Low Labeling Index Research Articles

Multiple Myeloma Patients Who Are Chemosensitive and Present with Plasma Cells Lacking Phosphorylated Retinoblastoma and/or Cyclin A Expression Benefit From High Dose Therapy and Autologous Stem Cell Transplantation More.

AbstractAbstract 1345 Background and aim:Multiple Myeloma (MM) plasma cells are known to posses low replicative potential and plasma cell labelling index is generally accepted as a strong prognostic factor. TC classification of MM defines categories of myeloma cases expressing different cyclins (Fonseca et all, Leukemia 2009). Although cyclin dependent kinase inhibitors (CDKI) are important for cell cycle, they are not included in this molecular classification. Patients and Methods:With an aim to define high and low proliferative myeloma cases antibodies detecting cyclins (Cyc) A, D1, D2, D3, phosphorylated retinoblastoma (Rb), p16, p21, p27, Ki67 were applied to tissue sections obtained from either marrow plasmacytoma of 106 consequtive myeloma patients (median: 59, 32–79 years) diagnosed between 1998–2007. 100 patients <65 years (n:51) were evaluable for treatment outcome and received an induction of VAD (4-6 cycles), followed by a novel agent containing regimen (2-4 cycles) or autologous stem cell transplant (ASCT) depending on the response to induction. Postransplant consolidation or subsequent transplants were performed when < VGPR was obtained. Elderly patients (n:49) received induction for at least a year. Novel agents (Thalidomide: 66.7%, Bortezomib: 27.5%) were given in 94.2% of patients as ≥2 line treatment. Mann Whitney U, Kaplan-Meier or log rank analysis were performed using the SPSS version 15.0. Results:Loss of CDKI's were detected in 55.4–88.5 % of the cases. Cyclin D1D2D3 or Cyclin A expression was detected in 29.2, 21.7, 5.7 or 19.8 % of the cases. Rb was detected in 25%. Among the CDK's or CDKI's only Cyc D2 was correlated with Ki67 percentage (p=0.027). Loss of all CDKI's was observed in 31.1 %. The majority of p16 (-) cases were Cyc A (-) too (p=0.001). Loss of p16 and p 27 was observed more frequently than loss of p 21 (88 and 85% versus 54%). Patients were either both CycA and p21 (-) (50%) or both CycA and p21 (+) (31 %) (p=0.025). However there was no patient expressing Rb and p16. On the contrary, the majority patients were both Rb and p16 (-) (60%) (p=0.037). Groups of patients with high proliferative protential group 1 (Cyc D+ p16-), group 2 (CycA+ p21-), group 3 (CycA+ Rb+) and low proliferative protential group 4 (Rb- p16+), group 5 (CycA- Rb-) were analyzed. These were observed 42.5 %, 5.5%, 11.7%, 15.4% and 57.1%. No correlation could be found between the CDKI/Cyc defined risk groups and ISS, B2MG, LDH, CRP, OS. However among p16 (-) cases Rb (-) ones (n:39) had longer OS than Rb (+) patients (n:18) (49 vs 39 months). All p16 (+) patients were Rb (-) too and had a shorter OS (42 vs 84 months, p=0.006). Response to initial treatment was ≥ VGPR 16.8 %, ≥ PR 52%, refractory 21.4 %. High proliferative group 1 or patients with high LDH values had higher initial response rates (82.1 vs 53.8 %, p: 0.016 and 67.6 % vs 36 % p= 0.018). The deepest response was observed with initial treatment among 47.3 % and was improved following ASCT (18.3%). ASCT also improved OS among all (58 vs 34 months, p=0.0) but more strongly among Rb and/or CycA (-) patients (Table). The response to initial treatment did not influence OS. However if response is not upgraded following subsequent treatments OS deteriorated (5 year OS: 34 vs 44 months p=0.022) (Figure). These suboptimal responding patients could not be predicted by ISS, age, B2MG, high/low proliferating group definition. Although LDH was high in 72.7 vs 54.5 % of poor-responders this comparison was not significant. Conclusion:While depth of response to therapy did not, improvement of response with subsequent therapy ie novel agents or ASCT prolongged OS (34 vs 44 months, p=0.022) and was associated with lower LDH values. Compared to gene expression profilling, tissue array using monoclonal antibodies is a method which can be more widely applicable and in our study was able to define new prognostic parameters. Overall ASCT extended OS. This effect was stronger among patients lacking Rb and/or Cyc A, This finding echoes the need for better treatment modalities for patients with poorer prognosis ie Rb and/or Cyc A positivity. Multivariate analysis results will be presented during the congress.TablePresence of cell cycle related markers and overall survival (months)nmonthsASCT (-)Cyc A (+)831,6Cyc A (-)4171Rb (+)1658,2Rb (-)3255,6Cyc A (+) Rb (+)329Cyc A (-) Rb (-)2330,1ASCT (+)Cyc A (+)1261,9Cyc A (-)3991,3Rb (+)1154,4Rb (-)4091,5Cyc A (+) Rb (+)646Cyc A (-) Rb (-)3456,9 [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Abstract 1473: Fra-2 and JunB are overexpressed in precancerous bile ducts and primary/transplanted hamster cholangiocarcinomas with a strong positive correlation

