自制根管润滑剂与不同浓度次氯酸钠液组合对粪肠球菌感染根管壁的清洁效果

目的:观察自制根管润滑剂与不同浓度次氯酸钠液组合对体外建立的粪肠球菌感染根管模型的清洁效果.方法:80个新鲜拔除的单根管牙随机分为8组,逐步深入法预备根管,预备过程中各组样本分别使用自制根管润滑剂配合不同浓度次氯酸钠液(5 g/L、25 g/L、52.5吕/L)3组、Glyde凝胶配合不同浓度的次氯酸钠液3组(为阳性对照)、过氧化氢液加生理盐水、蒸馏水(均为阴性对照)润滑、冲洗根管.通过扫描电镜观察根管壁玷污层、碎屑及细菌去除情况,并将样本根中1/3分为10个区采用单盲法分别评分,结果进行统计学分析.结果:除过氧化氢液配合生理盐水组,单纯蒸馏水组外,各润滑剂与次氯酸钠液配合组均能完全去除根中根管壁表面的粪肠球菌,两种润滑剂配合不同浓度次氯酸钠液与过氧化氰液+生理盐水组和单纯蒸馏水组相比,均能有效地去除碎屑和玷污层(P<0.05);两组润滑剂在去除玷污层方面组间相差不显著(P>0.05),次氯酸钠液各浓度之间亦无显著性差异,但对碎屑的去除效果则随次氯酸钠浓度升高效果越好,不同浓度间相差显著(P<0.05).结论:本实验室研制的根管润滑剂与52.5 g/L次氯酸钠液组合能有效去除根中根管壁玷污层和细菌.     

Carisolv应用于根管冲洗对根管内粪肠球菌抗菌作用的体外研究

目的：分析Carisolv祛龋凝胶作为根管冲洗剂对粪肠球菌（E．faecalis）离体牙根管感染模型的抗菌效果。方法：60颗牙体完整、发育正常的人单根管前磨牙,用ProTaper手用器械将建立根管感染模型的离体牙根管预备至F2后,建立粪肠球菌体外根管感染模型。随机分为5组,分别使用Carisolv凝胶、5．25％次氯酸钠溶液、EDTA凝胶、2％氯亚明溶液、0．9％氯化钠溶液进行根管冲洗,对根管冲洗前后的根管进行菌落计数,比较各种冲洗剂的抗菌作用。结果：5种冲洗剂在根管预备冲洗后细菌数量都显著性低于冲洗前（P＜0．001）。Carisolv组、氯亚明组、次氯酸钠组冲洗后根管的细菌数量没有显著性差异（P＞0．05）,低于EDTA组和氯化钠组（P＜0．01）。结论：Carisolv凝胶作为根管冲洗剂具备抗菌作用,其抗菌作用强于EDTA凝胶和0．9％氯化钠溶液。     

EDTA与次氯酸钠联合应用对根管内粪肠球菌清除效果:体外研究

目的:评价17%EDTA联合1%次氯酸钠对根管内粪肠球菌的清除效果.方法: 将40 个离体前牙的感染根管标本随机分为5组,器械预备时分别辅以冲洗液冲洗根管.其中A、B、C组分别用5.25%、2.5%及1%次氯酸钠冲洗,D组用17%EDTA联合1%次氯酸钠冲洗,E组用0.9%NaCl冲洗.冲洗前、后计数根管内的细菌量,并观察72 h细菌复苏情况.结果: 5 组根管内的细菌量均显著下降(P<0.01),其中D组与A、C组间差异有显著性(P<0.05),与B组间差异无显著性(P>0.05).结论: 17%EDTA联合1%次氯酸钠可有效清除根管内粪肠球菌, 其效果优于1%次氯酸钠,与2.5%次氯酸钠相近似.     

