Whole-Slide Imaging of Pap Cellblock Preparations Is a Potentially Valid Screening Method.

OBJECTIVE: To date, the impact of digital imaging on routine cytology remains far from perfect. Cellblock (CB) preparations from Pap samples have been shown to be diagnostically valuable. We evaluated the validity of utilizing whole-slide imaging (WSI) prepared from Pap CBs as a screening tool. STUDY DESIGN: A total of 1,110 CB slides prepared from residual Pap samples were analyzed - 563 normal, 282 atypical squamous cells of undetermined significance (ASCUS), 12 atypical squamous cells-cannot exclude high-grade squamous intraepithelial lesion, 188 low-grade squamous intraepithelial lesions (LSIL), 36 high-grade squamous intraepithelial lesions (HSIL), 25 atypical glandular cells of undetermined significance, 1 adenocarcinoma in situ, 2 invasive adenocarcinomas, and 1 squamous cell carcinoma. Virtual slides were obtained using the Aperio system. Test performance characteristics of liquid-based samples and WSI from CB samples were compared. RESULTS: Average sensitivity and specificity of the five WSI reviewers was 58.3 and 85.1% for ASCUS, respectively, 54.1 and 93.9% for LSIL, and 51.8 and 98.8% for HSIL. Overall WSI sensitivity and specificity for detecting lesions was 82.1 and 86.2%, respectively. Agreement (kappa values) between WSI reviewers was 0.56 for ASCUS, 0.69 for LSIL, 0.67 for HSIL, and 0.74 for negative samples. CONCLUSIONS: WSI of CB preparations is a feasible method to achieve high-quality specimen preparations. It is as sensitive as liquid-based methods and appears to be highly specific for the detection of LSIL and HSIL.

Grading invasive ductal carcinoma of the breast: advantages of using automated proliferation index instead of mitotic count.

Breast carcinomas are graded according to the "Nottingham modification of the Bloom-Richardson system" (SBR). The system is hindered, however, by lack of precision in assessing all three parameters including nuclear grade, mitosis, and tubular formation, leading to an element of subjectivity. Our objective was to evaluate a new grading system [the nuclear grade plus proliferation (N+P) system] for subjectivity, ease, and better representation of tumor biology. Its components are nuclear grade and automated proliferation index. Invasive ductal carcinomas, consisting of 137 SBR grade I, 247 grade II, and 266 grade III, were re-evaluated by the N+P system. The two systems were compared with each other and correlated with patients' overall survival, tumor size, angiolymphatic invasion, lymph node status, and biomarker status including estrogen receptor, progesterone receptor, p53, epidermal growth factor receptor, BCL-2, and Her-2. Although there was an agreement between the two systems with histologic and prognostic parameters studied, there was 37% disagreement when grading individual tumors. Fifty-three percent of SBR grade II tumors were "down-graded" to N+P grade I, and 7% were "up-graded" to N+P grade III. Distinction among the different histologic grades for overall survival curves was better indicated by the N+P than the SBR system.

Cell Block Preparation versus Liquid-Based Thin-Layer Cervical Cytology: A Comparative Study Evaluating Human Papillomavirus Testing by Hybrid Capture-2/Cervista, in situ Hybridization and p16 Immunohistochemistry.

OBJECTIVE: Cell block (CB) preparations from residual liquid-based Pap samples have been shown to be of diagnostic value. In this study we evaluated human papillomavirus (HPV) in situ hybridization (ISH) and p16 immunohistochemistry (IHC) on CB preparations and compared the results with the primary diagnosis and standard HPV tests. DESIGN: In total, 197 HE-stained CB slides prepared from CBs from residual Pap samples (152 ASCUS, 2 ASC-H, 32 LGSIL, 4 HGSIL, 1 AGUS and 6 normal) were analyzed. Hybrid Capture-2 (HC-2)/Cervista testing and HPV ISH and p16 IHC were performed on the CB samples. The test performance characteristics were compared with HPV and p16 assay performances. RESULTS: The cellular architecture was well maintained in CBs with excellent consistency. HPV ISH testing had an excellent concordance with the HC-2/Cervista methods (85%) with high sensitivity (82.6%; 95% CI 75.9, 89.4) and specificity (89.3%). Of all the p16 tests, 38% were positive (60 out of 159 samples). The overall concordance between p16 and HC-2/Cervista (64%), or between p16 and ISH (68%), was lower than the concordance between ISH and HC-2/Cervista (85%). CONCLUSIONS: HPV ISH and p16 IHC testing is feasible, cost effective and practical. A combination of the two tests would ultimately improve diagnostic accuracy, leading to better therapeutic decisions.

