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Telomerase activity in human pleural mesothelioma
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1 Telomerase activity in human pleural mesothelioma Karl Dhaene, Roland Hübner, Samir Kumar-Singh, et al. Thorax : doi: /thx Updated information and services can be found at: References alerting service These include: This article cites 12 articles, 5 of which can be accessed free at: Article cited in: Receive free alerts when new articles cite this article. Sign up in the box at the top right corner of the online article. Topic collections Articles on similar topics can be found in the following collections Respiratory cancer (167 articles) Environmental issues (2879 articles) Notes To order reprints of this article go to: To subscribe to Thorax go to:
2 Thorax 1998;53: Rapid communication Telomerase activity in human pleural mesothelioma Karl Dhaene, Roland Hübner, Samir Kumar-Singh, Barbara Weyn, Eric Van Marck Department of Pathology, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk/Antwerp, Belgium K Dhaene R Hübner S Kumar-Singh B Weyn E Van Marck Correspondence to: Dr E Van Marck. Received 20 May 1998 Returned to author 25 June 1998 Revised manuscript received 24 July 1998 Accepted for publication 27 July 1998 Abstract Background Gradual telomere erosion eventually limits the replicative life span of somatic cells and is regarded as an ultimate tumour suppressor mechanism, eliminating cells that have accumulated genetic alterations. Telomerase, which has been found in over 85% of human cancers, elongates telomeres and may be required for tumorigenesis by the process of immortalisation. Malignant mesothelioma is an incurable malignancy with a poor prognosis. The disease becomes symptomatic decades after exposure to carcinogenic asbestos fibres, suggesting the long term survival of pre-malignant cell clones. This study investigated the presence of telomerase in pleural malignant mesothelioma, which may be the target for future anti-telomerase drugs. Methods Telomerase activity was semiquantitatively measured in extracts from 22 primary pleural mesotheliomas, two benign solitary fibrous tumours of the pleura, four mesothelioma cell lines, and six short term mesothelial cell cultures from normal pleura using a non-isotopic dilution assay of the telomeric repeat amplification protocol. Results Twenty of the 22 primary mesotheliomas (91%) and all tumour derived mesothelioma cell lines were telomerase positive. DiVerent levels of enzyme activity were observed in the tumours of diverent histological subtypes. Telomerase activity could not be detected in the six normal mesothelial cell cultures or in the two mesotheliomas. Both benign solitary fibrous tumours showed strong telomerase activity. Conclusions Telomerase activity is found in a high proportion of mesotheliomas and anti-telomerase drugs might therefore be useful clinically. The results are consistent with the hypothesis that telomerase activity may be a feature of carcinogenesis in mesotheliomas and possibly in many other cancers. (Thorax 1998;53: ) Keywords: telomerase; mesothelioma; TRAP assay Telomeres are stretches of repetitive noncoding DNA sequences (TTAGGG) associated with specific DNA binding proteins at the ends of eukaryotic chromosomes. Human telomeres progressively shorten during cell division because DNA polymerase cannot replicate the end of a linear template, and critical shortening is believed to limit the cellular life span. A possible explanation for the attrition of human telomeric buvers is the somatic repression of telomerase. This ribonucleoprotein reverse transcriptase can synthesise telomeric repeats at chromosomal ends using its catalytic subunit (htrt) and a short template element of an integral RNA (htr). The telomeric repeat amplification protocol (TRAP) assay has been developed to detect the activity of the enzyme. Telomerase activity has been found in germ cells and in stem cells, and in a variety of cell lines and malignant tumours, but only weak or no activity was found in mortal somatic cell cultures and in benign tumours. 1 It has recently been shown that forced expression of htrt in telomerase negative cells resulted in reconstitution of telomerase activity, elongation of telomeres, and extension of replicative life span. 2 These data indicate a causal relationship between telomere erosion and cellular replicative senescence and suggest that this mechanism acts as an ultimate powerful tumour suppressor system which creates a barrier to any cell that has accumulated genetic alterations and escaped normal growth control by its environment. The data further lead to the speculation that telomerase may be required for the eycient growth of neoplastic cells and that inhibition of telomerase activity could represent a new therapeutic approach for cancer. 3 Since it still is unclear whether telomerase activity commonly occurs in most tumours, we investigated telomerase activity in malignant mesothelioma. This cancer is a highly aggressive malignancy, thought to arise from mesothelial cells, which is resistant to treatment. It is characterised by an extremely long latency period (20 40 years) between exposure to asbestos, a major contributory factor in the development of this cancer, and the onset of disease symptoms, which suggests the long term survival of genetically altered cell clones. 4 In this study we have investigated whether mesothelioma shares telomerase activity with other cancers and we further report the
3 916 Dhaene, Hübner, Kumar-Singh, et al absence of telomerase activity in somatic mesothelial cell cultures. Methods TISSUES AND CELLS Specimens of 22 mesotheliomas (nine epithelial, four sarcomatous, nine mixed types) and of two benign solitary fibrous tumours of the pleura were snap frozen in liquid nitrogen at the time of pleuroscopy and stored at 80 C. Short term mesothelial cell cultures were established from stripped pleura from necropsied non-cancerous patients after trypsin disaggregation. Mesothelial cells were grown as primary cultures on fibronectin coated culture flasks in RPMI 1640 supplemented with 20% heat inactivated fetal bovine serum (Gibco BRL, Belgium) and 10 ng/ml hydrocortisone (Upjohn, Belgium). The cells were identified as mesothelial cells by several criteria including immunocytochemical staining with antikeratin antibodies (Dako, Glostrup, Denmark), cell morphology (cobblestone), and the presence of long microvilli observed by transmission electron microscopy. Third passage confluent cultures grown at 37 C and 5% CO 2 were harvested for TRAP analysis. Malignant mesothelioma cells were obtained from four human mesothelioma cell lines (UIA-MM 1, 2, 5 and 7) which were isolated from pleural fluid as described elsewhere. 