不同根管冲洗方式对粪肠球菌生物膜清除效果评价

目的 比较不同根管冲洗方式对粪肠球菌感染根管预备、冲洗的清除效果,评价不同根管冲洗液残余药量的抗菌效果,为临床上选择根管冲洗方法提供参考。方法 将32颗离体牙（前磨牙、单根管）消毒,接种粪肠球菌60 d,随机分为4组（第1组：生理盐水;第2组：生理盐水+超声1 min;第3组：1%NaOCl;第4组：1%NaOCl+超声1 min）,使用ProTaper器械按照冠向下法进行根管预备,并在预备冲洗前及预备至F2进行冲洗后分别取样、计数,进行统计学分析。将预备好的离体牙再次消毒,随机分为2组,分别浸泡生理盐水和1%NaOCl;浸泡30 min后取出,置于接种粪肠球菌标准菌株（ATCC33186）的培养液内,分别在培养2、6、24、48 h时取样、计数。采用SPSS19.0软件包对数据进行统计学分析。结果 4组在预备至F2冲洗结束后,根管中的细菌数量均不同程度降低,其中,使用1% NaOCl结合超声冲洗1 min,几乎可完全去除根管内的细菌。经1% NaOCl浸泡的牙根样本内的细菌,在培养48 h时后总量少于生理盐水。结论 1%NaOCl是有效的根管冲洗液,用于根管化学预备后的残余液体,也可发挥有效的抗菌效果。联合使用超声器械,可以使其抗菌效果最大化。     

根尖终末工作宽度对根管内细菌清除效果的体外研究

目的:比较不同的根尖预备终末工作宽度对根管内细菌清除效果的影响。方法:将已感染粪肠球菌的48颗离体上颌磨牙的远中颊侧根管标本随机分为6组,实验组分别预备根管至25#(A组)、30#(B组)、35#(C组)、40#(D组);对照组E、F组不进行机械预备而只进行冲洗。采用ProTaper机用镍钛锉和不锈钢K锉冠向下法预备根管。A、B、C、D、E组用1%次氯酸钠和17%EDTA冲洗根管,F组仅用生理盐水冲洗。根管预备前和预备后分别取样作细菌学培养计数,并做统计学分析。结果:预备冲洗后6组根管内的粪肠球菌量均显著下降(P<0.05),其中D组下降最明显。A、B组间差异无统计学意义,其余各组间比较均有显著性差异(P<0.05)。在实验组A、B、C、D组中,根管预备后阴性培养的百分率分别为0%、12.5%、37.5%和75%;对照组E、F组中均未获得阴性培养。结论:增大根尖预备终末工作宽度能提高根管内细菌清除的效果。但上颌磨牙远中颊侧根管预备至40#配合抗菌性冲洗液,尚不能完全清除根管内粪肠球菌。     

Er:YAG激光联合次氯酸钠对粪肠球菌杀菌效果的研究

目的:比较掺铒钇铝-石榴石脉冲(Er:YAG)激光联合不同浓度次氯酸钠对根管内粪肠球菌杀菌效果的研究。方法:选择新鲜拔除的人单根管离体前磨牙建立粪肠球菌根管感染模型75个,随机分为5组,每组15个,分别进行如下处理:A组:5.25%次氯酸钠单独冲洗;B组:Er:YAG激光+5.25%次氯酸钠冲洗;C组:1%次氯酸钠单独冲洗;D组:Er:YAG激光+1%次氯酸钠冲洗。E组:空白对照。采集5组根管内的细菌进行培养计数并对结果进行卡方检验(Chi-square test)。结果:A、B、D三组均能完全杀灭根管内的粪肠球菌;C组未能完全杀灭根管内的粪肠球菌。15个培养皿中有11个可见粪肠球菌菌落。A、B、D三组间杀菌效果无差异且均显著优于C组。结论:高浓度次氯酸钠可以完全杀灭根管内的粪肠球菌,低浓度次氯酸钠不能完全杀灭根管内的粪肠球菌;Er:YAG激光可以显著增强低浓度次氯酸钠的杀菌作用。     

五种冲洗液对根管内粪肠球菌抗菌效果的体外实验

目的评价五种冲洗液对根管内粪肠球菌的杀菌效果。方法建立粪肠球菌根管内感染模型,将40个感染根管标本随机分为5组,每组8颗牙。器械预备时分别采用5.25%NaClO（A组）、2.5%NaClO（B组）、1%NaClO（C组）、17%EDTA（D组）、17%EDTA＋1%NaClO（E组）冲洗根管。冲洗前、冲洗后即刻及冲洗后72h分别取样,37℃下CO2孵育箱中培养,48h后计数菌落数（CFU/ml）。结果冲洗后5组根管内的粪肠球菌量均显著下降（P〈0.05）,其中A组与其余各组间差异均有显著性（P〈0.01）;B组与C组、D组间差异有显著性（P〈0.05）,与E组间差异无显著性（P〉0.05）;C组与D组、E组间差异有显著性（P〈0.05）;D组与E组间差异有显著性（P〈0.05）。根管冲洗后培养72h均有细菌生长。结论5.25%NaClO抗菌效果最强;17%EDTA＋1%NaClO的抗菌效果优于1%NaClO,与2.5%NaClO抗菌效果相近似。     

