Journal of Oral Biosciences最新文献

{"title":"Androgen suppression protects against hyposalivation and salivary gland damage in mice with type 2 diabetes","authors":"Ana Lilia García-Hernández,&nbsp;Nancy Cruz-Mendoza,&nbsp;Gerardo Arturo Rueda-Cortez,&nbsp;Saúl Ernesto Cifuentes-Mendiola","doi":"10.1016/j.job.2025.100646","DOIUrl":"10.1016/j.job.2025.100646","url":null,"abstract":"<div><h3>Objective</h3><div>Hyposalivation is one of the most common oral complications of type 2 diabetes (T2D). Sex hormone levels, which have been associated with hyposalivation, salivary gland atrophy, and inflammation, can be altered in T2D. However, the relationship between androgen levels and hyposalivation in the context of T2D is unknown. Therefore, this study investigated the role of gonadal androgen suppression on the function and histomorphometry of salivary glands in mice with T2D.</div></div><div><h3>Methods</h3><div>Four-week-old male C57BL/6 mice were divided into four groups: control, orchiectomy (ORQx), T2D, and ORQx-T2D. Orchiectomy was performed at eight weeks of age, and T2D was induced using a high-calorie diet and low-dose streptozotocin. At 20 weeks of age, the blood glucose levels, saliva secretion and quality, and serum testosterone were measured. The parotid and submandibular glands were retrieved, processed for histology, and sections were stained with hematoxylin and eosin, Sirius Red or immunohistochemically stained for α-amylase, interleukin (IL)-1, IL-6, IL-10, IL-17, and tumor necrosis factor-α.</div></div><div><h3>Results</h3><div>Mice with T2D exhibited decreased saliva secretion and quality, reduced α-amylase expression, and the number of acini. They also developed glandular fibrosis and acinar hypertrophy, along with increased in proinflammatory cytokines in both salivary glands. Androgen suppression in mice with T2D reduced hyperglycemia, normalized saliva secretion, decreased glandular fibrosis and acinar hypertrophy, increased α-amylase expression, and reduced proinflammatory cytokine expression in both glands.</div></div><div><h3>Conclusions</h3><div>Androgen suppression in mice with T2D reduces the development of hyposalivation and histomorphometric changes in the parotid and submandibular glands by modulating the inflammatory microenvironment.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 2","pages":"Article 100646"},"PeriodicalIF":2.6,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143587680","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":"Letter to the editor, “Mirror-polished ceria-stabilized zirconia/alumina nanocomposite enhances gingival junctional epithelial cell adhesion” [J Oral Biosci, Volume 67, 2025. DOI: 10.1016/j.job.2024.100593]","authors":"Nicholas G. Fischer","doi":"10.1016/j.job.2025.100647","DOIUrl":"10.1016/j.job.2025.100647","url":null,"abstract":"","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 2","pages":"Article 100647"},"PeriodicalIF":2.6,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143562257","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":"Antimicrobial effects of great salt lake mineral salts on oral pathogenic bacteria: Implications for oral care","authors":"Inori Inui ,&nbsp;Atsushi Iwatsuki ,&nbsp;Yoshie Yoshioka ,&nbsp;Manabu Habu ,&nbsp;Wataru Ariyoshi ,&nbsp;Ryota Yamasaki","doi":"10.1016/j.job.2025.100633","DOIUrl":"10.1016/j.job.2025.100633","url":null,"abstract":"<div><div>This study investigated the antimicrobial effects of purified natural mineral salts from the Great Salt Lake on oral pathogenic bacteria. Salts rich in sodium, calcium, potassium, and magnesium effectively inhibit the growth and biofilm formation of cariogenic and periodontopathogenic bacteria at lower concentrations than does sodium chloride. These findings suggest the potential applications of these salts in oral care products such as toothpaste and mouthwash.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 2","pages":"Article 100633"},"PeriodicalIF":2.6,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143548746","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":"Reactivation of Epstein-Barr virus by n-butyric acid from Pseudoramibacter alactolyticus induces inflammatory cytokines in periapical granulomas","authors":"Taiki Miyata ,&nbsp;Osamu Takeichi ,&nbsp;Kenichi Imai ,&nbsp;Masayuki Okano ,&nbsp;Seiya Inoue ,&nbsp;Takuya Yasukawa ,&nbsp;Yusuke Suzuki","doi":"10.1016/j.job.2024.10.001","DOIUrl":"10.1016/j.job.2024.10.001","url":null,"abstract":"<div><h3>Objectives</h3><div>This study investigates whether latent Epstein-Barr virus (EBV) can be reactivated by n-butyric acid from <em>Pseudoramibacter alactolyticus</em>, and if such reactivation induces expression of interleukin (IL)-1β and IL-6 in periapical granulomas.