5 The Jurkat cell line served as a positive control. NON-ISOTOPIC TRAP ASSAY Lysate preparation and the TRAP assay were performed as described by Kim et al 6 with minor modifications. Briefly, homogenates of frozen sections (100 mm 3 ) and cell pellets (10 6 cells) were prepared with disposable pestles in 200 µl ice cold lysis buver. After 30 minutes of incubation on ice the lysates were centrifuged at g for one hour at 4 C and the protein concentration of the supernatant measured according to Bradford (Bio-Rad Protein Assay, Bio-Rad Laboratories GmbH, Germany). Aliquots of a dilution series of each lysate (6 µg, 0.6 µg, and 0.06 µg of total protein) were used for the TRAP assays. After initial incubation at 23 C for 30 minutes telomerase products were amplified using TS and ACX primers (35 cycles at 94 C for 30 s, 53 C for 30 s, and 72 C for 30 s). Assay specificity was confirmed by inclusion of an RNase pre-incubation control step. The presence of Taq polymerase inhibitors in cases lacking the 6 bp ladder was assessed using the TSNT internal control oligonucleotide and the NT internal control primer, generating a 36 bp control band. PCR products were separated by electrophoresis on a 12.5% non-denaturing polyacrylamide gel (19:1). The gel was stained with ethidium bromide or SYBR green I nucleic acid gel stain (FMC, BioProducts, Rockland, Maine, USA) and analysed by the CCD camera coupled Gel Doc 1000 Molecular Analyst Software package (Bio-Rad). Telomerase activity was graded as negative (6 bp ladder not detected), weakly positive (ladder detected with 6 µg protein), moderately positive (ladder detected with 6 or 0.6 µg protein but not with 0.06 µg), or strongly positive (ladder detected at all three protein concentrations). Results In a preliminary experiment the sensitivity of our ethidium bromide based procedure was compared with an assay in which we stained the amplified telomerase products with SYBR green I that is known to be as sensitive as classical isotopic TRAP assays. 7 One hundred telomerase positive Jurkat cells were found to be suycient for the detection of telomerase activity in the study using SYBR green. Similar sensitivity was observed in the ethidium bromide based assay (data not shown). Representative results for serial dilutions of extracts of the analysed materials are shown in fig 1. The TRAP assay was performed with the Figure 1 Representative results of non-isotopic TRAP assays for serial dilutions of extracts from mesothelioma tumour samples (MM4, MM11 and MM16), mesothelial cells (MC1), and one solitary fibrous tumour (SFT1). Jurkat cell extracts (J) served as positive control (a). For every assay a 36 bp internal control band is visible (arrow). Each sample analysis contained an RNase A treated control (+ at top) showing loss of signal. Lysis buver alone (LB) was applied as negative control (g). PCR products were separated by electrophoresis on a non-denaturing 12.5% polyacrylamide gel, stained with ethidium bromide, and visualised on a UV transilluminator. The assay result is indicated at the bottom of the figure. Telomerase activity (TA) was semi-quantitatively estimated and was graded strong (b), moderate (c), weak (d) or absent (e).
4 Telomerase activity in human pleural mesothelioma 917 Table 1 Telomerase activity profiles of individual mesotheliomas and solitary fibrous tumours in relation to histological subtype Tumours Histological subtype 6* 0.6* 0.06* MM1 Epithelial + MM2 Epithelial MM3 Epithelial + MM4 Mixed + MM5 Mixed + MM6 Sarcomatous + MM7 Sarcomatous + MM8 Epithelial + MM9 Epithelial + MM10 Mixed + MM11 Epithelial + + MM12 Mixed + MM13 Mixed + MM14 Mixed + MM15 Epithelial + MM16 Mixed MM17 Sarcomatous + + MM18 Mixed + + MM19 Mixed + MM20 Epithelial + MM21 Sarcomatous MM22 Epithelial SFT1 Benign SFT2 Benign MM = malignant mesothelioma; SFT = solitary fibrous tumour. *Amounts of protein extracts (in µg) applied per TRAP assay reaction. use of 6 µg protein extracts and varying levels of telomerase activity were observed in 20 (91%) of 22 mesotheliomas, in all four cancer cell lines, and in both solitary fibrous tumours. Signals were abolished after pre-incubation with RNase A. Because the signal intensities varied among samples, we estimated telomerase activity by dilution assay using 10-fold (0.6 µg) and 100-fold (0.06 µg) diluted extracts. Five of 20 telomerase positive mesotheliomas retained signals after the 10-fold dilution (panels b and c) and two of these retained signals after the 100-fold dilution (panel b), indicating that these tumours expressed high levels of telomerase activity. High levels of activity were also observed after 10-fold and 100-fold dilution of the extracts of both solitary fibrous tumours (panel f). The dilution assay could not detect any telomerase activity in the mesothelial cells (panel e) nor in mesothelioma cases MM2 and MM21. False negativity caused by Taq polymerase inhibitors was excluded since 36 bp control products were amplified in all TRAP assays. Table 1 summarises the results of the dilution assays of mesotheliomas and solitary fibrous tumours according to signal intensity and histology. No statistical analysis was performed because of the small size of the study. Discussion The finding of telomerase activity in over 90% of mesotheliomas is in agreement with the reported activity in a variety of other cancer types. This result is consistent with the hypothesis that telomerase activity may be a feature of carcinogenesis in mesothelioma and also, perhaps, in many other cancers. Carcinogenesis is recognised as a multistep process resulting from the accumulation of sequential genetic alterations in a cell. The very long latency period characteristic of mesothelioma suggests that multiple cumulative genetic, cytotoxic, and proliferative events occur during the tumorigenic conversion of the progenitor cell. Of the known changes that we and others have investigated in mesothelioma, the most frequent are in the p16 and NF2 genes. 8 According to the telomere senescence model, such genetically altered pre-malignant cells may finally be eliminated before they can ever develop a full blown malignant phenotype. Conversely, the presence of telomerase activity in mesothelioma may indicate that these cells can overcome the restraint of their finite life span and are no longer hampered in their clonal evolution towards a more malignant phenotype. In view of the finding of telomerase activity in mesothelioma, anti-telomerase drugs might be useful clinically. However, telomerase activity could not be detected in the lysates of two mesotheliomas. It has been reported that some tissue samples contain an inhibitor of Taq polymerase and thus give a false negative result. 