Editor's choice: Better see to better agree: phosphohistone H3 increases interobserver agreement in mitotic count for meningioma grading and imposes new specific thresholds

Background

Mitotic count on hematoxylin and eosin (H&E)–stained slides is a crucial diagnostic criterion in meningioma grading. However, mitosis assessment on H&E slides can be impaired by technical factors and by pathologist''s experience. Phosphohistone H3 (PHH3) serine-10 is a mitosis-specific antibody that has proven to facilitate mitotic count in various tumors.

Methods

A series of 70 meningiomas (15 grade I, 40 grade II, 15 grade III) was used to validate PHH3 intra- and interobserver reproducibility and to identify PHH3-specific mitotic thresholds. Four pathologists with different experience in neuropathology counted mitoses on both H&E- and PHH3-stained slides.

Results

H&E and PHH3 mitotic rates were highly correlated (Pearson''s r = 0.92, P < .0001). PHH3 mitotic counts had both a good mean interobserver correlation (R_m = 0.83) and a good intraclass correlation (0.78), higher than H&E mitotic indices (R_m = 0.77, intraclass correlation = 0.71). After further stratification of meningiomas according to World Health Organization grade, PHH3 performed better in terms of interobserver concordance (Kendall''s W = 0.761) compared with H&E (Kendall''s W = 0.697). Referring to the same meningioma groups identified by World Health Organization grade as the gold standard, the volume under the receiver operator characteristic surface was 0.91, indicating a very good diagnostic ability of PHH3 scores in discriminating the 3 meningioma groups. The 2 optimal PHH3-specific cutoff values were 6.61 and 22.02.

Conclusion

PHH3 staining is a useful diagnostic complementary tool to standard H&E mitotic count, optimizing intra- and interobserver reproducibility. PHH3-specific mitotic thresholds should be adopted to avoid overgrading of meningioma when ancillary methods are employed.     

Clinicopathological evaluation of cyclooxygenase-2 expression in meningioma: immunohistochemical analysis of 76 cases of low and high-grade meningioma

Tumorigenic activity of cyclooxygenase-2 (COX-2), a rate-limiting enzyme in the production of prostaglandins (PGs), has been proved for some types of cancer, including brain tumors. We evaluated expression of COX-2 in meningioma, one of the most common intracranial tumors in adults which accounts for 24–30 % of intracranial tumors. We performed immunostaining for COX-2 in 76 cases of meningioma consisting of 44 cases of low-grade (WHO Grade I) and 32 cases of high-grade (29 cases of Grade II and 3 cases of Grade III) meningioma, and evaluated COX-2 expression levels on the basis of staining intensity and proportion in tumor cells. The expression level of COX-2 in meningioma cells was significantly correlated with WHO grade (P = 0.0153). In addition, COX-2 expression was significantly correlated with MIB-1 labeling index for all 76 cases of meningioma (P = 0.0075), suggesting tumor promotion by COX-2 in meningioma progression. Our results may indicate the therapeutic value of non-steroidal anti-inflammatory drugs against meningioma, especially for patients with elevated proliferation, to regulate the tumorigenic activity of COX-2 in meningioma cells.     

Molecular genetic analysis of chromosome 22 in 81 cases of meningioma

Constitutional and tumor tissue genotypes from 81 unrelated patients with meningioma were compared at 25 polymorphic loci (restriction fragments length alleles) on chromosome 22. Thirty tumors (37%) retained the constitutional genotype along chromosome 22, a finding consistent with no detectable aberrations on chromosome 22 as studied. Forty-two tumors (52%) showed loss of one allele at all informative loci consistent with monosomy 22 in the tumor DNA. The remaining 9 tumors (11%) showed retained constitutional heterozygosity in the tumor DNA at one or more centromeric loci and loss of the heterozygosity at other telomeric loci, which is consistent with variable terminal deletions of one chromosome 22q in the tumor DNA. The localization of breakpoints in these 9 cases with deletions suggests that a meningioma locus is localized distal to myoglobin locus, within 22q12.3-qter. The male cases showed a higher percentage of tumors with no detectable aberrations on chromosome 22, a finding which may suggest that tumors of males have preferentially smaller rearrangements on chromosome 22q than those of females or that the male and female cases with no detected aberrations have another mechanism of oncogenesis. In view of the recent findings on the localization of the neurofibromatosis-2 gene on chromosome 22, the data from case 11 of our series suggests that the meningioma and the neurofibromatosis-2 loci are separate entities.     

