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High-throughput chemosensitivity testing of low-passage cancer cell lines can be used to prioritize agents for personalized chemotherapy. However, generating cell lines from primary cancers is difficult, because contaminating stromal cells overgrow the malignant cells.
We produced a series of hypoxanthine phosphoribosyl transferase (hprt)-null immunodeficient mice. During growth of human cancers in these mice, hprt-null murine stromal cells replace their human counterparts.
Pancreatic and ovarian cancers explanted from these mice were grown in selection media to produce pure human cancer cell lines. We screened one cell line with a 3,131-drug panel and identified seventy-seven FDA approved drugs with activity, including two novel drugs to which the cell line was uniquely sensitive. Xenografts of this carcinoma were selectively responsive to both drugs.
Chemotherapy can be personalized using patient-specific cell lines derived in biochemically selectable mice.
Selecting the most appropriate chemotherapy for a given patient has historically been based on histopathology and past studies demonstrating that specific drugs are generally active against that malignancy. However, drugs deemed active against a particular type of cancer, are most commonly active in only a subset of patients with those cancers. Further, chemotherapeutic agents are generally more toxic and expensive than most other medications in widespread use. Accordingly, an alternative strategy is to test functionally whether a given patient's cancer is likely or not to respond to specific drugs. In vitro chemotherapy sensitivity and resistance assays (CSRAs), a natural extrapolation of antimicrobial susceptibility testing, was first reported in 1957 (1). Some 50 years later, however, the official position of the American Society of Clinical Oncology has been that this type of testing is still not ready for routine clinical use (2). Problems with CSRA include the many different formats for this testing (2), and results from clinical trials, comparing CSRA guided therapy to conventional treatment, have varied from no significant difference to highly significant benefit (3). Finally, most groups test cancer cells immediately after resection, and these may be sick or dying from hypoxia, anesthetic drugs, or overnight shipping, and so any toxicity to these cells may reflect synergistic toxicity of the drugs tested with any of these effects.
We hypothesize that low-passage cell lines might better represent their respective tumors and therefore more accurately predict in vivo chemosensitivity. Isolating cancer cell lines, however, can, counterintuitively, be difficult, especially from solid primary tumors(4). To date, the success rate for the generation of cell lines is only 10–40% for many solid tumors(4–8). The most significant barrier to routine cancer cell line production is that when tumors are explanted into tissue culture, fibroblasts and other stromal cells proliferate, overgrow and eliminate the malignant cells.
We report production of nude-, SCID- (severe combined immunodeficiency) and NSG- (Non obese diabetic [NOD], SCID, Interleukin-2 receptor Gamma [IL2Rγ]), hprt-null mice. During growth of implanted human cancers in these mice, biochemically defective mouse stromal cells replace the human stromal cells from the original tumor(9, 10). After explanting these xenografts into culture, mouse cells can be eliminated by selection in hypoxanthine, aminopterin, and thymidine (HAT) containing media(11, 12) to isolate pure human cancer cells. Using this system, we isolated a total of 6 pancreatic ductal adenocarcinoma (PDA) cell lines and one ovarian cancer cell line. One of the PDA cell lines was isolated from the corresponding surgically resected sample and tested for in vitro chemosensitivity using a 3,131-drug panel(13). In comparison to other PDA cell lines, this cell line in vitro was differentially more sensitive to digitoxin and nogalamycin, which correlated with in vivo response in mice, where the same two drugs demonstrated selective activity against xenografted tumors. These data suggest a possible novel paradigm for functional personalized chemotherapeutic selection by isolating low-passage cancer cell lines and screening them with large drug panels.
Primary tumors from patient-derived resection specimens or xenografts from standard nude mice were harvested with informed consent and IRB approval. They were implanted in anesthetized standard B6 nude and hprt-null B6 nude mice. Following a small skin incision, a 5mm3 sample coated with Matrigel Matrix (BD Biosciences, Bedford, MA) was implanted subcutaneously in each upper flank, the incision was closed using staples, and the site was disinfected with Betadine (Purdue Frederick, Stamford, CT). Among the 6 pancreatic cancers, 3 were selected because they were familial, and 3 because they have undergone full exomic DNA sequencing as xenografted samples(14).
When tumors reached approximately 1 cm in diameter, mice were sacrificed, and tumors harvested. In a laminar flow biosafety cabinet, tumors were finely minced and incubated in DMEM containing 750 units/ml of type IV collagenase (Invitrogen) and 500 units/ml of hyaluronidase (Sigma-Aldrich Inc., St. Louis, MO) for 1 hour at 37°C, and pipetting up and down 20 times to mechanically fragment the tumor. The cell solution was plated at 0.5, 1 and 2 mls per well on 6-well plates coated with or without type I rat tail collagen (BD Biosciences) in MEM with 20% fetal bovine serum, supplemented with standard concentrations of penicillin/streptomycin, L-glutamine, 5 ng/ml of human recombinant EGF (all Invitrogen) and 0.2 U/ml of human insulin (Sigma-Aldrich). Following two days recovery (designated day 0), 1X HAT (100uM sodium hypoxanthine, 400nM aminopterin, 16nM thymidine, Invitrogen) media was fed to half of the cultures. Cells are fed with or without HAT media every 2–3 days. All cell lines were confirmed as patient in origin by DNA fingerprinting with the powerplex 1.2 kit per manufacturers instructions (Promega, Madison, WI). They were also stained with anti-cytokeratin (Ventana, Tucson, AZ, 760-2595) and anti-smooth muscle actin (Ventana, 760-2835) by immunohistochemistry and demonstrated to be tumorigenic in nude mice.
Cancer cells were screened for chemosensitivity using the Johns Hopkins Drug Library (JHDL) consisting of 3,131 drugs arrayed in 96-well plates. This includes 1,907 United States Food and Drug Administration (FDA) approved drugs, 570 drugs approved in other countries, 654 other drugs in various phases of development(13). All drugs are maintained at minus 70 at a stock concentration of 200 microMolar in PBS containing 2% DMSO and 10% FBS.
Tumor cells were subcultured, counted, and inoculated into 96-well plates at 5,000 cells/well in complete media and allowed to adhere overnight. Drugs were then added to the plates in duplicate wells at a final concentration of 10 μM for a total of 48 hours. After the first 24 hours in drug, 1 microCi [3H]-thymidine was added, and incubated for an additional 24 hours (in both drug and [3H]-thymidine). Cells were then trypsinized using 1x trypsin-EDTA (Invitrogen). The suspended cells were transferred onto FilterMat-A glass fiber filters (Wallac, Turku, Finland) using the Harvester-96 cell harvester (Tomtec, Hamden, CT). The glass fiber filters were washed five times with water to wash away unincorporated free [3H]-thymidine. On each plate, the first and last columns were used as no-drug controls. Incorporated thymidine was read on a MicroBeta plate reader (Perkin Elmer), and the mean % inhibition was calculated relative to no-drug control wells. A histogram of the % inhibition for the drugs in the JHDL was established using 2.5% size bins, and normal distribution curve fitting was performed by the least squares method using GraphPad ver. 5.02 (GraphPad Software Inc., LaJolla, CA). The mean +/− 3 SD encompasses 3,014 drugs. Using Gaussian statistics and 3 SD as the cutoff (40.4% inhibition), then we expect ~4 drugs to be false positives among the drugs demonstrating 40–100% inhibition (3014 × 0.13=3.9).
IC50s of selected drugs were determined from % growth inhibition of ten different concentrations of the drugs obtained in the same experimental design described above. IC50s of each drug for each cell line were calculated using four-parameter logistic equation with three replicates results by GraphPad ver. 5.02.
Tumor xenografts were generated by subcutaneous injection of 10 × 106 cells bilaterally into 6 weeks athymic nude-Foxn1nu mice (Harlan, Indianapolis, IN). Tumor volumes were measured and when the tumors reached approximately 100 mm3 in size, the mice were stratified (day 0) to treatment and non-treatment groups with 5 mice per group so that each group was equivalent based on tumor volume. Tumor volume is obtained by the formula: length x (width)2 × 0.5, where length is the longest diameter and width is the shortest diameter perpendicular to the length. The mice received treatment with daily intraperitoneal injection of nogalamycin (0.2 and 1 mg/kg in vehicle), digitoxin (0.4 and 2 mg/kg in vehicle), or vehicle (0.9 % sodium chloride with 1 % DMSO) control for 30 days (day 1-day 30). Tumor volumes were measured twice a week. At the completion of the study, mice were euthanized and tumors were measured, harvested and weighed. The tumor volume index (TVI) was determined from a ratio of the tumor volume on a given day divided by the tumor volume of day 0. The harvested tumors were then weighed and means and standard deviations calculated. The normalized tumor weight of treatment group was calculated by dividing the treatment values by the control group for each cell line (i.e. the mean tumor weight of control group for each cell line is 100). Statistical analysis was performed using the unpaired Student's t test on Graph Pad Prism ver. 5.02.
A series of hprt-null immunodeficient mice, including nude hprt-null (Supplemental Fig. S1), SCID hprt-null (Supplemental Fig. S2) and NOD-SCID Interleukin2γ(NSG) hprt-null, were produced by breeding immunocompetent hprt-null mice with the appropriate immunodeficient mice (see Supplemental Information). Mice that are homozygous or hemizygous for the hprt-null allele and homozygous nu/nu appear to have no phenotypic variation from standard Foxn1nu/nu mice (Fig. 1A). To confirm genotyping (Fig. 1B, Supplementary Fig. S3), we harvested tail cuttings from mice II and III (Fig. 1A), trypsinized them and grew the cells in tissue culture. These samples were grown in the presence of HAT or 6-thioguanine (6TG) to select for or against HPRT function, respectively (Fig. 1C). As predicted from their genotype (Fig. 1B), cells from the hprt-null mouse died in HAT media and survived in 6TG containing media. The HPRT proficient heterozygous hprt-null/wt mouse cells gave the opposite result, as expected.
A PDA xenograft (Panc410) was implanted into these mice and the tumors were harvested for cell culture. After plating and 2 days of recovery, cultures of Panc410 (Day 0) showed a mixture of cancer cells clustering together to form "islands", and scattered fibroblasts and other stromal cells (Fig. 2A and 2D). Cultures explanted from the hprt-null mouse and treated with HAT selection media typically yielded fibroblast free cancer cell lines after 2–3 weeks (Fig. 2A–C). During this time, HAT selection eliminated the fibroblasts and other stromal cells, allowing for the small cancer cell "islands" to expand and eventually coalesce. In contrast, a sister culture, grown in the absence of HAT, was heavily contaminated with fibroblasts (Fig. 2D–F). These stromal cells surrounded cancer cell islands (Fig. 2E), overgrew them (Fig. 2F), and precluded producing cancer cell lines from these cultures. The resultant cell line matched the patient's microsatelllites and was tumorigenic in nude mice. To confirm its epithelial origin, the cell line was stained for cytokeratin and smooth muscle actin, which stained positive and negative respectively, as expected (data not shown).
One potential application of rapid cell line isolation is personalized chemosensitivity testing. To demonstrate utility of the mice for this purpose, we first implanted SCID hprt-null mice with a surgically resected PDA and isolated the cell line Panc502 (Supplementary Fig. S4). We then screened this cell line against the JHDL (13) consisting of 3,131 drugs (Supplementary Fig. S5). The library includes 1,907 US FDA approved drugs, 570 drugs approved for clinical use in other countries and 654 other drugs in various phases of development. A histogram of the number of drugs, graphed as a function of the percent growth inhibition (Fig. 3A), showed that most drugs fit under a Gaussian distribution (mean 5.0%, SD 11.8%), which we interpreted as inactive drugs for this cancer cell line. The mean plus or minus 3 standard deviations encompasses 2817 drugs, or ~90% of those tested. More importantly, 314 drugs (~10%) showed activity above the normal distribution, and these were considered potentially active drugs (Supplementary Table 1). These 314 drugs included 77 drugs that are currently FDA approved and 12 drugs previously approved, but currently discontinued (combined ~2.8% of drugs in the library).
The FDA approved drug, digoxin, showed 95% growth inhibition, consistent with its recent discovery as a potential anti-tumor agent(15). We also identified two terpenoid drugs, pristimerin and triptonide, isolated from Chinese medicinal plants, as potent inhibitors of growth(16–18). Among the 125 antineoplastic drugs tested (Supplementary Table 2), only a third (49 drugs, 39.2%) showed activity above the bell-shaped curve. Nine drugs demonstrated greater than 99% growth inhibition, including 7 nucleic acid-targeting chemotherapeutics (actinomycin C, actinomycin D, chromomycin A3, camptothecin, doxorubicin, gemcitabine, and mitoxantrone) and 2 terpenoid natural products (pristimerin and triptonide).