Evidence for coordinated interaction of cyclin D3 with p21 and cdk6 in directing the development of uterine stromal cell decidualization and polyploidy during implantation.

Uterine decidualization, characterized by stromal cell proliferation, and differentiation into specialized type of cells (decidual cells) with polyploidy, during implantation is critical to the pregnancy establishment in mice. The mechanisms by which the cell cycle events govern these processes are poorly understood. The cell cycle is tightly regulated at two particular checkpoints, G1-S and G2-M phases. Normal operation of these phases involves a complex interplay of cyclins, cyclin-dependent kinases (cdks) and cdk inhibitors (CKIs). We previously observed that upregulation of uterine cyclin D3 at the implantation site is tightly associated with decidualization in mice. To better understand the role of cyclin D3 in this process, we examined cell-specific expression and associated interactions of several cell cycle regulators (cyclins, cdks and CKIs) specific to different phases of the cell cycle during decidualization in mice. Among the various cell cycle molecules examined, coordinate expression and functional association of cyclin D3 with cdk4 suggest a role for proliferation and, that of cyclin D3 with p21 and cdk6 is consistent with the development of polyploidy during stromal cell decidualization.

Prognostic significance of ploidy, MIB-1 proliferation marker, and p53 in renal cell carcinoma.

BACKGROUND: Pathologic stage is currently the best prognostic factor for predicting outcomes in renal cell carcinoma. The objective of this study was to evaluate the role of DNA ploidy, p53, and Ki-67 (MIB-1) as individual and combined prognostic factors for survival in patients with renal cell carcinoma (RCC). STUDY DESIGN: From 1995 to 2004, 117 patients (78 men and 39 women; mean age 57.34 years), undergoing partial (n = 22) or radical (n = 95) nephrectomy for renal cell carcinoma were retrospectively analyzed. Analysis of MIB-1, p53, and DNA ploidy was performed. Disease-free and overall survival was calculated using Cox proportional hazard models. A combined score was given to incorporate p53, MIB-1, and ploidy as a single variable. RESULTS: On univariate analysis, tumor size, nuclear grade, MIB-1 <10% (p = 0.0059), ploidy (p = 0.0124), pathologic stage group, metastasis at time of operation, and combined score (p = 0.0024) were markedly associated with disease-free survival. On multivariate analysis, only metastasis and pathologic stage were pronounced. For overall survival, size, nuclear grade, MIB-1 <10% (p = 0.0167), pathologic stage group, metastasis, and combined score (p = 0.0456) were pronounced on univariate analysis. Only metastasis was pronounced on multivariate analysis. CONCLUSIONS: We incorporated a combined score to evaluate MIB-1, ploidy, and p53 as a single variable. A combined score is able to give a stronger predictive value of the cellular characteristics of each tumor. Individually and combined with p53, MIB-1, and ploidy were of prognostic significance on univariate analysis. Pathologic stage and presence of metastasis remain the best predictors of disease-free survival.

Transgelin: a potentially useful diagnostic marker differentially expressed in triple-negative and non-triple-negative breast cancers.

