genes that relate to telomere maintenance in gastric cancer.</p></div><div><h3>Methods</h3><p>We used transcriptomic and proteomic data from normal and gastric cancer patients to decide the relationship between genes and proteins. We ranked the gene-protein pairs with the Spearman correlation and subjected the differently ranked genes for functional analysis. We performed the co-expression analysis to select the genes from functional analysis. Then we assessed the selected genes for mutations and thereafter neoantigen analysis.</p></div><div><h3>Results</h3><p>We identified 20 genes through differential ranking based on Spearman correlation of gene-protein pairs. Among these, ERCC1, MRE11A, RAD50, TP53BP1, and YWHAE were linked to critical functions like DNA repair, apoptosis, and stress response. Gene prioritization focused on these five genes, and from them through gene co-expression, RAD50, TP53BP1, and YWHAE were selected. Mutational analysis revealed prevalent C &gt; T and G &gt; A mutations. These genes were associated with elderly patients, specific histologic grades, and types of gastric cancer. Importantly, these mutations generated neoantigens with potential for immunotherapeutic targeting.</p></div><div><h3>Conclusion</h3><p>Our extensive research shows the complex genetic makeup of gastric cancer and its potential effects on diagnosis and treatment. The identified genes and the mutations linked to them offer important information on the molecular processes behind gastric cancer progression. These results set the foundation for additional research and immunotherapeutic applications in the field of gastric cancer.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"10 ","pages":"Article 100025"},"PeriodicalIF":0.0,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590124923000068/pdfft?md5=9d115915f82ec87beae4dbf57cf84944&pid=1-s2.0-S2590124923000068-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135654955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Revealing the genotypic variants and proteomic mutations in Turkish SARS-CoV-2 driving evolution and virulence","authors":"Ragothaman Prathiviraj ,&nbsp;K. Prinsha ,&nbsp;Saqib Hassan ,&nbsp;S. Hari Krishna Kumar ,&nbsp;George Seghal Kiran ,&nbsp;Joseph Selvin","doi":"10.1016/j.meomic.2023.100024","DOIUrl":"https://doi.org/10.1016/j.meomic.2023.100024","url":null,"abstract":"<div><p>Turkey is a transcontinental nation with much of its territory in western Asia and southeast Europe, and it is the 6th most affected country by SARS-CoV-2 globally, so far, many mutations have occurred within this short period of time. The present study mainly focused on identifying genotypic variants and proteomic mutations, along with the folding rate and virulence action of Turkey SARS-CoV-2 isolates. The phylogenomic study reveals that the first origination of SARS-CoV-2 isolates (2020) in Turkey has more chance of an occurrence of mutation rate during evolution/new origination. 53 genotypic- and 45 proteomic variants among 83 isolates were found. The genotypic and proteomic variants from 2022 isolates have six times higher mutation rates than 2020 isolates. The highest number of mutations occurred in the 2022 isolate (ERAGEM-OM-1104/2022), with 60 mutations in the G53 variant, followed by 44 mutations in the G49 variant (ERAGEM-DEL-1103/2021) and 42 mutations in the G50 variant (Ank_DLT/2021). Also, we identified 38 novel mutations among 45 proteomic variants. The predicted virulence mechanism of some ORFs such as Nsp1, Nsp3, Nsp4, Nsp5, Nsp8, 2′O-ribose, Spike, Orf3a, Envelope, Orf7b, Orf8, and Nucleocapsid shows high virulence and transmission rate. Hence, our findings would help the researchers to develop new anti-virals against new variants from Turkey’s SARS-CoV-2 isolates.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"9 ","pages":"Article 100024"},"PeriodicalIF":0.0,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50188321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gene co-expression analysis uncovers the different effects of cathelicidin in lung squamous cell and in invasive breast carcinomas","authors":"Fabiano Pinheiro da Silva","doi":"10.1016/j.meomic.2023.100023","DOIUrl":"https://doi.org/10.1016/j.meomic.2023.100023","url":null,"abstract":"<div><p>Antimicrobial peptides are multi-functional peptides that possess a broad range of mechanisms of action. They have been investigated as novel therapeutic options in the field of infectious diseases, but an increasing number of publications suggest that they also play an important role in the pathology of cancer.</p><p>Here, using an in silico approach, we investigated the pathways associated with LL-37, the only human member of the cathelicidin family of antimicrobial peptides, in lung squamous cell and in invasive breast carcinomas.