Abstract The transciption factor complex AP-1 plays important roles in cell proliferation, differentiation and apoptosis of which dysregulation is closely associated with cancer development. Fra-2 and JunB, components of AP-1 family proteins, had been found overexpressed in certain of tumors, suggesting their possible implication in carcinogenesis. In the present study, we investigated differential expression patterns of those two AP-1family proteins during hamster cholangiocarcinogenesis and aggressive transplanted cholangiocarcinomas (ChCs). First, we prepared appropriate liver tissue samples with precancerous lesions and ChCs from hamster ChC model as well as normal liver. The transplanted ChCs were produced by syngenic transplantation of the primary ChCs. We then performed immunohistochemistry (IHC) for Fra-2 and JunB using ABC methods. The typical nuclear staining for Fra-2 and JunB was determined as positive immunoreactivity and the ratio of positive cells was evaluated by counting 800-3000 cells, followed by statistical analysis. The correlation of Fra-2 and JunB expression was analyzed by scattergram. Our IHC indicated significantly increased frequency of Fra-2 and JunB positive cells according to the tumor progression, i.e., respectively in the early dysplatic bile ducts (10∼20%) and late dysplastic bile ducts (40∼50%), primary ChC (40∼60%) and transplanted ChC (higher than 70% positivity), while normal and simple hyperplastic bile duct epithelia were negative or indicated very low labeling index for both Fra-2 and JunB (under 1% positivity). The expression of Fra-2 and JunB showed a strong correlation (r=0.93) and the immunoreactive cells for Fra-2 and JunB were mainly localized in the invading areas adjacent to normal surrounding tissue. In summary, we found in the present study the followings 1) Expression of Fra-2 and JunB progressively increased as bile duct cells become carcinogenic, 2) Fra-2 and JunB have a high correlation in the expression patterns in the biliary cell populations including cancers, 3) In the transplanted biliary cancers, overexpression of Fra-2 and JunB (especially JunB) were localized in the neoplastic cells which are very aggressive to invade into adjacent normal tissue. Based on our results, Fra-2 and JunB could play significant roles in cholangiocarcinogenesis, possibly by collaboratively participating in dysplastic changes and acquisition of tumorigenicity of candidate bile duct cells for ChC. Also, those two AP-1 family proteins could contribute to invasiveness of biliary cancer cells. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1473.

Minichromosome maintenance‐7 and geminin are reliable prognostic markers in patients with oral squamous cell carcinoma: immunohistochemical study

Minichromosome maintenance (MCM) proteins act as the origin licensing machinery that regulates initiation of DNA replication. Geminin is a licensing repressor and prevents reinitiation of DNA replication by blocking reloading of MCM proteins at replication origins. Recent studies have proposed that MCM7 and geminin may be sensitive proliferative and prognostic markers of various malignant tumors. This study aimed to analyze the expression of MCM7 and geminin to clarify pathobiological significance in human oral squamous cell carcinomas (OSCCs). We performed immunohistochemical analysis on 10 specimens of normal oral epithelia, 50 lesions with dysplasia and 113 OSCCs. Labeling indices (LIs) for MCM7, geminin and Ki-67 were evaluated, comparing with clinicopathological profiles. The mean LIs for MCM7 were 29.2% for normal epithelia, 32.2% for dysplasias, and 51.1% for OSCCs; the value was significantly higher in the last than in the former two (P < 0.01). The mean LIs for geminin were 6.8% for normal epithelia, 9.2% for dysplasias, and 21.3% for OSCCs; the value was significantly higher in the OSCCs (P < 0.01). The MCM7 LIs were correlated with the histological grade of OSCCs, in which the highest LIs were noted in the poorly differentiated type (P < 0.01). The survival rate was significantly lower in patients with a higher MCM7 LI (>49.5%) than in those with a lower LI (P < 0.05) at stage III-IV. However, the survival rate in the patients with a higher geminin LI (>19.5%) was significantly higher than in those with a lower LI (P < 0.05) at stage IV.

