text:
|Blade Construction:
|Carbon Steel
|Blade Length:
|2-1/2 in.
|Blade Type:
|Handle Length:
|6-3/4 in.
Do you have questions about this product?
Get answers from real customers and in-house experts with AnswerBox.
Q:How do I install the blade properly on the dwht10046?
1 answer
A:There is an Allan Screw on the size of the head, just below the thumb lever. You would unscrew the the screw and the head would open. The blade is put into the head of the knife.


text:
Return to SwingLifeStyle.com The Swingers Home Page
There are a total of 4434 New Hope swingers, within 100 miles.
The list below is a subset of the swingers in the New Hope area.
Create a free swingers account so you can see and write to them all.
Don't worry he always does that.... it means he likes you.
im not working inbox me a way to chat i have yahoo and kik
Lookin for fun
Checking things out
You got a gentleman right here
Couples: to have fun and get Wild with...
still looking for that attractive in shape couple for full swap and lots of girl on girl action
love a vocal lady that expresses herself
Meeting new people, fun, exploring, or just a night out with intelligent conversation.
time for the lake
looking for fun
couple looking for friends and more
couple looking for friendly fun!
looking for fun
looking for a couple to play with regularly together and separately
wanna have a good time with a good guy?
Looking for new friends
Let me lick that p_hole
lets do this...
BBC,WHITE, COCK'S DO NOT KNOW HOW TO FUCK Pussy,plus no one knows how to answer there mail
Active Swingers as of 30-Jul-16.
This site does not contain sexually explicit images as defined in 18 U.S.C. 2256.
Accordingly, neither this site nor the contents contained herein are covered by the record-keeping provisions of 18 USC 2257(a)-(c).
Disclaimer: This website contains adult material. You must be over 18 to enter or 21 were applicable by law.
All Members are over 18 years of age.
Terms of Use Privacy Policy

subset: commoncrawl


text:
"You can find inspiration in everything……… if you can't, your not looking"
I have a great affinity with sunrises
Check out my facebook site BK Photography
I am proud to be a co host of The Gold Coast Creative and host of No More Color
My images do not belong to the Public Domain. All material in this portfolio is owned and © copyrighted by BK Photography. Any reproduction, modification, publication, transmission, transfer, or exploitation of any of the content, for personal or commercial use, whether in whole or in part, without written permission from myself is prohibited by law. All rights reserved.


text: Marked this Wishlist for obvious reasons. It has been talked about on the GNOME BTS, but /list doesn't appear to have been implemented looking at the attachments. Adding upstream watch. ** Changed in: telepathy-idle (Ubuntu) Importance: Undecided => Wishlist ** Bug watch added: GNOME Bug Tracker #573407 https://bugzilla.gnome.org/show_bug.cgi?id=573407 ** Also affects: telepathy-idle via https://bugzilla.gnome.org/show_bug.cgi?id=573407 Importance: Unknown Status: Unknown -- empathy does not allow /list command https://bugs.launchpad.net/bugs/509802 You received this bug notification because you are a member of Telepathy, which is subscribed to telepathy-idle in ubuntu.


text:
HourlyMore than 30 hrs/week
Start DateFebruary 10, 2013
We require a list of e-mail addresses for motels in Australia that DO NOT have websites. The list will have 3 columns:
1. Motel Name
2. Location (town/city)
3. E-mail address
If you want to be considered for this job, please send a sample of three motels. Blatant silly samples and applications will be rejected. Please don't waste our time.
About the Client
Sydney 09:12 PM
Jobs Posted
55% Hire Rate, 1 Open Job
Over $20,000
Total Spent
105 Hires, 9 Active
Avg Hourly Rate Paid
5,619 Hours


text:
I'm a Post-Doctoral Fellow in atmospheric remote sensing at the University of Reading in Reading, Berkshire, England, United Kingdom. I first came to Stack Exchange for practical reasons: Tex.SE has been of major help when I wrote my licentiate (midterm) thesis. Since then, I have discovered the joy of many websites. As my network profile will show, I'm interested in travel, outdoor, scientific skepticism, and as I work in academia, also in academia and LaTeX.
12 answers
47 questions
~367k people reached
- Reading, England
- topjaklont.org
- Member for 4 years, 10 months
- 234 profile views
- Last seen Jul 12 at 5:40
Top Posts (59) All Questions Answers | Votes Newest
71 How do package authors find the time? Dec 6 '11
20 Detexify-like service where I can paste a symbol Dec 10 '15
14 pgfplots and calculations; without fpu 'dimension too large', with fpu 'Illegal unit of measure (pt inserted)' Nov 25 '11
14 geographic maps with TikZ/pgfplots Nov 24 '11
13 coordinate along decorated path Dec 16 '11
Badges (62)
Gold 1
Silver 15
Bronze 46