</p><p>We found that LL-37 is implicated in different and even opposing cell responses, depending on the type of cancer, and participates in intriguing molecular processes, such as platelets activity, coagulation and extracellular matrix organization.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"9 ","pages":"Article 100023"},"PeriodicalIF":0.0,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50188322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pomegranate (P. granatum) fruit juice protects against iron-induced oxidative testicular injury via amelioration of oxidative imbalance and modulation of metabolic indices linked to male infertility","authors":"Veronica F. Salau ,&nbsp;Ochuko L. Erukainure ,&nbsp;Kolawole A. Olofinsan ,&nbsp;Bukola R. Omotoso ,&nbsp;Md. Shahidul Islam","doi":"10.1016/j.meomic.2023.100021","DOIUrl":"https://doi.org/10.1016/j.meomic.2023.100021","url":null,"abstract":"<div><p>Male infertility is a challenging clinical condition that threatens global reproductive health. Majority of the issues found in testicular dysfunction are associated with oxidative stress. The present study investigated the protective effect of pomegranate fruit extract against Fe²⁺- mediated oxidative testicular injury using <em>ex vivo</em> experimental models. Testicular oxidative injury was induced by incubating testes tissues in Krebs buffer and FeSO₄.7H₂0 solution, and treated by co-incubating with different concentrations (30–240 μg/mL) of pomegranate fruit extract for 90 min (37 °C), with gallic acid serving as the standard antioxidant. Oxidative injury induction led to significant (p &lt; 0.05) elevation of testicular malondialdehyde and LDL-cholesterol concentration, acetylcholinesterase, ATPase, fructose-1,6-bisphosphatase, glycogen phosphorylase and lipase activities. These were concomitantly accompanied by depleted levels of reduced glutathione, glycogen, total cholesterol, triglyceride and HDL-cholesterol and activities of superoxide dismutase, catalase, ENTPDase and 5′Nucleotidase. It also led to distorted testicular histological architecture. Treatment with the fruit extract markedly reversed these levels and activities and preserved testicular morphology. Octadecenoic acid, 1,2,3-propanetriyl ester, (E,E,E)-; squalene and cholesterol, while concomitantly generating 6-Octadecenoic acid, (Z)-; (2Z)-2-Octenoic acid; methyl tetradecanoate; methyl 14-methylpentadecanoate; octadecanoic acid, 2-[(1-oxohexadecyl)oxy]ethyl ester and cholestan-3-ol, 2-methylene-, (3.beta.,5.alpha.)-. This was accompanied by the deactivation of alpha linolenic acid and linoleic acid metabolism, and steroidogenesis. Treatment with pomegranate depleted the oxidative-generated metabolites, while concomitantly generating arachidonic acid, cholesterol margranate and ethyl lithocholate, and reactivating the inactivated pathways. The results indicate the protective effect of pomegranate fruit juice against oxidative testicular injury.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"8 ","pages":"Article 100021"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50194473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular diagnostics and potential therapeutic options for mycobacterium tuberculosis: Where we stand","authors":"Zafran Khan ,&nbsp;Daniya Ualiyeva ,&nbsp;Khalid Jamal ,&nbsp;Babar Ali ,&nbsp;Fayaz Ahmad ,&nbsp;Sanjeep Sapkota ,&nbsp;Obed Boadi Amissah ,&nbsp;Petuel Ndip Ndip Bate","doi":"10.1016/j.meomic.2023.100022","DOIUrl":"https://doi.org/10.1016/j.meomic.2023.100022","url":null,"abstract":"<div><p><em>Mycobacterium tuberculosis<!--> </em>(Mtb) is a causative agent of the infectious disease tuberculosis (TB), which has hampered susceptible populations in modern-day societies for years. Additionally, the emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR)<!--> <!-->Mtb<!--> <!-->strains has intensified this global health issue. Each year the global death tally spikes by 1.6 million, and additional 10.6 million new cases of TB. Researchers are trying to find a treatment, mainly emphasizing the anti-tubercular drugs and/or vaccines that could potentially control the spread of TB to manage the associated health issue. To date, more than 20 anti-tubercular drugs have been developed and used at various stages of TB. The current review aims to discuss/describe useful information about available TB treatments and anti-tubercular resistance. Moreover, we also highlighted diagnostic approaches and current drug reserves for drug-resistant TB (DR-TB).