The correlation of TRPM1 (Melastatin) mRNA expression with microphthalmia‐associated transcription factor (MITF) and other melanogenesis‐related proteins in normal and pathological skin, hair follicles and melanocytic nevi

Melastatin (TRPM1), a.k.a. transient receptor potential cation channel, subfamily M, member 1 (TRPM-1) regulates melanocyte differentiation and proliferation. TRPM1 is transcriptionally regulated by the essential melanocyte transcription factor MITF (microphthalmia-associated transcription factor). For the most part, MITF expression is preserved during melanoma progression, while TRPM1 mRNA expression decreases or is completely lost. The loss of TRPM1 is associated with melanomas that are more aggressive. To assess the relationship between TRPM1 mRNA expression and the expression of MITF and nine other markers of melanocytes and melanin-related proteins by immunohistochemistry in normal skin, scars, hair follicles and ordinary melanocytic nevi. Samples of normal skin (n = 102; from tumor excisions and plastic procedures), scars (n = 5; from re-excision specimens) and compound melanocytic nevi (n = 4) were evaluated for the presence of TRPM1 mRNA transcripts as detected by chromogenic in situ hybridization (CISH). Immunohistochemical techniques were used to detect melanin-related proteins including: MITF, S100 protein, Mart-1, tyrosinase, Mel5, HMB45, tyrosinase-related protein-1 (TRP1), TRP2 and alpha-melanocyte stimulating hormone (alphaMSH). The labeling index (LI) was defined as the number of intraepidermal cells expressing mRNA or protein per one hundred basal keratinocytes. A wide range of LI was found for all markers (0-33 positive cells/100 keratinocytes). When these LI were compared, no significant differences in the expression of MITF, S100, Mart1, tyrosinase proteins and TRPM1 mRNA were identified. The LI for TRPM1 mRNA expression ranged from 74% of that for MITF to 86% for tyrosinase. The LI for TRP-1, TRP-2 and Mel5 was similar to that of TRPM1, while HMB-45 had a significantly lower LI than all other markers. TRPM1 mRNA correlated most tightly with MITF and tyrosinase expression (r = 0.81 and 0.68, respectively, both p = 0.0001). Likewise, the strongest correlation among all the melanin-related proteins existed between tyrosinase and MITF (r = 0.79, p = 0.0001). There was variable expression of melanin-related proteins when LI were analyzed by anatomic site, patient age, extent of sun-damage and proximity to a melanocytic tumor. Anogenital skin showed the highest and acral skin the lowest LI for TRPM1, MITF, S100 protein, Tyrosinase, Mel5 and HMB45. Advanced age (> 60 years) was associated with decreased TRPM1 expression. Sun-damaged skin exhibited significantly increased LI as measured by MITF, S100 protein, Mart1, tyrosinase and HMB-45, but no differences for TRPM1. However, the MITF-TRPM1 differential (i.e. MITF LI-TRPM1 LI = MITF+TRPM1--melanocytes) was significantly increased in site-matched skin (4.6 +/- 4.4 vs. 1.5 +/- 2.5, p = 0.01). There was a suggestion of reduced LI in normal skin in the proximity of melanoma (from melanoma re-excision specimens) for S100, HMB45 and TRPM1 mRNA. TRPM1 LI was significantly decreased in scars compared to normal skin (5.6 +/- 1.4 vs. 9.7 +/- 4.3, p = 0.02), this was reflected in an increase in the MITF-TRPM1 differential (9.6 +/- 7.5 vs. 3.2 +/- 3.1, p = 0.0001). MITF LI were consistently higher than MSLN LI at all levels of the hair follicle; notably, MITF was expressed by isthmic-bulge cells. In ordinary melanocytic nevi, MITF and TRPM1 expression decreased with melanocyte descent: there was more signal for both markers in superficial epithelioid type A melanocytes than deeper type C melanocytes. By CISH, TRPM1 mRNA expression is specific for melanocytes and strongly associated with MITF and tyrosinase expression, the latter implicating a mature melanocyte phenotype. However, in normal skin, TRPM1 mRNA expression appears to be dynamic, labeling most but not all melanocytes, with variable expression ostensibly related to local environmental factors.

Oncocytic choroid plexus carcinoma: case report

Herein, we report an unusual choroid plexus carcinoma with extensive oncocytic transformation. A 13-month-old girl presented with acute lethargy which quickly progressed to coma. A CT scan of the head revealed impending herniation due to hemorrhage within an intracranial tumor. An MRI scan showed a large, partly cystic and highly vascular left lateral ventricular mass. A near total resection was achieved. Microsections revealed a WHO Grade III choroid plexus carcinoma with extensive oncocyti c transformation. A minor portion of the moderately to poorly differentiated tumor exhibited classical microscopic features of choroid plexus carcinoma, including marked nuclear atypia, brisk mitotic activity (78/10 HPF), a high MIB-1 labeling index (44%) and zones of necrosis. In contrast, the large, eosinophilic, cytologically malignant but granular-appearing oncocytes comprising the majority of the lesion showed scant (1/10 HPF) mitotic activity and only a low MIB-1 labeling index (5%). A subsequent recurrence at 1 year consisted entirely of non-oncocytic tumor. Choroid plexus carcinoma with oncocytic transformation has not been previously reported. The remarkable extent of this alteration and its clinical significance remains to be determined.