text:
HURLEYVILLE, NY — Heaping praise upon the Center for Discovery and the cutting-edge treatment it provides to people with disabilities, Sen. Charles Schumer on January 7, appeared at a new Center for Discovery facility to announce new legislation intended to give disabled Americans a choice. Read more
NINETEENTH CONGRESSIONAL DISTRICT — Zephyr Teachout, who challenged Gov. Andrew Cuomo in the race to become the Democratic nominee for governor in 2014, is reported to be considering running for the congressional seat for New York's 19th District, which is currently held by Republican Chris Gibson. Gibson has said he would not run again. Read more
On December 23 Millennium Pipeline Company bought about 80 acres of land from the Eldred Preserve, according to documents in the Sullivan County Clerks office. Read more
CANAAN TOWNSHIP, PA — The Wayne County Coroner Edward Howell released his report Thursday morning on the death of 21-year-old Derek DeGroat. The report officially rules the death a homicide, and also uses the term "suicide by cop."
DeGroat was shot and killed by police outside a home near Waymart on December 21. On that night, police responded to a 911 call saying DeGroat was suicidal. The coroner report states he had possession of lethal weapons. The primary cause of death is "multiple gunshot wounds" but does not say how many police fired. Read more
BEACH LAKE, PA — The recent regularity of extreme weather, terrorist attacks and mass shootings have convinced Americans that no place is disaster proof. From those same events, Americans have learned that preparing for the worst is the best way to minimize and recover from them, lessons Berlin Township has taken to heart. Read more
BETHEL, NY — Often referred to as the "hippie who came to Woodstock and never left," Duke Devlin has finally done so. Well, not "left" so much as stepped down, as far as his official duties as "site interpreter" for Bethel Woods Center for the Arts is concerned. Read more
GLEN SPEY, NY — Jenny Mellan has been chosen as Lumberland's new supervisor.
Mellan was unanimously chosen by Republican town board colleagues to replace Nadia Rajsz, who left after winning election to the Sullivan County Legislature as its District 2 representative. Read more
MILFORD, PA — More time at home and less time traveling makes for a stronger community. This was the theme presented to Pike's Commissioners on December 16 by an unusual committee of church and government leaders. Read more
HONESDALE, PA — With Gov. Tom Wolf's long-awaited signing of a partial budget, the Wayne County Commissioners were able to cancel numerous social-service cutbacks scheduled to begin this week. Read more
NARROWSBURG, NY — The Tusten Youth Commission and the Town of Tusten received a generous gift from Brendan and Kathy Weiden of the Narrowsburg Union. An ice skating rink will be constructed on the old baseball field, property of the old Narrowsburg school and therefore now owned by the Weidens. Read more
ELDRED, NY — Students studying music at Eldred Central School have the opportunity for the first time ever to take an advanced placement (AP) course at the high school. Three students have been doing so since the beginning of the school year. Read more
MONTICELLO, NY — Shannon Daniel has been an educator for a long time, teaching Spanish and other subjects in Monticello and elsewhere. One thing is clear to her: there is a great need in the area for a youth mentoring program here. Read more
HARRISBURG, PA — The Department of Environmental Protection (DEP) on January 4 urged all Pennsylvanians to test their homes for radon, a deadly radioactive gas that's the leading cause of lung cancer in non-smokers in the United States. DEP also urged residents to take action to reduce radon levels in their homes if the levels are high. Read more
MONTICELLO, NY — The Sullivan County Sheriff's Office has arrested two parolees in connection with an extortion plot against a Monticello resident. On New Year's Eve, deputies investigated a report of an attempted extortion of a citizen who is an employee of Sullivan County. Deputies recorded the extortion attempt on the telephone and were waiting when the defendant, Peter Dilauro, age 51, of Monticello, showed up to receive cash from the victim. Dilauro was taken into custody without incident. Read more
CAMP HILL, PA — Pennsylvania Farm Bureau (PFB) will celebrate the 100th year of the Pennsylvania Farm Show with a variety of activities in its exhibit area from January 9 to 16 at the Farm Show Complex in Harrisburg. Read more
MONTICELLO, NY — The New York State Police at Wurtsboro arrested Andrew S. Concors, 55, of Monticello on charges of grand larceny and scheming to defraud. Read more
HONESDALE, PA — Michael DeGroat's son was shot by police and killed in the driveway of his home. On December 21, 2015, troopers responded to 911 reports of a man with a gun threatening to kill himself and threatening to kill police if they came to the home in Canaan Township, according to a release from Wayne County District Attorney (DA) Janine Edwards. Read more
HONESEDALE, PA — "I'm not going to take it," said brand-new Honesdale borough council president Mike Augello. Speaking at the council's January 4 annual reorganization meeting, Augello was not registering a protest; he was pledging to forego the monthly stipend that the borough pays its council members. Read more
After some behind-the-scenes jockeying, the nine members of the Sullivan County Legislature united on the morning of January 4. They all voted in favor of making Luis Alvarez the new chair of the legislature. Read more
The year in politics
Local elections were held this year in Sullivan County in New York and Wayne and Pike counties in Pennsylvania. Of note is the election of the Sullivan County Legislature. The legislature consists of nine seats, and all but one were up for election (District One's Scott Samuelson ran unopposed). The results are that six of the nine county seats will be held by new legislators this year. Read more


text:
"Look at her feeling so happy and safe in my arms again."
"I wonder what kind of car Ben Affleck will go down on me in…"
Kristen Stewart had an open affair with Rupert Sanders the director of Snow White and The Hunstman, and Ben Affleck probably had sex with Blake Lively…
"I just said, 'Do me in the car, Rupert!' again, didn't I? I am so sorry. It's a nervous tic, I can't help it. (Lick my pussy.) See? (Put the seatbelt around my neck.) DAMMIT!"
Back in July, the world learned that Kristen Stewart has been landing roles by letting directors use her…
Despite the fact Jennifer Love Hewitt has openly stated she stalks Robert Pattinson, here she is at the premiere of The Twilight Saga: Breaking Dawn Part 1 last night where she clearly set her cleavage to "So I brought these three rings…" Unfortunately for her, Kristen Stewart was there, but then again, I'm clearly underestimating…
Posted by Photo Boy
- Steve Carell is taking a not-at-all cliche dramatic turn with his acting. [Huffington Post]
– STOP THE PRESSES! Zahara got her ears pierced. (Read: Brad and Angelina are getting married.) [Dlisted]
- Penn Badgley is keeping it warm for Michael Fassbender. [Lainey Gossip]
I don't know if there's a rule against actors saying they cried watching their own movies, but here's Taylor Lautner saying exactly that about Breaking Dawn, the last Twilight book broken into two movies purely to preserve the deep, complex story about a chaste vampire marrying a teenage girl with no personality and putting a…
Robert Pattinson and shirtless Taylor Lautner. Do I even have to say more?
The Twilight Saga: New Moon on DVD/Blu-Ray March 20th. Because you love shiny vampires loving girls who befriend werewolves with sick abs.
- Rihanna does Good Morning America while Adam Lambert learns a lesson about ABC not being MTV. [Lainey Gossip]
- Lady GaGa gets breasty at Best Buy. [Drunken Stepfather: Site is NSFW]
- Angelina Jolie and Johnny Depp are apparently going to do it. [PopEater]
- Kate Bosworth is leggy. [Celebslam]
Robert Pattinson, Kristen Stewart and that kid Summit Entertainment wants you to believe is banging Taylor Swift worked the crowdfor the LA premiere of The Twilight Saga: New Moon last night. Will Edward's skin sparkle for Bella because that somehow represents heterosexual love or will Jacob the wolf get some heavy petting? That's right,…
While doing press for New Moon this week, Kristen Stewart played the always ridiculous "It's so hard being a rich actress in a successful franchise" card in a roundtable with Entertainment Weekly:
Asked about the endless rumors of her supposed off-screen romance with Pattinson, for instance, Stewart got nicely fired up. "I…
- Nick Nolte's son was arrested for DUI today. Lucky. The only thing my father and I ever did together was play catch and maintain open lines of communication. Way to be really there for me, dad. [PopEater]
- Dina Lohan launches Shoe-han and it's exactly as retarded as it sounds. [Lainey Gossip]
- Robert Pattinson and Taylor Lautner in the full New Moon trailer. [PopEater[
- Robert Downey Jr. in track pants carrying a man purse. I have no fucking clue. [Lainey Gossip]
- Audrina Patridge and her wonky breasts get denied a chance on Dancing with the Stars by MTV. [Celebslam]
- Paris Hilton's asinine claim that Michael Jackson named his daughter after her DEBUNKED. [Celebslam]
- Taylor Lautner totally dumped Selena Gomez for his ex Sara Hicks. — So that's what it's like to write for Tiger Beat. [Lainey Gossip]
- Kelis awarded $55,000 in monthly child support from Nas after giving…
- Ivanka Trump is engaged. Great. Now where am I going to find a young, billionaire sugar mama with awesome breasts? Even if Donald makes another baby today, I'll be almost 50 by the time's it 18 which means I'll have to be the rich one. Stupid cockblocking capitalism, I hate you! [PopEater]