Triple negative (TN) (estrogen receptor [ER], progesterone receptor [PR] and HER2-) are highly aggressive, rapidly growing, hormone-unresponsive tumors diagnosed at later stage that affect younger women with shorter overall survival. Most TN tumors are of the basal type. For the remainder, identification of target markers for effective treatment strategies remains a challenge. Transgelin (TGLN) is a 22-kd actin-binding protein of the calponin family. It is one of the earliest markers of smooth muscle differentiation. TGLN has been shown to have important biologic activities including regulating muscle fiber contractility, cell migration, and tumor suppression. We examined TGLN expression in the different molecular subtypes of breast cancer. TGLN expression was examined as a function of tumor size, grade, histologic type, lymph node status, patients' age and overall survival, ER, PR, HER2, and Ki-67 in 101 tumors that included 35 luminal A, 28 luminal B, 4 HER2, and 34 TN types. TGLN positivity (defined as 2+ or 3+) was associated with more aggressive tumors (10% of grade I/II tumors were TGLN+ versus 53% of grade III tumors; P < .001), high Ki-67 count, and low ER and PR expression (P < .001) but not with tumor size, age, or lymph node metastasis. TN (n = 34) tumors were 7.7 times more likely to be TGLN+ than non-TN (n = 67) tumors (77% versus 10%, respectively; P < .001). TGLN may be an excellent diagnostic marker of TN tumors and could be useful in stratification of patients. TGLN may also prove a potential target for future treatment strategies.

A comparison of prognostic tumor markers obtained on image-guided breast biopsies and final surgical specimens.

BACKGROUND: This study was initiated to determine whether tumor markers obtained on image-guided breast biopsy specimens provide accurate prognostic information for women with invasive breast cancer. METHODS: Prognostic tumor markers on preoperative image-guided biopsy and final surgical specimens were compared in 44 patients with invasive breast cancer. RESULTS: Progesterone receptor (PR) discordance was 18%. In 87% of PR discordant cases, the image-guided biopsy was positive and the final specimen was negative (P = 0.03). Tumor grade was discordant in 36% of patients Discordance for estrogen receptor (ER) = 2%; MIB-1 = 18%; Her2/neu = 9%; EGFR = 10%; p53 = 9%; and bcl-2 = 0%. The discordance for these markers was random and did not reach statistical significance. CONCLUSION: Image-guided core needle biopsies provide reliable information for the majority of prognostic tumor makers. A positive progesterone receptor is significantly more likely to be determined by core biopsy rather than the final surgical specimen. Tumor grade should be based upon the final surgical specimen whenever possible.

Psychological, neuropsychological, and electrocortical effects of mixed mold exposure.

The authors assessed the psychological, neuropsychological, and electrocortical effects of human exposure to mixed colonies of toxigenic molds. Patients (N = 182) with confirmed mold-exposure history completed clinical interviews, a symptom checklist (SCL-90-R), limited neuropsychological testing, quantitative electroencephalogram (QEEG) with neurometric analysis, and measures of mold exposure. Patients reported high levels of physical, cognitive, and emotional symptoms. Ratings on the SCL-90-R were "moderate" to "severe," with a factor reflecting situational depression accounting for most of the variance. Most of the patients were found to suffer from acute stress, adjustment disorder, or post-traumatic stress. Differential diagnosis confirmed an etiology of a combination of external stressors, along with organic metabolically based dysregulation of emotions and decreased cognitive functioning as a result of toxic or metabolic encephalopathy. Measures of toxic mold exposure predicted QEEG measures and neuropsychological test performance. QEEG results included narrowed frequency bands and increased power in the alpha and theta bands in the frontal areas of the cortex. These findings indicated a hypoactivation of the frontal cortex, possibly due to brainstem involvement and insufficient excitatory input from the reticular activating system. Neuropsychological testing revealed impairments similar to mild traumatic brain injury. In comparison with premorbid estimates of intelligence, findings of impaired functioning on multiple cognitive tasks predominated. A dose-response relationship between measures of mold exposure and abnormal neuropsychological test results and QEEG measures suggested that toxic mold causes significant problems in exposed individuals. Study limitations included lack of a comparison group, patient selection bias, and incomplete data sets that did not allow for comparisons among variables.

Whole Slide Imaging of Pap Cell Block Preparations versus Liquid-Based Thin-Layer Cervical Cytology: A Comparative Study Evaluating the Detection of Organisms and Nonneoplastic Findings.

