Growth fractions in malignant non-Hodgkin's lymphomas (NHL) as determined in situ with the monoclonal antibody Ki-67

The proportion of proliferating cells in malignant non-Hodgkin's lymphomas (NHL) was determined in situ by immunostaining with the monoclonal antibody Ki-67, which reacts with a nuclear antigen that is present only in proliferating cells. A highly significant correlation could be demonstrated between the proportion of Ki-67 positive cells and the classification into high and low-grade NHL according to the Kiel classification. 93.8 per cent of high-grade and 88.5 per cent of low-grade malignancies were correctly allocated to these groups using the percentage of Ki-67 positive cells as discriminant parameter. On the basis of the medians, the degree of proliferation also paralleled the succession of entities within the Kiel classification. Within most of these different entities, however, the ranges of Ki-67 positive cells varied considerably, indicating that the growth fractions within these groups are rather heterogeneous. Thus it might be useful to determine the growth fraction of each individual case of NHL, because this might be of prognostic value.     

Cell kinetic analysis of non-Hodgkin's lymphomas using in vivo iododeoxyuridine incorporation and flow cytometry

The aim of this study was to analyse dynamic cell proliferation parameters in non-Hodgkin's lymphomas. Sixty-one patients with newly diagnosed or with recurrent disease were given iododeoxyuridine (IdUrd) intravenously near 4 h prior to tumour biopsy. After staining with an IdUrd reactive antibody and propidium iodide, S-phase fraction (SPF), labelling index (LI), S-phase duration time (Ts) and potential tumour doubling time (Tpot) were determined by flow cytometry. Thirty-eight samples, 15 low grade (LGM) and 23 high grade (HGM) malignant lymphomas, were possible to evaluate. Twenty-three cases were excluded due to aneuploidy, insufficient amount of material or technical problems. Tpot values varied between 0.8–32.9 days (mean 7.0 days). HGM lymphomas had shorter mean Tpot times than LGM lymphomas (4.8 versus 10.4 days, p=0.05). For Ts the range was 4.2–20.1 h (mean 9.1 h), and a difference between the two histological groups was demonstrated with a longer mean Ts for HGM compared with LGM cases (10.0 versus 7.8 h, p=0.04). Tpot showed a negative correlation with SPF (P=0.003), and Ts demonstrated a positive correlation to SPF (p=0.02). The clinical significance of the dynamic cell proliferation parameters studied remains to be clarified, but the interelationships between Ts/SPF and Ts/morphologic subtype might be factors of interest for future prognostic studies in malignant lymphomas.     

Quantitative study of KI-67 antibody staining in 46 T-cell malignant lymphomas using image analysis.

Image analysis with a SAMBA 2005 (ALCATEL-TITN, Co) was used to quantify the Ki-67 stained area percentage in 46 T-cell malignant lymphomas (T-ML), classified according to the updated Kiel classification. This parameter demonstrated correlation with the number of Ki-67-positive cellular profiles (r = 0.88, P less than 0.001) and was more reproducible than cell counting. A significant difference was found between low and high grade T-ML (mean values +/- SEM respectively of 10.20 +/- 1.82 per cent and 25.63 +/- 3.15 per cent). The most interesting findings were that: (1) AILD-type T-ML showed an intermediate proliferation rate (15.55 +/- 2.72 per cent) between pleomorphic T-ML with medium and with large cells (respectively 12.53 +/- 3.64 per cent and 22.43 +/- 3.46 per cent), both of which belong to the high grade malignancy group. This finding is in accordance with the poor prognosis of this subtype despite its classification in the low grade malignancy group. (2) Subclassification of the pleomorphic MLs according to the predominance of small, medium or large cells, demonstrated significant differences between these three subtypes. However, the great overlap of values between pleomorphic T-ML with medium and with large cells, seems to indicate that the subclassification of these two subtypes is less valid. (3) A wide range of values with overlap was observed in AILD-type ML, in pleomorphic with medium or large cells and in lymphoblastic T-ML: for these T-ML with variable survival courses, the Ki-67 area percentage, one parameter of proliferative activity, appears worth studying as a prognostic factor.     