6 The successful co-amplification in our cases of the TRAP internal control excluded this possibility. At present we therefore cannot exclude the possibility that some mesotheliomas have telomere stabilising mechanisms that are independent of telomerase, or are not immortal at all. This certainly would weaken the eycacy of future antitelomerase drugs. We did not detect telomerase activity in mesothelial cells. There is considerable evidence that mesothelioma originates from surface mesothelial cells rather than from a multipotential subserosal stem cell, although this has not been established beyond all doubt. 4 Assuming the former scenario is correct, the absence of detectable telomerase activity in mesothelial cells, in contrast to the presence of activity in mesotheliomas, could fit with the interpretation that most adult tumours develop from telomerase negative precursors after telomerase re-activation. 9 On the other hand, others may interpret these results as evidence for the above mentioned stem cell origin of mesothelioma, arguing that these telomerase positive tumours do, indeed, arise from telomerase positive stem cells without the need for a re-activation step. 10 The strong telomerase activity detected in both benign solitary fibrous tumours may support the hypothesis of their origin from telomerase positive, CD34 positive fibroblastic stem cells or suggest unpredictable clinical behaviour. Similarly, Umbricht et al 11 found 19% of benign follicular tumours of the thyroid with detectable telomerase activity and argued that some histologically benign lesions may be precursor lesions of follicular carcinomas. Our observations may also indicate that the use of telomerase activity as a marker for malignancy of serosal lesions needs further validation. It is obvious that larger series need to be studied, which should also include unequivocal benign, mesothelium derived tumours such as adenomatoid tumours. The classic model states that telomerase upregulation is forced by critical telomere erosion beyond the point where cell multiplication normally stops. An in vitro situation is seen during continuous culture of SV40 large T
5 918 Dhaene, Hübner, Kumar-Singh, et al antigen transformed telomerase negative human cells which eventually undergo crisis the condition in which cellular chromosomes are characterised by ultrashort telomeres and that coincides with telomerase activity. 12 At present it is not known whether SV40-like viruses, for which DNA sequences have been found in some mesotheliomas, are responsible for an in vivo equivalent of crisis during mesothelioma carcinogenesis. 13 The co-selection hypothesis states that telomerase is indirectly re-activated as part of a package of changes in gene expression that occurs after some other genetic event. 14 It remains speculative whether asbestos induced DNA damage represents such a genetic event. Similarly, others have suggested that telomerase activation is a mere side evect of dediverentiation. 15 We observed diverent levels of enzyme activity in the tumours of diverent histological subtypes. Because of the limited size of the study we did not perform statistical analysis to find any correlation between the level of telomerase activity in the tissue lysates and the histological subtype. In any case, statistical analysis may be inaccurate because of weaknesses in the TRAP assay. Indeed, both heterogeneity of tumour diverentiation within a tissue section and its morphology are lost during an in vitro biochemical assay, indicating the need for alternative methods for detecting telomerase activity at the cellular level. In situ hybridisation techniques with probes directed against htrt mrna and htr, together with immunohistochemistry using antibodies to the various subunits of the holo-enzyme, may help in the further investigation of the expression of telomerase in benign and malignant pleural lesions. The authors would like to thank Petra Boukamp of the German Cancer Research Center, Heidelberg, Germany for constructive comments on an earlier version of the manuscript. This study was supported by a grant from the Belgian Sports against Cancer Society, Shay JW, Bacchetti S. A survey of telomerase activity in human cancer. Eur J Cancer 1997;33: Bodnar AG, Ouellette M, Frolkis M, et al. Extension of lifespan by introduction of telomerase into normal human cells. Science 1998;279: delange T. Telomeres and senescence: ending the debate. Science 1998;279: Craighead JE. Current pathogenetic concepts of divuse malignant mesothelioma. Hum Pathol 1987;18: Kumar SS, Segers K, Rodeck U, et al. WT1 mutation in malignant mesothelioma and WT1 immunoreactivity in relation to p53 and growth factor receptor expression, celltype transition, and prognosis. J Pathol 1997;181: Kim NW, Wu F. Advances in quantification and characterization of telomerase activity by the telomeric repeat amplification protocol (TRAP). Nucleic Acids Res 1997;25: Kyo S, Kanaya T, Ishikawa H, et al. Telomerase activity in gynecological tumours. Clin Cancer Res 1996;2: Lechner JF, Tesfaigzi J, Gerwin BI. Oncogenes and tumoursuppressor genes in mesothelioma: a synopsis. Environ Health Perspect 1997;105(Suppl 5): Shay JW, Wright WE. The reactivation of telomerase activity in cancer progression. Trends Genet 1996;12: Greaves M. Is telomerase activity in cancer due to selection of stem cells and diverentiation arrest? Trends Genet 1996; 12: Umbricht CB, Saji M, Westra WH, et al. Telomerase activity: a marker to distinguish follicular thyroid adenoma from carcinoma. Cancer Res 1997;57: Counter CM, Avilion AA, LeFeuvre CE, et al. Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 1992;11: Carbone M, Rizzo P, Pass HI. Simian virus 40, poliovaccines and human tumours: a review of recent developments. Oncogene 1997;15: Kipling D. Telomere structure and telomerase expression during mouse development and tumourigenesis. Eur J Cancer 1997;33: Wynford TD, Kipling D. Telomerase. Cancer and the knockout mouse. Nature 1997;389:551 2.
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Women in the chief executive seat of FTSE 350-listed companies have fallen by the wayside this year, with the departures of Anglo American's Cynthia Carroll, publisher Pearson's Dame Marjorie Scardino and stationer WH Smith's Kate Swann all stepping down.