From the drugs demonstrating 75% or higher inhibition at 10μM final concentration, we selected 10 drugs to study further (Table 1), after excluding drugs with well-documented activity against pancreatic cancer, and those with relatively high toxicities (i.e. low LD50 values in mice). Only one of these drugs is FDA approved. We generated dose-response curves to confirm activity and establish IC50 values for Panc502, Panc410 and another low-passage PDA cell line, Panc486 described below. From these, we selected two drugs because they showed significant variability in their IC50s among the three low passage PDA cell lines (Fig. 3B). The anthracycline, nogalamycin, was approximately 11x more effective against Panc502 compared to either Panc410 or the non-transformed pancreatic duct cell line, HPDE (data not shown). Finally, the cardiac glycoside digitoxin was substantially more toxic to Panc502 than Panc410, while Panc486 had an intermediate sensitivity. We also generated dose response curves using a panel of topoisomerase inhibitors, showing that topoisomerase I inhibitors were generally more effective than topoisomerase II inhibitors (Supplementary Fig. S6).
To address the hypothesis that in vitro response could predict in vivo response, we then raised xenografted tumors from Panc410 and Panc502 cell lines, and treated the mice harboring these xenografts with nogalamycin, digitoxin or control for 30 days. We measured the size of tumors twice a week during this time (Fig. 3C). Both nogalamycin and digitoxin demonstrated more activity against Panc502 than Panc410, supporting the notion that in vitro sensitivity does predict in vivo response, at least with these 2 drugs in these cell lines, as judged by tumor size (Fig. 3C), weight of the tumors after completing the treatment (Fig. 3D), and by visual inspection of the residual tumors after treatment (Supplementary Fig. S7).
To test whether we could use this system to routinely generate cell lines from solid cancers, we also isolated a cell line from an ovarian cancer, another highly lethal cancer. The ovarian cancer cell line, FM108, was established from an existing xenograft (Supplementary Fig. S8). We also isolated an additional cell line from a surgically resected PDA. Panc486 was isolated from a xenograft from a patient with a family history of pancreatic cancer (Supplementary Fig. S9). In each case, cell lines were documented to be tumorigenic in athymic mice, sequenced to document oncogenic mutations in the Kras2 gene, and DNA fingerprinted to confirm their patient origin (information available on request). During this time, we isolated 3 additional cell lines from other PDAs, but were unable to produce cell lines from two other PDA xenografts using similar approaches (total 7 cell lines, out of 9 attempts, 78%). This suggests that the in vitro propagation of the 2 failed cell lines is dependent on one of a number of factors such as factors intrinsic to the carcinoma, factors related to the non-neoplastic stromal cells, putative paracrine growth factors, or critical media components that were missing and remain to be identified.
We report successful production of nude, SCID, and NSG immunodeficient mice whose hprt-null cells are biochemically selectable in vitro. Using this system, we were able to isolate 3 PDA and 1 ovarian cancer cell line. The Panc502 familial PDA cell line was isolated directly from the tumor surgically resected from a patient, and we screened it against a panel of 3,131 drugs. Approximately 314 drugs demonstrated potential activity, where the vast majority of them (265, 84%) were not designated as antineoplastics. Of 125 anti-neoplastic drugs, only 49 (39%) demonstrated activity. Two novel drugs demonstrated selective activity in vitro, and this correlated with in vivo response. We also identified 2 terpenoid drugs with >99% growth inhibition.
In this paper, we describe an alternative strategy to traditional CSRA. In contrast to traditional CSRA, the approach described herein uses: i) pure low-passage human cancer cells, ii) cells that are viable and well-adapted to in vitro growth, and iii) larger drug panels. The use of pure cancer cells, means that any signal detected is due to the drug's effect on the malignant cells rather than stromal cell toxicity. Because the cancer cells are growing and healthy, measured effects are a combined effect of the drug and the cancer cells and not due to synergistic effects of the drug with anoxia, starvation, overnight shipping, etc. Finally, because the cells are expanded into a low-passage cell line, drug panels with thousands of drugs can be screened, rather than the 5–10 typically tested in traditional CSRA. This permits one to test all FDA approved anti-neoplastics, FDA approved drugs for other indications, late phase clinical trial drugs, herbal drugs and drugs approved in other countries for clinical use. The idea of screening drugs using cancer cell lines is well established (reviewed in (19)), and the personalized approach has recently been validated for one patient with a rare type of pancreatic cancer, acincar-cell carcinoma (20).
The approach describe herein differs from the xenograft-based approach demonstrated by Hidalgo's group (21, 22). The advantage of the low-passage cell line approach is that more drugs can be screened under highly controlled conditions. A possible advantage of the xenograft approach is that xenografted tumor response may be more biologically predictive of response in patients. We are eager to test whether low-passage personalized cell line responses can predict patient response. This may be best tested with a malignancy for which several alternative first-line chemotherapeutic choices are available.
Drugs with activity in vivo may not be limited to those that demonstrate >95% growth inhibition. While digitoxin demonstrated 95.1% growth inhibition in vitro, nogalamycin was chosen in part because it only had 75.4% growth inhibition in vitro. Finally, it is possible that the drugs identified could be synergistic with those currently in clinical use such as gemcitabine, although this remains to be demonstrated. In the future, the combination of the patient's germline SNPs, somatic mutations, mRNA expression and epigenetic changes in the cancer, may play an increasing role in the choice of chemotherapy. Despite this, one can also imagine a role for functional CSRAs. This is currently being attempted for various cancers(23–26), although it is controversial whether this testing is truly predictive of in vivo response(2, 3). This initial sample took about 8 months from surgical resection to full drug panel testing, however we estimate that, with additional optimization, this testing could be accomplished much shorter time. The shorter time could be achieved by implanting more mice, with additional experience, and most importantly by using robotics for the drug screening. Certain clinical settings, such as identifying drugs after a Whipple procedure for PDA, might be especially amenable to this time frame. Applying a similar approach to a cohort of genetically defined cell lines, could define a standardized core set of drugs to be tested on all resected PDAs.
Cell lines are useful for many other studies of cancer biology since they: i) can be expanded indefinitely, ii) contain all of the gene mutations present in the patient's primary cancer, iii) can be manipulated in vitro by adding or eliminating genes, iv) can be implanted in mice to test the effects of these manipulations on the ability to form tumors(21, 27), and v) carry few if any additional genetic changes from the primary cancer(28). Cancer cell lines are also important for other in vitro functional studies, molecular imaging, and possibly isogenic cancer vaccine production. With increasingly powerful tools such as whole genome DNA sequencing, RNAi libraries, and high-throughput drug screens, one can anticipate that the need for cell lines from human cancers will likely increase(13, 29–31).
We envision several applications of this system for cancer cell line production. First, there are no cell lines representing some malignancies, such as oligodendrogliomas and pheochromocytomas, and in others, such as esophageal, thyroid, and salivary gland tumors, many cell lines are contaminants (32). For others, such as prostate, ovarian and breast cancers, the number of available cell lines cannot fully represent the full spectrum of disease (e.g. most existing prostate cancer cell lines are hormone refractory). Additionally, some cancers contain unique genetic defects, and successful cell line production is essential to identify pathway members, and perhaps most importantly, for high-throughput drug screening.
There are some limitations of this system. For some cancers, stromal cells from the tumor microenvironment may provide paracrine growth stimuli to the malignant cells, and this may explain why we were unable to isolate two of the cell lines. It is generally believed that all of the human stromal cells are replaced by mouse stromal cells in a single passage in immunodefective mice. While we generally agree with this assessment based on phase microscopy of HAT treated cultures (unpublished data), this has never been formally quantified. The replacement of stromal cells may be slightly less than 100% based on the extremely faint residual PCR product from DNA isolated from xenografts of cancers containing homozygous deletions(33). In this regard, it is possible that some tumors may need to be passaged through additional mice to get complete replacement of the human stromal cells. It is also possible that some cancers require unique cytokines that are not currently components of tissue culture medium.
In addition to cell line production, one can envision other situations where an animal model is required to study a biological process, but where recovery of the exogenous cells is subsequently required. For example, the mouse may be useful for isolation of organ-specific spontaneous metastases or small numbers of dormant malignant cells refractory to chemotherapeutic agents. Finally, high-throughput functional chemosensitivity profiling may be especially warranted for those patients who have failed conventional chemotherapeutic options.
Cancer cell lines are essential for functional studies of cancer. When explanted tumors are grown in vitro, however, non-cancer fibroblasts paradoxically overgrow the malignant cells. Here we report production of a series of immunodeficient hprt-null mice. When human tumors are expanded in these mice, the resultant tumor is composed of human cancer cells and biochemically defective mouse stromal cells. When these tumors are explanted into culture and grown in HAT media, human cancer cell lines can be readily isolated. We demonstrate proof-of-principle of the mice for personalized cell line isolation and chemosensitivity testing. This approach may be used to guide chemotherapy selection in the future.
We dedicate this work to the memory of Dr. Dawn Audi. We thank Dr. Scott Kern for the suggestion to use hprt as the selectable marker (PCT/US01/31219), and Dr. Ming-Sound Tsao for generously providing the HPDE cell line. We acknowledge Drs. Yoshihisa Matsushita, Bert Vogelstein, Cynthia Zahnow, Julie Watson, Curt Civin, Cory Brayton, Don Price, Phil Wong, Craig Henke, Mehtab Khan and Rajni Sharma for helpful discussions. This work was funded in part from NIH grants CA130938 (JRE), CA62924 (Drs. Scott Kern, RHH, AM, JRE), CA122581 (RBSR), The Sol Goldman Pancreatic Cancer Research Center, The Stewart Trust Fund, The Lustgarten Foundation, the Mary Lou Wootton Pancreatic Cancer Research Fund, The Michael Rolfe Pancreatic Cancer Foundation and the HERA Foundation (RBSR).
Notes: The hprt-null NSG mice are available from Jackson Labs (Bar Harbor, Maine) as mouse #012480 (NOD.Cg-Prkdc<scid> IL2Rγ<tm1Wjl> Hprt<b-m3>/EshJ).
Author Contributions: AM and JRE conceived the project. HK, SR, JSS, CAK, GF, LH, MK, FWS, MTL, RMB, BK, HL, MEM, GM conducted the experiments. MH, EJ, RHH, HAJ, RBSR, AJ, JOL provided essential reagents and critical oversight.
Potential Conflict of Interest: Drs. Rauenzahn, Maitra and Eshleman may receive royalty payments if the mice are licensed; a patent licensed to Myriad Genetics (Drs. Hruban and Eshleman); and Advisory Board membership in Roche Molecular Diagnostics (Dr. Eshleman). No potential conflicts of interest were declared by other authors.
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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Looks like 3 Bedroom House is no longer available on Rent.com which means this information is likely out of date.
House on corner lot w/ pond in back
Nice spacious 2300+ SF house for rent. 3 bedroom 2.5 bath, Big
$1400 plus $1200 Security Deposit. It will be available 10 Dec 06.
Two Story Home with an attached 2 Car Garage
The home was built in 2002
Central Air
Located on the main drive
Corner of Westover and Charring Cross
Special Faux finish paint through out the house
Located on same corner of school bus stop
Has fire hydrant on same corner
Pond located in back yard
It is not fenced
No basement
First Floor
Kitchen Features (all items are energy efficient)
It has a walk in pantry
Water Heater is located in a Kitchen closet
Gas four burner Range
Built in microwave with fan located above Range
Dishwasher located next to sink
Garbage Disposal
Standard Fridge with an ice maker and water line
Maple Cabinets with basic laminate countertops
There is an area to place a 5ft table and chairs
Living Room
Gas fire place
It is open to the second floor
Ceiling Fan with remote
Dining Room
Large enough to accommodate
My 8 person table
Two Curios
Half Bath
A water closet is located on the first
2nd Floor
Bedroom # 1
Bedroom one is 11' 10" x 10' 1"
It has "California Closet System"
Bedroom #2
It is 11' 10" x 11' 2"
One closet with sliding Doors
Full Bath
Standard shower in tub
No doors (shower curtain)
New Floors in June
Washer Dryer Closet
Front load washer & pedestal
Front load dryer & pedestal
Master Bedroom
19" x 15'
Vaulted ceiling
Ceiling Fan with remote
8' x 6' Walk in closet
Attic is accessible through walk in closet
New Floors are to be installed 1 Dec 06
Master Bath
Garden soaking tub
Stand up Shower
Large Vanity with 2 sinks
No Pets
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subset: commoncrawl