H3K27M和Ki-67在小儿脑胶质瘤中的表达及相关性

目的:探讨H3K27M和Ki-67在小儿脑胶质瘤组织中的表达及意义。方法:收集2013年11月至2018年6月清华大学玉泉医院38例小儿脑胶质瘤和10例正常脑组织,采用免疫组织化学检测H3K27M和Ki-67表达并结合相关统计学方法进行分析。结果:H3K27M阳性表达与性别、年龄、肿瘤部位无显著性差异(P>0.05),但与肿瘤病理级别有显著性相关(P<0.05)。Ki-67在胶质瘤中阳性表达率为100%,Ki-67高标记指数与性别、年龄无显著性相关(P>0.05),与病理级别、肿瘤部位有显著性相关(P<0.05)。患儿的性别、年龄与生存时间之间无显著性差异(P>0.05),病理分级、肿瘤部位、H3K27M蛋白阳性、Ki-67高标记指数在患儿生存时间之间有显著性差异(P<0.05)。结论:小儿脑胶质瘤中H3K27M、Ki-67的表达水平对小儿脑胶质瘤的病理分级、预后具有提示意义,且H3K27M、Ki-67蛋白在胶质瘤组织中的表达具有相关性,为研究胶质瘤的发生发展及靶向治疗提供了线索。     

Distinctive clinical features between lymphoblastic and immunoblastic non-Hodgkin's malignant lymphomas (Kiel's classification) in a series of 141 patients

Out of a series of 427 NHML based on the Kiel classification and without previous treatment, 186 were of high grade malignancy and among them were 83 lymphoblastic (LB) and 58 immunoblastic (IB) lymphomas which were included in this study. The LB and IB, which represent the majority of high grade NHML, were compared regarding their clinical features and courses. Eight main criteria were significantly different between these two groups: age of patient, immunological history, mediastinal and pleural involvement, primary or secondary involvement of the bone marrow, secondary leukaemia, secondary meningeal involvement, and involvement of the facial region. Some of these differences (mediastinum and meningeal involvement) are related to subtypes within LB. The differences between these two groups suggest that different treatment programs may be warranted.     

Association between Ki-67 Labeling index and Histopathological Grading of Glioma in Indonesian Population

Background: Gliomas are the most frequent primary brain tumors. According to World Health Organization guidelines, gliomas are graded into four groups (Group I-IV). This histological grading will determine prognosis and treatment of the patient. Morphological criteria are not always accurate. Tumor proliferation index is a potent quantitative marker for tumor behavior and prognosis, also it’s the basis of gliomagenesis. Ki-67 immunohistochemistry examination for determining proliferation index has been suggested as an ancillary marker in deciding the definitive grading of glioma. Objective: To analyze the correlation between Ki-67 labeling index and histopathological grading of glioma in Indonesian population. Methods: One hundred and six formalin fixed-paraffin embedded tissue of glioma patients were collected from 4 different hospitals. Expression of Ki-67 was detected using immunohistochemistry staining and the labeling index was counted. The association between Ki-67 labeling index and histopathological grading was analyzed. Results: Age range of patient were 1-73-years old, with male predominance (55.70%). Glioblastoma was the most common diagnosis accounting for 41.51% of all samples. Ki-67 labeling index cut point of 6.35% was obtained and significantly sensitive and specific for determining low- or high-grade glioma (p<0.001). Conclusion: A significant association between Ki-67 labeling index and histopathological grading in Indonesian glioma patients has been revealed. The result of this study may be used to improve diagnostic and grading accuracy of glioma cases in Indonesia, especially in small biopsy specimens.     

Determination of proliferative fractions detected by immunohistochemistry with the monoclonal antibodies to bromodeoxyuridine and Ki-67 in malignant lymphomas

The stainability of proliferating cells with monoclonal antibodies to both bromodeoxyuridine (BrdU) and Ki-67 were compared in 22 malignant lymphomas and six related conditions. A significant correlation was observed (r = 0.78) between the positivities of proliferating cells determined using these two monoclonal antibodies. The growth fractions determined with the antibodies were usually related to the histologic classification, but labelling index was more varied in diffuse large cell type lymphoma. BrdU labelling was well correlated with the Working Formulation, and more useful as a prognostic indicator. On the other hand, Ki-67 was useful for broad recognition of cell proliferative ability.     

Proliferative activity in gastric cancer determined with cell cycle-related monoclonal antibodies Ki-67 and p105: analysis by flow cytometry.