</div></div><div><h3>Methods</h3><div>We analyzed periapical granulomas and healthy gingival tissues to detect the presence of EBV and <em>P. alactolyticus</em>. The concentration of n-butyric acid in <em>P. alactolyticus</em> culture supernatants was measured. BZLF-1 luciferase assays were conducted with or without these supernatants. Immunohistochemical detection of ZEBRA-, IL-1β-, and IL-6-expressing cells was performed in the tissue samples. Additionally, mRNA expression levels of BZLF-1, IL-1β, and IL-6 were quantified and statistically analyzed for correlation. The expression of these mRNAs was also measured in Daudi cells treated with or without the culture supernatants.</div></div><div><h3>Results</h3><div>Both EBV and <em>P. alactolyticus</em> were detected in periapical granulomas, but not in healthy tissues. The concentration of n-butyric acid in the culture supernatants was ∼3.58 mmol/L. BZLF-1 luciferase activity in the presence of the culture supernatants was comparable to that of commercially available butyric acid, whereas no activity was detected without the supernatants. Cells expressing ZEBRA co-expressed IL-1β and IL-6. The mRNA levels of IL-1β and IL-6 in periapical granulomas were correlated with BZLF-1 mRNA levels. Daudi cells treated with the culture supernatants expressed BZLF-1, IL-1β, and IL-6 mRNA, while those without the supernatants did not.</div></div><div><h3>Conclusions</h3><div>The study concludes that EBV can be reactivated by n-butyric acid produced by <em>P. alactolyticus</em>, leading to the induction of IL-1β and IL-6 expression in periapical granulomas.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100569"},"PeriodicalIF":2.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142477313","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":"Decline in stimulus responsiveness of secretory granules in salivary glands with age","authors":"Miyuki Toda ,&nbsp;Megumi Yokoyama ,&nbsp;Osamu Katsumata-Kato ,&nbsp;Junko Fujita-Yoshigaki","doi":"10.1016/j.job.2024.100583","DOIUrl":"10.1016/j.job.2024.100583","url":null,"abstract":"<div><h3>Objectives</h3><div>Secretory granules produced by salivary acinar cells accumulate if secretory stimulation is suppressed. Aged and deteriorated secretory granules can cause tissue damage because of abnormal secretion and/or intracellular leakage. To elucidate the deterioration process, we characterized the changes in the stimulus responsiveness of secretory granules using HaloTag technology.</div></div><div><h3>Methods</h3><div>We established a system in which HaloTag-fused cystatin D, a salivary protein, was transported to the secretory granules of rat parotid acinar cells in primary culture. HaloTags can be labeled with cell-permeable ligands conjugated to fluorescent dyes in living cells. To observe the new and old secretory granules, the cells were labeled with two HaloTag ligands conjugated to different fluorescent dyes at different times. We measured the secretion rates of newly synthesized and old HaloTag-fused proteins in the absence and presence of isoproterenol, a β-adrenergic agonist, at 3 and 6 h after initial labeling.</div></div><div><h3>Results</h3><div>Sequential labeling of HaloTag-fused proteins with two different dyes enabled the discrimination between new and old secretory granules. The secretory responsiveness of the proteins synthesized within 3 h to isoproterenol was higher than that of proteins synthesized earlier. However, there was no significant difference in the responsiveness between the new and old proteins at 6 h after initial labeling.</div></div><div><h3>Conclusion</h3><div>New secretory granules have a higher sensitivity to stimulants than older ones and that their response declines over time.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100583"},"PeriodicalIF":2.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142569540","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":"ED-71 promotes osseointegration of titanium implants in a rat model of GIOP by alleviating the effects of dexamethasone on bone remodeling in a SIRT1-dependent manner","authors":"Chunying Li ,&nbsp;Pengfei Xue ,&nbsp;Guanglin Duan ,&nbsp;Ailing Song ,&nbsp;Runbing Zhai ,&nbsp;Jie Ma ,&nbsp;Minqi Li","doi":"10.1016/j.job.2024.10.003","DOIUrl":"10.1016/j.job.2024.10.003","url":null,"abstract":"<div><h3>Objective</h3><div>Glucocorticoid-induced osteoporosis (GIOP), a common complication of glucocorticoid usage, plays a critical role in the success of dental implant restoration by affecting osseointegration. Eldecalcitol (ED-71) prevents GIOP; however, its role in the osseointegration of implants under GIOP conditions remains elusive.</div></div><div><h3>Methods</h3><div>Dexamethasone was used to establish a rat model of GIOP. Subsequently, mini-implant surgery was performed on the femur. GIOP rats were administered ED-71 via gavage to assess its role in the osseointegration of titanium implants under GIOP conditions. MC3T3-E1 and RAW264.7 cells were utilized to explore the molecular mechanism of ED-71 in ameliorating disorder of bone remodeling caused by dexamethasone.