Depth of Response with Stem Cell Transplantation and Outcome for Multiple Myeloma in the Era of Novel Agents.

Abstract 1228Poster Board I-250 BackgroundIt is generally believed that a ‘deeper' response in multiple myeloma is associated with an improved outcome, including time to progression (TTP) as well as overall survival. Before the advent of novel agents such as IMiDs or bortezomib, complete responses (CR) were rare in the absence of stem cell transplantation (ASCT). In that era, the depth of response achieved before ASCT tended to be dwarfed by the impact of the conditioning used for stem cell transplant. Hence, the time to progression in patients achieving CR before ASCT was not different from those who achieved CR after ASCT. With novel agents, CR rates are higher and again raises the question of whether patients who achieve CR before ASCT benefit from consolidation with ASCT. The purpose of this analysis was to determine whether CR achieved before ASCT still benefit when consolidated with ASCT MethodsAfter approval from the Institutional Review Board at Mayo Clinic Rochester, the transplant dysproteinemia database was searched for all patients treated with novel agents and patients who achieved CR before or after ASCT were identified. In addition, we identified patients with IMiDs induction therapy and achieved CR but did not proceed with ASCT. Relevant demographic, clinical and laboratory characteristics were determined at the time of ASCT. Comparisons between groups were performed with non-parametric tests. Overall survival and TTP were determined from the time of ASCT using the Kaplan-Meier method. ResultsA total of 354 patients who underwent ASCT within a year from diagnosis of multiple myeloma were identified. Of these, 24 achieved CR before proceeding to ASCT while another 100 patients achieved CR after ASCT. There were no differences between the two cohorts with respect to age (58 vs 57.5 yr, p=0.14), gender, number of treatment regimens (1 vs 1, p=0.83), time to ASCT (6.5 vs 6.3 months, p=0.80), b2M (2.41 vs 2.26, p=0.58) and ISS (p=0.23). Patients who achieved CR before ASCT had a lower plasma cell burden (1.0 vs 5.0%, p<0.0001) and lower labeling index (0 vs 0, p=0.0007). Aneuploidy was present in none of the patients with CR before transplant compared to 10% in patients who achieved CR after ASCT (p=0.11). The median TTP for patients with CR before ASCT has not been reached (71% at 70 months) compared to 30 months for patients who achieved CR after ASCT (p=0.07). After a median follow up of 70 months, 21 of 24 patients (88%) with CR before transplant are alive compared to 76 of 100 patients with CR after ASCT (p=0.44). In contrast, for patients who achieved CR with IMiDs but did not proceed with ASCT, 55% have not progressed at 51 months and 71.7% are alive at 51 months (Gay et al ASH, 2009). ConclusionOur results suggest that patients who achieve CR before ASCT benefit from high-dose therapy and experience prolonged TTP without the need for maintenance therapy. ASCT after achieving CR can result in a deeper response with improvement in disease free and overall survival. DisclosuresLacy:celgene: Research Funding. Gertz:celgene: Honoraria; genzyme: Honoraria; millenium: Honoraria; amgen: Honoraria.

Intracranial kaposiform hemangioendothelioma: proposal of a new malignant variant

Kaposiform hemangioendothelioma (KHE) is a rare vascular tumor of infancy and early childhood that occurs mainly in soft tissue and the retroperitoneum. The pathological characteristics of a KHE are fascicles of spindleshaped endothelial cells and slitlike vascular channels with irregular tumor margins. In spite of benign features such as rare mitoses and a low MIB-1 labeling index, a KHE is categorized as an intermediate malignancy because of local invasiveness into adjacent lymph nodes or organs. Resection is the treatment of choice for this type of tumor and adjuvant medical therapy is prepared for the tumor remnants and the associated Kasabach-Merritt syndrome. In this paper the authors report a case of a KHE without Kasabach-Merritt syndrome arising from the right tentorium cerebelli in a 13-month-old male child. The biological behavior of the tumor, including its pathological traits and clinical course, was malignant in nature. This case may be not only the first intracranial extraaxial KHE, but also the first truly malignant variant. Kaposiform hemangioendothelioma should be considered in the differential diagnosis of intracranial and extraaxial tumors.