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MISC 2009 conference to tackle policy-making in Canada
Do big crises generate good or bad policy? Do they allow government to act more efficiently, or simply react with little time for analysis or reflection? Where do innovative public policy ideas come from? Does evidence matter? Do media and public opinion?
On March 26 and 27, The McGill Institute for the Study of Canada (MISC) will present its 2009 conference, Public Policy in Crisis? Understanding Policy-Making in Canada, at McGill University's Faculty Club (3450 McTavish Street).
Practitioners and academics will come together to address issues central to current policy debates: how policy-making decisions are made in a time of crisis, the influence of public opinion and mass media on the policy-making process, the role of evidence based research, and the influence of non-governmental actors. It promises to be a dynamic exchange of ideas and opinions that will yield new perspectives on the processes and players that shape public policy in Canada, and, as a consequence, impact the lives of its citizens.
"The ongoing financial crisis has prompted national governments around the world to cobble together sweeping economic policies in very little time," said Antonia Maioni, Director of MISC and conference chair. "Does this type of urgency bring out the best or the worst of the policy-making process? It leaves hardly any time for consultation, but how much input is actually sought under more usual circumstances, and from what sources? These are some of the issues Public Policy in Crisis? will address, and some of the questions we hope to answer."
The conference is presented in partnership with the McGill-Max Bell Innovation and Implementation in Public Policy research project.
Confirmed participants include:
The Rt. Hon. Joe Clark, former prime minister of Canada.
Philippe Couillard, former Minister of Health, Government of Québec; Senior Fellow in Health Law, McGill Research Group on Health and Law.
Kevin Lynch, Clerk of the Privy Council and Secretary to the Cabinet.
Nik Nanos, President and CEO, Nanos Research.
Ian Brodie, former chief of staff to Prime Minister Stephen Harper; Visiting Scholar, McGill Institute for the Study of Canada.
Sandra Buckler, consultant; former Director of Communications for the Office of the Prime Minister of Canada.
Mel Cappe, President, Institute for Research on Public Policy; former Clerk of the Privy Council and Secretary to the Cabinet.
Sharon Manson Singer, President, Canadian Policy Research Networks.
Alain Dubuc, journalist, La Presse.
Pearl Eliadis, human rights lawyer.
Matthew Mendelsohn, former Deputy Minister of Intergovernmental Affairs and Deputy Minister for Democratic Renewal, Government of Ontario.
Peter H. Russell, Professor Emeritus, Department of Political Science, University of Toronto.
Bruce Doern, Distinguished Research Professor, School of Public Policy and Administration, Carleton University.
Leslie A. Pal, Chancellor's Professor of Public Policy and Public Administration, Carleton University.
B. Guy Peters, Maurice Falk Professor of American Government, Department of Political Science, University of Pittsburgh.
Mary Simon, President, Inuit Tapiriit Kanatami.
Wendy Thomson, Director, School of Social Work, McGill University.
Christopher Waddell, Carty Chair in Business and Financial Journalism and Associate Director, School of Journalism and Communication, Carleton University.
Since 1995 MISC has hosted large-scale annual conferences which foster informed, non-partisan discussions of issues affecting Canadians, ranging from Quebec-Canada relations, Aboriginal issues, citizenship and health-care to Canadian media, the Charter of Rights and Freedoms, food, and cultural policy.
On the Web: http://www.mcgill.ca/policyincrisis/