</div></div><div><h3>Results</h3><div>The administration of ED-71 promoted the formation of newly formed woven bone and the resolution of inflammation around titanium implants. In vitro experiments indicated that ED-71 ameliorated dexamethasone-induced dysfunction of osteoblasts and osteoclasts by increasing the expression level of sirtuin 1 (SIRT1). Inhibition of SIRT1 by selisistat counteracts the regulatory effects of ED-71 on dexamethasone-induced disorder of bone remodeling. Molecular docking and Western blotting revealed that the neurogenic locus notch homolog protein and nuclear factor kappa B signaling pathways are essential for the effects of ED-71 on dexamethasone-induced disorder of bone remodeling.</div></div><div><h3>Conclusion</h3><div>ED-71 promoted implant osseointegration in a rat model of GIOP by alleviating the effects of dexamethasone on bone remodeling in a SIRT1-dependent manner.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100571"},"PeriodicalIF":2.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142477312","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":"MANDI-code: A coding system for the human mandible","authors":"Doha Abualhija ,&nbsp;Julieta Gómez García-Donas ,&nbsp;Simon Shepherd ,&nbsp;Ranya Al Ghazi ,&nbsp;Scheila Manica","doi":"10.1016/j.job.2024.100584","DOIUrl":"10.1016/j.job.2024.100584","url":null,"abstract":"<div><h3>Background</h3><div>The mandible provides valuable insights into its biological identity. However, the existence of several terminologies for mandibular measurements and inconsistent language can lead to misinterpretation, confusion, and miscommunication.</div></div><div><h3>Highlight</h3><div>A standardised set of anatomical points, planes, and measurements would assist with these issues and ensure reproducibility and comparability.</div></div><div><h3>Conclusion</h3><div>The proposed coding system offers a comprehensive approach for professionals and researchers in dentistry, archaeology, and anthropology.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100584"},"PeriodicalIF":2.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142565260","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":"Conditional heterozygous loss of kit receptor tyrosine kinase in neural crest cell lineage is associated with midline cleft lip and bifid nose deformity","authors":"Hitomi Aoki ,&nbsp;Hiroyuki Tomita ,&nbsp;Akira Hara ,&nbsp;Takahiro Kunisada","doi":"10.1016/j.job.2024.10.004","DOIUrl":"10.1016/j.job.2024.10.004","url":null,"abstract":"<div><h3>Objectives</h3><div>The receptor tyrosine kinase Kit is expressed in cells derived from the trunk neural crest (NC), such as melanocytes; however, its role in cranial NC cell development is not fully understood.</div></div><div><h3>Methods</h3><div>We investigated the effects of the heterozygous loss of <em>Kit</em> in NC cells during embryonic development by mating <em>Kit</em>^{<em>2lox/+</em>} mice with <em>Wnt1-Cre</em> mice to produce <em>Wnt1-Cre; Kit</em>^{<em>2lox/+</em>} embryos. In addition, <em>Wnt1-Cre</em> mice were mated with <em>Rosa26R-yellow fluorescent protein</em> (<em>YFP</em>) mice to visualize the tissue regions expressing Cre recombinase. Histological studies of the craniofacial regions of these mice were performed using samples from embryonic day (E) 12.5 and postnatal day (P) 1. Cellular apoptosis and proliferation were both analyzed through the immunostaining of tissue sections collected on E13.5 and E14.5 using anti-cleaved caspase 3 (CC3) to detect apoptosis and anti-Ki67 to detect proliferation. Cells from YFP-positive tissue regions of the facial areas of <em>Wnt1-Cre; Kit</em>^<em>+/+</em><em>; Rosa26R–YFP</em> embryos and <em>Wnt1-Cre; Kit</em>^{<em>2lox/+</em>}<em>; Rosa26R–YFP</em> embryos collected on E12.5 and E15.5 were cultured and evaluated for cell proliferation.</div></div><div><h3>Results</h3><div>Compared with control littermates, <em>Wnt1-Cre; Kit</em>^{<em>2lox/+</em>} embryos exhibited midline cleft lip and bifid nose deformities. Substantial early (P1) postnatal lethality was observed in <em>Wnt1-Cre; Kit</em>^{<em>2lox/+</em>} mice, with none surviving to 3 weeks of age. YFP-positive cells from the maxillary regions of <em>Wnt1-Cre; Kit</em>^{<em>2lox/+</em>}<em>; Rosa26R–YFP</em> embryos exhibited defective cell growth and self-renewal <em>in vitro</em>.</div></div><div><h3>Conclusion</h3><div>Conditional heterozygous loss of <em>Kit</em> in <em>Wnt1-Cre; Kit</em>^{<em>2lox/+</em>} embryos is associated with craniofacial dysplasia and exhibit defective NC development <em>in vitro</em> and <em>in vivo</em>.