不同冲洗方法对粪肠球菌感染根管的抗菌性研究

目的比较超声、Er,Cr：YSGG激光及Er：YAG激光辅助1%次氯酸钠（NaOCl）溶液冲洗对人离体牙根管表面粪肠球菌生物膜及根尖区不同深度牙本质小管内粪肠球菌的杀灭效果。 方法单直根管下颌前磨牙42颗,建立粪肠球菌生物膜感染的根管模型,采用随机字表法随机抽取2颗确定建成粪肠球菌生物膜感染根管模型,剩余40颗牙采用随机数字表法按随机对照原则分为5组,每组8颗。A组：5.25% NaOCl联合17%乙二胺四乙酸（EDTA）注射器冲洗;B组：0.9%氯化钠溶液注射器冲洗;C组：1% NaOCl溶液超声荡洗;D组：1% NaOCl溶液Er：YAG激光活化冲洗;E组：1% NaOCl溶液Er,Cr：YSGG激光活化冲洗。按分组进行处理后取样菌落计数,计算灭菌率。使用SPSS 17.0统计软件对实验数据进行方差齐性及正态性检验比较各组间和组内的灭菌率。 结果（1）组间比较：对根管壁表面,各组冲洗方法灭菌率比较,B组（5.74%）相似文献   

光活化消毒技术根管内消毒效果的研究

目的利用体外实验和临床试验评估光活化消毒技术（PAD）进行根管消毒的效果。方法体外选择30颗单根管牙根建立粪肠球菌ATCC29212感染模型,分为3组。用PAD技术进行根管消毒并取样,质量分数为2.5%的NaClO冲洗和生理盐水冲洗分别作为阳性和阴性对照组。临床选取50例慢性根尖周炎病例,随机分为2组。根管预备时质量分数为2.5%的NaClO溶液冲洗。预备后组1采用PAD消毒,组2用生理盐水冲洗。预备前后取样,所有样本接种于脑心浸液（BHI）培养基培养后记录菌落数。结果体外实验：PAD组细菌减少了99.86%,阳性对照组减少了100%,阴性对照组减少了96.94%。PAD组细菌回复10例均为阳性,阳性对照组6例,阴性对照组10例。临床试验：根管机械预备后细菌减少了99.98%。组1中PAD消毒后细菌培养均为阴性,组2细菌减少了36%。2组差异无统计学意义（P〉0.05）。结论 PAD单独使用可以降低根管内粪肠球菌的数量,但效果低于NaClO。临床PAD技术辅助根管预备可以进一步降低细菌的数量。     

次氯酸钠对根管内粪肠球菌杀菌效果的体外实验

目的评价次氯酸钠对根管内粪肠球菌的杀菌效果。方法将45个离体前磨牙的感染根管标本分为6组，1、2组用5．25％及2．5％次氯酸钠冲洗，3组用0．9％NaCl冲洗，4、5组在根管预备时辅以5．25％及2．5％次氯酸钠冲洗，6组在根管预备时辅以0．9％Nacl冲洗。冲洗前、冲洗后即刻及冲洗后72h分别取样培养。结果6组根管内的细菌量均显著下降。1、2组差异无统计学意义（P〉0．05），但均好于3组（P〈0．05）。4、5和6组差异均有统计学意义（P〈0．05）。根管冲洗后培养72h均有细菌生长。结论2．5％次氯酸钠基本可达到更高浓度的灭菌效果，但是经过机械预备和次氯酸钠化学消毒后的根管内仍有细菌残留。     

Effect of MTAD on Enterococcus faecalis-contaminated root canals of extracted human teeth