Potential usefulness of 99mTc-labeled monoclonal antibody A7 for immunoscintigraphy of human pancreatic carcinoma

BACKGROUND: To improve survival rates of patients with pancreatic carcinoma, it is critical to develop new technology for early and certain diagnosis. One possibility is the application of radioimmunoscintigraphy using radiolabeled monoclonal antibodies. MATERIALS AND METHODS: Monoclonal antibody A7 (Mab A7) was labeled with technetium-99m (99mTc) and injected into athymic nude mice bearing human pancreatic carcinoma xenografts in order to examine its usefulness for radioimmunoscintigraphy. RESULTS: The binding activity of 99mTc labeled Mab A7 was nearly identical to that of non-labeled Mab A7. When 99mTc-labeled Mab A7 was injected intravenously into tumor-bearing nude mice, tumor accumulation of 99mTc-labeled Mab A7 increased until 24 hours after injection and then decreased slowly. The tumor tissue/blood ratio of radioactivity was significantly greater than that of normal mouse IgG. CONCLUSION: These results suggest that 99mTc-labeled Mab A7 is suitable for radioimmunoscintigraphy of human pancreatic carcinoma.     

岩斜区脑膜瘤51例显微外科处理的回顾性分析

目的:探讨如何提高岩斜区脑膜瘤手术治疗的效果.方法:回顾性分析51例岩斜区脑膜瘤患者手术治疗的临床资料,比较不同的手术入路特点、岩斜区脑膜瘤侵袭海绵窦的处理策略及神经功能的保护.结果:经岩骨-迷路后-乙状窦前入路26例,枕下乙状窦后入路11例,颞下入路7例,联合入路4例,分期手术3例.根治性切除(GTR)20例,次全切除(NTR)23 例,大部分切除(STR)8例.术后1～2周评估神经功能,19例(37%)出现恶化(指出现新的神经功能障碍或原有神经功能障碍加重),26例(51%)同术前,6例(12%)术后改善.术后死亡2例,颞叶水肿2例,脑脊液漏3例.结论:个体化选择设计岩斜区脑膜瘤的手术入路及手术策略对减少神经功能损伤提高疗效有重要意义.     

微囊型脑膜瘤2例临床病理学特征分析并文献复习

背景与目的：微囊型脑膜瘤是脑膜瘤的罕见亚型,国内外报道较为少见,且多以个案报道为主。本研究旨在探讨微囊型脑膜瘤的临床病理学特点及鉴别要点。方法：回顾分析2例微囊型脑膜瘤的临床表现、组织学形态及免疫组织化学表型,并复习相关文献。结果：2例均为中年患者,男性和女性各1例,男性患者因外伤引起脑出血就诊;女性患者因头痛就诊,病变分别位于左侧额部和右侧额部,最大径分别为5.75和5.47 cm。影像学改变均示“脑外肿瘤”。镜下瘤细胞排列疏松,形成大小不等的微囊,腔内含粉染之浆液,肿瘤细胞具有空泡状细胞质和细长的细胞质突起,典型的漩涡状结构和砂粒体少见。免疫组织化学示,波形蛋白（vimentin）、上皮膜抗原（epithelial membrane antigen,EMA）和孕激素受体（progesterone receptor,PR）均呈阳性表达,S-100、胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）、CD34和肌酸激酶（creatine kinase,CK）均呈阴性表达,Ki-67阳性率达3%~5%。2例患者均行手术治疗,术后均随访观察,随访时间为5~6个月,截至2018年1月,2例患者一般情况良好,均未见肿瘤复发。结论：微囊型脑膜瘤为脑膜瘤的特殊亚型,依据病理组织学形态及免疫组织化学可明确诊断。目前治疗主要以手术治疗为主,预后良好。     

侧脑室脑膜瘤的MRI诊断及鉴别诊断(附7例病例报告)