OBJECTIVE: Cervical cancer is one of the most common malignancies worldwide, yet it is preventable by population screening. In a previous study, we confirmed the feasibility of utilizing whole slide imaging (WSI) of cell block (CB) preparations to overcome the limitations of digitizing cytologic samples. In this study, we evaluated the accuracy of WSI in identifying various organisms and nonneoplastic findings. STUDY DESIGN: A total of 335 WS images from Pap CB preparations were analyzed using the Aperio system. The test performance characteristics of ThinPrep (TP) and WSI samples were compared for adequacy, for the presence of bacterial vaginosis (BV), fungi, Trichomonas vaginalis (TV) and herpes simplex virus (HSV) and for nonneoplastic findings. RESULTS: The WSI samples contained optimal material from all preparations. BV was diagnosed in 33 WSI versus 36 TP samples. Budding yeasts and/or pseudohyphal forms were noted in 18 WSI versus 19 TP samples. TV organisms (10 of 11 samples) and 1 HSV case were accurately identified in the WSI and TP samples. Squamous metaplasia, keratosis and reactive/reparative and inflammatory changes were easily identified by WSI. CONCLUSIONS: The concept of WSI from Pap CB preparations is potentially feasible for adoption. Digital remote web-based technology eliminates the need for an individual on site, saving time and resources.

Ki-67 expression in axillary lymph node metastases in breast cancer is prognostically significant.

Several studies have documented the prognostic significance of cell proliferation in breast cancer and its positive relationship with tumor grade, size, mitotic activity, hormonal and Her-2 status, and tumor progression. The Ki-67 antigen provides an accurate measure of the growth fraction of a tumor. Ki-67 expression in 103 primary breast carcinomas and their corresponding axillary lymph node metastases was correlated with age, tumor grade, size, estrogen receptor (ER), progesterone receptor (PgR), p53, epidermal growth factor receptor (EGFR), Bcl-2, Her-2 status, and patients' overall survival. Median Ki-67 expression in primary and metastatic tumors was 20% and 15%, respectively. Although there was no difference in overall survival (P = .65, log-rank test) between primary tumors with less than or at least 10% Ki-67 expression, there was significantly better overall survival when Ki-67 expression in lymph nodes was less than 10% (P = .040). For patients whose primary tumors exhibited Ki-67 expression less than 10%, most of their metastatic lesions had a similar low Ki-67; these patients had a favorable outcome. A small subgroup was noted to have a nodal Ki-67 of 10% or more and worse survival (P = .047). For patients whose primary tumors had a Ki-67 of 10% or more, most of their metastatic lesions had similar high Ki-67 values; however, a group of 12 patients had lymph node Ki-67 less than 10% and had a better overall survival (P = .092). Our results showed that measurement of Ki-67 in lymph node is superior to its evaluation in primary tumors. Identification of subgroups of patients in whom Ki-67 expression in lymph nodes differs from expression in primary tumor may assist in the selection of therapeutic options.

Prognostic significance of Bcl-2 in invasive mammary carcinomas: a comparative clinicopathologic study between "triple-negative" and non-"triple-negative" tumors.

Bcl-2 is a tumorigenic protein that is expressed in 25% to 50% of breast cancers. Although its expression has been widely accepted as a favorable prognostic marker, its protective mechanism of action remains unclear. "Triple-negative" tumors are an aggressive subgroup known to carry a poor prognosis. Studies documenting prognostic significance of Bcl-2 expression in triple-negative in comparison to non-triple-negative breast cancers are limited. Bcl-2 expression was correlated with tumor size, grade, histologic type, lymphovascular invasion, lymph node status, patients' overall survival, estrogen receptor, progesterone receptor, Her-2, p53, and epidermal growth factor receptor in 124 triple-negative and 458 non-triple-negative tumors. There were significant differences between triple-negative and non-triple-negative tumors in their relationship to Bcl-2 expression (81% versus 29%, respectively) and tumor aggression. As previously reported, in non-triple-negative tumors, Bcl-2 positivity correlated with less aggressive tumors (94% of grade I tumors were Bcl-2+ versus 62% of grade III tumors, P < .011) and overall survival (P = .008). However, the opposite was true in patients with triple-negative tumors, where Bcl-2 positivity was associated with poorer survival (P = .64). In triple-negative tumors, Bcl-2 positivity was not associated with any of the aforementioned parameters except for a lower incidence of lymph node metastasis. Moreover, by Cox regression analysis of all variables, in patients with triple-negative tumors, lymphovascular invasion (P = .009) and Bcl-2 expression (P = .028) were predictors of poor survival. In conclusion, there are major clinicopathologic differences between breast cancer phenotypes. Our results establish the value of using Bcl-2 in prognostic stratification of patients and its potential therapeutic implications in selecting patients for treatment.