So when Angela Spindler took over at fashion retailer N Brown, she gave a much-needed boost to aspiring businesswomen hoping to run some of the UK's major companies.
Just three months into the role, she has spent every day learning the ropes of the company, which spans brands from lingerie firm Figleaves to plus-size clothes stores Simply Be and Jacamo.
Beaming with pride and the chance to address the balance, she explains that she hopes to use her influence and position to inspire future businesswomen.
She says: "I feel pretty proud of myself and feel I'm flying the flag. I think those of us that have made it into senior positions have a responsibility to help other women who are on the same path, so I'm delighted to be here."
The former Deben-hams managing director certainly picked the right time to join the new boardroom, as profits continue to rise for the company that can trace its roots back to 1859 but came to prominence in 1999 when its biggest brand Simply Be launched as an online and catalogue-only business. Yesterday, N Brown revealed sales jumped 8 per cent to £409.6m and pre-tax profits rose 7.1 per cent to £45m in the six months to the end of August. More than 4 million customers used the websites, which also include brands JD Williams, Marisota and High and Mighty.
The best-performing brands were Simply Be and Jacamo, which recently signed up former England cricketer Andrew Flintoff to front its advertising campaigns.
Ms Spindler should have no problem working with celebrities at her new company, having persuaded Wayne Rooney's wife, Coleen, to become the face of George at Asda, where Ms Spindler had been managing director.
During a 10-year career at Asda, she worked her way up through the ranks and helped make George one of the biggest fashion brands, pioneering supermarket fashion, with rivals Tesco and Sainsbury's playing catch-up. She now believes Simply Be and Jacamo could fill the same empty space she saw Asda dominating with its George range.
"George at Asda democratised fashion and brought affordable, leading-edge fashion to the supermarket, when there wasn't that there before," she says. "Now we're democratising fashion in plus sizes, when it didn't really exist before. It's very hard to market a credible plus size fashion brand. The fashion industry has at best ignored it and at worst alienated the market, so it's really great to be a part of it. What's delighted me in the past three months is to understand how to fit and flatter larger people."
Part of her plans to become the dominant plus size fashion retailer include a major high street expansion, trebling its current roster of stores, and she has one eye on the US. There are currently seven Simply Be and Jacamo stores, with ones in Leeds' White City and Derby's Westfield due to open by the end of the year. But within the new few years Ms Spindler wants to expand to 25 stores, with London a target.
It is understood she wants to use the knowledge she gained from her last job as chief executive of the Factory Shop, where she opened stores in "under-served" areas, as a template for choosing the new Simply Be and Jacamo sites.
However, no timetable has been agreed, with Ms Spindler saying: "We'd like to open them as quickly as we can… but because there is so much competition for space, we can't put a time scale on it."
Tapping into the demands of the modern shopper, she hopes to offer a multichannel business where customers can seamlessly slip from the websites to the stores and back again.
If the UK rollout proves successful, attentions will swiftly move to the US, where in her short tenure Ms Spindler has overseen a series of focus groups to gauge reaction.
"They have loved it there, so the question now is can we step up the pace of making it happen?"
N Brown by numbers
+8 per cent Sales jumped to £409.6m in the six months to the end of August as pre-tax profits climbed 7.1% to £45m.
4 million The number of people who visited the company's websites. Jacamo was the best-performing brand.
N Brown: The brands
Plus-sized clothes for men, it signed up former England cricketer Andrew Flintoff as its brand ambassador in 2011, who still appears in its adverts today
Simply Be
Opened its first store in 2011 in Liverpool, after more than a decade of success online and through its catalogue, selling fast fashion for women sized 14 to 32
One of the biggest online-only lingerie retailers in the UK, it turned a profit for the first time last year and also launched a discreet erotic section
Similar to Simply Be, the company sells women's fashion in plus sizes up to 32 and is particularly popular for party frocks and shoesReuse content

subset: commoncrawl


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Police: 1½ year-old Child Found Unresponsive in Pool
JACKSONVILLE - Jacksonville Police say a 1½ year-old child was found unresponsive in a pool Saturday.
At approximately 6:35 p.m., officers responded to a home on Commons Drive in Jacksonville.
Police say the young boy was transported to Onslow Memorial Hospital and then airlifted to Vidant Medical Center in Greenville.
The incident remains under investigation. On Monday, police said their investigation indicates no evidence of foul play.
"No charges are anticipated in this tragic accident," stated Jacksonville Police Chief Mike Yaniero.


text:
Lovely House and Garden with Privacy and Clean Pond for Swim(4 Reviews)
The owner has been emailed.
This owner doesn't offer online booking through VRBO's checkout. You won't be eligible for our Book with Confidence Guarantee.
Never use instant money transfer services such as Western Union and MoneyGram. Learn more
- Minimum stay:7 nights
- Sleeps:2
- Bedrooms:1
- Bathrooms:2
- Property type:House
- Internet:Yes
- Pets considered:Yes
- Wheel chair accessible:N/A
About the Property
House, 1 Bedroom, 2 Baths, (Sleeps 2)
This is a lovely home with a European flair, private and idyllic with its own 200 feet lake front and clean water for swimming. From the house and garden you have beautiful views and complete privacy from neighbors. It is a ranch, built in 1964, has cathedral ceiling in living room and A/C in bedroom. On hot summer days a nice breeze usually comes up from the lake. Avid hikers can walk to JackKnife beach.
Riverbay is a residental area, approx. 1-2 miles to beaches and downtown Chatham. Stop and Shop and nearest restaurant about 1 mile from house. Danish and English guests come here every year. Barbeque in the garden on gas or charcoal while the moon rises over the lake. Sunbathe in privacy on the deck with a book and a drink. This is called a little paradise by many repeat guests.
Keywords: Cape Cod Ranch
Before making a payment, always call the owner at the number listed on our website to make sure your payment instructions are accurate.