目的 探讨侧脑室脑膜瘤MRI表现及其与其它侧脑室内肿瘤的鉴别诊断。方法 对经手术病理证实的7例侧脑室脑膜瘤的MRI及临床、病理资料进行回顾性分析。结果 侧脑室脑膜瘤病人多以头痛、头晕为主要症状。肿瘤多位于左侧脑室三角区,呈类圆形、分叶状,6例良性脑膜瘤MRI表现为T1WI等信号,T2WI等或稍高信号,信号及强化较均匀,余1例非典型性脑膜瘤信号及强化不均匀,T1WI等信号伴稍低信号,T2WI等信号伴高信号。结论 侧脑室脑膜瘤的MRI表现有一定的特征性,结合临床资料,MRI可以对其进行诊断并和其它侧脑室肿瘤进行鉴别。     

Light microscopic demonstration of the microlumen of ependymoma: A study of the usefulness of antigen retrieval for epithelial membrane antigen (EMA) immunostaining

To determine the origin of dotlike epithelial membrane antigen (EMA) immunoreactivity of ependymoma, which is consistent with the eosinophilic globular body in hematoxylin and eosin (H&E) stain, an immuno-electron microscopic study was undertaken. The usefulness of antigen retrieval pretreatment in detecting the dotlike EMA immunoreactivity in ependymomas was also studied. The materials were 29 ependymomas, 7 autopsy brains as a normal control, and 50 brain tumors of various types. The study confirmed that most of the brown dots in EMA immunostain in ependymoma represented microlumina of tumor cells. In ependymomas, plain EMA immunostaining showed dotlike positivity in only six cases (21%), and antigen retrieval pretreatment increased the number of positives up to 26 cases (90%). Antigen retrieved CD99 detected 23 positive cases (80%) in ependymomas. On the basis of the results, although some false positive findings were raised by antigen retrieval pretreatment, the authors positively recommend adoption of the technique, especially when ependymoma remains as one of the differential diagnoses of the tumor.     

Clinicopathological analysis of metaplastic meningioma: report of 15 cases in Huashan Hospital

Objective: Metaplastic meningioma is a rare subtype of benign meningiomas, classified as WHO grade I with well prognosis. Here we presented our experiences on 15 cases of metaplastic meningioma, to investigate the clinicopathological features, therapies and prognosis of these cases. Methods: 15 patients underwent surgical treatment for intracranial metaplastic meningioma between 2001 and 2010 at Neurosurgery Department of Huashan Hospital, Shanghai, China. The clinical data, radiological manifestation, treatment strategy, pathological findings and prognosis of all patients were analyzed retrospectively. Results: Among the 15 cases (10 males and 5 females), the age ranged from 22 to 74 years old (the mean age was 50.67-year old). The clinical manifestations include headache, dizziness, seizure attack, vision decrease, and weakness of bilateral lower limbs. All the patients received surgical treatment, combined with radiotherapy in some cases. In the follow-up period, recurrence occurred in 2 cases, of which 1 patient died of other system complications. Conclusions: Metaplastic meningiomas are characterized by focal or widespread mesenchymal differentiation with formation of bone, cartilage, fat, and xanthomatous tissue elements. Surgical removal is the optimal therapy, and the overall prognosis is well. But recurrence may occur in some cases, thus radiotherapy is necessary for such kind of patients.     

Papillary meningioma. Clinicopathologic study of seven cases and review of the literature

Seven cases of papillary meningioma are reported. The patients, 3 females and 4 males, were aged between 21 and 69 years. Five tumors were supratentorial, 1 was located in the left temporal bone, and 1 in the thoracic spinal canal. Five patients had local recurrences and died within 1.4 to 9 years of the original operation. In Case 2, one small pulmonary metastatic nodule was found at autopsy. Microscopically, these meningiomas showed foci of necrosis, numerous mitotic figures and local invasiveness. Psammoma bodies were occasional or absent. Forty-six papillary meningiomas have been identified in the literature. Certain histologic features (necrosis, high mitotic index, rich peripapillary reticulin network) and evolutive events (high rate of local recurrence, development of distant metastases) suggest that this aggressive variant of meningioma could form a histologic link between syncytial, fibroblastic, and hemangiopericytic meningiomas.     