Grading ductal carcinoma in situ of the breast using an automated proliferation index.

Tumor grade, size and margin status are the most significant factors in predicting the behavior of ductal carcinoma in-situ (DCIS). The inclusion of necrosis and nuclear grade in the grading of DCIS has demonstrated a fair but suboptimal agreement between pathologists. The grading of DCIS was studied and compared to the Van Nuys (VN) system, by using our newly proposed unifying "nuclear grade + proliferation index (N+P) grading system for invasive carcinomas. 162 DCIS tumors were studied including 49 VN I, 31 VN II, and 82 VN III cases. The VN and N+P systems were compared with each other and correlated with tumor size, ER, PR, p53, Her-2, EGFR, Bcl-2, p27 and p21 status. The two systems demonstrated similar frequencies for the different grades and an agreement with each other for all of the biomarkers studied. The greatest difference between the two systems was observed for those tumors initially classified as VN II (94% being down-graded to N+P I) and VN III (80% being down-graded to N+P II). These results suggest that the N+P system, combining nuclear grade with automated MIB-1 count, is a potentially valid and reproducible grading system for both non-invasive and invasive mammary carcinomas. It is automated, less subjective in assessing mitotic activity and necrosis and correlates with other prognostic biomarkers.

Clinicopathological characteristics of triple-negative invasive mammary carcinomas in African-American versus Caucasian women.

African-American (AA) women are more likely to have late stage, aggressive, rapidly growing, and less hormone-responsive breast tumors. An aggressive subtype of cancer, known as "Triple-Negative" (TN), that is negative for Her-2 and for estrogen and progesterone receptors (ER and PR), is reported to be more common in AA women. We examined the clinical, histopathologic, and prognostic features of TN tumors in AA and Caucasian women. Tumor size, grade, histologic type, lymphovascular invasion (LVI), lymph node status, patient survival, ploidy status, and expression of ER, PR, p53, epidermal growth factor receptor (EGFR), MIB-1, Bcl-2, Her-2, p27, and p21 were evaluated. The TN tumors (75%) were high grade, large, aneuploid tumors that occurred in younger women and were more likely to have a high rate of LVI, elevated MIB-1 score, and nodal metastases. Patients with TN tumors showed poorer overall survival. There was no difference in overall or disease-free survival (p = 0.46) in the AA versus Caucasian women. LVI was a significant predictor of overall survival in AA but not in Caucasian women. There were minor differences in histopathologic features, biomarker expressions, and survival in AA and Caucasian women with TN tumors. The absence of LVI in AA patients predicted an excellent probability of survival.

A newly proposed semi-automated method of grading invasive lobular carcinoma: a unifying concept and correlation with prognostic markers and patient survival.