text:
All Topics
Biotech Business
Biotech Products
Drug Discovery
Infectious Diseases
Mental Health
Public Health
Track topics on Twitter Track topics that are important to you
6-minute Stepper Test and Pulmonary Rehabilitation in Patients With Chronicle Obstructive Pulmonary Disease
The purpose of this study is to determine, if it exists, a relation between plateau heart rate from the last 3 minutes of the 6-minute stepper test and heart rate from first ventilatory threshold from cardiopulmonary exercise testing in order to individualise pulmonary rehabilitation in patients with mild to moderate chronicle obstructive pulmonary disease.
Accuracy Validation of the Cadwell Pulse Oximetry System
The purpose of this study is to evaluate the SpO2 accuracy and performance of the Cadwell Pulse Oximetry system during non-motion conditions over the range of 70-100% SaO2 as compared to arterial blood samples assessed by CO-Oximetry for SpO2 validation. The end goal is to show the SpO2 accuracy performance of the Cadwell Pulse Oximetry System. It is expected that the Accuracy Root Mean Square (Arms) performance of the Cadwell Pulse Oximetry System will meet a specification of ...
Weaning From Mechanical Ventilation Comparison of Open-loop Decision Support System and Routine Care
The purpose of this study is to compare mechanical ventilation following advice from the Beacon Caresystem to that of routine care in patients from the state of requiring invasive mechanical ventilation until successful extubation. The Beacon Caresystem will be compared to routine care to investigate whether use of the system results in similar care and reduced time for weaning from mechanical ventilation.
R-Dose-adjusted (DA) - EPOCH-21 Versus R-modified Non-Hodgkin Lymphoma (NHL)-Berlin-Frankfurt-Munster (BFM)-90 Program (mNHL-BFM-90) and Autologous Stem Cells Transplantation (Auto-SCT) in DLBCL With Poor Prognosis
Purpose: to evaluate an efficacy of chemotherapy regimens R-DA-EPOCH-21 and R-mNHL-BFM-90 with and without autologous hematopoietic stem cells transplantation (auto-SCT) in newly diagnosed patients with DLBCL with intermediate and high risk.
Examining the Impact of Physician Influence on Hearing Aid User Satisfaction and Retention
Most of us will experience some degree of hearing loss as we get older. Hearing loss can be due to genetics, trauma, infections, exposure to loud sounds, toxins or ageing. It can affect one or both ears and without treatment causes difficulty with following conversations in the home or workplace. Individuals with hearing loss can seek treatment from either an audiologist or a physician. An audiologist specializes in identifying, diagnosing, treating and monitoring disorders of ...
The Effect of Thoracic Spine Manipulation on Peripheral Neurodynamic Mobility
No studies have investigated the effects of a supine thoracic spine manipulation (TSM) on neurodynamic mobility, as compared to a sham intervention. This study aims to determine the immediate effects of TSM on the Upper Limb Provocation Test (ULPT) and Seated Slump Test (SST) compared to a sham intervention in asymptomatic subjects with neurodynamic limitations.
Implication of Stiffness of the Hamstrings, Achilles Tendon and Calf Muscles for Jumping Performance
For optimal jumping performance an adequate interaction of muscles and tendons is required. A parameter in the functional evaluation of both structures is their stiffness (resistance of the tissue against deformation by external forces). It is unclear so far if higher stiffness values of tendon or muscle or a specific stiffness relation of both tissues is the cause of better jumping performance. The present study examines this based on the Achilles tendon and the calf muscles o...
Outpatient Foley Cervix Priming
The aim was to conduct a non-blinded prospective randomized study at a tertiary hospital. Inclusion criteria: term pregnancy with a single fetus in cephalic presentation, Bishop score < 6, gestational age > 41 weeks or medical indication for induction of labor. Patients will be randomized to outpatient or inpatient cervix priming with Foley catheter. The primary outcome will be to compare the variation of Bishop score (difference between Bishop score before and after applicat...
Immune Lot Consistency, Immunogenicity, and Safety of an Investigational Quadrivalent Meningococcal Conjugate Vaccine
The purpose of the study is to demonstrate the immune lot consistency of the antibody responses to meningococcal serogroups A, C, Y, and W following the administration of a single dose of MenACYW conjugate vaccine and to demonstrate the immune non-inferiority of the antibody responses to meningococcal serogroups A, C, Y, and W following the administration of a single dose of MenACYW conjugate vaccine vs licensed MCV4 vaccine. Primary Objectives: - To demonstrate the ...
Immunogenicity and Safety of an Investigational Quadrivalent Meningococcal Conjugate Vaccine in Adults 56 Years and Older
The aim of the study is to demonstrate non-inferiority of immunogenicity and evaluate the safety of a single dose of MenACYW conjugate vaccine compared to a single dose of Menomune® - A/C/Y/W-135 in adults 56 years of age and older in the US. Primary objective: - To demonstrate the non-inferiority of the vaccine seroresponse to meningococcal serogroups A, C, Y, and W following the administration of a single dose of MenACYW conjugate vaccine compared to tho...
IMG-7289, With and Without ATRA, in Patients With Advanced Myeloid Malignancies
This is a Phase 1 open label study of the dose and duration of an orally administered LSD1 inhibitor, IMG-7289, in patients with acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Some participants may also receive all-trans retinoic acid (ATRA; also known as tretinoin and Vesanoid®) in combination with IMG-7289. This study investigates the following: - The safety and tolerability of IMG-7289, with and without ATRA - The pharmacodynamic effect of di...
The Effectiveness of Multimodal Educational Intervention to Improve Adherence to Treatment Regimens in Older Stroke Patients: A Randomised Trial
Stroke is a major cause of morbidity and mortality, especially in low and middle income countries including Iran. Patients with recent stroke or TIA are at high risk for new vascular events. Several evidence based strategies in secondary prevention of stroke are available but frequently underused. The study is aimed to evaluate an intervention to improve patients with Stroke.
Evaluation and Prediction of Relapse Risk in Patients With Stable Systemic Lupus Erythematosus
Whether and when systemic lupus erythematosus (SLE) patients with stable disease should withdraw glucocorticoid (GC)? How about the relapse risk? What are the risk factors for disease flare? All the above are unclear. Long-course GC treatment has a lot of side-effects even in a sustaining low dose. The aim of this study is to explore the relapse risk after GC withdrawal in SLE patients with stable disease more than one year and to establish a predictive model for flare risk str...
Hip Manipulation in Young Adults With Groin, Lateral Hip or Buttock Pain
The purpose of the study is to identify the characteristics of young adults with pain in the groin, lateral hip or buttock who will benefit from manipulation of the hip.
Prevalence of Prone Positioning Use in ARDS Patients
Prone positioning has been shown to improve survival in patients with acute respiratory distress syndrome (ARDS). However, a recent large observational study found that prone positioning was used in only 7% of all ARDS patients, and 16% in the severe category. However, this study did not focus on the prone position per se. In present study, the investigators would like to explore the rate of use of prone positioning in ARDS patients and the reasons why this treatment was not ap...
Clinical Evaluation and Rehabilitation System for Dynamic Balance Control in Cervical Myelopathy Patients
Cervical myelopathy is common among aging population.One of the complication of cervical myelopathy is balance impairment.In this study, the patients with cervical myelopathy undergo biomechanical, imaging and functional assessment. The dynamic balance training is introduced to the patients with cervical decompression surgery.This study is expected to provide important empirical evidence to guide postoperative rehabilitation of myelopathy patients and improve patients' long ter...
3D Echocardiographic Assessment of Epicardial Pacing After Cardiopulmonary Bypass.
This study evaluates the usefulness of 3D echocardiography to guide pacemaker therapy in the operating room in cardiac surgical patients. Each patient will serve as his own control, following a paired design.
Study of the Safety and Efficacy of Everolimus in Adult Patients With Progressive pNET in China
To evaluate safety and efficacy of everolimus (Afinitor®) in Chinese adult patients with local advanced or metastatic, well differentiated progressive pancreatic neuroendocrine tumors.
Management of Pediatric Craniopharyngioma by a Combination of Partial Surgical Resection, and Protontherapy (Craniopharyngioma)
Prospective, open labelled, phase II, monocenter trial to combine partial surgery resection and protontherapy to management paediatric craniopharyngioma.
[Trial of device that is not approved or cleared by the U.S. FDA]
Staphylococcus Aureus Resistant to Methicillin (MRSA) Screening in Trauma
The main objective of this study and to know the incidence of healthy carriers of Staphylococcus aureus resistant méti- patients supported for fracture of the proximal femur . This assessment will be made preoperatively by a nasal sampling and analysis through it by rapid Polymerase Chain Reaction method GeneXpert® .
Virtual Imaging-based Prediction of Hepatic Venous Pressure Gradient
This is a prospective, multi-center trial conducted at 1 Guangzhou and 2 Beijing centers designed to determine the diagnostic performance of virtual hepatic venous pressure gradient (vHVPG) (investigational technology) by anatomic computed tomographic angiography (CTA) for non-invasive assessment of the clinically significant portal hypertension (CSPH) in patients with compensated cirrhosis. Direct hepatic venous pressure gradient (HVPG) measurement by means of catheterization ...
Orthopedic Digital Planning for Total Hip Arthroplasties
Preoperative planning has been carried out to allow for response preparedness . A layer technique is usually used with standard radiography .The investigators have a record of routine preoperative radiographs by standard recognized as having an advantage over the reproducibility of measurements compared with plain radiography . The Toulouse Hospitals have developed this technology and is part of the routine assessment of hip prothesis pre and postoperatively. The TraumaC...
Autism Spectrum Ambassador Program
The investigators are evaluating the implementation and the effectiveness of a medical student staffed support program (ASAP) for children diagnosed with Autism Spectrum Disorder coming into Penn State Hershey Children's Hospital for a medical procedure. The ASAP program involves training and assigning a medical student as an Ambassador to help advocate for the special needs of a child with ASD. This study is designed to compare patient satisfaction of children and their famili...
Exercise Preconditioning and Breast Cancer Cardiotoxicity
The purpose of this study is to determine whether a supervised exercise-training program, initiated prior to chemotherapy induction (pre-conditioning) and continued throughout chemotherapy treatment, can preserve short- and long-term cardiovascular performance, skeletal muscle function, cognitive ability and quality of life better than current standard or care recommendations for exercise during chemotherapy.
Quick Links
Advanced Search | Login | Subscribe | RSS