Galectin‐3 Upregulation During Tumor Progression in Head and Neck Cancer

To examine the level of expression of galectin-3 in relation to neoplastic progression of hypopharyngeal squamous cell carcinomas (HSCCs) and laryngeal squamous cell carcinomas (LSCCs). Retrospective study. Using a polyclonal antibody against galectin-3 without cross-reactivity to other galectins, we analyzed the presence of galectin-3 using quantitative immunohistochemistry in i) a series of 79 HSCCs compared with 16 normal epithelia, 20 low-grade dysplasia (Low_D) and 25 high-grade dysplasia (High_D) and in ii) a series of 58 LSCCs compared with 34 normal epithelia, 12 Low_D, and 18 High_D. In parallel, galectin-3 expression was studied using Western blotting on a series of 19 fresh biopsies from patients presenting a head and neck tumor. Western blotting excluded a notable degree of proteolytic truncation of galectin-3 in situ. Immunohistochemical galectin-3 positivity expressed as percentage of cells was significantly higher in LSCCs and HSCCs than in Low_D (P = .01) or High_D (P = .0002), respectively. Increased expression of galectin-3 in HSCCs was accompanied by a shift from the cytoplasmic compartment to the nucleus (P = .007). In intertumor-type comparison, laryngeal carcinomas presented nuclear presence of galectin-3 only rarely (1 of 58 cases in laryngeal cancer vs. 27 of 79 cases in hypopharyngeal cancer, P = .00006) and a comparatively low labeling index (P < 10(-6)). Our data reveal an association between level of presence of galectin-3 and neoplastic progression of HSCCs and LSCCs.

Clear cell odontogenic carcinoma: case report with immunohistochemical findings adding support to the challenging diagnosis

Clear cell odontogenic carcinoma (CCOC) is a rare odontogenic tumor associated with aggressive clinical behavior, metastasis, and low survival. We report a case of CCOC affecting the mandible of a 39-year-old man. The tumor presented a biphasic pattern composed of clear cell nests intermingled with eosinophilic cells and separated by collagenous stroma. Immunoreactivity to cytokeratin (CK), specifically AE1/AE3 and CK 8, 14, 18, and 19 was found, as well as to epithelial membrane antigen (EMA). The tumor cells were negative for S100 protein, CK 13, vimentin, smooth muscle actin, laminin and type IV collagen. Low labeling indices for the proliferation markers Ki-67 and proliferating cell nuclear antigen and to p53 protein might predict a favorable prognosis for the lesion. A surgical resection was performed, followed by adjuvant radiotherapy. A 2-year follow-up has shown no signs of recurrence. The significance of histochemical and immunohistochemical resources in the correct diagnosis of CCOC is analyzed.

非機能性膵内分泌腫瘍の１切除例

We report a rare case of a very large nonfunctioning endocrine tumor of the pancreas without malignant histological features. A 63-year-old woman referred for appetite loss and general fatigue was found to have a tumor in the pancreas head. Computed tomography demonstrated a well-defined pancreatic tumor 45mm in diameter with hypervascular staining in the pancreas head. Angiography showed a hypervascular tumor of the pancreas head and a dilatation of the anterior superior and posterior superior pancreaticoduodenal arteries. The preoperative diagnosis was an endocrine tumor of the pancreas, with undeniable malignancy. Pylorus-preserving pancreaticoduodenectomy was performed. The histopathological diagnosis was a benign nonfunctioning endocrine tumor of the pancreas based on immunohistochemical staining for Chromogranin A, Synaptophysin, and NSE, but not for hormones. The tumor revealed a low labeling index (<2.0%) of Ki-67 indicating its benign character. No tumor recurrence has been identified in the 18 months since surgery.

Nipple adenoma found in a mastectomy specimen: Report of a case with special regard to the proliferation pattern

We report a case of nipple adenoma incidentally found in a mastectomy specimen, and describe its unique histological appearance and the immunohistochemical distribution of Ki-67 positive tumor cells. A 45-year-old woman with no symptoms or sign related to the nipple had a left mastectomy for invasive breast cancer. A small nipple adenoma, 7 mm in size, was incidentally recognized in the nipple of the resected breast. Histologically, the tumor in the nipple was composed of numerous proliferative ducts with a tubular and florid papillomatous appearance. Many demarcations between squamous cells of the epidermis and tumor cells were recognized in the summit as well as the lateral wall of the nipple. A high Ki-67 labeling index (20.3%) was recognized in the tumor cells in the superficial region, and a low labeling index (0.7%) was seen in the deeper region of the tumor. Based on these proliferative patterns, the symptoms and clinical signs related to the nipple that are often found in patients with nipple adenoma are thought to be associated with the destruction of the epidermis of the nipple by the invasion of benign tumor cells with high proliferative activity.