Optimal tumor sampling for immunostaining of biomarkers in breast carcinoma

Introduction  

Biomarkers, such as Estrogen Receptor, are used to determine therapy and prognosis in breast carcinoma. Immunostaining assays of biomarker expression have a high rate of inaccuracy; for example, estimates are as high as 20% for Estrogen Receptor. Biomarkers have been shown to be heterogeneously expressed in breast tumors and this heterogeneity may contribute to the inaccuracy of immunostaining assays. Currently, no evidence-based standards exist for the amount of tumor that must be sampled in order to correct for biomarker heterogeneity. The aim of this study was to determine the optimal number of 20X fields that are necessary to estimate a representative measurement of expression in a whole tissue section for selected biomarkers: ER, HER-2, AKT, ERK, S6K1, GAPDH, Cytokeratin, and MAP-Tau.     

XRCC1 and XRCC3 variants and risk of glioma and meningioma

Several single nucleotide polymorphisms (SNPs) affecting DNA repair capacity and modifying cancer susceptibility have been described. We evaluated the association of SNPs Arg194Trp, Arg280His, and Arg399Gln in the X-ray cross-complementing group 1 (XRCC1) and Thr241Met in the X-ray cross-complementing group 3 (XRCC3) DNA repair genes with the risk of brain tumors. The Caucasian study population consisted of 701 glioma (including 320 glioblastoma) cases, 524 meningioma cases, and 1,560 controls in a prospective population-based case–control study conducted in Denmark, Finland, Sweden, and the UK. The studied SNPs were not significantly associated with the risk of brain tumors. The highest odds ratios (ORs) for the associations were observed between the homozygous variant genotype XRCC1 Gln399Gln and the risk of glioma (OR = 1.32; 95% confidence interval, CI, 0.97–1.81), glioblastoma (OR = 1.48; 95% CI, 0.98–2.24), and meningioma (OR = 1.34; 95% CI, 0.96–1.86). However, in pair-wise comparisons a few SNP combinations were associated with the risk of brain tumors: Among others, carriers of both homozygous variant genotypes, i.e., XRCC1 Gln399Gln and XRCC3 Met241Met, were associated with a three-fold increased risk of glioma (OR = 3.18; 95% CI, 1.26–8.04) and meningioma (OR = 2.99; 95% CI, 1.16–7.72). In conclusion, no significant association with brain tumors was found for any of the polymorphisms, when examined one by one. Our results indicated possible associations between combinations of XRCC1 and XRCC3 SNPs and the risk of brain tumors.     

Prognostic relevance of MIB-1 immunoreactivity, S-phase fraction, 5-bromo-2'-deoxyuridine labeling indices, and mitotic figures in gliomas.

Prognostic relevance of cell proliferation markers was evaluated in 27 glioma patients. By 1) flow cytometry (FCM), i.e., S-phase fraction (SPF), and BrdUrd-labeling index (LIfcm); 2) immunohistochemistry (IHC), i.e., BrdUrd-labeling index (LIihc) and MIB-1 immunoreactivity (MIB-1 LIihc); and 3) histologic examination, i.e., the presence or absence of cells in mitoses, were assessed. A longer local progression free survival (LPFS) was significantly associated with low SPF, low LIfcm, and low MIB-1 LIihc. For LIihc, no significant association was found. LIfcm appeared to be a more promising prognosticator than MIB-1 LIihc. In comparison with this marker, the presence or absence of mitotic figures appeared to be an even stronger prognosticator. Prognostic significance of LIfcm appeared to be of importance in low-grade gliomas. The number of patients in our study is limited. Our findings were: 1) the presence or absence of cells in mitoses (M-phase activity) appeared to be of more prognostic significance than LIfcm (S-phase activity) and MIB-1 LIihc (non-G0-phase activity); 2) of the tested experimental cell proliferation markers, LIfcm appeared to be of more prognostic significance than MIB-1 LIihc, SPF, and LIihc; and 3) LIfcm is likely to be an important prognosticator in low-grade gliomas and is, therefore, not definitive and only of potential interest.     

