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Analytically validated assays to interrogate biomarker status in clinical samples are crucial for personalized medicine. PTEN is a tumor suppressor commonly inactivated in prostate cancer that has been mechanistically linked to disease aggressiveness. Though deletion of PTEN, as detected by cumbersome fluorescence in situ hybridization (FISH) spot counting assays, is associated with poor prognosis, few studies have validated immunohistochemical (IHC) assays to determine whether loss of PTEN protein is associated with unfavorable disease.
PTEN IHC was validated by employing formalin fixed and paraffin embedded isogenic human cell lines containing or lacking intact PTEN alleles. PTEN IHC was 100% sensitive and 97.8% specific for detecting genomic alterations in 58 additional cell lines. PTEN protein loss was then assessed on 376 prostate tumor samples, and PTEN FISH or high resolution SNP microarray analysis was performed on a subset of these cases.
PTEN protein loss, as assessed as a dichotomous IHC variable, was highly reproducible, correlated strongly with adverse pathologic features (e.g. Gleason score and pathological stage), detected between 75% and 86% of cases with PTEN genomic loss, and was found at times in the absence of apparent genomic loss. In a cohort of 217 high risk surgically treated patients, PTEN protein loss was associated with decreased time to metastasis.
These studies validate a simple method to interrogate PTEN status in clinical specimens and support the utility of this test in future multi-center studies, clinical trials and ultimately perhaps for routine clinical care.
Prostate cancer is the most common solid organ tumor in American men and the second most common cause of cancer deaths (1). Despite improvements in early detection, we still lack molecular markers to effectively distinguish men with high risk disease from the indolent majority. Identification of molecular aberrations that contribute to the development of lethal disease has the additional benefit of providing specific targets for therapy. Work over the last decade has firmly established that loss of the PTEN tumor suppressor (phosphatase and tensin homologue on chromosome 10) is one of the most common somatic genetic aberrations in prostate cancer and is frequently associated with high risk disease (2-13). Despite these findings, however, prostate cancer specimens are not yet routinely interrogated for PTEN loss in any clinical setting.
The majority of previous work on PTEN loss in prostate cancer has focused on genomic deletions of the PTEN locus at 10q23. Such deletions, most commonly identified by fluorescence in situ hybridization (FISH), occur in 10-70% of prostate cancer cases depending on the study population examined (2, 4, 6, 8, 10-15) and are associated with poor prognosis (13, 16-20). Interestingly, heterozygous PTEN deletions far outnumber homozygous deletions in prostate cancer and may also result in poor outcomes (11-13, 15-17, 20). There has been much debate about whether this may be explained by haploinsufficiency for PTEN or whether inactivation of the second allele in these cases has occurred but is not detectable by FISH (21, 22). While a number of early studies suggested that the rate of PTEN mutation and epigenetic modification in prostate cancer was relatively high, it is now known that many of these initial estimates may have been falsely elevated due to the existence of a PTEN pseudogene (2, 3, 23). Recent studies have suggested that alternative mechanisms of PTEN post-transcriptional down-regulation may play an important role in prostate cancer (24-28). To date, however, the relative frequency of PTEN inactivation by mechanisms other than genomic deletion in clinical prostate cancer specimens remains unclear (29).
For clinical settings, the evaluation of allelic loss of PTEN by FISH is cumbersome, requiring counting of the number of fluorescent signals relative to control signals in partially sectioned interphase cells. Thus, reliable detection of PTEN protein status by immunohistochemistry (IHC) in routinely processed clinical formalin-fixed and paraffin-embedded (FFPE) pathology specimens could prove highly useful for the implementation of PTEN status as a clinically relevant prognostic biomarker. Importantly, development of such an assay may also detect cases where PTEN inactivation occurs by mechanisms other than genomic deletion. As more therapies targeting various components of the PI3K signaling cascade become available, a robust assay to determine PTEN status will likely have an important role in clinical care. To address this need, we have developed and validated a simple and robust IHC assay to detect PTEN protein in paraffin-embedded specimens using a commercially available rabbit monoclonal antibody. Since our approach to analytical validation of PTEN staining can be ported easily to other laboratories, if our findings can be validated in additional large cohorts, this robust IHC assay may prove to be useful in a number of clinical arenas.
Ten tissue microarrays (TMA) were constructed from formalin-fixed paraffin-embedded prostate, lymph node or distant metastasis tissue collected from a total of 376 patients with prostate cancer who underwent radical prostatectomy or surgical resection of metastatic lesions at our institution. Between 1 and 4 (average = 3) 0.6 mm cores of tumor tissue for each patient were arrayed. For most patients, surrounding benign prostate tissue was included in the TMA as a control. Of these 376 patients, 56 patients were enrolled in an adjuvant trial of docetaxel treatment due to high risk pathologic features (30). An additional 217 patients were from a previously described cohort with biochemical recurrence following radical prostatectomy for clinically localized prostate cancer (31).
Finally, an additional group of 53 patients with concurrent high resolution single nucleotide polymorphism (SNP) microarray analysis for genomic copy number alterations in their prostatic carcinoma were also included. Detailed results of the genome-wide study from these patients will be reported elsewhere (J Xu, WB Isaacs et al., manuscript in process). For these cases, a single representative histologic section that was adjacent to the tumor lesion harvested for genomic DNA was analyzed for PTEN protein loss.
Three different human cell lines in which PTEN alleles were subjected to targeted disruption using homologous recombination were used in this study as controls for immunohistochemistry, including MCF-10A mammary epithelial cells (32; gift of Kurtis E. Bachman), and the DLD-1 and HCT116 human colorectal cancer cell lines (33; gift of Todd Waldman). Wild type (PTEN+/+) heterozygous null (PTEN+/−), and homozygous null (PTEN−/−) cell variants were maintained as previously described (32,33), harvested and subjected to formalin fixation and paraffin embedding as described (34). The human prostate cancer cell lines LNCaP, DU145, and PC-3 were acquired from the American Type Culture Collection (Manassas, VA). LNCaP cells harbor one deleted allele of PTEN and one mutated allele of PTEN and do not express PTEN protein (7). PC3 cells harbor a homozygous deletion of PTEN and do not express PTEN protein (7, 10). DU145 cells harbor an apparent coding SNP in PTEN, but are otherwise wildtype and express PTEN protein (10). We also obtained 56 additional cell lines of the NCI-60 panel of cell lines from the Developmental Therapeutics Program at the NCI to evaluate PTEN staining. Cell lines were used to generate cell line microarrays (analogous to TMAs) (35) using a recently updated method that will be described in detail elsewhere (D. Esopi, S Yegnasubramanian, AM De Marzo, manuscript in process).
As an additional positive control we obtained a molecular clone of PTEN that is driven by the CMV promoter (PTEN NM_000314 Human cDNA Clone, Origene; Rockville, MD) and transfected this construct into HCT116 PTEN−/− and DLD-1 PTEN−/− cells using Oligofectamine according to the manufacturer's directions (Invitrogen; Carlsbad, CA). Mock or PTEN transfected cells were harvested after 48 hours and prepared for cell blocks as described above.
Fluorescence in situ hybridization (FISH) was performed on two TMAs (145 patients, 361 spots representing a spectrum of lesions including benign prostate, PIN, primary tumors and lymph node metastases) using the Vysis LSI PTEN (10q23) / CEP 10 Dual Color Probe (Abbott Molecular Inc.; Abbott Park, IL) according to the manufacturer's directions. Briefly, a 4 μm paraffin section was baked at 56°C for 2 hours, then de-waxed and rehydrated using xylene and graded ethanol, respectively. The TMA sections were pretreated using Paraffin Pretreatment Reagent Kit III (Abbott Molecular Inc.; Abbott Park, IL). TMAs and the PTEN/CEP10 FISH probes were co-denatured at 94°C for 5 min and hybridized overnight at 37°C in a humid chamber (StatSpin ThermoBrite; IRIS Inc, MA)
FISH scoring was conducted using a Nikon 50i epifluorescence microscope equipped with X-Cite series 120 illuminator (EXFO Photonics Solutions Inc., Ontario, CA) and a 100x/1.4 NA oil immersion Neofluar lens. Grayscale images were captured for presentation using Nikon NIS-Elements software and an attached Photometrics CoolsnapEZ digital camera, pseudo-colored and merged.
FISH interpretation was performed by a pathologist (BG) blinded to the IHC results. To establish normal cutoffs for scoring and to ensure that FISH signal truncation seen in tissue sections was not scored as a false positive, the PTEN/centromere 10 (CEP10) signals were tallied in 50 non-neoplastic normal-appearing prostate specimens contained on the same TMAs as the experimental specimens, and the mean and standard deviation (SD) for PTEN/CEP10 ratios and percentage of epithelial cells containing 0 or 1 PTEN probe were calculated. After scoring 30 cells, a tumor was considered to have PTEN loss if the percentage of cells showing 0 or 1 PTEN probe signal was greater than 2 SD above the mean established from the normal tissue and the PTEN/CEP10 ratio was more than 2 SD below the mean established in normal tissue. A case that was represented by more than one tissue core on the TMA was considered to have PTEN loss if any TMA spot in the case showed PTEN loss.