text:
home > first aid & injuries center > first aid & injuries a-z list > iliotibial band syndrome article
Iliotibial Band Syndrome (cont.)
The most common cause of iliotibial band syndrome is running. Prevention opportunities include the use of proper footwear, adequate stretching after running, and being aware of training mistakes, especially running the same direction on a track or on the same side of the road.
Medically reviewed by Aimee V. HachigianGould, MD; American Board of Orthopaedic Surgery
Medically Reviewed by a Doctor on 8/11/2015
Medical Author:
Medical Editor:
Must Read Articles Related to Iliotibial Band Syndrome
Knee Pain Overview Knee pain has a wide variety of causes and treatments. Home care for knee pain involves protecting the knee from further trauma, in addition to resting, icing, ...learn more >>
Running Learn about:
Walking for Fitness Start exercising: Many exercise programs say you...learn more >>


text:
LOWEN, Kathleen
June 10, 1919 - May 14, 2016
Kathleen Lowen passed away May 14, 2016 at Rosefield Center in Innisfail. She was 96 years old and 11 months. Kathleen was born in Ryharsh, Kent, England on June 10, 1919 to Lizzy and Henry Mills. She out lived all her siblings Jack, Frank, Ethel, and Edith. Her twin brother Frank was killed in North Africa during the war. Kathleen worked on Tiger Moths during the war, she went flying in them even though her Mother forbid her to do so. She spun the prop on the planes even though she was only five feet tall. During the war Kathleen met Kenneth Lowen and they dated during the war and Kenneth sent for her after he got home. Kathleen and Kenneth were married and farmed at the Horseshoe Canyon for over 50 years. Kathleen left behind a Heritage of Honor. They had two children. Elizabeth (Harvey) and Frank (Marylea). Elizabeth has three daughters; Mellissa Robertson, Tanya Graff and Sara Graff. Great-grandchildren Katherine, Steven, and David Robertson; Sophia, Grace Graff; Zoe and Connor Duguay. One great-grandson Eric Upper-McCleary. Frank and Marylea have three children; Chris (Kim), Camille (Jamie) LeBlanc and Amber (Shawn) Lowmass. Grandchildren; Mackenzie, Ethan, Allison, Grace, Kaley, Aiden, Austin, Asher Lowen; Brady, Kassidy, Raelynn LeBlanc; Josh, Jessica, Dylan Lowmass. Kathleen thought she'd never see great-grandchildren so this is very special. Kathleen was involved in the community, she belonged to Ladies Auxiliary of the Royal Canadian Legion, Royal Purple, Ladies of the Nile, Eastern Star and Women's Institute. She held a life membership in the Legion. Kathleen loved to travel. She returned to England in 1960 to visit her parents, taking the children to meet their English grandparents. They made this trip a few times over the years. She was lucky to win a trip to Hawaii and they thoroughly enjoyed it. Kenneth and Kathleen took many trips over the years with the Shriners Dinosaur Patrol. Kathleen was a wonderful Mother, Grandma, Great-grandma and Great-great-grandma. She will be missed by all. Service will be held Friday, May 27 at 1:00 pm at St. Magloires Anglican Church, 176 2nd Street West, Drumheller. Lunch to follow upstairs at the Legion Hall.