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Lodge House PlansSeptember 1st, 2011
An Idaho log-and-timber beauty showcases an innovative design that blends seamlessly into the rugged mountain landscape. Lodge plans, lodge home plans, and lodge style home plans are all offered at Stockton Design. Makers of superior quality milled log wall packages with pre-cut and pre-drilled thru-bolt attachment system. Find details on Elegant Lodge Style Craftsman (HWBDO14533) at BuilderHousePlans. FREE SHIPPING ON EVERY PLAN + Call or email us about special summer promotion. Search quality Lodge home plans, house plans, one & two story floor plans from Associated Designs. Rustic mountain style house plan with family room, lodge room, observation loft and covered porches.
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Devonshire Green Medical CentreTel: 0114 272 0255
Hanover Medical CentreTel: 0114 276 2248
Following recent re-inspection our practice has been upgraded to an overall Good rating.
From 26 June 2017 the Electronic Prescription Service (EPS) will be offered at our practice.
EPS makes it possible for your prescriptions to be sent electronically straight to the pharmacy of your choice. If you have regular prescriptions the EPS may be able to save you time and saving unnecessary trips to the surgery.
Choosing a pharmacy to process your EPS prescription is called a 'nomination'. This means, you will no longer need to collect a paper repeat prescription from the surgery, instead you can go straight to the nominated pharmacy to pick up your medication.
For more information visit the NHS England website.
This is a new service, and there may be the occasional problem while we try to get to grips with this new procedure, we would ask that you bear with us and we will try to sort any issues that arise.
On Friday 3 March we will be trialling a new telephone system.
If you call the surgery and the lines are busy you will receive a message telling you where you are in the queue, you can then make the decision to either call back later or continue hold. As soon as a Receptionist is free they will take your call.
We are going to see how this works for patients and staff for a three month period and then review this system at the end of May. Any feedback would be greatly appreciated.
As you will know we have had unusually high demand on booked and drop-in appointments over the past six weeks. To manage patients safely we have decided to change the arrangements regarding number of patients to be seen at drop-in. This change will start from 1st of May 2016.
There will be 18 appointments available to book at each surgery on the same day for drop-in, by attending at reception from 8.30am, as before. When 18 appointments are full, you will be asked if your problem is 'medically urgent for today'.
If it is medically urgent a doctor will speak to you to decide when you can be seen. Children under 5 years old will also be assessed by a doctor to decide when they can be seen.
If you have a minor illness, please check with your local pharmacist if they are able to treat this under the 'Minor Ailment Scheme'. You can also be seen at the Walk-In Centre on Broad Lane.
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So I started making the card yesterday (stamped the images, cut background papers) but did not finish it until this morning. I woke up early to bid hubbs goodbye as he went to work…plus could not wait to complete the card before running some errands.
So here is the card:
Loved the pattern paper (which came in the kit)…and aren't those envelopes cute??? Most of the materials are from the Kit…. except the card base, the sentiments "Sending" (Your Next Stamp) and "On your Birthday" (Stampin' Up! Retired) and Oval punch (Stampin' Up!)
I stamped and hand cut the envelopes…. added hearts (included in the set), gave it some shine with Wink of Stella Clear Glitter Pen and added some Glossy Accents. These are popped up, so is the greeting at the bottom.
Absolutely love this card…. so cute and sure to bring a smile on the receivers face 🙂
Until next time,
Happy Krafting,
Cheers =)