A number of commercial antibodies were evaluated to identify those most highly sensitive and specific in IHC cell line staining experiments. These antibodies included the following: Zymed/Invitrogen (Carlsbad, CA) rabbit polyclonal cat No 51-2400; Epitomics (Burlingame, CA) rabbit monoclonal clone Y184; Cascade BioSciences (Winchester, MA) mouse monoclonal clone 6H2.1; Santa Cruz Biotechnologies (Santa Cruz, CA) mouse monoclonal clone 28H6; and Cell Signaling Technology (Beverly, MA) rabbit monoclonal D4.3. Using the PTEN positive and negative cell lines described above we found that the rabbit monoclonal antibody from Cell Signaling Technology performed the best overall at the following conditions: Antigen unmasking was performed by steaming in EDTA buffer (pH 8.0) for 45 minutes. Endogenous peroxidase activity was quenched by incubation with peroxidase block for 5 minutes at room temperature. Non-specific binding was blocked by incubating in 1% bovine serum albumin in Tris-HCl pH 7.5 for 20 minutes at room temperature. Slides were incubated with a 1:100 dilution of rabbit monoclonal anti-PTEN antibody (clone D4.3, #9188, Cell Signaling Technologies, Beverly, MA) overnight at 4°C. A horseradish peroxidase–labeled polymer (PowerVision Poly-HRP anti-Rabbit IgG; Leica Microsystems, Bannockburn, IL) was then applied for 30 minutes at room temperature. Signal detection was performed using 3,3′-diaminobenzidine tetrahydrochloride (DAB) as the chromagen. Slides were counterstained with hematoxylin, dehydrated, and mounted. In a number of experiments we also evaluated another rabbit monoclonal antibody from Cell Signaling Technology (clone 138G6) and found it to perform similarly to the D4.3 clone.
PTEN protein was visually scored using a dichotomous scoring system by two urologic pathologists (TLL and AMD). IHC scoring was blinded with respect to FISH and SNP array results, pathologic stage and final Gleason score at radical prostatectomy, as well as patient outcome. Using this system, each spot of tumor tissue was scored as negative or positive for PTEN protein by comparing staining in malignant glands with that of adjacent benign glands and/or stroma which provided an internal positive control within each tissue core. Staining was classified as negative if the intensity was markedly decreased or entirely negative across all tumor cells compared to the surrounding benign glands and/or stroma. A given spot was dropped from the analysis if these benign areas lacked PTEN staining (this occurred in <5 TMA spots total).
PTEN IHC was performed on an additional 53 prostate tumors for which PTEN copy number data was available from high resolution single nucleotide polymorphism (SNP) microarray analysis (Affymetrix 500K). Further details regarding the genome wide copy number analyses will be provided in a separate publication (J Xu, WB Isaacs et al., manuscript in process).
Pearson chi-squared and Fisher's exact tests were used to determine the association of PTEN protein loss with pathologic variables and PTEN genomic loss. To investigate univariable associations between PTEN loss and time to distant metastasis and prostate cancer-specific death, Kaplan-Meier survival curves were calculated and compared by log-rank tests. Multivariable-adjusted associations were investigated using Cox proportional hazards regression; models included terms for pathologic stage, Gleason grade, preoperative PSA, and surgical margin status. Statistical analyses were performed using SAS version 9.1.
We first validated our immunohistochemical (IHC) assay analytically for PTEN protein loss using formalin fixed paraffin-embedded cell blocks created from 3 distinct isogenic human cell lines with and without PTEN deletion. In wild-type HCT-116, MCF-10A, and DLD-1 cells, a variable degree of cytoplasmic and nuclear PTEN protein immunostaining was evident, while isogenic lines which had undergone two rounds of somatic homologous recombination (resulting in disruption of both alleles of PTEN showed complete absence of PTEN protein by IHC (Figure 1). In MCF-10A cells that were heterozygous for PTEN disruption (PTEN+/−), there were intermediate levels of staining between wild-type (PTEN+/+) and homozygous mutant variants (PTEN−/−) (Figure 1). In contrast, PTEN−/− HCT-116 or DLD-1 cells transiently transfected with an expression vector containing a cDNA encoding full length human PTEN showed a number of cells with greatly increased PTEN protein by IHC (Figure 1). Taken together, these isogenic cell line experiments verify the specificity of the Cell Signaling Technologies rabbit monoclonal antibody as well as our IHC protocol. For additional validation, we also stained cell pellets that were obtained from PC3, LNCaP and DU145 human prostate cancer cell lines that have known PTEN genomic status. As expected, both PC3 cells and LNCaP were completely negative for staining, whereas DU145 cells showed moderate levels of both cytoplasmic and nuclear staining for PTEN (not shown). To further assess the performance of the assay we next examined 55 of the remaining cell lines from the NCI-60 cell line panel, since PTEN genomic status has been reported from these lines (Table S1; 36, 37). The sensitivity for IHC staining to identify cell lines with predicted oncogenic alterations in PTEN was 100% (13 of 13 lines with mutant PTEN showed loss of IHC staining), with a specificity of 97.8% (44 of 45 cell lines without PTEN mutation did not show PTEN protein loss by IHC).
PTEN protein loss by IHC was scored on paraffin-embedded prostate tissues from a total of 376 patients on 10 tissue microarrays (TMAs). PTEN protein was expressed in all (100%) cases of benign prostate tissue present on tissue microarrays. While there was some variability in staining intensity, PTEN protein was diffusely expressed in the cytoplasm and nucleus of luminal and basal epithelial cells, as well as in the surrounding stromal tissue (Figure 2). In fact, virtually all cell types present (including endothelial cells, smooth muscle cells, inflammatory cells, and peripheral nerves) were positive for PTEN staining. This pattern of staining was also found on 53 standard tissue slides (see below).
To score PTEN protein expression in prostatic carcinoma, we devised a binary scoring system wherein malignant glands were scored as positive or negative for cytoplasmic PTEN protein relative to the internal control of surrounding benign glands and/or stroma (Figure 2). We considered cases in which the majority (>90%) of cells retained easily discernible levels of PTEN staining to be positive for PTEN protein (e.g. Figure 2C, left panel). Cases considered negative for PTEN protein either showed a complete absence of PTEN staining or extremely weak intensity staining in >10% of cells. In the latter cases, however, the overall staining was always strikingly reduced as compared to surrounding tissue, particularly in nuclei which appeared blue only without discernible brown staining. Interobserver reproducibility for this simple scoring system was high. On an array with 117 tumor spots scored independently in a blinded fashion by two pathologists (TLL and AMD), there was 95% (111/117 spots) agreement on the presence or absence of PTEN protein by IHC in malignant glands (kappa = 0.87 or "almost perfect" agreement). In this set, 26-27% (depending on the observer) of the spots showed PTEN loss in tumor cells.
A fraction of tumors arrayed on the TMAs showed prominent intra-tumoral heterogeneity for PTEN expression, with some spots positive for PTEN while other spots from the same index tumor were negative. Overall, intra-tumoral heterogeneity in PTEN expression was observed in 9% (20/220 cases) of primary tumors. Occasionally, this heterogeneity of PTEN protein expression was seen even within a given TMA spot, with some malignant glands expressing PTEN while adjacent glands (or even adjacent cells within a malignant gland) were entirely negative (Figure 2D and 2E). For the purposes of this study, a case was scored as PTEN-negative if any tumor spot from that case showed greater than 10% of tumor cells with markedly decreased PTEN protein. Similarly, a total of 53 prostatic tumors were scored for PTEN expression using single tumor sections on standard histologic slides rather than TMAs, and 11% (6/53) of these cases showed two clear populations of tumor cells where one population had PTEN loss and the other did not. In this setting, as above, a case with PTEN protein loss in greater than 10% of the sectioned tumor was considered to have PTEN loss for the purposes of data analysis.
Overall, PTEN protein was scored by IHC in 397 tissue samples collected from 376 individual patients. Pathologic stage data was available on 263 patients (66%) and Gleason grade was available on all tissues. In these samples, PTEN protein loss by IHC was highly correlated with increased pathologic stage (p=0.003) and Gleason grade (p<0.0001, Figure 3). Overall, 41% (17/41) of pT3bN0 cases showed PTEN loss, compared to 34% (39/112) of pT3aN0 cases and 14% (6/44) of pT2N0 cases. Similarly, 45% (65/144) of Gleason 8-10 cases showed PTEN protein loss compared to 39% (61/155) of Gleason 7 cases and 20% (20/98) of Gleason 5-6 cases.
Consistent with the correlation with pathologic stage and grade, PTEN loss by IHC was also highly correlated with the tissue diagnosis (Figure 3). High grade prostatic intraepithelial neoplasia (PIN) showed the lowest rate of PTEN loss (12% or 3/25 cases). In contrast, pelvic lymph node metastases and distant metastases showed the highest rates of PTEN loss (46% or 20/43 cases and 57% or 4/7 cases respectively). Primary prostatic tumors showed an intermediate rate of PTEN loss when considered as a group (38% or 119/308).
To determine whether the PTEN IHC assay was sensitive or specific for detecting PTEN allelic loss in clinical prostate cancer specimens, we evaluated PTEN protein by IHC in a series of prostate tumors for which we also had available PTEN genomic status by concurrent FISH (66 cases, of which 56 were from the high risk group enrolled in an adjuvant trial of docetaxel) or high resolution copy number SNP microarray analysis (53 cases). For the cases with concurrent PTEN FISH data, presence or absence of PTEN protein was highly correlated with presence or absence of PTEN loss of heterozygosity (Figure 4, Table 1, p = 0.0044 by Fisher's exact test). Of 66 primary prostate tumors with available PTEN FISH data, 24 (36%) had loss of at least one PTEN allele. Of these, 75% (18/24) had loss of PTEN protein by IHC, indicating that PTEN IHC is sensitive for the detection of genomic PTEN loss. For the cases with concurrent SNP microarray copy number analysis, PTEN IHC was also highly correlated with PTEN genomic status (Table 2, p = 0.00026 by Fisher's exact test). Of 53 cases with SNP microarray data, 41% (22 cases) showed loss of at least one allele of PTEN. Of these cases, 86% (19/22) showed PTEN protein loss by immunohistochemistry. Of the cases with SNP microarray data and heterogeneous intra-tumoral PTEN immunohistochemical staining, 33% (2/6 cases) showed no evidence of genomic PTEN loss, 50% (3/6 cases) showed loss of one PTEN allele, and 17% (1/6) showed homozygous loss of both PTEN alleles.