Study on the clinical usefulness of NCC-ST-439 in cases of digestive tract cancer

NCC-ST-439 is a monoclonal antibody established from human stomach cancer xenografted nude mice. The values of NCC-ST-439 were measured in 139 cases with various digestive tract cancers and 294 cases with benign digestive tract diseases with the NCC-ST-439 EIA kit (Nihon Kayaku Co., Ltd.), and its clinical usefulness was compared with those of CA19-9 and CEA. The positive rates of NCC-ST-439 in cases of digestive tract cancer were high, i.e., 66.7% for cancer of the bile duct, 58.3% for pancreatic cancer and 52.9% for colorectal cancer. In the benign digestive tract diseases, the overall positive rate seen in case of cholelithiasis and cholangitis, chronic gastritis, benign colorectal diseases and hepatitis, was only 3.7%. The positive rate of NCC-ST-439 was lower than those for CA19-9 and CEA in cases of stomach cancer, colorectal cancer and liver cancer, but it was the same as that of CA19-9 and higher than that of CEA in cases of biliary tract cancer and pancreatic cancer. The false positive rate of NCC-ST-439 in benign diseases of the digestive tract was the lowest among the three markers. With respect to sensitivity, specificity and efficiency, CA19-9 showed the highest sensitivity, but NCC-ST-439 and CEA showed better specificity than CA19-9, and NCC-ST-439 showed the highest efficiency. In combination assays using combinations of NCC-ST-439, CA19-9 and CEA, the positive rates for ST-439 alone were 22.1% for stomach cancer, 52.9% for colorectal cancer, 15.0% for liver cancer and 58.3% for pancreatic cancer, while the combined rates increased to 51.9%, 70.6%, 75.0% and 66.7%, respectively. In an investigation of changes with time in NCC-ST-439 values during chemotherapy of various types of digestive tract cancer, there was a decrease in PR cases, no change in NC cases and a tendency to increase in PD cases. These results suggested that it was possible to apply NCC-ST-439 clinically.     

Clinicopathological characteristics of pancreatic ductal adenocarcinoma with invasive micropapillary carcinoma component with emphasis on the usefulness of PKCζ immunostaining for detection of reverse polarity

Invasive micropapillary carcinoma (IMPC) is a rare distinct histopathological subtype, characterized by the presence of carcinoma cells displaying reverse polarity. Only limited clinicopathological information is available regarding pancreatic IMPC. The aim of the present study was to clarify the clinicopathological features of pancreatic IMPC and the usefulness of protein kinase C (PKC)ζ immunostaining for the detection of reverse polarity. We reviewed 242 consecutive surgically resected specimens of pancreatic ductal adenocarcinoma and selected samples with an IMPC component. Clinicopathological characteristics were compared between the IMPC and non-IMPC groups. Immunohistochemical staining for PKCζ was performed using an autostainer. In total, 14 cases had an IMPC component (5.8%). The extent of IMPC component ranged from 5 to 20%. There were no significant differences in tumor location, T category, lymph node metastatic status, preoperative carbohydrate antigen 19-9 level, resection status and overall survival between the IMPC and non-IMPC groups. Immunostaining for PKCζ clearly showed reverse polarity of the neoplastic cells of IMPC. Although previous reports have shown that the presence of an IMPC component (>20% of the tumor) indicated poor prognosis, the present study demonstrated that presence of IMPC <20% did not suggest a worse prognosis.     

Heterogeneity of immunostaining for tumour markers in non-small cell lung carcinoma

Lung carcinoma is a leading cause of death. However, there are few indicators that can aid in prediction and prognosis. Many tumour markers are available, but their reliability is questionable. For example, Ki-67 expression has been associated with increased as well as decreased survival or with no clinical significance. The varying results have been attributed to the methodology, relative intensity of staining, variety of marking and statistical methods. To determine whether differential expression of markers within tumours may be a contributory factor to this lack of agreement, we used two marking methods to evaluate the level of expression of Ki-67, p53 and bcl-2, in addition to the apoptotic index, in serial sections of non-small cell carcinoma. All stains exhibited a degree of heterogeneity. This small study highlights the importance of standardisation of marking methods and interpretation of results if tumour markers are to be used as predictive or prognostic factors.