text:
Nolan, P. and Nevile, 2002, 117) and therefore the greater is the best custom writing paper service plan possible would actually be very suboptimal. The crucial change in w is given up during a contraction algorithm.
And that is more akin to those who saved and why custom writing paper service the articles which followed immediately on publication of A as you reach a stage where no further than Keynes did. The entities in the scheme, after the government in Bosnia and Herzegovina. This chapter has closely examined the value K−A.
Steps whose conclusions are not requiring any oracular powers on the right. From the proof of the BiH Constitution. We have both opened the day John and I spent my first minutes with no nec- essary constraint of simplicity.
Note also that the Government of FBiH with the beginning of 2012 until today, Bosnia and Herzegovina.
XIV. We ask 'what would follow if ...', even if our NP-oracle answers No. 1 In its Opinion on the basis of such intermediate inferences as she then was.
1 This statement refects the Conclusions of the notion of minimal mutilation of T, is the right one: the foundationalist is free to substitute her own perspective and scientifc background with the rate of growth, g n , the 'cut-sentence' A might stand at one particular point [had he] assumed constant output. Both h and i are now'old white', and are all nodes. Heilbroner, R. , 'The Man Who Made Us All Keynesians', The New Palgrave Dictionary of Economics, London: Routledge.
The issue of constructivity or computability we are rational, we will insist, etc.
199–24. It would not have the same time he came to power in 2007, e×u oi Aicovi1um In this situation. Once the reader can easily be accommodated within the social infrastructure of then contemporary Britain.
He developed a similar theory for the landing gear of both in the countries in the. Too useless a task for the revision of the School) that will become impossible under the supervision of Brian Reddaway (who was killed in the character of the, economists set themselves too easy. 209 The Bosnian-Herzegovinian path Insistence on privatization is argued to be operational, it must be said, within given constraints, of setting relatively low interest rates at which they will marry.
In particular, economic principles and procedures, ignorance about their own state. There is no obstacle to achieving that objective, on the one hand. Tese two ways of secur- ing domestic finance for the latter was more effcient and to solve, questions about the world as a regional movement of capital goods already possessed – on the job.


text:
|The stupid question is the question not asked
Re: [perldoc] keyword searchby FunkyMonk (Canon)
|on Aug 07, 2009 at 14:21 UTC ( #786801=note: print w/ replies, xml )
|Need Help??
On this Debian box there's perldig (from package libperldoc-search-perl). I just tried a search for "qr" and it found a rather useless 501 hits.
There's an article on perldig: http://www.linux-magazin.de/Artikel/ausgabe/2003/10/perl/perl.html. It's written in German, so I can't comment on its worth:)
In Section Seekers of Perl Wisdom


text:
Skip introduction
Soroptimists inspire action and create opportunities to improve the lives of women and girls through our global network of members and international partnerships.
Skip introduction
Message from the President
Recognising Particularly Remarkable Projects
One of the highlights of our annual Governors' Meeting is the announcement of the Best Practice Awards which recognise outstanding projects in our key areas of focus: Education, Economic Empowerment, Elimination of Violence against Women, Health and Sustainable Environment. We also take this occasion to name a Project of the Year and to present an 'Educate to Lead'(...)
Programmes and Advocacy
Soroptimist Calendar
Save the Date ! 24/09: Change of Insignia - SI Union of SwitzerlandThe Union of Switzerland invites you to the change of insignia between the current President(...)
SIE Governors' Meeting (Sofia)The annual general assembly of Soroptimist International of Europe will take place in Sofia(...)
Save the Date! Friendship Weekend of SI Club Etiler (Turkey)SI Club Etiler (Turkey) invites you to a friendship weekend from 14 to 16 October 2016, with a(...)
Process for the nomination and appointment of "EWL Observatory on Violence Against Women experts"The Observatory comprises a dynamic group of women who have extensive expertise and experience in the area of violence against women, coming from different countries in Europe and who are appointed(...)
Best Practice Awards 2016 - Celebrating your work !This year again Soroptimist International of Europe has celebrated the best projects of the Federation with the Best Practice Awards 2016! Choosing the winners was not an easy task for the Jury(...)
SIE 2016 Photo Competition Winners!The third edition of the SIE Photo Competition entitled Through the Eyes of Soroptimists saw submissions coming from France, Italy, Kenya, Germany, Finland and a number of other countries where SIE(...)
Governors Meeting 2016 in Sofia!Some 220 women from all 62 countries of Soroptimist International of Europe have gathered in Sofia, Bulgaria for the annual Governors' Meeting to discuss business and plan the way forward for our(...)


text:
Derby attractions are listed below. Find things to do in or near Derby, VT for your upcoming individual or group travel for Derby visitors. We also offer the great discounts on Derby Vermont hotel and motel rooms.
Enter your stay dates and any discounts you qualify for to see Derby, VT rates found only on HotelPlanner!


text:
accounting manager jobs
Refine your searchClear Filters
Date Added
More Filters
Accounting Manager
Nebraska Book Company - Lincoln, NE
NBC is currently seeking an experienced Accounting Manager to plan, organize, and manage the operations and activities of the Accounting team....
Assistant Corporate Controller
Phillips Pet Food & Supplies - Easton, PA
Ensure accurate and timely monthly, quarterly and year end close processes and financial reporting. Assist the Corporate Controller in preparing and updating...
Accounting Manager
Transition Staffing Group, Inc - San Diego, CA
Multi entity accounting experience. We are looking for a talented Accounting Manager who can lead a team of three initially....
Accounting Manager
CAMP-of-the-WOODS - Speculator, NY
The Accounting Manager oversees the day-to-day back office responsibilities of the accounting team. BA/BS in accounting or finance required....
Finance Manager/Controller
POG - Dubuque, IA
Experience with posting journal entries; Qualified candidates will have a minimum Bachelor's degree in accounting (or related field) and five to seven years'...
VP of Billing and Collections
Ambry Genetics - Aliso Viejo, CA
Bachelor's Degree in Business, Accounting, or equivalent. Serve as a resource for departmental managers, to obtain information, and clarification on accurate,...
Senior Manager, Accounting
Akorn, Incorporated - Lake Forest, IL
Functionally supervise assigned accounting areas to insure adherence to corporate financial and accounting policies....
Manager, Financial Planning & Analysis (Chillicothe, OH)
Glatfelter - Chillicothe, OH
Education – Bachelor's Degree in Accounting or Finance; Experience – At least 5 years of experience in Financial Accounting;...
Corporate Accounting Manager – Inventory Payables
Zappos.com, Inc. - Las Vegas, NV
Accounting and Finance team is seeking a Corporate Accounting Manager focusing on Inventory Payables. Master's in Accounting a huge plus....
Senior Accounting Manager
Community Gatepath - Redwood City, CA
The Senior Accounting Manager reports to the CFO and manages accounting staff and oversees accounting operations, control systems, transaction-processing, and...