PTEN immunohistochemistry was as sensitive for the detection of cases with loss of one PTEN allele (heterozygous deletion) as it was for the detection of cases with loss of both alleles (homozygous deletion) (87% or 13/15 vs 86% or 6/7 respectively, Table 3). Importantly, of cases with PTEN protein loss by immunohistochemistry, 45% (15/33) and 37% (11/30) did not have any PTEN genomic loss detectable by FISH or SNP microarray, respectively.
A subset of the RRP specimens are part of a previously described retrospectively-studied high risk surgical cohort of patients who underwent radical prostatectomy (RRP) by a single surgeon at the Johns Hopkins Hospital between 1986 and 1996 and subsequently developed biochemical recurrence (31) (Table 3). Of note, none received neoadjuvant chemotherapy or hormonal therapy prior to development of metastatic disease (31). Median patient follow-up was 16 years and tissue was present for TMA construction from 217 of the original 304 cases in this cohort. At RRP, 22% (49/217) had a Gleason score (GS) of 6 or less, 51% (110/217) had GS of 7 and 27% (58/217) had a GS of 8-10. 83% (181/217) had extracapsular extension, 34% (74/217) had seminal vesicle involvement and 27% (58/217) had lymph node metastases. 36% (78/217) had positive surgical margins. By definition in this cohort, 100% (217/217) of patients had biochemical recurrence during the follow-up period, with 25% (52/210) showing evidence of local recurrence and 60% (124/208) with distant metastases during follow-up. 38% (79/207) of patients died of prostate cancer during the study period. In Kaplan-Meier analysis, loss of PTEN immunostaining was significantly associated with decreased time to metastasis (p=0.03, Figure 5). There was a correlation between PTEN protein loss and decreased time to prostate-specific death, although this did not reach statistical significance (p=0.06). In multivariable Cox regression analysis, PTEN protein was not associated with time to metastasis or prostate cancer death (Table 3).
PTEN genomic loss was first identified as a molecular aberration common in prostate cancer nearly 15 years ago (2, 37). In early studies using microsatellite analysis, loss of heterozygosity (LOH) at the PTEN locus was reported in 10-55% of primary tumors from surgical cohorts (2, 4-6, 8, 9). In more recent studies using FISH, loss of at least one PTEN allele has been reported in as few as 17% of patients with tumors incidentally discovered on transurethral resection (TURP), however PTEN allelic loss is present in 17-68% of primary tumors from surgical cohorts and up to 77% of hormone-resistant primaries discovered on TURP (11-13, 15-17, 20, 38). In general, PTEN loss is more common in prostate cancer metastases than in primary tumors, with rates of loss reported near 50% in three independent studies using different methods of detection (6, 15, 19). Despite the variations in reported rates of genomic PTEN loss, a nearly universal finding in the FISH studies is that loss of one PTEN allele is significantly more frequent than loss of both PTEN alleles in surgical cohorts. However, rates of homozygous loss also vary by the cohort examined. While homozygous loss of PTEN hovers around 0-10% of primaries, it is closer to 50% of metastatic or hormone-resistant cases (11-13, 15-17, 20).
Overall, the wide range in reported frequency of PTEN genomic loss in prostate cancer likely reflects the close association of PTEN loss with high risk pathologic features and possibly an association with androgen-insensitive disease. Thus, the frequency of PTEN loss is higher in cohorts enriched for aggressive disease, with increased pathologic stage and Gleason grade (18). Despite the close association with pathologic variables, at least two separate studies have found that PTEN genomic loss is independently correlated with decreased time to biochemical recurrence (13, 16, 20). However, at least three separate studies performed in TURP and surgical cohorts have not found an association between PTEN genomic loss and survival (16-18).
Interestingly, the association between PTEN genomic loss and biochemical recurrence has most commonly been documented for hemizygous deletions, likely because they are much more frequent than homozygous deletions. Although several studies have demonstrated higher hazard ratios for homozygous compared to hemizygous PTEN loss, the correlation between loss of only one allele of PTEN and decreased time to biochemical recurrence remains significant (13, 16, 20). This suggests that either PTEN is a haploinsufficient gene, or in cases of hemizygous loss, the second allele is commonly inactivated by additional mechanisms which are not detected by FISH. While there is some evidence for PTEN haploinsufficiency in the mouse, data to support this hypothesis are lacking in humans (21, 22). Thus the correlation between loss of a single PTEN allele and features of aggressive disease in prostate cancer strongly suggests that alternative epigenetic or perhaps non-genomic mechanisms of PTEN inactivation play an important role in prostate cancer progression (29). This also suggests that FISH may be systematically underestimating the frequency of PTEN loss in prostate cancer.
The possibility that PTEN FISH may fail to detect some cases of prostate cancer with PTEN inactivation strongly argues for the need for an alternative assay to detect PTEN loss. Although immunohistochemistry to detect PTEN protein levels is an obvious alternative, until recently, studies using this method have been impeded by the lack of reliable antibodies as well as by a paucity of dependable genetic controls. To our knowledge, there are at least 10 prior studies in the literature looking at the utility of PTEN immunohistochemistry for the detection of PTEN loss in prostate cancer (10-12, 14-16, 39-42). While some found an association between PTEN protein loss and Gleason grade, stage, or biochemical recurrence, many did not. Some of this variation may be due to antibody performance. While this manuscript was in preparation, a study evaluating commercially available PTEN antibodies for immunohistochemistry found that many antibodies resulted in nonspecific nucleolar staining in cell lines with known PTEN genomic loss (43). Importantly, this study independently corroborated our findings that recently available rabbit monoclonal antibodies to PTEN perform much more reliably than older clones or polyclonal antibodies. In addition to problems with older antibodies, earlier studies also employed widely variable and often complex scoring systems for PTEN protein, and did not account for interobserver variability in scoring. Only two studies have looked at the association between PTEN protein loss and disease progression and survival in prostate cancer, and while one study scored cytoplasmic staining intensity and extent, the other found an association only with nuclear PTEN staining (14, 16).
Given the wide variation of methodology and results in the literature, we set out to develop an immunohistochemical assay for PTEN that would be simple enough to allow routine use in clinical pathology specimens. One advantage of our study is that we had access to a number of isogenic cell lines both with and without PTEN genomic deletion. Using these cell lines, we optimized a staining protocol for a recently available rabbit monoclonal antibody, wherein all lines with PTEN deletion showed a complete absence of PTEN protein. With our staining protocol, background benign prostatic glands and stroma showed robust staining for PTEN. Because of the presence of an internal positive control in all samples, we were able to apply a simple dichotomous and highly reproducible scoring system for malignant glands, with cytoplasmic PTEN either present, or markedly decreased. Interestingly, Sangale et al. also applied a simple dichotomous scoring strategy in their approach to PTEN IHC scoring in clinical samples (43).
Perhaps one of the most important advantages of our study is that we were able to correlate PTEN protein expression with PTEN genomic status. We found that PTEN immunohistochemistry is highly sensitive for detection of PTEN genomic loss, detecting nearly 80% of cases with loss by FISH and over 80% of cases with loss by high resolution SNP array. Only three previous studies have validated their immunohistochemical assay in a similarly rigorous fashion. Yoshimoto et al reported PTEN levels as a product of the intensity and percentage of cytoplasmic and/or nuclear staining and although they did not provide data for each individual case, they found an overall correlation between PTEN protein levels as a continuous variable and PTEN genomic status (12). Verhagen et al employed a similar scoring system and found that 66% (10/15) of cases with PTEN deletion by FISH showed PTEN protein loss (11). Recently, Han et al did a similar study where they scored cytoplasmic staining intensity on a 0-2+ scale and correlated with FISH results (15). They reported that they detected 52% of cases with PTEN loss by FISH using immunohistochemistry if only cases with 0+ immunostaining were considered to be truly PTEN protein negative.
One of the most intriguing findings in the current study was that 45% and 37% of tumors with PTEN protein loss did not show genomic deletions detectable by FISH or high resolution SNP microarray, respectively. Although it is possible that FISH may not detect some small deletions in PTEN, the similar data obtained from the high resolution SNP microarray suggests that this is a less likely explanation for our findings. Additionally, we found that PTEN IHC was as sensitive for the detection of hemizygous loss by SNP array as it was for the detection of homozygous loss. Although even high resolution SNP microarrays can miss very small deletions depending on a number of factors, this data strongly suggests that in addition to genomic deletion, alternative mechanisms for PTEN inactivation likely exist. Interestingly, other authors have reported similar findings. Han et al reported that 35% (6/17) of their PTEN protein negative cases did not show genomic deletion by FISH and Verhagen et all reported that 33% (5/15) cases without PTEN protein showed no evidence of deletions by FISH (11, 15).
The frequency with which PTEN is inactivated by mutations, epigenetic modifications and/or non-genomic means remains unclear in prostate cancer. Although early studies reported a high rate of mutations and methylation in the PTEN promoter region, it is likely that some of these estimates were falsely elevated because of detection of a PTEN pseudogene that harbors a high rate of such changes (2, 3, 11, 15, 23, 37, 44-46). More recent studies have documented only rare cases in which PTEN is inactivated by point mutations or small insertions or deletions (indels) (19, 28, 47). Additionally, the functional consequences of PTEN promoter methylation for PTEN expression are unclear since recent findings that the PTEN promoter is shared with that of a p53-target gene (KILLIN) which may be a tumor suppressor in its own right (48). Recent studies have elucidated the role of microRNAs and pseudogene deletion in the regulation of PTEN protein levels (26, 27). Further, a separate study identified chromosomal translocations in MAGI-2, a membrane-associated guanylate kinase known to bind and stabilize PTEN protein (28, 49). Overall, these data strongly suggest that PTEN inactivation in prostate cancer occurs through a number of mechanisms, many of which have yet to be described. This, in turn, highlights the importance of an assay that will detect PTEN inactivation occurring via multiple mechanisms.