Fractalkine‐CX3CR1 axis regulates tumor cell cycle and deteriorates prognosis after radical resection for hepatocellular carcinoma

Fractalkine is the only CX3C chemokine, and its receptor, CX3CR1, is expressed on NK cells, CD8+ T cells, monocytes, and dendritic cells (DC). Although studies have reported that fractalkine regulates the host immune response, the roles of the fractalkine-CX3CR1 axis in tumor biology and the clinical results of hepatocellular carcinoma (HCC) remain unknown. Fractalkine and CX3CR1 expression in HCC were evaluated and compared with the clinicopathologic features, including tumor progression determined by proliferating cell nuclear antigen (PCNA) antibody and patient prognosis after surgery. Tumors with high expression of both fractalkine and CX3CR1 had significantly fewer intra- and extrahepatic recurrences, a low PCNA labeling index (PCNALI), and different histological grades. Patients with tumors that expressed both had a significantly better prognosis in terms of disease-free (DFS) and overall survival (OAS), and this finding was identified as one of the independent prognostic factors in the multivariate analysis. Our results suggest that the fractalkine-CX3CR1 axis plays a pivotal role in the prognosis of patients with HCC, which might arise from the known modulation of the host immune response, and that of the cell cycle in HCC.

Apoptosis Gene‐Expression Profiling of Ex Vivo Expanded Human Limbal Epithelial Cells

Ex vivo expanded human limbal epithelial cells (HLEC) used to treat patients with limbal stem cell deficiency can be preserved in a closed eye bank organ culture (OC) system. This study was conducted to investigate cell death due to apoptosis after eye bank storage of cultured HLEC. HLEC were expanded on intact amniotic membranes and stored for one week at 31°C and 23°C in OC medium or at 5°C in Optisol-GS. Cultures were fixed in formaldehyde and embedded in paraffin. Low labeling indices were observed under different storage conditions as assessed by immunostaining with caspase-3 (range 0.2%–1.2%) and a DNA fragmentation assay (TUNEL, range 1.0%–2.3%). Cellular results were confirmed at the gene expression level using the Human Apoptosis RT2Profiler™ PCR Array from SuperArray Bioscience, which profiles the expression of key apoptosis genes. The results showed that pro-apoptotic genes were significantly down regulated and anti apoptotic genes were significantly up regulated in all three storage conditions. This study demonstrates that eye bank storage of cultured HLEC is associated with minor cell death due to apoptosis at both the cellular level and the gene expression level. This work was supported in part by the Eastern Norway Regional Health Authority, the Norwegian Association of the Blind and Partially Sighted and the Blindmission IL.

The Predictive Value of CK19 and CD99 in Pancreatic Endocrine Tumors

Prediction of behavior in pancreatic endocrine tumors (PETs) is reliant on clinicopathologic features. However, there remains a cohort of PETs that behave aggressively despite showing indolent pathologic features. Recently, it has been suggested that CK19 and CD99 are sensitive ancillary markers that predict outcome in PETs. An analysis of 54 PETs and 2 resected liver metastases was undertaken to examine the relationship of CK19 and CD99 and the pathologic criteria in the WHO classification of PETs. CK19 was found to correlate with mitotic count (>5/50 high-power fields), an MIB-1 labeling index of > or =2%, lymphovascular/perineural permeation, lymph node involvement, and liver spread. Although not statistically significant, CK19-negative tumors tended to be smaller than the average tumor size in the series (2.5 vs. 3.6 cm). CD99 did not show any significant correlation with any of the WHO criteria. Tumors that are confined to the pancreas with low mitotic count and MIB-1 labeling index, tended to be CD99-positive. Both CK 19 (negative) and CD99 (positive) correlated with insulin-positive PETs. In conclusion, CK 19 may prove to be a useful ancillary diagnostic test in the routine work-up of PETs. CD99 does not appear to be as useful. There is no compelling evidence, from our study, to suggest that both these markers may be used in concert to predict the behavior of PETs.

Follicular carcinoma of the thyroid with aggressive metastatic behavior in a pregnant woman: Report of a case and review of the literature

Distant metastases as initial presentation of follicular carcinoma of the thyroid is rare, especially in young patients. We report the clinical and pathological features of a 33-year old pregnant patient with follicular carcinoma of the thyroid who presented with widespread bone and lung metastases at the time of diagnosis. the resected tumor had a focal insular component that showed extensive vascular invasion spreading beyond the thyroid capsule, and was associated with widespread bone and lung metastases. Despite its aggressive behavior, the tumor had low mitotic activity and Ki-67 nuclear labeling index. tumor cells showed high microvascular density and down-regulation of E-cadherin, a calcium-dependent trans-membrane epithelial protein molecule known to promote intercellular adhesion. We suggest that architectural differentiation of the tumor and cell proliferation rate are not reliable markers of metastatic behavior in this particular thyroid neoplasm. Microvascular density and down-regulation of E-cadherin expression in the tumor should be included among histologic hallmarks of metastatic potential. the role of pregnancy in the aggressive behavior of this tumor is discussed along with a literature review.