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Your search for "nox4" returned 1,942 TaqMan® SNP Genotyping Assays
NADPH oxidase 4
This gene encodes a member of the NOX-family of enzymes that functions as the catalytic subunit the NADPH oxidase complex. The encoded protein is localized to non-phagocytic cells where it acts as an oxygen sensor and catalyzes the reduction of molecular oxygen to various reactive oxygen species (ROS). The ROS generated by this protein have been implicated in numerous biological functions including signal transduction, cell differentiation and tumor cell growth. A pseudogene has been identified on the other arm of chromosome 11. Alternative splicing results in multiple transcript variants.[provided by RefSeq, Jan 2009]
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Running Time: 92 Min
Age Restriction: 18V
Synopsis: In Paris, a young employee in the office of the US Ambassador hooks up with an American spy looking to stop a terrorist attack in the city.
Link1 | Link2 | Link3
Copyright © 2017. All Rights Reserved by Queensmead Video.
Designed by Marius Strydom.


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|Reviews for Travel Games
| chloe chapter 3 . 7/1/2014
| Darkness chapter 1 . 5/5/2014
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Not bad
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| gitsfanatic117 chapter 1 . 1/4/2013
I read all of these and i must I loved them all I never really thought about sokka x zuko but now I love it Great works of art in my opinion :)
| TheGodlessAngelOfDarkness chapter 3 . 11/11/2012
chapter four. Splatter art ;D ... God this is like idk the 8th time i read this and i never get a enough...but now... i have an acccount and can comment. brava dear! keep it up and i hope/wish you wpould update a chaoter four...
forgive spelling errors...i am partly blind -_-
| TamiLove chapter 3 . 8/23/2012
lol! Great! I loved it!
| TamiLove chapter 1 . 8/23/2012
omg! laughing my butt off! this is sooo awkward! Sokka is waaaaay too okay with all this! lololl!
| NanaLoveChel chapter 3 . 8/16/2012
This was funny through and through. I like how you don't write the characters in a way that makes me hates them. There are some authors that have made me permanently bate an okay character, which is something you have successfully not donee! *busy*
| NanaLoveChel chapter 2 . 8/16/2012
This chapter was super funny! I was snickering pretty much the whole way through. Zuko is so cute! -_-
| NanaLoveChel chapter 1 . 8/16/2012
I must say I love you. Not only do you write better than decently, but you write gay porn your sisters request. I wish my sister would write fanfiction for me! A good first chapter! _
| Sir Shirkin chapter 3 . 7/24/2012
I have to say... this was quite amusing ;3 3 I thoroughly enjoyed it Good job 3
| She.Drowned.In.Venom chapter 3 . 7/8/2012
That was damn sexy! Love it!
| Ruby of Raven chapter 1 . 5/30/2012
Ha! What a nice game! Sarcasim intended. ;)