While several studies have shown an association between PTEN protein expression and the surrogate clinical endpoint of biochemical recurrence, only two prior studies have examined whether PTEN protein expression is associated with metastasis and death in prosate cancer. Halvorsen et al looked at PTEN protein expression in prostate tumors from 104 surgically treated patients and found that cytoplasmic PTEN expression was an independent predictor of locoregional recurrence in this cohort (14). McCall et al found that nuclear PTEN was independently associated with disease specific survival in a group of 68 tumors in TURP specimens (16). In our cohort of 217 patients with biochemical recurrence, we found that PTEN protein expression, as a single variable, was a significant predictor of decreased time to metastasis and correlated with decreased time to prostate-cancer specific death (although this didn't reach statistical significance). Given that this cohort was significantly enriched for high risk pathologic features and that PTEN protein loss is highly correlated with such features, it is likely that PTEN protein expression as a single variable prognosticator will perform even better in more typical surgical cohorts.
Ultimately, the IHC test for PTEN protein expression described herein will most likely be of use in the setting of prostate tumors diagnosed on needle biopsy for a number of reasons. First, this simple test, which is easier and cheaper to perform than FISH, could easily be routinely performed on needle biopsy specimens and compared to FISH, will likely identify additional cases lacking PTEN protein. Second, although we found PTEN protein loss was highly correlated with tumor grade and pathologic stage in radical prostatectomy cases (and thus not independently associated with clinical outcome), in the setting of needle biopsies, pathologic tumor stage is unknown and tumor grade is routinely underestimated in approximately 20% of cases. In this way, as a prognostic biomarker, loss of PTEN protein in a needle biopsy specimen may be useful for the identification of presumed low risk prostate cancer patients (such those on active surveillance) that are prone to disease progression and hence require treatment. Third, as a predictive biomarker, loss of PTEN may prove useful for the selection of appropriate patients for treatment with emerging PI 3-Kinase pathway-targeted therapies, a number of which are currently in clinical trials for prostate cancer. Finally, PTEN loss may also serve as a biomarker of hormonal therapy resistance in advanced prostate cancer (50).
PTEN is a tumor suppressor commonly inactivated in prostate cancer. Genomic deletions of PTEN are associated with poor prognosis, but are interrogated in tissue samples using relatively cumbersome fluorescence in situ hybridization (FISH). We demonstrate that PTEN protein expression evaluated by immunohistochemistry (IHC), using a new commercially available monoclonal antibody, is simple to perform and score after genetic validation and sensitively detects PTEN genomic loss. Further, PTEN IHC detects additional cases of PTEN protein loss compared to genomic methods. Finally, loss of PTEN by IHC correlated with adverse pathological features and decreased time to metastatic disease in a surgical cohort. This analytically validated assay may be used in additional studies to determine its efficacy as a clinical test, including as a prognostic biomarker in needle biopsies and as a predictive marker to identify patients who could benefit from emerging PI3K pathway-targeted therapies or who may be resistant to hormonal therapies.
Grant Support: Funding for this research was provided in part by the NIH/NCI Prostate SPORE P50CA58236 and the Patrick C. Walsh Prostate Cancer Research Fund of which AMD is the Peter J. Sharp Scholar.
Disclosure/Conflicts of Interest: The authors have no disclosures/conflicts of interest to declare.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Bought 2 posters to decorate Mom's room in a dementia care community. This poster has a beautiful fall scene with nice color and it is printed on a heavy poster board unlike the other poster that was printed on thin paper. Would recommend to others.
see comment below
By Sparrer
from Weste Australiarn
very pleased with my purchase but now have no more room for any size pics. So need to unsubscribe with regret.
(5 of 5 customers found this review helpful)
Vivid Colors
By Bamboo
from Overseas - Asia Pac
This canvas was purchased for a work colleague. She wanted something that would calm and sooth her. She particularly loves trees and fall foliage. The colors are so vibrant and lovely. She was thrilled! This is now her spot to relax when she has a stressful day. Thanks so much AllPosters.com!
Fall Picture in woods
By Doctor Dee
from Texas
I put it in a frame and it's on the Living Room WallVery nice to look at when you go into that room. Bright - Colorful - Relaxing look.
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World |
KABUL The Taliban said they and other militant groups were behind a coordinated assault in the Afghan capital, Kabul, on Sunday which included attacks on the British and German embassies in the heavily guarded, central diplomatic district.
(For Slideshow Taliban under siege, click here)
The Taliban also said they were carrying out attacks in several Afghan provinces.
(Reporting by Hamid Shalizi; Writing by Michael Georgy; Editing by Robert Birsel)
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Scott is a total alarm and he doesn't know it.
dodane przez Scott luty 15, 2005
Trending Now
Top Definition
The alarm describes a freaky asian girl's orgasm. When she continuously wails that high pitch, the sound resembles an alarm hence the name alarm.
I was fuckin this asian chick and i thought the fire alarm went off.
dodane przez jeffr kwiecień 27, 2008
An audible alert to wake you up in the morning.
He set Alarm to wake him up for work.
Plural: He set 2 larms to wake his tired ass up.
Plural: He set 2 larms to wake his tired ass up.
dodane przez Green31 lipiec 10, 2014


text: Google's Gonna Keep Riding This Search Thing OutKyle VanHemert5/26/10 1:20pmFiled to: BlockquoteGoogleYahooSearchEnginescarolbartzArringtonAroraNikesharora152EditInvite manuallyPromoteDismissUndismissHideShare to KinjaToggle Conversation toolsGo to permalink Recently, Yahoo CEO Carol Bartz forecasted trouble for Google because search accounts for "99.9% of their business." In a talk with Michael Arrington today, Google's President of Global Sales Operations Nikesh Arora leveled this response. Giddyup.When Arrington brought up Bartz's comments, Arora was quick to respond: If we are a one trick pony, we have a pretty good trick. And it's true—Google dominates search because they're good at it—but at the same time it's not: clearly the company's looking at Android as a future source of revenue, and more ads are showing up on YouTube seemingly every day. Advertisement When someone asked, as a follow-up question, if Google's "second trick" could be its cloud services, Arora said he thought it could be a billion dollar source of revenue within four years. I wonder what Bartz would have to say about that. [TechCrunch]Reply152 repliesLeave a reply


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My New Culti Cave
In the end, I decided to buy myself a culti cave. I figured this would be the most sensible option to get me started because it's inexpensive and easy to assemble. I'm really very impressed with it. It opened up like a pop up tent so I had to put almost no effort at all into its assembly. I haven't actually put any plants into the culti cave yet but I tbink I will initially use it for growing cherry tomatoes and basil and, if successful, expand to other veggies too. Tomatoes and basil are not only a winning taste combination but they also grow really well together. They are both robust and hungry plants that rape the minerals from the earth and can be damaging for other plants of grown in their vicinity. But together, they equal one another out with their strength and hunger. Read more
The Defiance of Apple
Hi guys, I felt compelled to write about the latest Apple scandal. In what is perhaps the most defiant act of data protection, Apple are contesting an FBI requested court order for the mobile phone information belonging to San Bernadino gunman Syed Rizwan Farook. Read more
The best small smartphones
The best quality/price ratio: Alcatel OneTouch Idol 3
Google fighting cyberterrorism
Today, an article on a very recent subject : google fighting cyberterrorism. Read more
Bad-boy Vacuum Cleaners for 2016
Normally when I'm on the market for technology and electronics, it's usually for something that's going to entertain me, like something I play or watch. OK, we did have to buy a new fridge last year and that got me looking at some of the ridiculous fridges available, but unfortunately, as we'd just moved house and things were a little tight, we couldn't go for one of those big ice-maker style ones with the built-in water compartment. We had to settle for a standard fridge-freezer just as a temporary solution until we can afford something better. Read more
Welcome back to my Technology blog. I'm a pretty big media junkie, I spend a lot of time watching my favourite movies and TV series, a lot of the time from the comfort of my bed or sofa. As there's other people in the household, I don't just get the TV to myself all the time and I don't always get the peace and quiet needed to fully enjoy what I watching. That's why it's important to have a good set of headphones on-hand that you can throw on whenever things get a little noisy and you need an uninterrupted viewing experience. Read more
My Passion for Lambretta
My Passion for Lambretta Read more
Samsung Galaxy Tab S
Samsung Galaxy Tab S Read more
All Things Recent TV
Like most things technological today, we live in an age where the tech around us can stun us over and over again. Everything evolves at such a ridiculous rate that every year almost exponential improvements are being made. The modern day TV is testament to that. Only 50 years ago people were mainly watching black and white TV shows and screens that were no bigger than a tablet of today, which wouldn't be so bad if you had HD quality pictures, but factor in the arial and analogue signal, you were screwed if it was raining or there was a bit of bad weather about. Without anything to record shows or movies, if you missed it once there was a possibility you might not see the same program again, that was until UK Gold and YouTube came onto the scene anyway! Read more
Sony TV
Evening TechLovers. Had a recent urge to buy a new TV, and thought I would let you all know about which one I chose. I wanted a huge screen, so I decided to go for a 75"!! And I chose my favourite TV brand, Sony. I opted in the end for a Sony KD-75X9405C. Read more
Now in 2015 we have seen a very significant landmark in the tech world… The worlds first drone delivery. It happened in the USA this summer. Now this "drone delivery" was only about a mile away from its source, but you can just imagine a future skyline full of drones dropping off deliveries to peoples homes. Amazon have a drone program in development, called 'Prime Air'. They say they eventually want orders to be delivered within 30 minutes of the order being placed. Wow. Facebook have developed a drone that is the size of a passenger plane, that will circle the earth at heights of up to 30 miles. The aim of this drone is to beam Internet signal to very isolated places of the globe. Read more
Blog Intro!