The purpose of this investigation was to compare the antimicrobial effect of MTAD (a mixture of a tetracycline isomer, an acid, and a detergent) with that of NaOCl with and without EDTA. Eighty-five extracted human teeth were contaminated with Enterococcus faecalis for 4 weeks. After biomechanical instrumentation using 1.3% or 5.25% NaOCl as root canal irrigant, the root canal and the external surface of each tooth were exposed to a 5-min application of MTAD, 1.3% NaOCl, 5.25% NaOCl or a 1-min application of EDTA followed by irrigation with 5 ml of 1.3% NaOCl or 5.25% NaOCl. Teeth or dentin shavings were cultured to determine presence or absence of the test bacteria. Fisher's exact test showed that the combination of 1.3% NaOCl as a root canal irrigant and MTAD as a final rinse was significantly more effective against E. faecalis than the other regimens. The chi2 test showed no difference between the other regimens.     

Comparison of the antimicrobial efficacy of 1.3% NaOCl/BioPure MTAD to 5.25% NaOCl/15% EDTA for root canal irrigation

The purpose of this study was to compare the antimicrobial efficacy of 1.3% NaOCl/BioPure MTAD to 5.25% NaOCl/15% EDTA for root canal irrigation. Twenty-six bilaterally matched pairs of human teeth were collected. The teeth were incubated with Enterococcus faecalis for 4 weeks. The teeth were divided into two experimental groups and one positive control group. The canals were instrumented and irrigated with either 5.25% NaOCl/15% EDTA or 1.3% NaOCl/BioPure MTAD. Bacterial samples were collected after instrumentation/irrigation and after additional canal enlargement. Statistical analysis of the data using the Wilcoxon Signed Rank test showed significant differences between the experimental groups. The first bacterial samples revealed growth in 0 of 20 samples with 5.25% NaOCl/15% EDTA irrigation and in 8 of 20 samples with 1.3% NaOCl/BioPure MTAD irrigation. Samples taken after additional canal enlargement revealed growth in 0 of 20 samples in 5.25% NaOCl/15% EDTA and in 10 of 20 samples in 1.3% NaOCl/BioPure MTAD group. This investigation showed consistent disinfection of infected root canals with 5.25% NaOCl/15% EDTA. The combination of 1.3% NaOCl/BioPure MTAD left nearly 50% of the canals contaminated with E. faecalis.     

Efficacy of various concentrations of NaOCl and instrumentation techniques in reducing Enterococcus faecalis within root canals and dentinal tubules

AIM: To evaluate the efficacy of 0.5%, 2.5% and 5.25% sodium hypochlorite (NaOCl) as intracanal irrigants associated with hand and rotary instrumentation techniques against Enterococcus faecalis within root canals and dentinal tubules. METHODOLOGY: A total of 180 extracted human premolar teeth were infected for 21 days with E. faecalis. The specimens were divided into 12 groups, as follows: group 1: 5.25% NaOCl + Hybrid technique (Valdrighi et al. 1998); group 2: 5.25% NaOCl + nickel-titanium (NiTi) rotary technique 4 mm shorter than the apex (by FOP-UNICAMP); group 3: 5, 25% NaOCl + NiTi rotary technique (Hero 642); group 4: 2.5% NaOCl +Hybrid technique; group 5: 2.5% NaOCl + NiTi rotary technique 4 mm shorter than the apex; group 6: 2.5% NaOCl + NiTi rotary technique (Hero 642); group 7: 0.5% NaOCl + Hybrid technique; group 8: 0.5% NaOCl + NiTi rotary technique 4 mm shorter than the apex; group 9: 0.5% NaOCl + NiTi rotary technique (Hero 642); group 10: sterile saline solution + Hybrid technique; group 11: sterile saline solution + NiTi rotary technique 4 mm shorter than the apex; group 12: sterile saline solution + NiTi rotary technique (Hero 642). Canals were sampled before and after preparation. After serial dilution, samples were plated onto brain heart infusion (BHI) agar, and the colony forming units (CFU) that were grown were counted. The teeth were sectioned into three thirds and dentine chips were removed from the canals with conical burs. The samples obtained with each bur were immediately collected into test tubes containing BHI broth, and were incubated at 37 degrees C and plated onto BHI agar. The CFU were counted and analysed. RESULTS: At all depths and thirds of the root canals and for all techniques used, 5.25% NaOCl was shown to be the most effective irrigant solution tested when dentinal tubules were analysed, followed by 2.5% NaOCl. No differences among concentrations in cleaning the canals were found. CONCLUSIONS: Especially at higher concentrations, NaOCl, was able to disinfect the dentinal tubules, independent of the canal preparation technique used.     