Invasive ductal carcinoma (IDC) of the breast is currently graded according to the Nottingham modification of the Scarff-Bloom-Richardson system (SBR). This system involves subjective evaluation of 3 morphologic features: tubule formation, nuclear pleomorphism, and mitosis. Our recently proposed semi-automated Nuclear and Proliferation Index [N+P] grading system for IDC has demonstrated agreement among grades and prognostic markers with better prediction of patient survival than the SBR system. Our present objective is to expand the utilization of the N+P system to grading invasive lobular carcinoma (ILC). Fifty-eight ILC cases were evaluated by the SBR and N+P systems. The 2 systems were compared in terms of correlation with patient survival, tumor size, grade, angiolymphatic invasion, lymph node status, ploidy status, and ER, PR, Her-2, p53, EGFR, and Bcl-2 staining. The N+P and SBR systems demonstrated overall agreement when correlated with clinical and prognostic parameters. Twenty-four of 30 tumors initially classified as SBR Grade II were down-graded to N+P I. Three of 26 tumors initially classified as SBR Grade I were up-graded to N+P II. Grading of ILC provides valuable predictive and prognostic information. The N+P grading system for ILC decreases the element of subjectivity for assessing mitotic activity and appears to be superior to the SBR system in predicting patient survival.

Comorbid anxiety corresponds with neuropsychological dysfunction in unipolar depression.

INTRODUCTION: Unipolar depressives seem apt to show neuropsychological impairment, particularly involving executive function and memory. Yet, not all depressed patients show such deficits. Major depressive illness shares a high rate of comorbid anxiety disorder, and anxiety disorders also tend to correspond with cognitive difficulties. Consequently, depressed individuals with comorbid anxiety disorders may be inclined to demonstrate greater neuropsychological dysfunction than those without anxiety disorders. METHOD: We compared nonpsychotic depressed inpatients with (n=22) and without comorbid anxiety disorders (n=30) to a group of control subjects (n=38) on a brief but broad battery of neuropsychological tests. Patients were tested during an inpatient admission, and data were collected retrospectively from available records. RESULTS: Both groups of depressed patients showed worse memory function than the controls. Yet, executive dysfunction and psychomotor slowing were specific to the depressed group with comorbid anxiety. The comorbid anxiety group also had more impaired scores than either the nonanxious depressed group or the control group. The depressed group without a comorbid anxiety disorder demonstrated no significant slowing compared to the control group. CONCLUSIONS: Major depressive disorder corresponds with significant memory impairment, regardless of comorbid anxiety disorder. Yet, presence of a comorbid anxiety disorder coincides with deficits involving executive function and psychomotor slowing. Clinical and theoretical relevance of the data is discussed.

Bleomycin treatment causes enhancement of virus replication in the lungs of SHIV-infected macaques.

Pneumonia is a major complication of human immunodeficiency virus (HIV) pathogenesis but it develops only after prolonged infection. We used the macaque model to explore a hypothesis that the disease is a two-stage process, the first stage being establishment of the viral infection in the lung and the second being amplification of virus replication by host factors induced by chemical agents or opportunistic pathogens in the lung. Bleomycin, a chemical known to induce diffuse alveolar damage and pulmonary fibrosis with accumulation of macrophages and a rich T helper type 2 (Th2) cytokine environment, was inoculated intratracheally into five of eight SHIV 89.6P-infected macaques and into one uninfected macaque. Three additional simian HIV (SHIV)-infected macaques without bleomycin treatment served as untreated virus controls. Although none of the animals became clinically ill, bleomycin induced classical host responses in the lungs of all the treated, virus-infected macaques. There was enhanced production of the chemokine, monocyte chemotactic protein-1 (MCP-1), that had previously been shown to cause enhanced replication of the virus. Four of the five treated animals developed more productive SHIV infection in the lungs compared with the infected untreated animals. Enhanced virus replication was found primarily in infiltrating macrophages. Enhanced replication of the virus in the lungs was associated with host factors induced by the drug and supported the hypothesis for a two-stage process of pulmonary pathogenesis.

Fixed-dose single agent pegfilgrastim for peripheral blood progenitor cell mobilisation in patients with multiple myeloma.