Tech tech tech lovers. Come and subscribe to my blog about all things tech. I'm fascinated by modern technology and gadgets and that is what this blog will be all about. Anything from phones to drones. It will be in here! Name is Tom by the way. Nice to meet you all!


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Airports in Taiwan
Browse 548 attractions, meet 40 travelers, 7 tour guides and discover 1,214 photos
Travelers are voting Kinmen Airport, Chiayi Airport and Hengchun Airport as the best of 14 airports in Taiwan. Also popular are Hualien Airport in Hualien, Kaohsiung Intl. Airport in Kaohsiung and Matsu Beigan Airport in Taiwan. Want more? Check out our map of airports in Taiwan or these 2 airports in Taipei.
Taipei Songshan AirportTourist Essentials AirportsSongshan, Taipei, Taiwan
Taipei Songshan Airport or Taipei Sungshan Airport is a midsize commercial airport and military airbase located in Songshan, Taipei, Taiwan. The…1
Taitung AirportTourist Essentials AirportsTaitung, Taiwan
Taitung Airport (Chinese: , Hanyu Pinyin: T The airport sits on 153.7 hectares of land began as a holding room facility in 1977 and…
Wangan AirportTourist Essentials AirportsWonan, Taiwan
Wang-an Airport is an airport in Wang-an, Penghu Islands, Republic of China (Taiwan). Construction of the airport started in January 1988, and…
Taiwan Taoyuan International AirportTourist Essentials AirportsTaiwan
Taiwan Taoyuan International Airport (IATA: TPE, ICAO: RCTP) (traditional Chinese: 臺灣桃園國際機場; simplified…
Lyudao AirportTourist Essentials AirportsTaiwan
Lyudao Airport (IATA: GNI, ICAO: RCGI) is an airport serving Lyudao (Green Island), in Taitung County, Taiwan, Republic of China.
Kinmen AirportTourist Essentials AirportsTaiwan
Kinmen Airport or Shang Yi Airport (IATA: KNH, ICAO: RCBS) is a civilian airport serving Kinmen, Fujian Province, Republic of China (Taiwan). It was…
Chiayi AirportTourist Essentials AirportsTaiwan
Chiayi Airport, commonly known as Shueishang Airport, is an airport in Chiayi County, Taiwan. The airport has two runways.
Hengchun AirportTourist Essentials AirportsHengchun Airport, Taiwan
Due to its location, the airport often experiences strong Katabatic wind and is forced to close. Because of this, the load factor of the flights…1
Hualien AirportTourist Essentials AirportsHualien, Taiwan
The airport was opened on May 16, 1962 for military and domestic civilian use. Prior to this time, Hualien was a military gravel airfield.
Kaohsiung Intl. AirportTourist Essentials AirportsKaohsiung, Taiwan
The Kaohsiung MRT Kaohsiung International Airport Station is a station on the Red Line in Siaogang District, Kaohsiung, Taiwan. The station refers…1


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|71 Reviews
|Property type
|Minimum Stay
|2 - 3 nights
When you book through the HomeAway checkout, your booking is backed by our Book with Confidence Guarantee™ offering comprehensive payment protection.Learn more
Stay in a beautiful guest house nearby all LA attractions -15 minutes will take you to Hollywood, the Beaches, Beverley Hills, Westwood UCLA, Downtown, Santa Monica, Culver City. You will feel the warmth that has been infused into making your charming, private guest house so comfortable and inviting. It's cute and cozy, on one level. All doors lead out to your private patio.smoking allow only in the patio !
The guest cottage has just been renovated. The living room has gorgeous new hardwood furniture. Both bedrooms feature new comfort mattresses.
Located in the heart of Los Angeles - quick and easy freeway access. Near it all!
Rudy and Angelina are very personable & customer-oriented. Long-time residents of Los Angeles, they know the city like the back of their hand and are more than happy to share their local tips about the best places to eat, sight-see, etc. They have just recently converted this personal home into a vacation rental and are so excited to share this wonderful home with you so that you, too, can enjoy the convenient location and the elegant home at an exceptionally low rate.
Rudy purchased this cottage in 2004
It is located in a quiet residential neighborhood, is spacious and has a private outdoor patio for the warm LA days and nights. Parking is off-street inside the property gates.
Is conveniently located close to most LA attractions. Hollywood, Beverly Hills, Westwood UCLA, USC, West LA, SMC, Santa Monica, Downtown, Culver City.
The property was just as pictured, comfortable, cozy, and had everything we needed to have a great vacation. We were able to make a couple a meals at home with what was provided and it was a quiet and relaxing place to come back to after long days out playing in the city or on the beach. It was close to everything and easy to get to. We would definitely stay there again!
The guest house is exactly the same of pictures. Rudy is a great owner. When I return to L.A., I will rent this guest house again!
I would definitely recommend this rental to anyone. Ruti is really accommodating. You are about 20 mins away from everything. It has its own little parking area which is great because it's really hard to find street parking. Thanks again for everything
This house had everything our family needed and was a cozy place to call home for our vacation. It was cleaned upon our arrival and the beds are comfy as ever. Rudy, the owner, is very accommodating and friendly! The area is not the nicest, but if you're looking for something cheaper and still very nice this is the house for you. Even with the area not being the greatest, we still felt safe in this home with cameras, gates, and the vacation homes being towards the back of the estate.
First time visiting the city and I absolutely loved it. Central to pretty much everything. Rudy was a great host. Would definitely rent this property for future visits.
The house and furnishings are just as pictured. Very cute and cozy. It was myself, my husband and our two adult daughters and it was a bit small for four, but we managed just fine. The place was clean and serviceable, except for a bit of mold in the bathroom and a couple electrical outlets that didn't work. The shower and the patio were the best things about the place. It was relatively cool outside, so the house was comfortable without needing the a/c, especially with the front door open, which you can leave open as there are no bugs and it opens onto the private patio. Excellent location, within reasonable driving distance of all things we needed/wanted. Definitely need to have a car, though not an SUV or even a large car, as the parking is way too tight for that. Trader Joe's, Ralph's and Whole Foods and as well as CVS all within a couple of miles, but needed to go out of the area for quality restaurants. Recommend Vito Restaurant in Santa Monica for high end Italian food and world-class service and Soi 7 downtown for wonderful Thai food. We never met Rudy, the owner, but he was courteous and accommodating on the phone.
15 minutes from Hollywood, Beaches, Most all of LA.
Golf: Several golf course nearby
Ski: Several ski resort nearby
|Rate Period
|Weekend Night
|Monthly *
High Season
May 1 2016 - Sep 15 2016
2 night minimum stay
|Notes: Rates are based on 3 guests; add $30 per night, per additional guest
My Standard Rate
3 night minimum stay
|Notes: Rates are based on 3 guests; add $30 per night, per additional guest
|Property Insurace
|cleaning fee
|Pet Fee
|Tax Rate
After three people, the charges are $30.00 extra per adult and kids each night. There is also a charge of $30.00 per night for each pet.
If you would like, these extra charges can be paid in cash on the arrival date.
Until confirmed, rates are subject to change without notice.
Book ahead at this low rate before others take away your dates.
* Approximate monthly rate. Actual rate will depend on the days of the month you stay.
Learn more about this vacation rental at the manager's website:


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Without gyms, people would resort to outdated fitness techniques, such as pushing Model Ts up slight inclines and jumping rope. Get a modern workout with this Groupon.
$49 for a Three-Month Family Membership (a $194.25 value)
Family memberships are valid for two adults aged 18 or older and any children 17 and under, or full-time students up to age 25 who live in the same household.
Lynn YMCA
At the Lynn YMCA, members can workout in the gym and then record a song about it. Thanks to recent upgrades made possible by the Boston Celtics Foundation, professional-grade music equipment aids aspiring artists in their musical endeavors. Elsewhere at the Lynn Y, the usual amenities abound, including a six-lane, 25-yard lap swimming pool, an indoor track, a basketball gym, and two racquetball courts. The expansive gym has men-only and women-only workout areas, as well as a coed Nautilus Center stocked with free weights and cardio equipment. Lynn Y also boasts an air-conditioned aerobics studio that holds more than 30 classes each week.


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Writing Down the Bones: Freeing the Writer Within Paperback – Dec 6 2005
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Product Description
"I'm convinced that none of the writers of my acquaintance can go another day without a copy of Natalie Goldberg's magical manual Writing Down the Bones."—Linda Weltner, The Boston Globe
"The secret of creativity, Natalie Goldberg makes clear, is to subtract rules for writing, not add them. It's a process of 'uneducation' rather than education. Proof that she knows what she's talking about is abundant in her own sentences. They flow with speed and grace and accuracy and simplicity. It looks easy to a reader, but writers know it is the hardest writing of all."—Robert Pirsig, author of Zen and the Art of Motorcycle Maintenance
About the Author
Natalie Goldberg is the author of ten books. Writing Down the Bones, her first, has sold over one million copies and has been translated into twelve languages. For the last thirty years she has practiced Zen and taught seminars in writing as a spiritual practice. She lives in New Mexico.
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Refugee centre for one
A multi-million-dollar refugee detention centre set up by Australia on an island in Papua New Guinea has housed just one asylum-seeker over the past six months.
But Immigration Minister Amanda Vanstone yesterday denied accusations from the opposition Labor Party that the camp, on remote Manus island, was a waste of money.
Since July it has held just one inmate - Aladdin Sisalem, a Palestinian who fled his adopted homeland, Kuwait, because he claimed he faced persecution.
The government is spending an estimated A$716,000 (HK$4.3 million) a month maintaining the centre, which was opened in 2001 as part of the so-called 'Pacific solution' of denying asylum-seekers access to the Australian mainland.
Senator Vanstone conceded the detention camp was expensive but insisted it would not be closed.