</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"8 ","pages":"Article 100022"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50194474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Physiological Pathway, diagnosis and nanotechnology based treatment strategies for ovarian Cancer: A review","authors":"Monika Gulia ,&nbsp;Suchitra Nishal ,&nbsp;Balaji Maddiboyina ,&nbsp;Rohit Dutt ,&nbsp;Prasanna Kumar Desu ,&nbsp;Rubina Wadhwa ,&nbsp;Vikas Jhawat","doi":"10.1016/j.meomic.2023.100020","DOIUrl":"https://doi.org/10.1016/j.meomic.2023.100020","url":null,"abstract":"<div><p>Ovarian cancer is a fatal disease requiring great attention from the scientific community to find novel ways for diagnosis and treatment. Most ovarian malignancies, or 90% of cases, are epithelial ovarian cancers. HGSC (High Grade Ovarian Cancer) is the most prevalent subtype, and the majority of women who are diagnosed with it eventually develop resistance to standard treatments. Expression of genes linked to these immune pathways and increased cytotoxic immune cell infiltration is primarily associated with HGSC cancer that shows DNA damage repair gene deficiency and high chromosomal instability. Currently, chemotherapy, radiation, and cytoreduction are the most common forms of treatment; nevertheless, in the absence of targeted therapies, patients experience unfavorable side effects and develop drug resistance. It is critical to understand the intricate biology of the disease and find relevant biomarkers in order to make an early diagnosis or anticipate how a patient will respond to a given treatment. Interferon activating medicines have been studied as a potential means of enhancing anti-tumor immunity in ovarian cancer. Chemical and biological nano-sensors have also been developed to detect different types of cancers, including ovarian cancer. Further, the application of nanotechnology for diagnostic and therapeutic purposes makes it a more convenient, target-specific, and side effect free delivery system for ovarian cancer treatment. A combination of nanotechnology with the physiological biomarkers and therapeutic agents created a novel system of nano theranostics, which have the potential of real-time monitoring and diagnosis and simultaneous delivery of the therapeutic agent for the treatment of ovarian cancer.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"8 ","pages":"Article 100020"},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50195101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolite profiling and anti-cancer activity of two medicinally important Euphorbia species","authors":"Sulaiman C.T. ,&nbsp;Deepak M. ,&nbsp;Praveen T.K. ,&nbsp;Lijini K.R. ,&nbsp;Salman M. ,&nbsp;Sadheeshnakumari S. ,&nbsp;Indira Balachandran","doi":"10.1016/j.meomic.2022.100018","DOIUrl":"https://doi.org/10.1016/j.meomic.2022.100018","url":null,"abstract":"<div><h3>Background</h3><p><em>Euphorbia thymifolia</em> L. and <em>Euphorbia hirta</em> L. are two medicinally important species used in <em>Ayurveda</em> and are known as <em>Laghudugdhika</em> and <em>Dugdhika</em> respectively. This study was carried out to evaluate the comparative metabolite profiling and anti-cancer activity of <em>E. thymifolia</em> and <em>E. hirta</em>. Chemical profiling was done by High Performance Thin Layer Chromatography (HPTLC) and quantitative estimations of a major class of phytochemicals were done using specified spectrophotometric methods. Detailed metabolite profiling was done by tandem mass spectroscopic evaluation by Q-TOF-LC-MS/MS analysis. Anticancer activity was evaluated using Ehrlich Ascites Carcinoma (EAC) model in mice with 5-fluorouracil as standard.</p></div><div><h3>Results</h3><p>HPTLC profile showed that most of the chemical contents are similar in both species. Major group of secondary metabolites such as phenolics, flavonoids and sterols were found to be in equal concentration in both species. LC/MS analysis and data processing based on the molecular ion fragmentation pattern led to the identification of 44 compounds from the selected species. Pharmacological evaluation showed that both species possess anti-cancer activity against ascites carcinoma in mice model, however, <em>E. hirta</em> showed a significant dose dependent anticancer activity against EAC induced periotoneal ascites in mice.</p></div><div><h3>Conclusion</h3><p>The study concluded that the selectd species comparised of variety of active phytoconstrtiuents with anti-cancer properties.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"7 ","pages":"Article 100018"},"PeriodicalIF":0.0,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50192430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immuno-modulatory drugs: A rapid way to combat the tuberculosis","authors":"Zafran Khan ,&nbsp;Daniya Ualiyeva ,&nbsp;Ubaid Ahmad ,&nbsp;Buhari Yusuf","doi":"10.1016/j.meomic.2022.100016","DOIUrl":"10.1016/j.meomic.2022.100016","url":null,"abstract":"<div><p>Drug-resistant tuberculosis (DR-TB) epidemics pose a potential threat to global health and are defined by higher morbidity and mortality, sequelae, cost, and complexities. Previously, it has been evidenced that establishing a new drug regimen containing 2–3 new effective drugs may take around 30 years and will be cost-effective. Due to these limitations, drug repurposing has become critical, and the repurposing of existing drugs, licensed for other diseases is the broad alternative for DR-TB treatment. Thus, immunomodulatory drugs are an alternative approach to combat the deadly infectious disease, caused by<!