High-dose chemotherapy and autologous peripheral blood progenitor cell transplantation is an effective treatment for multiple myeloma. Progenitor cells are generally mobilised into the peripheral blood by administration of filgrastim. Pegfilgrastim is a covalent conjugate of filgrastim with a longer half-life. The results of a phase II study of pegfilgrastim, administered as a single injection to mobilise autologous peripheral blood progenitor cells in patients with multiple myeloma, is reported. All patients (n = 19) received 12 mg of pegfilgrastim. Leukaphaeresis was started when the peripheral blood CD34(+) count was >0.015 x 10(9)/l. Daily, leukaphaeresis was performed until the target progenitor cell dose was obtained. The median number of leukaphaeresis procedures required to collect the target CD34(+) cell dose was 2 (range 1-5). A median of 8.4 x 10(6) CD34(+) cells/kg (range 4.1-15.8) was collected. The most common toxicity was bone pain/myalgia. Sustained haematological recovery occurred in all the patients who underwent high-dose chemotherapy followed by autologous peripheral blood progenitor cell transplantation with pegfilgrastim-mobilised cells. A single fixed dose of pegfilgrastim was effective in mobilising adequate peripheral blood progenitor cells in patients with multiple myeloma. The efficacy and toxicity profile was similar to that described with filgrastim treatment.

Phyllodes tumor of borderline malignancy: seven year follow up with immunohistochemical study.

The patient was 80 years old when she initially presented with a left breast mass. Originally, a needle biopsy showed benign stromal and ductal cells. Five years later, the breast mass increased in size and a core needle biopsy showed a biphasic intraductal papillomatous tumor with cellular stroma. Eighteen months later, another biopsy was taken from the breast mass, revealing a well-developed phyllodes tumor (PT) of borderline malignancy. One month later, a simple mastectomy was performed for this 87-year-old woman. Histolopathologic and immunohistochemical studies, including estrogen and progesterone receptors, Ki-67 and p53, performed on tissues from the different biopsy specimens confirmed the progressive transition of the tumor in a 7 year period. An increase in mitotic activity was noted in the later samples. Similarly, percentages of p53- and Ki-67-positive cells were much higher in the stromal and ductal cells of the later samples compared to the original specimen. These findings support the notion that Ki-67 and p53 immunohistochemical staining may be used as simple and practical markers for the evaluation of the malignant potential of PT.

Biphasic acute myeloid leukemia with near-tetraploidy and immunophenotypic transformation.

This report describes a case of acute myeloid leukemia (subtype M1) with biphasic morphology. The bone marrow biopsy showed 2 distinct regions of blasts, one containing large cells and the other small cells. Morphometric and DNA ploidy analysis showed that the mean nuclear area and mean DNA index for the large cell region were 2-fold higher than those for the small cell region. Cytogenetic analysis showed an abnormal near-tetraploid clone. The tumor relapsed following aggressive therapy. The cells from the relapse specimen were similar to the original small cell region with respect to nuclear area and DNA index; however, there was immunophenotypic transformation with gain of CD7 and gain of CD56. Cytogenetically, the relapse specimen showed no evidence of the near-tetraploid clone, but instead had a previously unidentified abnormal clone containing 46 chromosomes and structural abnormalities of 2q and 7q. Biphasic morphology in acute myeloid leukemia may be predictive of a near-tetraploid subclone and immunophenotypic transformation.

HB-EGF directs stromal cell polyploidy and decidualization via cyclin D3 during implantation.

Stromal cell polyploidy is a unique phenomenon that occurs during uterine decidualization following embryo implantation, although the developmental mechanism still remains elusive. The general consensus is that the aberrant expression and altered functional activity of cell cycle regulatory molecules at two particular checkpoints G1 to S and G2 to M in the cell cycle play an important role in the development of cellular polyploidy. Despite the compelling evidence of intrinsic cell cycle alteration, it has been implicated that the development of cellular polyploidy may be controlled by specific actions of extracellular growth regulators. Here we show a novel role for heparin-binding EGF-like growth factor (HB-EGF) in the developmental process of stromal cell polyploidy in mice. HB-EGF, which is one of the earliest known molecular mediators of implantation in mice and humans, promotes stromal cell polyploidy via upregulation of cyclin D3. Adenoviral delivery of antisense cyclin D3 attenuates cyclin D3 expression and abrogates HB-EGF-induced stromal cell polyploidy in vitro and in vivo. Collectively, the results demonstrate that the regulation of stromal cell polyploidy and decidualization induced by HB-EGF depend on cyclin D3 induction.