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After much speculation Derry City have confirmed to the Derry Journal that they will play their first round Europa League qualifier in Sligo Rovers ground 'The Showgrounds'. The news comes as the clubs agreed a fee of around €5,000 for the use of the ground which previously was 10-15k. Dundalk had offered
Sligo Rovers
Ian Baraclough is set to be named as the new Northern Ireland u21's manager
Former Sligo Rovers boss Ian Baraclough is set to be named the Northern Ireland u21's manager in the next few days. According to a report in The Times, the current Oldham assistant manager was interviewed by current Northern Ireland boss Michael O'Neill for the role. The Leicester man led Sligo Rovers to
Sligo Rovers appoint Gerard Lyttle as new manager
Sligo Rovers are pleased to announce the appointment of Gerard Lyttle as new manager. Lyttle comes to The Showgrounds following two years in charge of Danske Bank Premiership club Cliftonville. He has today signed a contract to the end of the 2018 season. He led the Belfast club to a fourth-place finish
Former Togo & Trinidad & Tobago manager applies for Sligo Rovers role
Dave Robertson and Sligo Rovers part ways
Following a review of the season to date Sligo Rovers and their manager Dave Robertson have mutually agreed to part on an amicable basis. Chairman Martin Heraghty thanked Dave Robertson for his contribution and total professionalism with the club over the past eighteen months: "Everyone at Sligo Rovers would like to thank
Shane Keegan determined to get three points against Sligo Rovers
O'Connor stoppage time penalty propels Harps to sixth
Connor McGinty Ciaran O'Connor is very quickly becoming a fans favourite on the Ballybofey terraces despite only being on loan from Dundalk. With another brilliant display and a goal to add to his current tally it's evident why with a great energetic performance every game. Sligo had the first real chance
Limerick show their class and breeze past Sligo Rovers
2017 League of Ireland Premier Division predictions
With the 2017 season nearly upon us, we at LeagueofIreland.ie wanted to give our season predictions for the Premier& First Divisions this season. This article will feature the Premier Division predictions and as you can see below we feel that the title will head to Leeside this season. The re-signing of Sean
Sligo Rovers defeat Finn Harps in pre-season friendly
Connor McGinty A fast and physical introduction with a long ball tactic being adopted by both sides early on. Sligo's mixed play found space in a tight harps defence however nothing came of several over the top through balls in what was a rather uneventful opening 10 minutes. Several players on trial with


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QUI ou QU'Y ?
QUI ou QU'Y ?
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Ambassador John Bruton
Head of Delegation of the European Commission in the United States
Ambassador Bruton spoke about how a bi-directional partnership between the EU and California is evolving and what chances there are for future regulatory cooperation, learning, and emulation between them. California has been a regulatory trendsetter at the national and international levels. Recently the European Union has become a global regulatory leader, while California has become both a launching pad for American versions of European regulation and an innovator that influences Europe.


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Knowing the code is all well and good, but if you don't know how to use it, there's no point. Pretty much everything in the tutorial I linked above is to do with the design and not the code. Coding a website (overly complex ones aside) is only about 20% of the importance. The ability to use CSS/HTML is commonplace nowadays on the web, however the ability to design is much more sought after. Start with that page, look up some other tutorials on Google for design advice.
Thanks for the link, there is just so much crap that has to be taken into account when designing. If you ask me its more complicated than programming. Atleast in programming you have a goal, you know what you have to do and you have a few ways to get thre, or sometimes you just have the direct way. Either way picking up this design stuff is going to be harder than I thought. I wish I had the money to take a good design course..
I really dont understand how people can expect you to do both. The programming on a site can get pretty complicated, then they want you to design their site too? SMH.. oh well I'll get this.
EDIT: I am reading some articles that made me realize I'm going about the design process all wrong, my main problem is im trying to rush/force it..


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Rebel (The Starbuck Chronicles #1)
By Bernard Cornwell
When Richmond landowner Washington Faulconer snatches younger Nate Starbuck from the grip of a Yankee-hating mob, Nate is either thankful and awed by way of his idealistic rescuer. To pay off his generosity, he enlists within the Faulconer legion to struggle opposed to his domestic, the North, and opposed to his abolitionist father. whilst the regiment joins up, able to march into the ferocious conflict at Buff Run, the boys are ready to begin a struggle . . . yet they are not prepared for the way they—and the nation—will be endlessly replaced by means of the oaths they've got sworn for his or her cherished South.
Quick preview of Rebel (The Starbuck Chronicles #1) PDF
Simply kill him for me. " Sally paused. "It ain't that I brain being the following, Nate, in truth it's most likely the simplest position for me, yet I hate that son of a complain for telling me lies, and that i hate that he thinks he obtained away with telling me lies, and that i wish that son of a whinge lifeless and that i wish the very last thing he hears in this earth to be my identify so he don't ever disregard why he's long past to hell. Will you do this for me? " expensive God, Starbuck idea, yet what percentage sins have been right here introduced into one foul package deal? what percentage entries may the Recording Angel be making within the Lamb's ebook of existence?
A few shells, fired too excessive, crashed throughout the branches above the accomplice line, scaring birds out of the bushes and showering twigs and leaves down onto the bandsmen and chaplains and servants and scientific orderlies who crouched within the rear. A regiment of normal U. S. military troops marched out from the timber, deployed from column into line, mounted their bayonets, and complex uphill with the recent Yorkers and New Englanders. Colonel Evans had galloped again to his line's heart the place Colonel Sloan's South Carolinians have been crouching on the wood's aspect to make themselves right into a tough goal for the enemy artillery.
Starbuck requested a bit too urgently. "I'll let you know at dinner," Anna stated. "Now why don't you end dressing, Mister Starbuck, then tie your drained horse to the again of the cart and experience domestic with me. you could carry the parasol and inform me all concerning the raid. i would like to listen to every little thing. " Ethan Ridley took Sally Truslow to Muggeridge's material and Millinery in alternate Alley the place he obtained her a parasol in published calico to compare her light eco-friendly linen cambric costume. She was once additionally donning a fringed paisley scarf, yarn stockings, a wide-brimmed hat trimmed with silk lilies, white ankle-length boots and white lace gloves.
The sergeant started jogging back, asserting not anything, yet simply gripping the silver ring as though it was once a talisman for all his destiny existence. He didn't communicate back until eventually they have been a number of yards from the fires of a firm, after which he placed a hand at the blood-hardened textile of Starbuck's sleeve. His voice, while he spoke, used to be unwontedly meek. "So how is she, Captain? " "She's satisfied. strangely satisfied. She was once handled badly, yet she got here via it and she's chuffed. yet she sought after you to have the hoop, and she or he sought after me to take it from Ridley.
Ja, Colonel? " "Give Boston a few barrelito. One cup. " It appeared that Evans had selected the identify "Boston" for Starbuck's nickname, simply because the curious stone barrel at the German orderly's again was once known as "barrelito. " the large German rode his piebald horse with regards to Starbuck whereas one other guy, hurrying to obey Evans's orders, reduce the rope at Starbuck's wrists. Starbuck all started massaging the rope burns, then observed that the emotionless German orderly had reached at the back of his again to govern a small wood faucet that was once permit into the bottom of the stoneware barrel.