The proliferative activity of gastric cancer cells was determined by DNA flow cytometric (FCM) analysis and labeling rates of Ki-67 and monoclonal antibodies and proliferation-associated nuclear antigen (p105) autoantibodies in 28 patients with fresh human gastric cancer cells. By setting the cutoff line at the level as used in a negative control study without primary antibody in the same sample, the Ki-67 and p105 labeling rates were calculated by the dual fluorescence analysis. A total of 43 experiments was performed on FCM analysis for each antigen: 28 with Ki-67 and 15 with p105. The mean Ki-67 labeling rate of gastric cancer cells was 45.1% (13.9-76.3%). The Ki-67 labeling rates were significantly higher for larger size tumor, peritoneal metastasis, and advanced clinical stage. A significant correlation was found between Ki-67 labeling rate and p105 labeling rate (P < 0.05). Bivariate FCM may be an easy method for obtaining useful information of cell kinetics.     

Flow cytometric analysis of DNA ploidy in large cleaved cell lymphomas

The results of nuclear DNA content analysis of a series of 28 patients with large cleaved follicular centre cell non-Hodgkin's lymphomas (NHL) are reported. DNA aneuploidy was found in 11 (39 per cent) cases. The DNA indices of the DNA-aneuploid peaks ranged from 1.14 to 2.28. Seven (25 per cent) cases were tetraploid. DNA ploidy was not associated with prognosis. The percentage of S phase cells (SPF) ranged from 2.0 to 30.5 per cent (median 5.1 per cent). Lymphoma patients with SPF higher than 9.7 per cent had a worse survival rate than patients with lymphoma with less than 9.7 per cent S phase cells but the difference did not reach statistical significance. The results of the DNA ploidy and SPF were comparable to those of intermediate and high grade malignancy NHLs.     

IMP3和Ki-67在人脑胶质瘤中的表达及其相关性

目的:探讨胰岛素样生长因子Ⅱ mRNA结合蛋白3（insulin-like growth factor-Ⅱ mRNA-binding protein 3,IMP3）和Ki-67在脑胶质瘤组织中的表达及意义。方法:收集神经外科手术切除后经病理证实的胶质瘤标本126例,另收集脑外伤行内减压术患者的正常脑组织20例作为对照组,采用免疫组化SP法检测IMP3、Ki-67在不同级别胶质瘤组织和正常脑组织中的表达水平;应用Spearman秩相关分析IMP3和Ki-67在胶质瘤组织中表达水平的相关性。结果:IMP3、Ki-67在胶质瘤组织中阳性率为53.17%、56.35%;IMP3、Ki-67在Ⅲ、Ⅳ级胶质瘤中阳性率均高于Ⅱ级(P＜0.05)。免疫组化半定量评分结果显示:正常脑组织和Ⅰ级胶质瘤中IMP3、Ki-67均为阴性表达;Ⅱ级胶质瘤中IMP3、Ki-67阳性表达多为弱阳性;Ⅲ、Ⅳ级胶质瘤中IMP3、Ki-67多数表达为弱阳性、中度阳性和强阳性;Ⅲ 级与Ⅱ级、Ⅳ级与Ⅱ级胶质瘤中IMP3、Ki-67阳性表达率比较,差异有统计学意义(P＜0.05);IMP3、Ki-67阳性表达均与胶质瘤的病变程度密切相关(P＜0.05)。胶质瘤组织中IMP3、Ki-67阳性表达率在不同性别、年龄、肿瘤大小、肿瘤位置和病理类型之间差异均无统计学意义(P＞0.05);IMP3和Ki-67在胶质瘤组织中阳性表达率呈正相关(P＜0.05)。结论:IMP3、Ki-67分别可以有效地反映胶质瘤的病变程度,两者联合诊断可以客观反映胶质瘤病变程度,两者联合检测可能成为胶质瘤生物标记的新指标。     