--> <em>Mycobacterium tuberculosis.</em> <!-->These repurposed drugs target many pathways, and also reduce the risk of acquiring resistance. In this review, we discussed some of the immunomodulatory drugs, and their effectiveness to combat TB.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"5 ","pages":"Article 100016"},"PeriodicalIF":0.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590124922000049/pdfft?md5=b33f37dc3bdc232e37d2341f48c892b5&pid=1-s2.0-S2590124922000049-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73469068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An in vivo assessment of inflammatory and oxidative stress responses in Echis ocellatus-venom induced cardiotoxicity","authors":"Babafemi Siji Ajisebiola ,&nbsp;Ayomikun Busayo Fawole ,&nbsp;Olubisi Esther Adeyi ,&nbsp;Akindele Oluwatosin Adeyi","doi":"10.1016/j.meomic.2022.100017","DOIUrl":"10.1016/j.meomic.2022.100017","url":null,"abstract":"<div><p><em>Echis ocellatus</em> is one of the many viper species that accounts for severe pathophysiological alterations in tissues of organs after envenoming. However, limited information regarding the potential cardiac toxicity due to viper envenoming is available. This current study investigated cardiotoxicity associated with <em>E. ocellatus</em> envenoming in rat model. Twenty (20) male Wistar rats weighing between 140 and 180 g were divided randomly into two groups (n = 10). Rats in group 1 (control) were injected with saline while rats in group 2 were envenomed intraperitoneally with 0.055 mg/kg⁻¹ (LD_6.25) of <em>E. ocellatus</em> venom. The rats were envenomed on day 1 with a repeated dose administered on day 15, afterwards the animals were monitored till day 30. The venom caused significant (P &lt; 0.05) reduction in body and heart weights including the heart index of envenomed rats compared to the control. Levels of malondialdehyde significantly (P &lt; 0.05) increased with decrease in glutathione concentration and catalase activity in heart tissues of envenomed rats. <em>E. ocellatus</em> venom elevated pro-inflammatory cytokines response as levels of tumor necrosis factor-alpha and interleukin1-beta significantly (P &lt; 0.05) increased in cardiac tissues of the envenomed rats compared to control. The venom induced severe morphological defects in the heart tissues of envenomed rats indicating that <em>E. ocellatus</em> venom could actuate cardiotoxicity post envenoming.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"5 ","pages":"Article 100017"},"PeriodicalIF":0.0,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590124922000050/pdfft?md5=a55f21302a90a612d5865e6a22aa6a8e&pid=1-s2.0-S2590124922000050-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79071219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genome editing: An essential technology for cancer treatment","authors":"Muhammad Tufail","doi":"10.1016/j.meomic.2022.100015","DOIUrl":"10.1016/j.meomic.2022.100015","url":null,"abstract":"<div><p>When genetic engineering was first developed in the 1970s, it opened new possibilities for genome editing. The life sciences have reaped numerous gains from gene-editing technology. The CRISPR/Cas9 system makes adding or removing sequence-specific amounts of DNA from the genome with ease achievable. CRISPR/Cas9 targeting efficiency improves, and concerted efforts reduce off-target effects. Cancer biology and oncology utilise CRISPR/Cas9 applications to do robust site-specific gene editing, making the technology valuable for research and therapeutic use. CRISPR/Cas9 is quickly evolving into several variations and uses. Helped by CRISPR-based techniques, scientists have developed a simple and inexpensive tool for developing powerful cancer therapies. CRISPR-based gene-editing technologies in oncology, gene editing for personalised medicine and overcoming resistance to cancer therapies, targeted therapies, optimisation of gene-editing technology and future cancer therapeutics are discussed in this review.</p></div>","PeriodicalId":100914,"journal":{"name":"Medicine in Omics","volume":"4 ","pages":"Article 100015"},"PeriodicalIF":0.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590124922000037/pdfft?md5=c37edaf04ce0822f7184dd8c8730f1d6&pid=1-s2.0-S2590124922000037-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83460365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}