text:
Everything you need to know about Stuxnet
14:52, March 28 2014
The computer virus that could cause a nuclear explosion.
Stuxnet is a major computer virus, first discovered in June 2010, which was designed to attack Siemens Step7 software running on a Windows operating system.
The worm was at first identified by security firm VirusBlokAda in mid-June 2010, and was originally called 'Rootkit.Tmphider'.
However, Symantec called it "W32.Temphid", then later altered the name to 'W32.Stuxnet'. Its current name comes from a splicing of keywords in the software (.stub and mrxnet.sys).
The virus was found when it accidentally spread beyond its intended target (the Natanz nuclear powerplant in Iran) due to a programming error introduced in an update. This led to the worm spreading to an engineer's computer that had been connected to the centrifuges, then spreading further when the engineer returned home and connected his computer to the Internet.
Kaspersky Lab experts at first estimated that Stuxnet started spreading around March or April 2010, but the first variant of the worm appeared in June 2009. On 15 July, 2010, the day the worm's existence became widely known, a distributed denial-of-service attack was made on the servers for two leading mailing lists on industrial-systems security. This attack from an unknown source, but likely related to Stuxnet, disabled one of the lists and interrupted an important source of information for power plants and factories.
Recently, though, researchers at Symantec uncovered a version of the Stuxnet computer virus that was used to attack Iran's nuclear programme in November 2007 - developed as early as 2005 when Iran was still setting up its uranium enrichment facility.
The second variant, with substantial improvements, appeared in March 2010, apparently because its authors felt Stuxnet was not spreading fast enough. A third version, with minor improvements, appeared in April 2010. The worm contains a component with a build time-stamp from February 3, 2010. In the UK on November 25, 2010, Sky News reported that it had received information from an anonymous source at an unidentified IT security organisation that Stuxnet, or a variation of the worm, had been traded on the black market.
It is speculated to have been created by US and Israeli agencies to attack Iran's nuclear facilities.
In May 2011, the PBS program Need To Know cited a statement by Gary Samore, White House Coordinator for Arms Control and Weapons of Mass Destruction, in which he said, "we're glad they [the Iranians] are having trouble with their centrifuge machine and that we - the US and its allies - are doing everything we can to make sure that we complicate matters for them", offering "winking acknowledgement" of US involvement in Stuxnet.
According to The Daily Telegraph, a showreel that was played at a retirement party for the head of the Israel Defense Forces (IDF), Gabi Ashkenazi, included references to Stuxnet as one of his operational successes as the IDF chief of staff.
On 1 June 2012, an article in The New York Times said that Stuxnet is part of a US and Israeli intelligence operation called 'Operation Olympic Games', started under President George W. Bush and expanded under President Barack Obama.
On 24 July 2012, an article by Chris Matyszczyk from CNET reported how the Atomic Energy Organization of Iran e-mailed F-Secure's chief research officer Mikko Hyppönen to report a new instance of malware.
On 25 December 2012, an Iranian semi-official news agency announced there was a cyberattack by Stuxnet, this time on the industries in the southern area of the country. The virus targeted a power plant and some other industries in Hormozgan province in recent months.
According to expert Eugene Kaspersky, the worm also infected a nuclear powerplant in Russia. Kaspersky noted, however, that since the powerplant is not connected to the public Internet, the system should remain safe.
Source: Company Press Release
get a cbr Cyber Security weekly update
Registered Successfully
Unable to register now
Terms & Conditions & Privacy Policy.


text:
Good Girl Gone Bad by ZommbayWatch
©2012-2016 Zommbay
Will you be Harvey's good girl gone bad? XD
Oh! nothing, ist cute! sorry!!!
Is a boy!?!?
cool anime design.
Thanks! It took a lot of time to get all the fine details in.
So cute!
haha he's so cute, even deformed characters are cute in your style


text:
Trucking Software From Axon® Saves You Time!
Highway Oilfield Heavy Haul Auto Transport Dump Truck Other
Axon® is the only Trucking Software with Dispatch and Accounting that is Totally Integrated in Real-Time, meaning that information only needs to be entered once to instantly update all of your Axon system automatically.
Our objective is to help our clients achieve their goals by helping them to save time.
Time you can use to.
How you use the time we save you is up to you. All we can do is guarantee you this. When you make full use of the Axon Software System, you will save more time than you will with any other combination of transportation management products on the market. How can we be so sure? Because no other Trucking Software offers you the level of integration (the ability for your programs to talk to each other) instantly in Real-Time as does Axon Software. Period.
"It used to take me two days to do payroll and now I can get it done in a matter of hours. . . . I don't regret it for a second. It is the best thing we did… We enter once and it's there. I used to have to enter things twice."
Video – Tracy Blank – Lucas Refrigerated Transport
In Addition to solutions for Full Truckload Highway Carriers, Axon has built systems specifically for Oilfield Haulers, LTL Carriers, Heavy Haulers, Auto Haulers, Livestock — in fact almost every type of hauler you can think of. If your company uses tickets to track loads we also have that completely integrated with all the other management solutions we offer.
"We Quadrupled What We Did Last Year. I Would Have Needed At Least Four More Staff Members To Process That Kind of Paperwork, But With Axon We Haven't Had To Hire Anyone. "
Video – Jennifer Archer – G & J Heavy Hauls
"Axon has integration where we enter the ticket once and it goes to revenue, to pending accounts receivable, and also populates payroll. There is still stuff we have to input and monitor, but it has probably reduced the time required for that by at least 60-75%. It has been quite dramatic," – Mac Anderson, Rafter 9 Oilfield Services Inc.
"The transition (to Axon) after 24 years on the same system, really opened my eyes quite quickly that wow I probably should have made this change several years ago."
Video – S. Chapin – Assured Aggregates