Ki-67与p53在肺硬化性血管瘤中的表达及其临床意义

 【摘要】　目的　分析Ki-67与p53蛋白在肺硬化性血管瘤（PSH）表面被覆细胞和间质多边形细胞中的表达以及Ki-67、p53蛋白与PSH的生物学行为的关系。方法　随机数字表法抽取21例PSH患者手术切除标本,应用免疫组织化学双标法检测21例标本Ki-67和CK8／18蛋白表达,免疫双荧光组织化学染色技术检测9例p53和AE1／AE蛋白的表达。结果　CK8／18和AE1／AE3定位于细胞膜,表达于表面被覆细胞; Ki-67与p53定位于细胞核,在两种细胞均见表达。其中Ki-67在表面被覆细胞中表达阳性率＜1 %,在间质多边形细胞中阳性率为1 %～10 %;p53主要表达于间质多边形细胞（单信号6例）,仅个别被覆细胞阳性（双信号3例）。结论　PSH中的间质多边形细胞和表面被覆细胞具有形态和免疫表型的差异,间质多边形细胞的增殖指数和基因突变均高于被覆细胞,PSH的生物学行为可能主要决定于间质多边形细胞。     

Inclusion of rituximab in treatment protocols for non-Hodgkin's lymphomas and risk for progressive multifocal leukoencephalopathy

Objectives. Rituximab is an anti-CD20 monoclonal antibody that promotes better treatment outcomes in patients with non-Hodgkin's lymphoma (NHL). Case series of progressive multifocal leukoencephalopathy (PML) in patients receiving rituximab within polychemotherapy regimens have led to the introduction of a black box warning, but no risk estimation has ever been provided. Methods. We performed a retrospective, monocentric cohort study on 976 NHL patients diagnosed in 1994-2008, including 517 patients who received at least one dose of rituximab. Results. Inclusion of rituximab into standard chemotherapy regimens for NHL caused a significantly higher incidence of PML cases (rate difference, 2.2 every 1,000 patient-years; 95% confidence interval, 0.1-4.3). Interpretation. Based on this finding, clinical surveillance of PML-related symptoms is recommended in NHL patients exposed to rituximab.     

Combination of shear wave elastography and Ki-67 index as a novel predictive modality for the pathological response to neoadjuvant chemotherapy in patients with invasive breast cancer

PurposeThis study evaluated shear wave elastography (SWE) and SWE combined with the Ki-67 index as novel predictive modalities for the pathological response of invasive breast cancer to neoadjuvant chemotherapy (NAC).MethodsThe prospective study recruited 66 eligible patients from July 2014 to November 2015. Tumour stiffness, which corresponds with tumour progression and invasiveness, was assessed by quantitative SWE 1 d before biopsy (time point t0, elasticity E0), 1 d before next NAC cycle (t1−t5, E1−E5), and 1 d before surgery (t6, E6). The relative changes in SWE parameters after the first and second NAC cycles were considered as the variables [ΔE (t1), ΔE (t2)]. The pathological response was classified according to the residual cancer burden (RCB) protocol. Correlations between RCB scores and variables were evaluated. The predictive diagnostic performances of SWE parameters, Ki-67 index, and the predictive RCB (predRCB) score determined by a linear regression model were compared.ResultsSome immunohistochemical and molecular factors and SWE parameters were significantly different among the three RCB groups. The ΔE_mean (t2) and Ki-67 had significantly better diagnostic performance than other parameters regarding predicting the pathological response (the RCB-I response and RCB-III resistance). However, the correlation between ΔE_mean (t2) and Ki-67 index was significantly weaker as a diagnostic predictor (r = 0.29). We generated a new predictive modality, predRCB, which is a multivariable linear regression model that combines ΔE_mean (t2) and the Ki-67 index. The predRCB modality showed better diagnostic performance than SWE parameters and Ki-67 index alone.ConclusionOur findings highlight the potential utility for adding the Ki-67 index to the SWE results, which may improve the predictive power of SWE and facilitate personalising the treatment regimens of patients with breast cancer. These results should be validated in the future by performing a multicentre prospective study with a larger cohort.     