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100572"},"PeriodicalIF":2.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142477311","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":"Remineralizing capacity of zinc oxide eugenol sealer following the addition of nanohydroxyapatite-tyrosine amino acid: An in vivo animal study","authors":"Rasha M. Al-Shamaa,&nbsp;Raghad A. Al-Askary","doi":"10.1016/j.job.2024.09.006","DOIUrl":"10.1016/j.job.2024.09.006","url":null,"abstract":"<div><h3>Objective</h3><div>Although several sealers have been developed, the zinc oxide eugenol (ZOE) sealer is still used in many private dental clinics. This study aimed to compare the biocompatibility and remineralizing capacity οf ZOE sealer with the addition of nanohydroxyapatite-tyrosine amino acid.</div></div><div><h3>Methods</h3><div>This study was conducted on Twenty rabbits. The rabbits were divided into four groups based on the test observation period (3, 7, 21, and 28 days) following surgical implantation. General anesthesia was administered to each rabbit, and a subcutaneous incision of approximately 1 ± 0.5 cm was made along the symphyseal area of the mandible of each rabbit. Four bone cavities were generated in the interdental space of the lower jaw between the central and molar teeth of each rabbit, with one longitudinal subcutaneous incision. The ZOE sealers were mixed according to the manufacturer's guidelines and directly inserted within the cavities generated at the end of each test period. The animals were sacrificed, and bone biopsy was carried out at the site of testing. The biopsy samples were obtained and subjected to histological analysis using a low-power light microscope (Olympus C ⅹ 21, Japan) and immunohistochemistry using Ki67 antibody.</div></div><div><h3>Results</h3><div>The collected information was examined by parametric statistical tests using SPSS software version ‘‘22.” One-way ANOVA and post-hoc Duncan's tests were used to measure the significance among various groups, with statistical significance set, when “P ≤ 0.01”.</div></div><div><h3>Conclusion</h3><div>The 20% mixed endodontic sealer displayed excellent outcomes compared to other experimental groups, as identified by higher new bone formation at every evaluation period.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100567"},"PeriodicalIF":2.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142477314","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":"Impact of organic, conventional, and stingless bee honeys on the antibacterial activity of gummy candies against oral bacteria","authors":"José Renato Silva ,&nbsp;Jerônimo Kahn Villas-Bôas ,&nbsp;Guilherme Biz ,&nbsp;Ricardo Sergio Couto Almeida ,&nbsp;Wilma Spinosa ,&nbsp;Sandra Helena Prudencio","doi":"10.1016/j.job.2024.100589","DOIUrl":"10.1016/j.job.2024.100589","url":null,"abstract":"<div><h3>Objectives</h3><div>This study aimed to investigate the impact of organic, conventional, and stingless honey on gummy candies, focusing on the effect of the cariogenic bacterium, <em>Streptococcus mutans</em> UA159, and total bacterial count in saliva from adolescents.</div></div><div><h3>Methods</h3><div>Antimicrobial compounds in three honey samples were identified, and the minimum inhibitory concentration against <em>S. mutans</em> UA159 was determined. The antibacterial activities of the three honey candy formulations were determined against <em>S. mutans</em> UA159 in artificial saliva and total bacteria in saliva collected from adolescents. The sensory acceptance of the candy formulations by children, adolescents, and adults was investigated.</div></div><div><h3>Results</h3><div>Candies prepared using conventional honey showed the highest antibacterial activity against <em>S. mutans</em> UA159 <em>in vitro</em> and total bacteria in human saliva. This effect was attributed to the higher levels of quercetin, myricetin, caffeine, and hydrogen peroxide in conventional honey.</div></div><div><h3>Conclusions</h3><div>Nicotinic, ferulic, and <em>p</em>-coumaric acids found in honey had low antibacterial activity against oral bacteria. Quercetin, myricetin, caffeine, and hydrogen peroxide are the main anticariogenic compounds in honey and exert antibacterial effects on adolescent saliva, despite added to candies. However, organic production does not necessarily improve the biological properties of honey. All candies were equally liked by sensory assessors (acceptance &gt;70%), facilitating the selection of honey with higher biological activities to formulate functional candies.</div></div>","PeriodicalId":45851,"journal":{"name":"Journal of Oral Biosciences","volume":"67 1","pages":"Article 100589"},"PeriodicalIF":2.6,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142689164","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}