'We would keep the option of Manus island even if there wasn't anybody there,' she said. 'If a country town in Victoria has an ambulance and the ambulance hasn't been used for 12 months you don't say 'let's get rid of it'. You've got to be prepared.'
Senator Vanstone said the centre acted as a deterrent to illegal refugees and the snakehead syndicates that tried to smuggle them into Australia.
'We shifted to offshore processing because we think that is what really kills the people-smugglers. When they can't get their customers to Australia, their future clients say 'we aren't going to give you any money because you won't get us to Australia',' she said.
But Labor MP Tanya Plibersek said the Pacific solution had been costly and cruel. 'It is absurd having one poor fellow sitting in the middle of the Pacific. He is our responsibility,' she said.
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Children's Literature - Children's LiteratureThis computer book will be a delight for homeschoolers, just beginning to incorporate writing into their program. It is based on Microsoft Works but the basic information can be transferred to any program. Computer basics are presented, so that everyone knows what a keyboard, or a mouse, is. Chapter One teaches kids basic words skills--rhyming words, similes, and adjectives are emphasized. Chapter Two builds toward sentence construction and teaches kids how to make a flipbook. Chapter Three brings all of these elements together to create a story. The Paint program is used to illustrate the story. This is a great interactive tool for the independent learner. Drawings and diagrams are bright and colorful. One of eight titles in the "Click It!" series. 2000, Millbrook Press, Ages 8 to 12, $21.90 and $5.95. Reviewer: Joyce RiceChildren's Literature
Computer Fun Writingby Lisa Trumbauer, Sydney Wright, Sydney Wright
View All Available Formats & Editions
Explains how to use a personal computer to complete activities that explore various aspects of Halloween.
Explains how to use a personal computer to complete activities that explore various aspects of Halloween.
Editorial Reviews
Product Details
- 9780761312895
- Publisher:
- Lerner Publishing Group
- Publication date:
- 04/01/2000
- Series:
- Click It Series
- Pages:
- 32
- Product dimensions:
- 8.50(w) x 10.97(h) x 0.15(d)
- Age Range:
- 6 - 9 Years
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PROTSMOUTH, Britain, June 5 (Xinhua) -- Peaceful southern British port city Portsmouth was not so peaceful 70 years ago and today, thousands of people across the country and from other countries gathered on the seafront to commemorate D-Day, known as Invasion of Normandy, on Thursday.
To remember people who lost their lives on D-Day and remark the 70th anniversary of the landings, a series of events have been held in Portsmouth, a small city on the side of the English Channel, where the other side is Normandy.
In the morning, Princess Anne paid tribute to those who involved in D-Day and inspected the parade. Drumhead ceremony and Cathedral Choir performed on the beach.
More than 200 veterans who took part in D-Day Landings or the following operations in World War II attended the event.
Amphibious demonstration operated by Royal Navy and Royal Marines displayed the landing, which involved the Albion-class assault ship HMS Bulwark, helicopters, troop carriers and armoured vehicles.
"We are from Winchester, My father-in-law John was landed in one of the five beaches, he has been telling stories to students in schools and we often tell the children about the D-Day to teach them remember the history," said Emily Bray, adding that as this year is the 70th annivesary of D-Day, she came to Portsmouth with her family to witness these big events.
Young soldiers 70 years before are now in their 80s and 90s, and those old soldiers who attended the event on Thursday have been seeing with gray and silver hair. Some of them have lost their hearing, others have their hands getting wrinkled and trembled.
Victor Tarry, 92, was a soldier who crossed the English Channel on June 5,1944. He has been awarded a medal after liberating a town in France, which made him very proud of.
He can still remember clearly the 10-feet seawall on the beach and bullets shot through the wall. He can still recall the French girls, whose hair had been shaved by German armies and forced to become prostitutes, as well as the debris scattering on the streets in Paris. However, he didn't remember how many injuries he got during the war.
"We didn't know where were we going, just know we were on an American boat that day," said Robert Piper, a 89-year-old veteran who departed from South Hampton on June 6, 1944.
"We need more tolerant with people, if people not tolerate they will fight. We are all together whether you are from China, India or wherever, we can live together no matter what religion you believe, we can live together and talk together," he said.
"Teachers should teach student the history and every child should go to France, stand in the middle of the cemetery, put their hands on the tombs which buried soldiers who was 18 or 19 years old, even not began their lives, they need to learn not to kill each other," he added.
In the afternoon, the Red Arrows of Royal Air Force performed a splendid aerobatic display, HMS Bulwark has set sail to Normandy this afternoon. While a sunset concert will be held on the beach for the heroes of war.
"It's a big event in Portsmouth, a lot of events will be held in the next five days, makes people involved have the ideas what happened those days. It will be a good memory in their lives," said Andrew Beecher, a local resident in Portsmouth, who invited his 89-year-old veteran uncle, Charles Beecher, to attend the significant event on Thursday.
"We learn the history of D-Day and World War II from school. The events today were great and I really love the amphibious demonstration. It's a good way to know the history," said Maleha Khan, a girl from a high school in Portsmouth.
Hundreds of pupils and high school students have been organized to participated in the events.
Portsmouth was the headquarters and main departure point for the military and naval units destined for Sword Beach on the Normandy coast.
The Invasion of Normandy established a crucial second front in the liberation of Europe from Nazi occupation, ultimately leading to victory for Allied Forces in 1945. British and Commonwealth casualties, including killed, wounded or missing, on D-Day numbered about 4,300 in total.


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A subspecialty certification by the Board of Internal Medicine. Practitioners treat diseases of the joints, muscles, bones, and tendons. Rheumatologists diagnose and treat arthritis, back pain, muscle strains, common athletic injuries, etc. They may work closely with other specialists such as orthopaedic surgeons and physical therapists.View More
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3000 N Halsted St Ste 820
Chicago, IL 60657
1900 W Polk St FL 7
Chicago, IL 60612
901 McClintock Dr Ste 201
Burr Ridge, IL 60527
890 Garfield Ave Ste 211
Libertyville, IL 60048
675 W North Ave Ste 305
Melrose Park, IL 60160
911 N Elm St Ste 115
Hinsdale, IL 60521


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Written by Jen Finn
Persistence and assistance win the day
From U.S. Coast Guard reports
One cold, windy January evening, an Alaska Coast Guard Communications Center received a mayday call. A 50-foot longliner was reporting uncontrolled flooding from an unknown source.
Aboard the fishing vessel, the skipper and a crew of four worked frantically to determine the source of the flooding. The boat's four bilge pumps couldn't match the deluge of incoming water, leading the skipper to instruct the crew to don their immersion suits and prepare their survival gear.
A Coast Guard helicopter lowered a self-contained P-100 pump (capable of discharging 100 gallons per minute) to the longliner.
Although wave action and the cumbersome immersion suits complicated the process, the crew moved the bulky dewatering pump into position. But the cold and the immersion suits' fingerless gloves made grasping the pump engine's pull-cord handle difficult, foiling repeated efforts to start the 10-hp engine.
With the vessel now down by the bow, the skipper issued the order to abandon ship. The rescue helicopter pilots checked their fuel and realized they might soon have to return to base.
The helicopter crew broadcast a radio message requesting assistance from vessels in the vicinity. A nearby longliner heard the message, stopped fishing and steamed toward the scene.
Meanwhile, in 35-knot winds, the men aboard the distressed vessel deployed and inflated their life raft. One by one the men jumped from their boat and struggled to enter the raft. A Coast Guard rescue swimmer was there to assist in case anyone dropped into the 40-degree water.
After the second crew member was retrieved, the helicopter winch operator alerted the pilots that the hoisting cable was frayed. Fearing the cable would part mid-hoist and that fuel levels were dangerously low, the pilots realized they had to leave the scene.
But as the helicopter left, a good Samaritan fishing boat was arriving. The boat's crew retrieved the three remaining fishermen and rescue swimmer, who were later transferred to a second Coast Guard rescue helicopter. The stricken longliner's entire crew was uninjured, but the boat was lost.
Lessons learned
After interviewing the crew, Coast Guard personnel determined a mechanical shaft seal that had been leaking prior to the incident likely caused the progressive flooding.
The skipper and crew quickly assessed their situation and did everything possible to ensure their own safety. The vessel owner's records showed proper maintenance of their survival equipment and regular abandon-ship drills. However, the leaking shaft seal should have been repaired.
Persistence and teamwork can turn the tide in a bad situation. The combined efforts of the Coast Guard and fishing vessels ready to provide assistance help ensure that fishermen will continue to fish safe!
Pat Fiorelli, the long-serving public affairs officer for the New England Fishery Management Council, will step down at the end of July.Read more...
The National Fish and Wildlife Foundation announced last week the sixth round of grant awards from its Fisheries Innovation Fund, a program launched in 2010 to foster innovations that support sustainable fisheries in the United States.
The goal of the Fisheries Innovation Fund is to sustain fishermen and fishing communities while simultaneously rebuilding fish stocks.Read more...


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- Top of page
- Abstract
- Introduction
- Patients and Methods
- Results
- Discussion
- Acknowledgements
- Conflict of Interest
- References
What's known on the subject? and What does the study add?
- Studies comparing adverse effects after different treatment methods are few and lack information on additional treatment and cancer relapse. Some US studies have provided population-based estimates on adverse effects, however, figures from Europe are most often based on mono-institutional experience or multicentre studies from high-volume university-affiliated hospitals with selected patient populations. Few studies have investigated the relationship between presence of urinary, bowel or sexual dysfunction and global quality of life (QoL).
- This population-based study investigated real-life practice with regard to prostate cancer in men who were potential candidates for curative treatment, based on high-quality national registry data. Treatment groups of recurrence-free men who completed their treatment or who received no cancer treatment were measured as to adverse effects, global QoL and the association between them. The study shows that each treatment method is associated with distinct patterns of urinary, bowel and sexual dysfunction and that irritative–obstructive urinary symptoms are associated with increased risk of low global QoL. This large survey also investigates the use of medication for erectile dysfunction and the relationship between such use and global QoL.