错配修复蛋白联合血清肿瘤标志物与Ki-67增殖指数对结直肠癌预后的临床价值

背景与目的：结直肠癌是消化系统常见的恶性肿瘤之一,近年来中国结直肠癌发病率显著升高。各项临床与病理学指标对于结直肠癌的诊断、临床分期及预后的判断提供了帮助。探讨错配修复蛋白表达联合血清肿瘤标志物与Ki-67增殖指数在结直肠癌中的相关性及对预后判断的临床价值。方法：收集复旦大学附属华东医院2014年7月—2018年6月收治的234例结直肠癌手术患者,分析其术前血清标本中肿瘤标志物癌胚抗原（carcinoembryonic antigen,CEA）、糖类抗原（carbohydrate antigen,CA）19-9、CA72-4、CA12-5水平及手术标本组织中的Ki-67增殖指数与错配修复蛋白的表达率,与结直肠癌临床病理学特征和预后的关系。结果：在234例结直肠癌患者术后标本中发生错配修复蛋白缺失表达（deficient mismatchrepair,dMMR）的共29例（12.4%）,错配修复蛋白正常表达（proficientmismatchrepair,pMMR）的有205例（87.6%）。在临床病理学指标的相关性分析中dMMR组与pMMR组在肿瘤原发部位、分化类型、分期、T分期、淋...     

JMJD3和Ki-67定量检测与乳腺浸润性导管癌分子分型及临床病理学特征的关系

背景与目的：对乳腺癌分子标志物的研究有助于乳腺癌患者的诊断及预后判断，探讨乳腺浸润性导管癌中JMJD3表达和Ki-67增殖指数与分子分型及临床病理学特征的关系。方法：采用免疫组织化学法检测河南中医药大学人民医院的57例正常乳腺组织、38例乳腺低级别导管内癌、52例乳腺高级别导管内癌及150例乳腺浸润性导管癌原发灶中JMJD3表达和Ki-67增殖指数，用图像分析软件对JMJD3表达和Ki-67增殖指数进行定量测试，分析JMJD3表达和Ki-67增殖指数与乳腺浸润性导管癌分子分型及临床病理学特征的关系。结果：JMJD3在正常乳腺组织、乳腺低级别导管内癌、乳腺高级别导管内癌和乳腺浸润性导管癌中表达依次降低，Ki-67增殖指数在上述组织中表达依次升高。JMJD3表达和Ki-67增殖指数与乳腺浸润性导管癌组织学分级、淋巴结转移状况、pTNM分期及分子亚型均有关（P<0.05）。受试者工作特征（receiver operating characteristic，ROC）曲线显示，JMJD3诊断阈值≤10.87，Ki-67诊断阈值≥8.08时，诊断乳腺癌的灵敏度和特异度均较高。Spearman相关分析显示，JMJD3表达与Ki-67增殖指数呈显著负相关（r _P =0.984，P=0.000）。结论：JMJD3和Ki-67在乳腺癌发生、发展中扮演着重要角色。JMJD3表达与Ki-67增殖指数呈负相关，提示JMJD3的高表达能够抑制乳腺癌细胞的增殖，其具体分子生物学机制有待进一步深入研究。     

Methylation of miR-155-3p in mantle cell lymphoma and other non-Hodgkin's lymphomas

Mantle cell lymphoma (MCL) is an aggressive B-cell non-Hodgkin''s lymphoma (NHL). In cancers, tumor suppressive microRNAs may be silenced by DNA hypermethylation. By microRNA profiling of representative EBV-negative MCL cell lines before and after demethylation treatment, miR-155-3p was found significantly restored. Methylation-specific PCR, verified by pyrosequencing, showed complete methylation of miR-155-3p in one MCL cell line (REC-1). 5-aza-2′-deoxycytidine treatment of REC-1 led to demethylation and re-expression of miR-155-3p. Over-expression of miR-155-3p led to increased sub-G1 apoptotic cells and reduced cellular viability, demonstrating its tumor suppressive properties. By luciferase assay, lymphotoxin-beta (LT-β) was validated as a miR-155-3p target. In 31 primary MCL, miR-155-3p was found hypermethylated in 6(19%) cases. To test if methylation of miR-155-3p was MCL-specific, miR-155-3p methylation was tested in an additional 191 B-cell, T-cell and NK-cell NHLs, yielding miR-155-3p methylation in 66(34.6%) including 36(27%) non-MCL B-cell, 24(53%) T-cell and 6(46%) of NK-cell lymphoma. Moreover, in 72 primary NHL samples with RNA, miR-155-3p methylation correlated with miR-155-3p downregulation (p = 0.024), and LT-β upregulation (p = 0.043). Collectively, miR-155-3p is a potential tumor suppressive microRNA hypermethylated in MCL and other NHL subtypes. As miR-155-3p targets LT-β, which is an upstream activator of the non-canonical NF-kB signaling, miR-155-3p methylation is potentially important in lymphomagenesis.     