Never use instant money transfer services such as Western Union and MoneyGram.Learn more
Member since: 2007
Property Type
- House
Accommodation Type
- Vacation Rental
- Guests Provide Their Own Meals
- Minimum Age Limit For Renters
Bedrooms: 1 Bedroom, Sleeps 2, Beds for 2
Other Amenities
- CD Player
- DVD Player
- Churches
- Cinemas
- Library
- Live Theater
- Marina
- Museums
- Restaurants
Local Services & Businesses
- ATM/Bank
- Fitness Center
- Groceries
- Medical Services
Leisure Activities
- Antiquing
- Boating
- Scenic Drives
- Shopping
- Sight Seeing
- Walking
- Whale Watching
Location Type
Lake View:
There is a beautiful view of Trout Pond and Pickerel Pond from the house and garden...moreless
Sports & Adventure Activities
- Cycling
- Deepsea Fishing
- Fishing
- Freshwater Fishing
- Golf
- Hiking
- Kayaking
- Paragliding
- Sailing
- Surfing
- Swimming
- Tennis
- Wind-Surfing
- Dining Area
- Air Conditioning:
- Clothes Dryer:
- Covered Parking:
- Free Long Distance:
- Garage:
Linens Provided:
Guests may use linens and towels if they wash and dry them before leaving, so they are ready for the next guests....moreless
- Parking Off Street:
- Telephone
- Cooking Utensils
- Dishwasher
- Refrigerator
- Deck / Patio
- Outdoor Grill
Pool / Spa
- Jetted Tub
RatesCurrency Conversion
|Weekend Night
|Monthly *
My Standard Rate
7 night min stay
Additional information about rental rates
Security deposit on reservation. Rent 30 days before arrival
Don't forget your vacation protection! Get protected now
Adding our Vacation Protection services can make sure your getaway goes smoothly, no matter what. We offer Cancellation Protection and Damage Protection so you can truly relax.
The property's calendar has not been updated in over 9 months. You can contact the owner for availability or look for properties with confirmed availability
Car: Recommended
There are lovely shops in Chatham and Orleans. There are good restaurants and theaters and wonderful salt water beaches nearby.
Owner info
Year Purchased: 1991
About the owner: I bought this house because of its location. The views are beautiful and the lake great for swimming. I grew up and was educated in Denmark, have worked both there and here as a social worker. When I married my late husband we retired to Nevada. Two of my daughters have lived here over two different periods, and for many years I have rented out to private guests and have had many come back year after year. Some day I expect to move back and live here permanently.
Why the Owner Chose Chatham, MA, USA:
The house was not bought for rental purpose, but because this is a nice place to live and have guests. They come from Denmark and England and from all over this country. It has central air-condition.
The Unique Benefits at this House:
Here is beauty and privacy. Yet it is close to beaches, to Chatham and Orleans, shops and theaters and golf. Pleasant bay is great for sailing and kayaking.
Contact us
Send email
- Secondary: +1 7752241810
- Click Here to see All My Rental Listings
From the Diningroom-kitchen
This listing was first published here in 2011.
Date last modified - Sunday, February 12, 2017
Partner Offers
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We appreciate your participation and assistance with our efforts to operate the most secure and trusted vacation rental marketplace in the industry
If you have any questions, please visit our Help Center.


text:
Attack On Titan Lightweight Scarf
Protect your neck
- Lightweight fabric features a chibi version of the Colossal Titan
- Looming over chibi Armin, Eren, and Mikasa
- A good scarf can make an outfit
Protect your neckMikasa and Levi have taught us many things. That humanity sometimes faces insurmountable odds. That bravery is doing a thing even when the odds are not in your favor. And that a good scarf can make an outfit.
This lightweight scarf is like giving other AoT/SnK fans the salute. Only in secret so you don't look silly doing it to all the other muggles. With vibrant art, it goes with almost anything. And remember: it may be the only piece of you to return home so wear it with pride.
Product Specifications
- Attack On Titan Lightweight Scarf
- Officially-licensed Attack on Titan merchandise
- Fabric features a chibi version of the Colossal Titan looming over chibi Armin, Eren, and Mikasa (above the logo)
- Materials: 100% polyester
- Care Instructions: Hand wash cold. Line dry.
- Imported
- Dimensions: 40" wide x 70" long
- Ages 14+
- WARNING - SMALL PARTS - Not intended for children under 3 years of age.
Wanna chat about it?
Have questions about Attack On Titan Lightweight Scarf or your order? We monitor these comments daily, but it may be faster to email us directly or call us at 1-888-GEEKSTUFF.


text:
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Senate President Bukola Saraki, who led a team of National Assembly leaders to visit President Muhammadu Buhari in London, said at the end of the visit: "there is no cause for alarm"
All Airtel users can now read News 24 Nigeria for free through a Facebook-led Internet.org initiative.
Leicester - Leicester City manager Claudio Ranieri looked
forward to "another fairytale" after the 5,000-1 Premier League
champions booked their place in the knockout phase of the Champions League.
Leicester are toiling in the Premier League, sitting two
points above the relegation zone, but Tuesday's 2-1 win over Club Brugge took
them into the Champions League last 16 with a game to spare.
Asked which of Europe's glamour clubs he would like to face
in the next round, a smiling Ranieri replied: "I don't know. It's not
important, believe me.
"I hope to be there in good condition. That's
important. The name is not important. It will be another fairytale. Win, lose,
it's not important. It's important to go there."
Leicester's win had the added bonus of guaranteeing them top
spot in Group G, meaning they will be paired with one of the second-place teams
in next month's draw.
A team like Real Madrid or Bayern Munich could provide the
opposition in Leicester's next home game in the competition and captain Wes
Morgan is relishing the prospect.
"It is a fantastic feeling to reach the last 16. It is
more than we could ever imagine," the Jamaica centre-back told BT Sport.
"For Leicester to be newcomers and finish group winners,
it is an amazing achievement. A great result and a great performance."
At the final whistle, Leicester's jubilant supporters
launched into a rendition of the chant: "We're all going on a European
Also read: Ranieri heartened by Leicester 'spirit'
But while the fans' thoughts are already turning to
springtime sojourns on the continent, Ranieri wants his players to plough all
their energies into arresting their domestic slump.