Clear cell Meningioma, an uncommon variant of meningioma: a clinicopathologic study of nine cases

Clear cell meningioma (CCM) is an uncommon variant of meningioma, which affect younger patients, occur more often in spinal or cerebello pontine locations and shows a higher recurrence rate. Only few case reports have been described in the literature. The study has been undertaken to document the clinicopathological features of nine cases of CCM, operated at All India Institute of Medical Sciences during 1998 to December 2005. Clinical information was retrieved from the records of our Neurosurgery Department. The cases were stained with H&E, periodic Acid Schiff (PAS) with and without diastase. Immunohistochemistry for pancytokeratin, epithelial membrane antigen, vimentin, glial fibrillary acidic protein, and MIB-1 was done in all cases. During a period of 8 years, nine cases of CCM were diagnosed. Age ranged from 10 to 65 years (median age 26.0 years) with female predominance. Most common location was posterior fossa (CP angle). Clinically most of the patients presented with history of headache and features of cranial nerve palsies. The duration of symptoms varied from 3 to 60 months (mean 16.7 and median of 4 months). Radiologically lesions showed homogenous enhancement and were isointense to brain parenchyma. Histopathologic examination revealed tumor cells to be arranged in sheets with clear cytoplasm and monomorphic nuclei. MIB-1 labeling index (LI) ranged from 2 to 12% with a mean of 9%. Follow up varied from 3 to 84 months (median 36 months) and recurrence was noted in two patients after 2 and 3 years of surgery, respectively, despite their low MIB-1 labeling indices. CCM is a rare variant of meningioma with poor outcome. Less than 50 cases have been described in the literature. Low rate of recurrence and recurrence despite their low MIB-1 LI are some of the features, which needs to be documented. Hence, larger number of cases with adequate follow-up data need to be studied further to establish the clinical significance of this variant.

Cell Proliferation Measurements by Bromodeoxyuridine or Thymidine Incorporation: Clinical Correlates.

T he thymidine or bromodeoxyuridine labeling index (LI) is measured by determining the incorporation of precursor by viable cells in DNA synthesis (S-phase) and is expressed as percent. LI relates closely to cell proliferative rate. More precise measurement requires determination of S-phase duration (Ts), currently not practical for clinical application. LI correlates with mitotic index (MI), 1,2 but LI is much easier to measure accurately, particularly when the proliferative rate is low, because the ratio of labeled to mitotic cells may approach 100 to i. 3 Measurements of LI with tritiated thymidine (3H-dThd) or 5-bromo-2'-deoxyuridine (BrdUrd) administered intravenously or used in vitro with fresh tissue slices are similar. 4 Labeled cells in S-phase are visualized by autoradiography (3H-dThd) or immunohistochemistry (BrdUrd) in tissue sections (Fig 1). Histological identification of cells circumvents problems that can be troublesome with flow cytometry. Our purpose is to describe LI distributions of human tumors that we have studied by a single, standardized technique of in vitro labeling of fresh tissue slices with 3H-dThd or BrdUrd. 5,6 Results show that certain cancers have characteristic high or low proliferative rates, while others have a wide spectrum. The latter probably represent several discrete tumor types that have not been adequately classified by histological criteria. For the former, cell kinetic characteristics can be construed from the histological classification. For the latter, kinetic measurements can provide useful data for subclassification. LIs of some normal tissues are listed in Table 1. Certain tissues are in continuous renewal and have high LIs, whereas others have very low LIs. Some of the latter can respond to cell loss with proliferation

Autologous stem cell transplantation for multiple myeloma in patients over 70 years: A matched comparison with patients under 65 years

Background: High dose therapy with autologous stem cell rescue has been shown to prolong survival in patients with multiple myeloma in randomized controlled trials. However, most of the prospective studies have included younger patients, usually 65 or less. It is important to have a better understanding of the outcome of transplantation in the older patients given the median age of onset of myeloma of 65 years. We retrospectively reviewed our institutions experience with high dose therapy for patients over 70 years. Methods: We identified 35 patients with multiple myeloma, from the transplant database, who were at or over the age of 70 at the time of their high dose therapy. We matched these patients to 70 patients (two matches for each patient), based on stage at transplant (primary refractory, plateau phase, relapse off therapy, or relapse on therapy), Durie Salmon stage, high or low labeling index, conventional cytogenetics (abnormal vs normal), presence or absence of circulating plasma cells at time of transplant, and whether cyclophosphamide was used as part of mobilization in that order of priority. Results: The median age of the two groups were 55.3 (Range 37.3–64.8) and 71.7 (Range 70–75.8) years at the time of transplant. The median time to transplant from diagnosis was similar (6.4 for the older patients compared to 6.9 months for the other, P = NS). Ten of the 35 older patients received reduced dose melphalan (140 mg/2)) compared to 3 patients in the control group; P Conclusions: High dose therapy and autologous stem cell transplantation is feasible in selected patients with multiple myeloma over 70 years. It is likely that these older patients were selected based on their overall performance status, a factor that is difficult to analyze in this retrospective review. Nearly 70% of the elderly patients received full dose melphalan for conditioning (200 mg/m2). The toxicity of transplant as well as the outcome appears to be very similar to the younger patients. Patients with multiple myeloma should not be excluded from high dose therapy solely on the basis of their chronological age.