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Very authentic Eastern Italian (Abruzzi/Pescara) cuisine. I thought the food was very good and very true to the area of Italy. I have eaten in Pescara and the restaurant could have been transplanted from there. I had a vegetarian sandwich and it was excellent and huge, but a business associate had the Mahi Mahi special and said it was very good as well.
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Now comes the time where most Atlanta Braves fans become so happy that one may believe that the team has won a World Series title. Late Saturday night, it became known throughout Twitter that second baseman Philip Gosselin was being called up to the major-league club, which sent Braves country into an absolute frenzy. The main thought, that Dan Uggla was being designated for assignment, excited people for some reason. However bad it is, though, that someone is losing their job, it is really best for the team if these rumors are true.
Then, when everyone woke up on Sunday, we found out that due to an internal matter, Uggla had been suspended for one game. Enter the most confused fanbase in all of baseball. There is a player who has made only two starts in the remote past that has been suspended from the team for one game for what is believed to be tardiness. A player who doesn't play, ever, gets suspended for being late? Weird.
However, this is quite likely a move to give the team some time to attempt to put Uggla in a trade package before the All-Star game. With this being the last game for four days, the team has plenty of time to try to hash out a deal for a bench bat or, more importantly, a good bullpen arm. Yet, it seems that if a trade doesn't happen soon, it is highly likely that Uggla will be released by the time the team returns from the break. The team would much rather trade Uggla and not have to eat his entire contract over the next two seasons, but the chances of that seem really slim. However, without him on the roster, it opens the possibility to put a better bat on the bench and to remove some of the drama from the team.
Steven Whitaker is an Atlanta Braves writer for Rantsports.com. Follow him on Twitter @CoachStevenWhit or add him to your circles on Google


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This Car Price: 1200, Year: 2000, Transmission Type: Automatic, Position: Québec, Mileage: 220000, Brand: Toyota Other,
Selling my car, the price is slightly negotiable, and selling because I'm looking to upgrade. Water pump needs changing soon, windshield has a crack, and the rear bushings need changing eventually, they make a slight rattle over some bumps. Other than that it runs like a Camry; very well. I have two sets of winters, one set on the white mags in the pics, one set on black steelies. I use the white rims as summers and the black steelies as winters, both in great shape. Bluetooth radio installed. http://www.kijiji.ca/v-autos-camions/ouest-de-lile-qc/2000-toyota-camry-solara-4cyl-automatic/1212423044
Really want to order 2000 Toyota Camry Solara 4cyl automatic? Remember! Meet face to face and never pay for something you have not seen.


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Saturday 4/13 Workshop - Stop learning Steps - Learn to navigate and improvise
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- Saturday April 13thArgentine Tango 2 hour WorkshopLearn to comfortably dance on a crowded floor by learning the various possibilities at any particular point. 90% of Argentine Tango dancing is navigating and, perhaps, 10% is dedicated to steps. If you want to dance Argentine Tango, you first need to learn to navigate and then, if there is space, you can execute a step.In just two hours you will be able to properly navigate, using the available space, and execute the step of your choice.This Workshop is intended for Argentine Tango dancers who have been taking lessons for one or two years and find themselves overwhelmed and unable to comfortably dance at a milonga.Come this Saturday, April 13th at 12:30, and learn the freedom Argentine Tango offers.The cost of the two hour workshop is only $30.Argentine TangoHave you been learning tango for a year or more and........ have learned a new step every week but don't know how to put them together?.... feel you don't know how to navigate?.... have been told to improvise but don't know how?.... you sit down when the music starts and don't know what to do.?Then this workshop is for you. In just two hours you will be able to put it all together and enjoy dancing Argentine Tango.
Chosen by Westchester Magazine
as the Best Argentine Tango Instructor
Carlos and Karen (Keni)
Certified Tango Instructors
sampelayo@... or tangodance@...
Dance Argentine Tango in Hawthorne, NY,SMITHS DANCING SCHOOL
26 Broadway Avenue, Hawthorne, NY
We teach using the fastest method to learn Argentine Tango
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Dentists in Colmar, PA
"At Dental Innovations we strive to be the place where patients can feel safe and confident that they are receiving the most attentive gentle and modern dental care they have ever experienced. We pride ourselves on developing good relationships with our patients and treating them as we would treat our friends. Our team is devoted to helping you achieve the best possible dental results in a comfortable and friendly environment.We accept most payment methods but do not accept state aid. Call 855-846-1749 now."
Office is near:Colmar, Pennsylvania 18915
Practice Focus
- 1 Hour Whitening
- Bonding
- Braces
- Brite Smile Whitening Process
- Crest Whitestrips
- DaVinci Veneers
- Day White
- In Office Tooth Whitening
- Interactive Patient Education System
- Invisalign
- Laser Whitening
- Lumineers
- Mac Veneers
- Nite White
- Non-surgical Bite Correction
- Opalescence
- Porcelain Veneers
- Retainers
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- Tres' White
- Waterlase


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You can add GIF, JPG and PNG files, up to 2MB each.
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All kinds! Besides draftest websites and logotypes, we've prepared a third, flexible option
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Just enter your e-mail address in order to regain access to all previously created draftests.
Go to the folder marked "spam". The email may have ended up there. If you still can't find it, please contact us. We'll help you out.
In the email that was sent to you after you created your draftest there's a link to project management.
Click on it to see the ratings and comments of people who have tried out your draftest.
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Subscribe to our Newsletter and receive information about In-Person and Virtual Programs, promotions, articles and resources.. delivered with just a touch of beauty & inspiration.
complimentary consultation
Your privacy & security are right alongside your safety as of our highest priorities. Exceptional care is taken to ensure how your personal and health information is managed & stored by core harmony, and the professional organizations we work with are evaluated and chosen to make sure their privacy policies are in alignment with ours. We never use or sell any of your personal information with any other company for any reason and your contact information is shared only as necessary to provide services for core harmony. Information and records are kept and stored in locked areas and under the guidelines & time limits required by law. Documents containing personal and health information are shredded and destroyed before disposal.
While personal testimonials of your experience and achievements are very beneficial in helping others to know first hand what to expect from classes and programs, many people come to core harmony because of the exceptionally high level of confidentiality we offer.
All health information is HIPAA compliant. As a private company that is not affiliated in any way with any other organization, we are one of the few lifestyle & wellness companies that can truly guarantee that your personal and health information will remain 100% confidential. In corporate settings where optional risk evaluations and testing are completed, aggregate data may be compiled to make general recommendations for wellness programs. As we do not release information to your employer or insurance company this guarantees not only your privacy, but that you alone will have the information necessary to make informed decisions about your health and that it will be never be used against you. Under certain medical conditions we are required to obtain medical clearance from your physician and wellness providers. If this is necessary, the release will be sent to you for proof and you can choose to have the release completed by your MD or we provide this service. Faxed documents return to our confidential fax line if you have given your MD approval to correspond directly with us for this or to discuss modifications necessary for your program. All information we receive on your behalf from your MD will be shared with you. All medical clearance related activities are completed to ensure your safety and are included as part of your program with no additional fees.
The foundation of all programs includes a non-denominational spiritual component that is based on the premise that a 'higher good' and benevolent force is always present as part of the coaching process. While your coaching program may have nothing to do with religion or spirituality in any way, this unseen force is always present to create a 'sacred' setting of emotional & energetic safety - and therefore a place where healing, deep & powerful insights, learning and growth can occur. Because the coaching environment offers a place for honesty at a core level, the privacy of your coaching program is protected much in the way other spiritual practices are – we do not share the content of your program or even that you are a client with anyone for any reason.
We are grateful for any referrals that are made to us, and your satisfaction with services provided is the highest compliment we can receive. If you initiate a referral, the decision if and what to disclose about the nature of our relationship is up to you. Please make us aware if you have shared details of your program with a referral so we can acknowledge appropriately. We do not discuss details of any other person's program or private information even if there is a relationship between referrals. We do not give or receive any type of compensation for giving or receiving referrals of any type.
In some cases, a small commission is received for products & music sold or advertised on the web site.
Secure online payment is available through our web site or in person. Verification of our business and the security of the web site for payments can be made via the SSL Certificate links on each page of the site through our hosting company, Network Solution/ Web.com. We do not take or store any credit card information - all payments are completed through our trusted and verified relationship with Paypal. Returned checks are subject to additional fees. Additional late fees are detailed in individual or corporate contracts.
We offer a confidential email address to each client and class participant to use for your program and to maintain privacy for correspondences around your health information. You will choose a username & a password. Access to your email can be set up or accessed via web mail from any computer. Messages can be encrypted to add an additional layer of privacy if you require this. In corporate settings we recommend using a personal email or one of ours vs. your company email address to support compliance by you and your employer to HIPAA regulations.