"Our minds must go on the Premier League because I want
to push my players to think about the Premier League," said the Italian.
"Now our job is done. We're through the group. I don't
know which team we play. It will be a fantastic team. We're very happy.
"But now we have to go back. We're very close to
relegation and we must play at the same level as the Champions League.
"When we want to do something, we achieve something. We
must concentrate on the Premier League now."
- 'Disaster' -
Shinji Okazaki's hooked finish in the fifth minute and Riyad
Mahrez's 30th-minute penalty, won by Marc Albrighton, looked to have set
Leicester up for a routine victory at the King Power Stadium.
But Jose Izquierdo reduced the arrears with a breakaway goal
early in the second half and there were some anxious moments for the hosts
before their passage was confirmed.
"We concede a goal on the counter-attack. Why?"
Ranieri asked in his post-match press conference.
"An Italian manager concedes a goal on a
counter-attack, I get crazy."
Leicester have already lost six games in the Premier League
-- twice as many times as last season -- and Ranieri conceded Brugge's goal had
stirred unwelcome recent memories.
"Of course (they are) lacking confidence because in the
Premier League when something happens, (the opposition) score a goal," he
"And tonight, the first chance (Brugge's goal), was the
first shot on goal. It's unbelievable. We have to change this mood."
Having lost all five of their Group G games to date, Belgian
champions Brugge can no longer claim even the consolation prize of a berth in
the Europa League.
When it was put to beaten coach Michel Preud'homme that his
side had produced a much-improved showing in the second half, he replied:
"Yeah, but there was the first half. That's the problem.
"In football at this level, you have to play 90
minutes. The first half was a disaster and the second half was our best
He also gave short shrift to a suggestion his team had shown
Leicester too much respect.
"Yes, of course," the former Belgium goalkeeper
said in a voice thick with sarcasm.
"It's my big problem in the Champions League. I'm
afraid and I play defensively."
Claudio Ranieri (Getty)
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With 355 preschools and child development centers operating in the state of Utah, the right daycare option is waiting for you. Whether you prefer a larger preschool with an innovative early childhood curriculum or the cozy personalization of smaller daycare centers, there are Utah childcare centers to fit every preference and budget. Check out the Quick Search box on the right hand side of the page in order to search for childcare providers by zip code or city/state. You can also click on your county and follow the links. You will be able to access information about scores of different childcare providers in your area, complete with reviews, business hours, a street view map, the age ranges the childcare providers service, the size of the service, and where the provider is located.
If you're looking for a family day care provider in Utah, click on Utah Home Daycare.
BumbleBee daycare 2923 childs ogden ut 84401 #801-644-6219 Mona Murillo open 24 hours monday-friday saturday & sunday 5:30am-5:30 pm transport to all schools in ogden north ogden and south ogden
Mini Monsters offers a smaller facility to give your child the individual attention they need. We offer before and after school drop offs. We are open 6 days a week. Tuition includes all meals and snacks.
Our brand new school offers beautiful, well organized, and developmentally appropriate learning classrooms: Science and Math, English and Social Studies, Spanish, and a Physical Education/Dance studio. Our classes are full of creative, academic and playful experiences that not only fascinate our students, but also effectively increase their level of elementary school readiness. Students rotate from each learning area to another, and our teachers are highly specialized in the subject they teach. Each classroom is equipped with high end furniture and materials. We are proud to present a higher quality early education program.
One of the biggest challenges parents face is finding affordable child care. It is usually incredibly expensive, often forcing parents to make some very difficult choices. Even for those who can affor...
Florida child care facility licensing requirements can be confusing, but the Florida Department of Children and Families has provided a wealth of information to both providers and parents. ...


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StartUpHire is the place to find BI Product Manager jobs. Review the list of BI Product Manager career opportunities to the left. These BI Product Manager jobs represent some of the most rewarding careers available.
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Location: Goleta, CAType: Full TimeMin. Experience: Experienced InTouch Health is a leading telehealth solution company enabling remote physician care anytime, anywhere. Our solutions are robust, secure, and provide excellent physician-patient experiences to maximize care and1/31/2017 Goleta, CA Save This Job
ABOUT THE ORGANIZATIONAbout Healthline Healthline.com is the fastest growing health information site on the planet, and the 2nd largest health site in the US (per comScore)! Every month, over 50 million people count on our talented teams to support, guide, and inspire them toward1/27/2017 San Francisco, CA Save This Job
Manager, Clinical Informatics FLSA: Exempt REPORTS TO: Director, Clinical Informatics LOCATION: White Plains, NY DEPARTMENT: Clinical Informatics PREPARED: 11/11/2016 POSITION SUMMARY: The Manager of Clinical Informatics will play a lead role in overseeing program portfolio for12/31/2016 White Plains, NY Save This Job
CredSimple is a New York-based startup laser-focused on improving healthcare infrastructure with revolutionary, digital technology. Our platform focuses on data about healthcare practitioners and facilities. We create data integrity, efficiency, and transparency between the many2/14/2017 Brooklyn, NY Save This Job
Senior Technical Product Marketing Manager? Help Please fill in the information below and use the navigational buttons to continue. Warning: Using your browser's Back and Forward buttons may produce undesirable results. The Role Are you a technical professional who loves to1/3/2017 Redwood City, CA Save This Job
at Prosper (View all jobs) San Francisco, CA We are looking for an experienced digital analytics professional to join our Marketing Analytics team. Senior Manager of Marketing Analytics is responsible for supporting Prosper?s borrower acquisition by building demand forecast12/26/2016 San Francisco, CA Save This Job
Sales Engineering Tampa, FL, United States Apply Send Jobvite The Territory Sales Engineer is responsible for the following prospect and customer-related activities: Act as the primary technical ?go-to? resource for the sales cycle associated with a given territory Facilitate2/15/2017 Tampa, FL Save This Job