Evaluation of the effectiveness of sodium hypochlorite used with three irrigation methods in the elimination of Enterococcus faecalis from the root canal, in vitro

The effectiveness of 4.0% sodium hypochlorite (NaOCl) used with three irrigation methods in the elimination of Enterococcus faecalis from the root canal was tested in vitro . Root canals contaminated with E. faecalis were treated as follows: (i) irrigation with 2 mL of NaOCl solution and agitation with hand files; (ii) irrigation with 2 mL of NaOCl solution and ultrasonic agitation; (iii) irrigation with NaOCl alternated with hydrogen peroxide. Contaminated canals irrigated with sterile saline solution served as the control. Paper points used to sample bacteria from the root canals were transferred to tubes containing 5 mL of brain heart infusion (BHI) broth. Tubes were incubated and the appearance of broth turbidity was indicative of bacteria remaining in the root canal. There were no statistically significant differences between the experimental groups. However, NaOCl applied by the three methods tested, was significantly more effective than the saline solution (control group) in disinfecting the root canal.     

Chemomechanical reduction of the bacterial population in the root canal after instrumentation and irrigation with 1%, 2.5%, and 5.25% sodium hypochlorite

Given the importance of bacteria in the development of periradicular lesions, the eradication of the root canal infection is paramount in endodontic treatment. This study evaluated the in vitro intracanal bacterial reduction produced by instrumentation and irrigation with 1%, 2.5%, and 5.25% sodium hypochlorite (NaOCl) or saline solution. Root canals inoculated with Enterococcus faecalis were instrumented and irrigated with the solutions tested. Canals were sampled before and after preparation. After serial dilution, samples were plated onto Mitis salivarius agar, and the colony-forming units grown were counted. Inhibitory effects of the three NaOCl solutions on E. faecalis were also evaluated by means of the agar diffusion test. All test solutions significantly reduced the number of bacterial cells in the root canal (p < 0.05). There was no significant difference between the three NaOCl solutions tested (p > 0.05). Nonetheless, all NaOCl solutions were significantly more effective than saline solution in reducing the number of bacterial cells within the root canal (p < 0.05). The three NaOCl concentrations showed large zones of inhibition against E. faecalis. The results of this study suggest that regular exchange and the use of large amounts of irrigant should maintain the antibacterial effectiveness of the NaOCl solution, compensating for the effects of concentration.     

Evaluation of the antimicrobial effect of Er:YAG laser irradiation versus 1% sodium hypochlorite irrigation for root canal disinfection

This laboratory study evaluated Er:YAG laser antibacterial action in infected root canals. Forty-eight maxillary central incisors were used. After canal preparation, the teeth were autoclaved and divided into four groups: (1) non-treated teeth (control group); (2) teeth treated with NaOCl; (3) teeth irradiated with Er:YAG laser (7 Hz, 100 mJ, 80 pulses/canal, 11 sec) to the working length; (4) teeth irradiated similarly to, but 3 mm short, of the apex. The root canals from Groups 2, 3 and 4 were inoculated with 4 bacteria: Bacillus subtillus, Enterococcus faecalis, Pseudomonas aeruginosa, and Staphylococcus aureus, together with Candida albicans, and maintained for 24 h at 37 degrees C. All suspensions were adjusted to tube 2 of the MacFarland scale. The intracanal material was then collected with sterile paper points, which were placed in the canals for 5 min and then immersed in 5 ml of BHI medium. This was then seeded onto agar and stained by Gram's method. The NaOCl solutions and the Er:YAG laser irradiation to working length were effective against all five micro-organisms; however, 70% of the specimens irradiated 3 mm short of the apex remained infected.     