- To provide population-based estimates of typical adverse effects (AEs), e.g. urinary, bowel and sexual dysfunction, in patients with non-metastatic recurrence-free prostate cancer (PCa) by curative treatment method, including no treatment.
- To describe associations between typical AEs and global quality of life (QoL) and to study patients' use of medication for erectile dysfunction (EDmed) and the relationship between such use and global QoL.
Patients and Methods
- In October 2006 a national population-based sample of PCa survivors diagnosed in 2004 was invited to a postal survey focusing on treatment-related AEs and global QoL, 12–32 months after treatment start. All had completed their initial treatment.
- In the present study, 771 compliers were categorized into four groups of localized or locally advanced PCa related to the treatment they completed: (i) no treatment; (ii) radical prostatectomy (RP); (iii) radiotherapy (RAD) without hormones; and (iv) RAD with hormone therapy of 3–24 months duration.
- Measurement of AEs was restricted to function, using selected items from the 50-item Expanded Prostate Cancer Index Composite and the Brief Sexual Function Inventory among others, whereas global QoL was measured with the 12-item short-form health survey.
- National prescription data enabled assessment of adjuvant hormone application and EDmed use.
- Men who had undergone RP reported more urinary incontinence (24%) than the other treatment groups, but had the lowest level of moderate/severe urinary irritative–obstructive symptoms. Men from the 'no treatment' group had the highest level of moderate/severe irritative–obstructive urinary symptoms. Men who had undergone RAD reported higher levels of irritative intestinal symptoms and faecal leakage compared with the RP group and the no treatment group.
- In all treatment groups, poor sexual drive and poor erectile function were common AEs, with men treated with RP reporting the highest prevalence of poor erectile function (89%).
- Presence of irritative–obstructive urinary symptoms and poor sexual drive were independently associated with low global QoL in multivariate analyses.
- Fifty percent of the study group had used EDmed after treatment start, but only 47% of them were still using EDmed at the time of the survey. Use of EDmed was not significantly associated with global QoL.
- PCa survivors after curative treatment, but also patients without any anticancer therapy, report high levels of urinary and sexual AEs.
- Irritative–obstructive urinary symptoms and poor sexual drive were significantly associated with low global QoL, whereas erectile function and use of EDmed were not.


text:
Market Update: The ECB Speaks And The Result - Another Dow Theory "Buy" Signal 12 comments
In a move many are saying is "radical," the ECB lowered its deposit rate below zero (to -0.10%) and cut its benchmark rate to 0.15% from 0.25%. This marks the first time the deposit rate has been negative, which effectively means banks in the region will be charged in order to maintain their reserves. These measures are being implemented to help the eurozone stave off deflation. In a press conference following the event, ECB President Mario Draghi discussed a plan to offer long-term loans at lowered rates for the next four years.
And so while it was widely anticipated it wasn't a "sell on the news" event as I originally though might happen .. Instead, with that headline the equity markets powered higher this past week. But it wasn't only the ECB headline that made a difference ..
There were plenty of good economic reports published this week.
On the global front ; China factory data hits 5 month high. As one of the first leading indicators gauging economic momentum. Reinforcing expectations that their economy is starting to regain strength in the second quarter after dipping to a 18-month low in the first three months.
The growth rate of global manufacturing production ticked higher in May, as companies raised output in response to rising levels of new business and international trade volumes. The effects of the upturn are still being felt in the labour market, with job creation registered for the tenth successive month. Improving demand and rising employment should position the sector well to achieve faster growth heading into the second half of the year."
Back here in the U.S. :
On Monday , The ISM Manufacturing report was miscalculated by using the wrong seasonal factors. The Institute for Supply Management initially reported a decrease in its manufacturing index in April, but economists spotted the error and the ISM soon corrected the results, which then showed a gain that was in line with expectations. In its report, the ISM used the wrong seasonal factors (putting the blame on "a software error"). The headline figure was initially reported as 53.2 (median forecast: 55.5), corrected to 56.0, and then re-corrected to 55.4. The components on new orders, production, and employment were higher than initially reported.
Markit U.S. Manufacturing PMI- final data
Sharpest rise in production levels since February 2011
Adjusted for seasonal influences, the final Markit U.S. Manufacturing PMI picked up to 56.4 in May, from 55.4 in April. The latest reading was well above the neutral 50.0 value and signaled a robust improvement in overall business conditions.
The latest survey pointed to a steep increase in production levels, with the rate of expansion accelerating for the second month running to the fastest since February 2011. Anecdotal evidence from survey respondents cited improving underlying demand, better economic conditions and greater investment spending among clients.
Robust auto sales in May - yet another positive regarding the consumer.. and an example of how the weather did in fact impact the first quarter as the May results shows a nice sharp rebound.
Thursday's ISM report on the Non-Manufacturing, or Services sector, went off without a problem. While economists were expecting a level of 55.5, the actual reading came in at 56.3, which was the highest level since August. Combining the ISM for the Services and Manufacturing sectors, May's ISM was 56.2, which is the sixth highest level of the current expansion.
Finally ;
U.S. Job Creation Index Hits New High as reported by Gallup and also bodes well as we move forward.
While attention has been focused on the recent poor GDP number, the recent economic data that has been put forth clearly indicates that Q1 was severely impacted by the harsh weather and it wasn't a mere "excuse" that was citied by the bulls to bury the negatives.
This past week I came upon a positive story. A fundamental Long term scenario that I have mentioned before as one catalyst in the secular bull theme that we are experiencing.
The story's title is, "Are We Underestimating America's Fracking Boom?" The byline reads, "Check Out Sasol's Energy Complex in Lake Charles, La." Now the company is going to build a plant in Louisiana and at $21 billion this would be the largest foreign investment project in U.S. history. The 3,000-acre complex will tap into America's cheap natural gas and convert it into high-quality diesel fuel, ethylene (used in plastics, paints, packaging, etc.), and other liquids. The article goes on to note:
This is engineering on a scale so large that it requires closing 26 public roads, buying out 883 public-property lots, and hiring 7,000 workers at peak construction. Some 100 additional trucks will be on the road each day once the complex is completed. Entrepreneurs have already begun construction of a "man camp" to house 4,000 temporary workers streaming into Lake Charles for this and other projects. In that way, Sasol is a metaphor for what we don't yet understand about America's gas boom. Most know what fracking has meant for oil and gas prices. But because much of the work hasn't started yet, few appreciate the true extent of the industrialization that's about to begin. So let's put it this way, we are building a Qatar on the Bayou. Companies are laying new cities of fertilizer plants, boron manufacturers, methanol terminals, polymer plants, ammonia factories and paper-finishing facilities.
According to the Greater Baton Rouge Alliance, when completed some 66 industrial projects will be breaking ground over the next five years, worth about $90 billion. Currently, Louisiana's entire GDP is $250 billion, so we are talking about a 36% increase in the state's GDP ... !!!!!
The implications of projects like this are "transformational" a word often used by Joel Kurtzman in his book, "Unleashing the Second American Century."
This is exactly what I have been talking about in prior missives here on this blog. Energy Independence, the American Industrial Renaissance, etc.
Joel cites the four forces for America's economic dominance are: soaring levels of creativity; massive new energy reserves; gigantic amounts of capital; and unrivaled manufacturing depth. As Joel states, "It's like taking America's industrial strength and layering in Saudi Arabia's energy on top." While most of the media and the naysayerrs ignore this, the stock market seems to be getting it.
This chart is quite impressive:
Look at the positive PMI & ISM economic data that has been reported and then highlighted here on this blog in the past 3 weeks.
The investment world is distracted with fears of the next great calamity, but heading into summer U.S. financial markets are enjoying a remarkably positive environment.
And yet there is the "r" word surfaced weekly and the talk of a bear market lurking around the corner.. all citing the 1st Q GDP as their backdrop.
Please ....... Bear markets are made of tight liquidity, significantly deteriorating fundamentals, and investor euphoria. NONE of these are present today. Although the Fed is starting to reverse course, there are no signs yet of a significant tightening of liquidity. Rather, the data are beginning to suggest that private sector credit growth is starting to replace the Fed as the provider of liquidity.
Prophecies of imminent economic catastrophe have become almost the "acceptable thinking" in recent weeks. Whether it is talk of Europe facing deflation, Japan's Abenomics experiment failing, China heading toward a real estate crisis, or Russia being on the brink of direct conflict with Ukraine, the narrative is similarly dark. The "imminent crisis" theme has become overplayed, but if experience has taught me anything it is that nothing is more tiresome than a fashionable consensus. The reality is that none of these regions seem likely to suffer a calamitous near-term crisis, thanks in part to the world being afloat in a sea of liquidity.
All these fears of a major crisis have spurred a flight to quality to U.S. Treasuries, which has been partly responsible for the recent bond market rally. With many investors fearful, the 10-year U.S. Treasury note broke out of its tight trading range in recent days, closing at 2.45 percent on Thursday -- its lowest yield since June of last year. Will rates head lower, just as the outlook for the U.S. economy is starting to brighten ? Hmmm ----not so fast....
In Europe, economic confidence has reached levels not seen since 2011 and economic growth is modestly improving. All of this means that a huge swath of the global economy is now growing, & add the China data I reported earlier, which should benefit struggling economies.
However as I commented on last week - Market highs again this week , with yet another Dow Theory Buy signal, but many still choose Bonds and money market funds. The equity market has advanced over 180% since the lows and many are gripped with the confusing signals, warnings about the complacency and low VIX environment that will lead to a market crash.. A correction of 10-15% -- ok, but a crash ?? - Not likely at this time..