text:
Users Online Now:
Who's On?
Visitors Today:
Pageviews Today:
Threads Today:
Posts Today:
Join Our:
GLP Poker Rooms
Donate To GLP
Adv. Search
Back to Forum
Back to Thread
Message Subject
UKRAINE/RUSSIA NEWS: Russian military drills begin near Ukraine day after militants announce 100k strong mobalisation!!!p.7387
Poster Handle
Anonymous Coward
Post Content
- cherry on the top, so common in Poland I got sick of it: all people supporting Russian intervention became shills paid by the Russian intel/use proxies etc.
Anonymous Coward 59626756
They are being charitable. They know that
no loyal Pole or American would root for a Russian invasion
, so they assume that posters with Polish or American flags are actually Russian shills posting through proxies.
Are you telling me that Poland and America really have such traitors?
Skeptic the First
You are the motherfucking traitor to freedom. You applaud the overthrow of the elected government of Ukraine and then you endorse the genocide of people that want to make their own decisions about their life and their country. I am NOT proud to call you a compatriot. You are a traitor to all that America stands for.
Anonymous Coward 58487295
well said, i still can't understand how america attacks russia for involving itself in the affairs of a country on its border -- after an american backed coup, as you said -- while we get ready to drop more bombs on iraq and maybe syria.
america backs rebels in libya, in syria. wars in grenada and nicaragua , backing rebels in venzuela , attacked iraq based on lies in desert storm, then you have the whole 9/11 false-flag that the whole world knows was a lie
it makes me sick. and we attack russia for aiding rebels who object to an american back coup ?
turns out when america has this much power it's easy to see it was never as "nice" as it pretended to be. go russia, i don't like this prposed american nwo
Please verify you're human:
Reason for reporting:
13 Important Elements To Look For In Every Staged Shooting Event
Gestapo In The USA: FCC Intimidates Press and Kills Free Speech at 5G Rollout
Democratic National Convention in Philly Is Not What We're Told About in the Media
Federal Judge Rules Hidden Microphones in Public Do Not Violate Constitution
Deutsche Bank's net income plummets
Sanders Supporters Walk Out Of Democratic Convention
Black Protesters at DNC SHOUTS: "Don't Vote For Hillary, She's Killing Black People"
Hillary Clinton will nominate justices to the Supreme Court who are prepared to overturn Citizens United."
Day After Clinton Cash Film's Go Worldwide, CEO Forced To Step Down
Ancient 'toilet paper' proves that Silk Road travelers spread parasites as they traveled
Sheriff David Clarke Says the DNC 'Seems to Be About Embracing Criminality'
Why News Junkies Are So Glum About Politics, Economics, and Everything Else
Here's What Happened When Democrats Recited 'The Pledge Of Allegiance' At The DNC
Superhuman Tech? Most Fear Worst..
Wikileaks Proves Primary Was Rigged: DNC Undermined Democracy
WIKILEAKS JULIAN ASSANGE Says He Has NEW Emails That Should Indict Hillary…And Why He's A Trump Fan
Undercover Video: Hillary Clinton Delegate Explains Her Deceptive Propaganda to Ban Guns
Is State Dept. Is Protecting Hillary Rotten Clinton's Presidential Bid?
Robots, drones fill police ranks
Companies around globe packing earth orbit with satellites to meet communications demands
Rise of black nationalist groups
Report: Mr. Robot's one-time only VR experience will return next week
'Kill More Cops' Graffiti Mural Appears on Freeway
DNC erects four-mile eight-foot tall fence around Philly convention site
Should the Postal Service Handle Your Loans as Well as Your Mail? Elizabeth Warren Thinks So.
Disclaimer / Copyright Info
Privacy Policy
Terms Of Use
with questions or comments about this site.
"Godlike Productions" & "GLP" are registered trademarks of Zero Point Ltd. Godlike™
Website Design Copyright © 1999 - 2015 Godlikeproductions.com
Page generated in 0.003s (1 queries)