A comparison of the antimicrobial efficacy of NaOCl/Biopure MTAD versus NaOCl/EDTA against Enterococcus faecalis

The purpose of this investigation was to compare the antimicrobial efficacy of irrigating with 1.3% NaOCl/Biopure MTAD versus irrigation with 5.25% NaOCl/15% EDTA in the apical 5 mm of roots infected with Enterococcus faecalis. Bilaterally matched human teeth were sterilized and inoculated with E. faecalis. After chemomechanical root canal preparation, the root-ends were resected and pulverized in liquid nitrogen to expose E. faecalis in dentinal tubules or other recesses away from the main root canal system. The number of colony forming units (CFU) of E. faecalis per mg was determined from the pulverized root-ends. No significant differences were seen (t = 0.70, p = 0.495) between the number of colony forming units of E. faecalis for teeth irrigated with 5.25% NaOCl/15% EDTA (mean 131 +/- 291 CFU/mg) versus those teeth irrigated with 1.3% NaOCl/Biopure MTAD (mean 187 +/- 237 CFU/mg). This study demonstrated that there is no difference in antimicrobial efficacy for irrigation with 5.25% NaOCl/15% EDTA versus irrigation with 1.3% NaOCl/Biopure MTAD in the apical 5 mm of roots infected with E. faecalis.     

Comparison of antibacterial and toxic effects of various root canal irrigants

AIM: To compare the antibacterial properties and toxicity of 5.25% sodium hypochlorite (NaOCl), 2% chlorhexidine gluconate and 0.2% chlorhexidine gluconate plus 0.2% cetrimide (Cetrexidin); Vebas, San Giuliano, Milan, Italy). METHODOLOGY: The antibacterial effects of the irrigants in vitro were examined after 5 min and 48 h in freshly extracted human teeth with single roots, whose canals were infected by Enterococcus faecalis ATCC 29212. In a separate in vivo study, bacterial culture samples were collected before treatment from the infected root canals of deciduous teeth containing necrotic pulp tissue. Irrigants were used to clean the canals which were then left empty for 48 h. Aerobic/facultative anaerobic and anaerobic bacterial growth were compared before and 48 h after irrigation. Finally, the toxic effects of the irrigants were assessed by injecting them into the subcutaneous tissues of rats. The inflammatory reactions that occurred 2 h, 48 h and 2 weeks after the injections were evaluated. RESULTS: In the laboratory study, the 2% chlorhexidine gluconate and Cetrexidin were significantly more effective on E. faecalis than the 5.25% NaOCl at 5 min (P < 0.05). Similarly, in the in vivo study, 2% chlorhexidine gluconate and Cetrexidin were significantly more effective on anaerobic bacteria than the 5.25% NaOCl at 48 h (P < 0.05). At the end of 2 weeks, the toxicity of the NaOCl solution was greater than that of the other irrigants (P < 0.05). CONCLUSIONS: Cetrexidin and 2% chlorhexidine gluconate were more effective, and had more residual antibacterial effects and lower toxicity than 5.25% NaOCl solution.     

Inhibitory activity of root canal irrigants against Candida albicans, Enterococcus faecalis and Staphylococcus aureus

The present study evaluated the antimicrobial activity of three root canal irrigants against Enterococcus faecalis, Candida albicans, and Staphylococcus aureus. These microorganisms were incubated in the presence of citric acid (6 and 10%), EDTA (17%), and NaOCl (0.5, 1.0, 2.5, and 5.25%). Agar diffusion tests were performed and redox indicator resazurin was used to evaluate the inhibitory effect of the irrigants on the metabolic activity of these microorganisms. The mean diameters of the inhibition zones for the C. albicans cultures were 11.6 mm (17% EDTA), 5.5 mm (0.5% NaOCl), 12.9 mm (1% NaOCl), 22.1 mm (2.5% NaOCl), and 28.5 mm (5.25% NaOCl). The mean diameters of the inhibition zones for E. faecalis were 2.8 mm (1% NaOCl), 5.4 mm (2.5% NaOCl), and 8.3 mm (5.25% NaOCl). For S. aureus, the mean values were 8.0 mm (17% EDTA), 3.0 mm (1% NaOCl), 8.8 mm (2.5% NaOCl), and 10.0 mm (5.25% NaOCl). Most of the irrigant solutions presented effective antimicrobial activity against C. albicans. A high inhibitory effect on the metabolic activity of E. faecalis was detected when the microorganisms were incubated with 17% EDTA. The same result was reached when S. aureus was incubated in the presence of > 2.5% NaOCl. Altogether, these results indicate that 2.5% and 5.25% NaOCl are microbicides against S. aureus while 0.5% and 1% NaOCl are only microbiostatic against the tested bacteria. The 6% and 10% citric acid as well as 17% EDTA did not affect the viability of any of the assayed microorganisms.