Autologous Stem Cell Transplantation for Multiple Myeloma in Patients over 70 Years: A Matched Comparison with Patients under 65 Years.

Background: High dose therapy with autologous stem cell rescue has been shown to prolong survival in patients with multiple myeloma in randomized controlled trials. However, most of the prospective studies have included younger patients, usually 65 or less. It is important to have a better understanding of the outcome of transplantation in the older patients given the median age of onset of myeloma of 65 years. We retrospectively reviewed our institutions experience with high dose therapy for patients over 70 years.Methods: We identified 35 patients with multiple myeloma, from the transplant database, who were at or over the age of 70 at the time of their high dose therapy. We matched these patients to 70 patients (two matches for each patient), based on stage at transplant (primary refractory, plateau phase, relapse off therapy, or relapse on therapy), Durie Salmon stage, high or low labeling index, conventional cytogenetics (abnormal vs normal), presence or absence of circulating plasma cells at time of transplant, and whether cyclophosphamide was used as part of mobilization in that order of priority.Results: The median age of the two groups were 55.3 (Range 37.3–64.8) and 71.7 (Range 70–75.8) years at the time of transplant. The median time to transplant from diagnosis was similar (6.4 for the older patients compared to 6.9 months for the other, P = NS). Ten of the 35 older patients received reduced dose melphalan (140 mg/2)) compared to 3 patients in the control group; P < 0.01. The median follow up from transplant was 10.1 months for the older patients compared to 18 months for the control group. The overall response rate was similar for the two groups (97.1% for the older patients compared to 95.5 for the control group). Eleven (31%) of the older patients and 17 (24%) of the control patients achieved a CR (P = NS). The post transplant progression free survival estimate at 1 year post transplant was 65.3% for the older patients compared to 66% for the control group (P = 0.3)The two year estimated overall survival from transplant was similar in the two groups; 58% for the older patients compared to 67% for the control group. The overall survival from diagnosis was similar for the two groups (P = 0.6). The median number of days hospitalized was 9 days for the older population compared to 5 days for the control group (P = 0.37). Four patients died within the first one hundred days, one (3%) among the older patient group and 3 (4.3%) in the control group.Conclusions: High dose therapy and autologous stem cell transplantation is feasible in selected patients with multiple myeloma over 70 years. It is likely that these older patients were selected based on their overall performance status, a factor that is difficult to analyze in this retrospective review. Nearly 70% of the elderly patients received full dose melphalan for conditioning (200 mg/m2). The toxicity of transplant as well as the outcome appears to be very similar to the younger patients. Patients with multiple myeloma should not be excluded from high dose therapy solely on the basis of their chronological age.

The prognostic value of p53, Ki‐67 and matrix metalloproteinases MMP‐2 and MMP‐9 in transitional cell carcinoma of the renal pelvis and ureter

To investigate the prognostic and predictive relevance of p53 protein, Ki-67 antigen, MMP-2 and MMP-9 in patients with transitional cell carcinoma (TCC) of the upper urinary tract. The expression of p53 protein, Ki-67 antigen, MMP-2 and MMP-9 was examined by immunohistochemistry in 69 patients with TCC of the upper urinary tract. Correlation of p53, Ki-67, MMP-2 and MMP-9 over-expression with conventional pathological parameters and patient survival was examined. p53 over-expression was significantly correlated with histological grade (P < 0.05), but not with pathological stage, vascular invasion, lymphatic invasion or lymph node metastasis. Ki-67 over-expression was significantly correlated with stage, grade, lymphatic invasion and vascular invasion (P < 0.05). In survival analyses, Ki-67 over-expression was a significant prognostic factor in the univariate analysis (P < 0.05), but it did not have a significant impact on survival in the multivariate analysis. Ki-67 labeling index was a significant prognostic factor in patients with a low p53 labeling index, but not in patients with a high p53 labeling index. Ki-67 over-expression is of prognostic value in TCC of the upper urinary tract, while p53, MMP-2 and MMP-9 are of limited value.