3-氯-4-二氯甲基-5-羟基-2(5氢)-呋喃酮诱导人胚胎肝细胞ras基因突变

背景与目的:研究饮水氯化消毒副产物3-氯-4-二氯甲基-5-羟基-2(5氢)-呋喃酮(3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone,MX)对体外培养的人胚胎肝细胞(L-02细胞)ras基因突变的诱导。材料与方法:MX染毒剂量为300μmol/L,以二甲基亚砜(DMSO)做溶剂对照,将L-02细胞连续染毒培养12d后,收获细胞提取基因组DNA,应用PCR-克隆测序法检测ras基因(K-ras、H-ras、N-ras)12、13、61密码子是否存在突变。结果:MX染毒组H-ras基因57密码子的GAT置换成GGT,未检测到K-ras、N-ras及H-ras12、13、61密码子突变,DMSO溶剂对照组相应的ras基因目的片段均未检测到突变。结论:MX可能诱导L-02细胞ras基因突变。     

3-氯-4-二氯甲基-5-羟基-2(5氢)-呋喃酮对L-02细胞增殖周期和凋亡的影响

背景与目的：研究饮水氯化消毒副产物3-氯-4-二氯甲基-5-羟基-2（5氢）-呋喃酮（3-chloro-4-（dichloromethyl）-5-hydroxy-2[5H]-Furanone,MX）对人胚胎肝细胞株（Human derived fetal hepatocytes,L-02）增殖周期和凋亡的影响。材料与方法：L-02细胞经MX（10、30、100、3009mol／L）染毒3h后继续培养0、3、6、9h,用吖啶橙／溴化乙啶染色法和流式细胞术分析凋亡和增殖周期。结果：L-02细胞在周期不同时相的分布与MX染毒剂量和染毒后继续培养的时间有关;染毒后继续培养0h时,30μmol／LMX诱导S期细胞显著增加,10、100、300μmol／L MX诱导G2期细胞显著增加（P〈0．01）;在染毒后继续培养的第9h,10μmol／L MX诱导G2期细胞显著增加,30、100、300μmol／L MX诱导S期细胞显著增加（P〈0．05）。MX各剂量组L-02细胞的凋亡率,随继续培养时间的延长,呈现先升后降的趋势,在6h时达最大值,与溶剂对照组相比,差异有统计学意义（P〈0．01）;30μmol／LMX剂量组L-02细胞的凋亡率最高。结论：MX可诱导L-02细胞增殖周期G2期、S期阻滞和凋亡,细胞周期阻滞的类型与MX染毒剂量和染毒后继续培养的时间有关。     

The role of aberrant crypt foci induced by the two heterocyclic amines 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ) and 2-amino-1-methyl-6-phenyl-imidazo[4,5-b]pyridine (PhIP) in the development of colon cancer in mice

Aberrant crypt foci (ACF) have recently been identified as early putative preneoplastic lesions which appear in the colons of experimental animals treated with colon carcinogens. In a recent study the two heterocyclic amines, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-1-methyl-6-phenyl-imidazo[4,5-b]pyridine (PhIP) were shown to be able to induce ACF in the colon of mice after, respectively, 4 and 10 weeks of exposure. In spite of the induction of ACF in colon of mice, IQ and PhIP have not been found to have colon as target organ in carcinogenicity studies. Therefore, one may question that ACF induced by IQ and PhIP in mice represent early stages of colon cancer. In order to investigate the possible role of PhIP- and IQ-induced aberrant crypt foci in the development of colon cancer in mice, colons from mice participating in other IQ- and PhIP-studies of much longer duration were analyzed for ACF. The results of these studies showed that the number of ACF increased statistically significantly over time, and that the small ACF were predominant (95–100%) at all time-points. In conclusion, this finding suggests that the detection of a high number of ACF with low crypt multiplicity (1–3 AC/Focus) in mice colon after IQ- or PhIP-treatment is not indicative for the end-point colon cancer, and thus supports the hypothesis that only the presence of a high number of ACF with high crypt multiplicity is predictive for tumor outcome.