Job ID 2015-1030 # of Openings 1 Job Locations US Posted Date 11/25/2015 Category Administrative More information about this job: Position Description: SUMMARY: Responsible to lead and manage multiple projects. Most of these projects will be for customers installing Jenzabar1/22/2017 Boston, MA Save This Job
Located in Santa Monica, CA. Society6 is a rapidly growing online marketplace powered by hundreds of thousands of artists from around the world. Society6 is an ecommerce platform that enables artists to easily upload and immediately offer their art for sale as art prints, iPhone2/18/2017 Santa Monica, CA Save This Job
Location: Redwood Shores, CA About Zyme Zyme is the global leader in the emerging channel data management (CDM) space, empowering the New Smart Channel?, a proven approach to creating visibility to actionable knowledge that manufacturers need to accelerate partner sales and12/31/2016 Redwood Shores, CA Save This Job
ABOUT US Television today spans far beyond the traditional TV set. It?s the last few seconds of a basketball game streamed to your iPhone ?and all of season two on your tablet. A leading video platform company, IBM Cloud Video enables TV networks, cable providers, and studios to10/30/2016 Duluth, GA Save This Job
Location: Boise, ID Jelli?s mission is to become the audio advertising platform globally. We love radio, audio and music. Join us as we transform an industry by introducing technology that helps radio advertising become as efficient and easy to buy as Digital. We have created2/7/2017 Boise, ID Save This Job
Maana is looking for a Senior Manager Product Marketing to join our marketing team in Palo Alto. Maana is the analytics platform that operationalizes big data insights into line-of-business applications. Maana makes it easy for enterprises to turn data from multiple silos into1/8/2017 Palo Alto, CA Save This Job
at ServiceSource (View all jobs) Nashville Job Title: Senior Business Intelligence Developer Job Location: Nashville, TN Job Description: Maintain or update business intelligence tools, databases, dashboards, systems, or methods; generate standard or custom reports1/9/2017 Nashville, TN Save This Job
Finance/Accounting San Luis Obispo, CA APPLY NOW Send Jobvite JOB SUMMARY: The Team Lead, Billing is a supervisory position that provides leadership and support to an assigned group of Billing Specialists, as well as secondary support to clients. The ideal candidate will have1/11/2017 San Luis Obispo, CA Save This Job
Mission To help customers drive decision making and business value through the use of our Population Manager using the Pentaho tool suite. This position is exempt. The position reports to the Manager of Information Management. Position Summary The candidate?s responsibilities2/4/2017 Philadelphia, PA Save This Job
Mission To support customers in all of their reporting; extract needs which can span from custom to regulatory compliant. This position requires a person with data analysis experience who can help ensure that the product is delivering maximum value to each customer based on2/4/2017 Philadelphia, PA Save This Job
Merchandising New York, NY, United States Apply Now Assistant Buyer Location: New York City Reporting Relationships: The Buyer is a leadership role within merchandising reporting to the divisional merchandise manager Company Description Hudson?s Bay Company is one of the2/2/2017 New York, NY Save This Job
Merchandising New York, NY, United States Apply Now Assistant Buyer Location: New York City Reporting Relationships: The Buyer is a leadership role within merchandising reporting to the divisional merchandise manager Company Description Hudson?s Bay Company is one of the2/2/2017 New York, NY Save This Job
Location: Johns Creek, GA Job Type: Full Time Employee Experience Required: 6 ? 10+ years Education Level: Bachelor?s Degree or equivalent experience Career Level: Experienced, Non-Manager GENERAL DESCRIPTION Movius is seeking Technical Support Engineers to join our1/16/2017 Johns Creek, CA Save This Job


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Recycled Funk
Recycled Funk Episode 7
For this episode I did something different than with any of the previous ones. After recording the set, I got in my car and took a drive while giving it a listen. I just wanted to see what it felt like to hear this collection of songs while on the go. I have it say…Read More
Recycled Funk Episode 06
Transitions, that's the mood of this latest installment of Recycled Funk. Here in NY we are transitioning into fall season. Musically, my taste is pretty eclectic and so I'm constantly transitioning from one style of music to another, but there's always a cohesiveness to the overall mix, and an underlying theme/vibe. That's what "Recycled Funk"…Read More
Recycled Funk Episode 05 (Got Money)
I originally conceived and recorded a different version of this mix a few years back. I decided to revamp it with a 2016 version exclusive to Brooklynradio.com It's not really possible to understate the importance of money in our daily lives. There are more than enough slang & slogans to go around, but I'm prone…Read More
Recycled Funk Episode 04
It's therapy for my mind while I light the bong, shit hurts but I feel like life goes on". That's a quote from Guilty Simpson's track "a man's world", produced by the late, great J Dilla. It's a featured track on this latest episode of "Recycled Funk". Guilty Simpson is referring to his father when…Read More
Recycled Funk Episode 03
Hip Hop has always carried the Funk torch, so this episode is dedicated to a Recycled Hip-Hop Funk theme. Some old and new school classics. Throw in a few funky r&b grooves for good measure and balance. And for some icing on the cake, my latest release "Stuck in the past" makes a cameo appearance…Read More
Recycled Funk Episode 02
In my music studio I have a couple of windows (not typical unless the studio is in your apt) and as I was sitting in my studio starring out of the window, I just starting appreciating the feeling of a seasonal change. In my part of the world, we're transitioning from winter, into spring. Which…Read More
Recycled Funk Episode 01
Since it's inception, Funk music has been very influential, and "recycled" in other forms of music. From Hip-Hop, to Disco and R&B, as well as House music. It's roots may be in America but it has no geographic borders. This first episode is an introduction, showcasing a little bit of that aesthetic to give the…Read More


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Filter Settings
Soviet Ural Mountains 1988 № 17
Newsreel, 1 part, Duration: 0:10:01.193G 6/2/2014
Scene 1 "Amnesty" Directions camera by bookshelves.
Storage Kurgan Regional Library.
Female librarian works with filing cabinets, hands are removed from the box card.