Don't lose sight of this fact ; the economy today is a much different place than it was back when these same signals were cited as cause for the market declines of that era.
The way I see the present mindset of investors; the opportunity to make money is being undermined by the fear of losing it. That's ok for those who stay calm and invested in equities; (many equity accounts are up over 200% since the March lows) ---not so much for those who got out or never get in. And therein lies the "fear of losing" mentality that is pervasive among many. That tune is louder now as the market has gone higher and drifted farther away from that crowd. The same ICI (Investment Company Institute) data that reflected last month's net outflow in domestic equity mutual funds also captured a big net inflow into taxable bond funds. One thing for sure , this data does not suggest a euphoric investment community that "has to be" or "has to get into" the market , even as the S & P makes new highs.
Perhaps Treasury yields will continue to fall, generating gains that barely keep up with inflation. But what if they don't? And what if stocks keep rising ?
.. Are you OK with those potential (and from my perspective likely) outcomes? Managing risk includes managing opportunity. At times like this, investors need to navigate, not leave, the market. There is a difference ...
Remember a "correction " is not a "crash" , and can be "navigated' successfully. Yet the mindset of many investors and "would be investors" seems to be fresh from the 2008 crisis that any downturn will be Armageddon.
While there is still little or no evidence a major market top is imminent, investors should recognize that we have had a nice run and as I mentioned all during 2014 that calls for more selective buying and for adjusting portfolios to account for changing market conditions. For example, while mid and small cap stocks were among the strongest performers during much of this bull market, the recent plunge in the mid and, especially, small cap indexes, suggest investor enthusiasm for these stocks has largely disappeared, -------- for the moment. More about the Russell 2000 recovery next week.
Stocks like (NYSE:AAP) up $10 since I wrote about the company is a name that falls into that 'selective' category.
(NYSE:LAZ) mentioned here in March, broke to another new high this week .
(NASDAQ:FFIV) powered higher again and is up $8 since my article on May 13th..
(NYSE:LVS) mentioned last week, has had some negative press lately regarding slower gaming growth (Only 9% in the last month ! , instead of double digit expectations ). Keep an eye on this one if it shows further weakness - as the "Japan" story may be the next catalyst to take the stock higher. There is a 2.7% yield now as the dividend was raised for the March '14 quarterly payout.
Just a few examples of how an investor can navigate the market with selective buying. Scaling into a position is always recommended.
If a company "beats" their earnings estimates and "raises guidance , and /or raises their dividend they are an immediate candidate for due diligence - regardless of how many articles we can read about any impending correction .. Take a look at Copa Holdings -CPA, as an example.
And there is nothing wrong with the Buy/Write strategy, I have mentioned repeatedly - buying a quality stock and selling a covered call for income..
Best of Luck to all !!
Disclosure: I am long AAP, FFIV, CPA, LAZ, LVS.
Additional disclosure: I am long numerous equity positions - all of which can be seen here on my Instablog."It is my intention to present an introduction to these securities and state my intent and position. It should be used as a 'Starting Point' to conduct your own Due Diligence before making any investment decision."Back To Fear & Greed Trader's Instablog HomePage »
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F&G, it's almost scary : ) too much good stuff happening, shouldn't we be worried about something?!
F & G: I would note that there are many more long term secular forces, in addition to the abundant and relatively inexpensive natural gas supplies, that will be supporting more U.S. manufacturing jobs over the next decades.
F&G, I think we are just getting started. If you are invested in the crazy stocks with unjustified valuation....then it's not a good time. They do still amaze me....(AMZN) is up today.
BlueSky: I just left a comment in the LMH Instablog about entering a sell order for CSX. I am merely attempting to reduce my stock allocation back to where I thought it was.
SG, (CSX) has a higher valuation than (UNP). I own both, but (UNP) has been the clear winner. (UNP) split 2 for 1 today. Honestly, if I'd just bought (UNP) and no (CSX) things would have worked out much better, especially when you compare the dividends. CSX is 2.11% while UNP is 3.58%.
Blue: I sold my 100 CSX this morning at $30.4 and realized a gain of $390.78 which I view as sufficient compensation for that horn honking at 3:00 A.M. for about three months. When I bought those shares last November, I included a snapshot of a rail crossing near my home where the train is legally required to tout that horn several times, though there is only one house whose occupants would ever be crossing the track at that point. That is one loud horn.
Good read as the data and points are clean. It Just has me off set as to after 5 years of just flushing money into the markets all over the world by central banks and still any time there is any weak data they have to do more easing to keep this going?
When do we let the economy's and markets work on there own ? How will this work the day we let interest rates normalize and all the cheap money is gone?
Just thinking out loud,as to say this is just getting started seams to be a little late and the easy money has been made.
$AAPl,$SBUX,$MCD,$NKE, "Everybody's Unhappy" I explain why in my weekly update http://seekingalpha.com/a/24dkc Oct 11, 2015
$DAL, a beneficiary of the current energy situation http://seekingalpha.com/a/218uu Aug 11, 2015
Check out the homebuilders on this down day $TOL,$WCIC- weekly analysis here http://seekingalpha.com/a/218uu Aug 11, 2015
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But then the whole thing turns nasty.
Nasty, because legitimate companies set up and financed by scrubbed criminal cash are fronted by some powerfully connected members of Parliament; nasty because several important London banks were being propped up by the laundered money, the "only liquid private investment" available during the Great Recession, and finally, nasty because the British intelligence community's interagency warfare converts Dima's promised gift into a threat.
Caught in a vortex of spellbinding dialogue, the reader is sucked into the conflicted souls that inhabit this story. John LeCarré is at his best here, capturing both the endearing qualities and flaws of his characters. There's Perry, the Oxford don whose overprotective chivalry nearly usurps Gail's choice to be involved. Gail, a rising star London lawyer who stumbles into a hidden and disturbing side of herself; Hector, the aging sidelined intelligence lion who seizes one last chance to try to do the right thing, and Dima, whose complex motivations careen wildly into one another.
The riveting narrative flows effortlessly until it falters for a moment. In order to explain the complexity of laundering, LeCarré uses an awkward device of listening to a recording of Dima's examples of his activity. At that point the story's intricate tango momentarily stumbles, but within a few pages the dance recovers.
In "Our Kind of Traitor," LeCarré continues his long exploration of the nature and power of large bureaucratic organizations and how they subjugate and betray individual hopes and dreams in order to achieve their ends. Charles Dickens would love LeCarré's grasping of dark moral issues threatening to destroy our human values, along with Hector's challenge to Perry -- and us: "Dima is holding out to you, as I am, an opportunity to do something instead of bleating about it."
Doug Wallace is an essayist on the philosophy and spirituality of LeCarré. He lives in Minneapolis.
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Celebrity Eclipse Cruise Review by zeldarob: Celebrity Eclipse met our expectations
Overall Member Rating
Celebrity Eclipse met our expectations
Destination: Transatlantic
Embarkation: Southampton
This was our first cruise on Celebrity, recommended by a family member who cruised with them and Aqua class. We were not disappointed. We at in the Blu dining room few times and the main dining room once. Food was good but sometimes service was slow. We enjoyed the buffet as we don't like to be too scheduled.
The drink package worked well but we are not sure we took full advantage of it.
There were some problems with the WIFI but that was because of the hurricane Sandy in NY while we were at sea. They were very good about refunding some of the packages. Entertainment was very good for the most part. WE really enjoyed the smoke free casino.
There was never a lack of places to sit either in the solarium or the main pool. plenty of activities if you wanted to participate. I really enjoyed the classes at the gym and Zumba with Miggy Mendoza was a wonderful treat. He was awesome, Kudos to Celebrity for having him on board.
Full Tilt the Acapella group was More very good, really enjoyed them and the Eclipse show cast.
the weather was as we expected, cool in the beginning but a few days out we were in bathing suits. would be nice to have a bargain laundry day when you are traveling so many days.
The ports, Le Havre ( Paris), Cherbourg ( Normandy) and Azores were wonderful .
We already have another cruise booked on Celebrity to Hawaii in April. Less
Published 11/14/12
Read more Celebrity Eclipse cruise reviews >>
Read Cruise Critic's Celebrity Eclipse Review >>
Cabin review: A21537 AquaClass 2
Eclipse cabin 1537 bed near balcony. great location even though across from crew space, we were not bothered by any noise. close to stairs and elevators. Nice balcony furniture. Storage could be better, space over the bed is hard to access for vertically challenged people.Read All AquaClass 2 (A2) Reviews >>
Port and Shore Excursions
Hey HAL... it could be worse!
Don't travel Fred Olsen.
Crossing QM2 June 16
QM2: Near Perfect, but not Qui...
Fabulous Eclipse Baltic Cruise...
What a fantastic cruise
Perfect Itinerary and Ship
Bucket List Fulfillment 10+
Love the Summit and the Staff
Great Cruise, and Great Servic...
Cruising With the Kids
Alaska, July 15
Sponsored Links


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June/July: School Snippets
News and information about conferences, residencies, pride points, and more from Pace's School of Education and the Lubin School of Business.
SOE's Making the Grade
- STEM-D Conference Helps Educators Share Strategies for Engagement
- Is There an App for That? SOE Professors' Research Collaboration Examines Use of Technology in the Classroom
- Pace Leads Westchester and NYC Schools in Inquiry Practices to Boost Student Achievement
Pace is pleased to announce the appointment of Jonathan H. Hill, DPS, as Dean of the Seidenberg School of Computer Science and Information Systems, effective July 1, 2016.
A New Dean for Seidenberg
Do you remember your transition to college life? Wish there was something someone had told you? Know a special Pace perk that could benefit the Class of 2020? Let's hear it!
In Hindsight
Dyson Biology Professor Erika Crispo shares how to study for success, why she can't be bothered with a career in law or politics, and what makes her so passionate about animals and her area of research.
The Professor Is In: Erika Crispo