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4 bedroom villa for sale
Spain - Andalucia, Malaga, Estepona
like this property?Call: 03339 392086
Key features:
- Pool
- Garden
- Heating
Full description:
The villa itself was fully renovated in 2015. There is a fully fitted, open plan kitchen and a separate laundry area.
There is a two car car port and the swimming pool is 8 x 15 metres.
The villa benefits from solar panels for water heating.
Nearest Airports*
- Gibraltar (North Front) (International), 22.5 miles
- Málaga (International), 39.8 miles
- Jerez (International), 55.2 miles
- Sevilla (San Pablo) (International), 79.4 miles
*Distances are straight line measurements. This feature is currently in beta.


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2017 Breakfast Dates
We look forward to supporting you and your business.
This offer for Annual Supporter is available for the 2017 Calendar Year and can be purchased online, follow the link below.
Renew Membership & Join Online


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Planes, Dog Sleds and Automobiles – New York Times
Planes, Dog Sleds and Automobiles
New York Times
… talk about flying)." A few weeks later, he introduced her to Leyla and Sheba. ROSALIE R. RADOMSKY. A version of this article appears in print on March 30, 2014, on page ST18 of the New York edition with the headline: Planes, Dog Sleds and Automobiles.
This Post sponsored by: Find Business for Sale on BizLocal.com
BizLocal.com a division of Inspetta, provides an online marketplace for individuals and brokers to list businesses for sale to locate persons seeking to buy a business. 1-800-785-2925
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Words ending with omasum, the most common words first
2 words found.
There are 2 less common words:abomasum
List all words ending with omasum: sorted by length, sorted alphabetically.
Some random words: uglier snib eluant kiang ocotillo old daiquiri
This is not a dictionary, it's a word game wordfinder.
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text: Shock Tactics. Available to all. Does long term suppression. Suppression lasts all turn. Retrieved from "http://pg2.wikia.com/wiki/PeG:Shock_Tactics?oldid=1227" Ad blocker interference detected! Wikia is a free-to-use site that makes money from advertising. We have a modified experience for viewers using ad blockers Wikia is not accessible if you've made further modifications. Remove the custom ad blocker rule(s) and the page will load as expected. Categories: PeG:Leaders Add category Cancel Save


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The combination was made using a free download programme entitled "CZM".
I wasn't 100% happy with the combination since there appeared to be a slight misalignment of the images. This is why I finished it off with Fractalius.
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HIL Simulation
- Details
- Created: 08 June 2012
On Wednesday, May 23 2012, Controllab and Bachmann electronic have organized a Hardware-in-the-Loop (HIL) Simulation event.
At this event live demonstrations have been given to show how quality improvement can be obtained with HIL simulation. Over 40 visitors were present at the event to attend an inspiring day.


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Join Kara on Tuesday, January 12 from 2-7 pm. $75 plus materials.
Learn to use pick up sticks to make a honeycomb textured scarf. This is a beautiful stitch that creates a slightly puckered 3-dimensional effect, great for all types of wovens. The design is best made using two colors, one for warp and a contrast for weft. A yarn with a good amount of stretch works especially well for the warp. The store sample was woven using Colinette Jitterbug for the warp on a 10 dpi reed and Baah Sonoma for the weft. Other great yarn choices would be Rowan Lima, Zara, or Artyarns Hand-Dyed with a 7.5 dpi reed. Misti Alpaca or Debbie Bliss Paloma on a 5 dpi reed would make a chunky version. Finer yarns such as the Baah Aspen would also be great using a 12.5 dpi reed. Use your own loom with at least one pick up stick or register early to reserve one of ours.
Advance preparation may be required depending on the class topic. Details will be provided one week in advance or upon sign-up. Each weekly class session includes two hours of hands-on instruction, and extra help outside of class if needed and our time allows (it almost always does!). Yarn for classes must come from Compatto Yarn Salon, and we are always happy to help you choose the best yarn for your class or project. If you have questions about this or any other class, please give us a call at 310-453-2130.


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|Peer-Reviewed Publications
|Desai, P. K., Tseng, H. & Souza, G. R. Assembly of hepatocyte spheroids using magnetic 3D cell culture for CYP450 inhibition/induction. Int. J. Mol. Sci. 18, 1085 (2017).
|Souza, G. R. et al. Magnetically bioprinted human myometrial 3D cell rings as a model for uterine contractility. Int. J. Mol. Sci. 18, 683 (2017).
|Tseng, H. et al. A high-throughput in vitro ring assay for vasoactivity using magnetic 3D bioprinting. Sci. Rep. 6, 30640 (2016).
|Hogan, M. et al. Assembly of a functional 3D primary cardiac construct using magnetic levitation. AIMS Bioeng. 3, 277–288 (2016).
|Lin, H. et al. Nanoparticle improved stem cell therapy for erectile dysfunction in a rat model of cavernous nerve injury. J. Urol. 195, 788-95 (2016).
|Tseng, H. et al. A spheroid toxicity assay using magnetic 3D bioprinting and real-time mobile device-based imaging. Sci. Rep. 5, 13987 (2015).
|Jaganathan, H. et al. Three-dimensional in vitro co-culture model of breast tumor using magnetic levitation. Sci. Rep. 4, 6468 (2014).
|Tseng, H. et al. A three-dimensional co-culture model of the aortic valve using magnetic levitation. Acta Biomater. 10, 173–82 (2014).
|Timm, D. M. et al. A high-throughput three-dimensional cell migration assay for toxicity screening with mobile device-based macroscopic image analysis. Sci. Rep. 3, 3000 (2013).
|Haisler, W. L. et al. Three-dimensional cell culturing by magnetic levitation. Nat. Protoc. 8, 1940–9 (2013).
|Tseng, H. et al. Assembly of a three-dimensional multitype bronchiole coculture model using magnetic levitation. Tissue Eng. Part C. Methods 19, 665–75 (2013).
|Daquinag, A. C., Souza, G. R. & Kolonin, M. G. Adipose tissue engineering in three-dimensional levitation tissue culture system based on magnetic nanoparticles. Tissue Eng. Part C. Methods 19, 336–44 (2013).
|Becker, J. L. & Souza, G. R. Using space-based investigations to inform cancer research on Earth. Nat. Rev. Cancer 13, 315–27 (2013).
|Souza, G. R. et al. Three-dimensional tissue culture based on magnetic cell levitation. Nat. Nanotechnol. 5, 291–6 (2010).