Books laid out on the table.
Leaf order the return of the books to the general fund.
The librarian tells of translating books from special funds for general use.
Plot 2 (Untitled) Black boy about 5 years, a woman teaches his Russian language.
Soviet Ural Mountains 1988 № 14
Newsreel, 1 partG 6/2/2014
THE BEST IN THE WORLD MOST. Sverdlovsk.
Special issue. 1 All-Russian festival of children's musical creativity.
Soviet Ural Mountains 1988 № 18
Newsreel, 1 partG 6/2/2014
UPI I.Atsygin graduate.
Commissioner dose unit in Pripyat B.Kulyabin.
Vice Rector of UPI, Professor V.S.Kortov.
SPRING UPI. Special issue.
Says Head.
Chair, Professor S.P.Raspopin.
Academician Month
Soviet Ural Mountains 1988 № 19
Newsreel, 1 part, Duration: 0:09:18.044G 6/2/2014
PNRM. the crowd in the square around the monument to Lenin.
Titer "Tyumen.
Lenin Square ... "PNRM. The building with columns.
The rally in the square.
Speakers with a megaphone.
Rally to protect the ecology of the district.
Poster "Bread - YES !, Nature - YES !, chemical plant - NO!" People sign the paper.
Soviet Ural Mountains 1988 № 20
Newsreel, 1 partG 6/2/2014
Special issue.
Students are protected from demolition monument.
Says the secretary of the urban society of protection of monuments VISmirnov.
Said the chief architect Belyankin
Activist Larisa Mikhaylova.
Commission Chairman V.I.Bulochnikov
V.G.Kusenko architect.
Writer B.Ryabinin
Soviet Ural Mountains 1988 № 22
Newsreel, 1 partG 6/2/2014
Strahagent Novitsky., A former researcher.
COMMUNIST IN THE STREET. Special issue dedicated to the movement of informals in Ufa.
The deputy B.V.Hakimov.
Chairman of the People's Control S.A.Shafikov.
Soviet Ural Mountains 1988 № 24
Newsreel, 1 partG 6/2/2014
LONG AND WILL BE kind ...
Special issue.
Disabled Veriskov want to live a full life.
He is actively involved in sports.
Soviet Ural Mountains 1988 № 25
Newsreel, 1 partG 6/2/2014
Fashion Theatre at the club?
Oilman "in Surgut.
Gorodischenskiy log "in Tyumen.
Rybkolhoz village Uvalovo Uvatskogo district of the Tyumen region.
Farm chairman - A.Kravtsov.
Soviet Ural Mountains 1988 № 26
Newsreel, 1 partG 6/2/2014
Forest fire.
Troopers Khanty-Mansiysk branch of Tyumen airbase.
Maholeta Toporov.
Khokhlovka dying village.
Perm region.
Soviet Ural Mountains 1988 № 27
Newsreel, 1 partG 6/2/2014
Folklore festival in Lower Sinyachikha.
Sverdlovsk region.
Museum director I.D.Samoylov
Celebrations in honor of the 1000th anniversary of Christianity in Russia.
Participates Archbishop of Omsk and Tyumen Theodosius.


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Music News - Page 12
Taylor Swift Chips a Tooth on Tour
Not only is Taylor Swift a phenomenal performer, she's also made of really tough stuff. The '22' hitmaker chipped a tooth onstage recently and didn't even stop the show.
Martina McBride Is Getting a Namesake
Martina McBride and Train frontman Pat Monahan gave two newlyweds the gift of a lifetime, performing 'Marry Me' during their onstage wedding at the 2012 ACM Awards. Now, that lucky couple -- Frank Tucci and Christina Davidson -- are giving a huge gift to McBride: a namesake.
Florida Georgia Line's 'Cruise' Sets All-Time Record on the Charts
Florida Georgia Line are on fire with their hit 'Cruise,' and they just set the all-time record for the most weeks at No. 1 on Billboard's Hot Country Songs chart. When they smashed through the 17-week record, everyone was wondering if they'd have enough momentum to set this new record, and they did -- easily.
Shania Twain Will Still Be the One With Las Vegas Return
Shania Twain will light up the the Colosseum's stage at Caesars Palace again soon, showing everyone that she's still the one.
Hear Carrie Underwood's 'Sunday Night Football' Intro
Carrie Underwood is the new voice of 'Sunday Night Football,' and everyone's been wondering how her version will sound -- especially since NBC said she was their first and only choice upon learning Faith Hill wouldn't return in 2013. Well, the intro is finally out, and fans can decide for themselves if it's a touchdown -- or a fumble.
Ronnie Dunn Releasing Second Solo Album This Fall
When Ronnie Dunn has a big announcement, he often takes to Facebook to make them. Almost casually, the country legend has revealed when we can expect his second solo effort.
Randy Travis Released From Dallas Hospital
Randy Travis is out of the hospital!!
The country crooner was discharged earlier today from Baylor Heart Hospital and has been relocated to a physical therapy facility.
Blake Shelton Says Miranda Lambert Is Sweet as Pie … Until You Cross Her
Miranda Lambert is known for her sweet smile and huge heart, especially when it comes to animals. But her husband Blake Shelton warns that the songstress truly is sweet as pie -- until you cross her.
New Country Music Releases – August 2013
Summer heats up with Luke Bryan, Brett Eldredge and many more putting out new country music releases in August, 2013. Bryan will put 'Crash My Party' in stores on August 13. It's the second of four very busy Tuesdays.
Taylor Swift Performs 'You're So Vain' Onstage With Carly Simon
Generations and genres collided when Taylor Swift invited '70s soft pop icon Carly Simon to perform with her at Gillette Stadium in Foxboro, Mass. on Saturday night (July 27). The ladies teamed up for an amazing duet of Simon's smash 'You're So Vain.'
Blake Shelton Addresses Rumors of 'Heart Scare'
Like he does with everything else in life, the good-natured Blake Shelton addressed a recent tabloid rumor about a terrifying heart scare with a sense of humor.


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