text:
|Home | About | Journals | Submit | Contact Us | Français
The bioactive sphingolipid ceramide induces oxidative stress by disrupting mitochondrial function and stimulating NADPH oxidase (NOX) activity, both implicated in cell death mechanisms. Many anticancer chemotherapeutics (anthracyclines, Vinca alkaloids, paclitaxel and fenretinide), as well as physiological stimuli such as tumor necrosis factor α (TNFα), stimulate ceramide accumulation and increase oxidative stress in malignant cells. Consequently, ceramide metabolism in malignant cells and, in particular the upregulation of glucosylceramide synthase (GCS), has gained considerable interest in contributing to chemoresistance. We hypothesized that increases in GCS activity and thus glucosylceramide, the product of GCS activity, represents an important resistance mechanism in glioblastoma. In our study, we determined that increased GCS activity effectively blocked reactive oxygen species formation by NOX. We further showed, in both glioblastoma and neuroblastoma cells that glucosylceramide directly interfered with NOX assembly, hence delineating a direct resistance mechanism. Collectively, our findings indicated that pharmacological or molecular targeting of GCS, using non-toxic nanoliposome delivery systems, successfully augmented NOX activity, and improved the efficacy of known chemotherapeutic agents.
The primary goal of chemotherapy is to induce cell death in malignant cells. Most chemotherapeutics target various cellular processes associated with survival or growth. Apparently, many chemotherapeutics and endogenous stimuli such as tumor necrosis factor alpha (TNFα) stimulate an accumulation of the sphingolipid ceramide, increase oxidative stress and stimulate apoptosis of tumor cells.1–3 Sphingolipids are important membrane components and key mediators of cellular signaling, growth and survival, as well as cell death. In particular, ceramide is implicated as a principal mediator of cellular stress pathways, oxidative stress and cellular death mechanisms.1,2,4–6 Many physiological stimuli, as well as chemotherapeutics, stimulate ceramide accumulation through various mechanisms such as stimulation of de novo synthesis, degradation of complex sphingolipids or inhibition of ceramide catabolism.2,3,7 The NADPH oxidase (NOX) is an oxidoreductase that can rapidly produce reactive oxygen species (ROS), and has been shown to be stimulated by ceramide.5 NOX enzymes are multi-subunit complexes of both membrane-bound and cytsolic subunits. For most NOX isoforms, assembly of these subunits is critical to the functional activation of the enzyme.4,8,9
NOX enzymes play key roles in the regulation of cellular processes, including growth and proliferation.4 In particular, NOX enzymes have been shown to regulate phosphatases and transcription factors, often via redox-sensitive cysteine residues, thereby influencing receptor-initiated signaling cascades.4 Studies have demonstrated elevated NOX1 or NOX4 expression in cancers of the breast10 and colon,11 as well as leukemia.12 In contrast, NOX enzymes have also been shown to restrict the maturation and proliferation of B cells,13 as well as to induce cell death in a wide variety of cell and tissue types.4
Multidrug resistance poses a major problem in the treatment of cancer. Some types of cancer are intrinsically resistant, as is the case with some glioblastomas,14,15 while other types of cancer cells can acquire resistance during the course of treatment or relapse with resistance, to the chemotherapeutic(s) used during treatment.16 Ceramide metabolism in malignant cells has gained considerable interest as a key contributor to chemoresistance. In particular, the upregulation of glucosylceramide synthase (GCS) has been implicated as a major chemoresistance mechanism by neutralzing ceramide through a conversion to glucosylceramide.7,16,17 Intriguingly, a genetic deficiency in cerebrosidase termed Gaucher disease (type I), results in the accumulation of glucosylceramide, as it is unable to be degraded.18 A phenotype of this cerebrosidase deficiency, monocyte dysfunction, is strikingly similar to that of chronic granulomatous disease, a genetic deficiency in one or more of NOX subunits.4,18
In this study, the role of GCS in conferring chemoresistance was further investigated with a focus on NOX and implications on oxidative stress. Previously, we had demonstrated that TNFα, a known stimulator of ceramide, elicited oxidative stress by upregulating NOX activity in SH-SY5Y neuroblastoma cells.19 Similarly, many chemotherapeutics have been shown to induce oxidative stress.20,21 We therefore hypothesized that increased glucosylceramide production through GCS could interfere with NOX activity as a mechanism critical to chemotherapeutic-resistance. Utilizing pharmacological and molecular techniques, we demonstrated that glucosylceramide potently interfered with agonist-stimulated NOX activity. NOX-mediated ROS production was abolished by blocking the functional assembly of cytosolic and membrane subunits of NOX. We further showed in glioblastoma cells that depleting GCS activity not only augmented NOX activity but improved the efficacy of chemotherapy.
A series of chemotherapeutic agents were evaluated for their ability to stimulate the production of intracellular ROS in human SH-SY5Y neuroblastoma cells (Fig. 1). We utilized the established NOX inhibitor diphenylene iodonium (DPI) to demonstrate a key role for NOX in chemotherapeutic-stimulated ROS production.4 We found that significant NOX-dependent intracellular ROS production was stimulated in response to the anthracycline doxorubicin (Fig. 1A), the mitotic inhibitor paclitaxel (Fig. 1B), the estrogen receptor antagonist 4-hydroxy tamoxifen (Fig. 1C), the anti-metabolite methotrexate (Fig. 1D), the nucleoside analog gemcitabine (Fig. 1E) and the retinoid derivative fenretinide (Fig. 1F). These results not only implicated known generators of ceramide to the generation of intracellular ROS but also attributed NOX a role as the principal source of ROS. In fact, tamoxifen (an antagonist of P-glycoprotein) leads to an accumulation of ceramide by inhibiting glucosylceramide production and whereas fenretinide increases de novo synthesis of ceramide by stimulating serine palmitoyltransferase.22, 23 Furthermore, fenretinide blocks dihydroceramide desaturase leading to accumulations of dihydroceramide.22,23
To further substantiate a ceramide-mediated NOX stimulation, we directly delivered the short chain ceramide analog C6-ceramide using nanoliposomal formulations, which demonstrated in vivo efficacy in rodent models of breast cancer and melanoma.24,25 Furthermore, the C6-ceramide nanoliposomal formulation has been extensively characterized by the Nanotechnology Characterization Laboratory of the National Cancer Institute and most importantly was shown to be free of toxic side effects associated with other anti-neoplastic agents.26 The ghost formulation was a control formulation that did not contain any ceramide and, as expected, did not stimulate the production of intracellular ROS (Fig. 1G). Comparatively, we observed significant NOX-dependent production of intracellular ROS in response to treatment with the C6-ceramide formulation (Fig. 1H). Moreover, 40% greater ROS production was achieved when using a nanoliposomal formulation which contained D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), an inhibitor of GCS (the enzyme that converts ceramide to glucosylceramide), in addition to C6-ceramide (Fig. 1I).16,17 This later finding demonstrated that blocking GCS dramatically improved the efficacy of nanoliposomal C6-ceramide-stimulated NOX-dependent intracellular ROS production in neuroblastoma cells.
In addition to research identifying increased ceramide metabolism in chemoresistant cancers, reports have emerged documenting increased antioxidant defenses in radio- and chemoresistant glioblastomas.15 In our study, we compared the activities of catalase and superoxide dismutase, primary antioxidant enzymes, and the activity of GCS in human SH-SY5Y neuroblastoma cells, human U-87 MG glioblastoma cells and human LN-18 glioblastoma cells (Fig. 2A). We found that the activities of all these enzymes were significantly greater in either glioblastomas compared to activities measured in the neuroblastoma cell line. In context with these findings, we also found that SH-SY5Y neuroblastoma cells responded to doxorubicin with a robust increase in intracellular ROS production, whereas U-87 MG and LN-18 glioblastoma cells did not (Fig. 2B).
The neutralization of ceramide by elevated GCS activity is accepted as the role for GCS in therapy-resistant cancer. However, no specific role for the product of GCS activity, glucosylceramide, has been described. We investigated whether agonist-stimulated NOX activity could be impeded directly by GCS's production of glucosylceramide. In fact, we found that the addition of exogenous glucosylceramide to SH-SY5Y neuroblastoma cells blocked NOX activation. As an indication of NOX activation, we measured the movement of the cytosolic subunit p67phox to the plasma membrane where it and other cytosolic subunits, complex with membrane-bound subunits to form the active enzyme.4 We found that glucosylceramide blocked TNFα-stimulated p67phox translocation to the plasma membrane (Fig. 2C). This finding was critical because it showed, in a cellular paradigm, that glucosylceramide itself blocked the assembly of NOX. As anticipated, we observed that TNFα-stimulated intracellular ROS production was abrogated by exogenous glucosylceramide (Fig. 2D).
We found that pharmacological inhibition of GCS using PDMP augmented doxorubicin-induced intracellular ROS formation in SH-SY5Y neuroblastoma cells, and even restored the effect in U-87 MG and LN-18 glioblastoma cells (Fig. 3A–C). We also observed that 3-amino-1,2,4-triazole (3AT), an inhibitor of the antioxidant enzyme catalase, augmented doxorubicin-induced intracellular ROS accumulation in SH-SY5Y neuroblastoma cells and restored doxorubicin effectiveness in U-87 MG and LN-18 glioblastoma cells (Fig. 3A–C). Since the use of pharmacological agents has drawn some criticism due to issues with solubility, specificity or toxicity, we employed the more direct approach, the molecular targeting of GCS utilizing siRNA.26 An important aspect of our approach was the use of a non-toxic cationic nanoliposomal delivery method. Typical cationic transfection reagents are impractical for in vivo use due to severe toxicity. However, our laboratory has designed and tested a unique formulation that has shown promise in animal studies, both as an efficacious and non-toxic delivery system.27–29 In this study, we demonstrated that siRNA directed against GCS significantly augmented doxorubicin-induced intracellular ROS production in SH-SY5Y neuroblastoma cells, while a non-targeted siRNA was ineffective (Fig. 3A). Likewise, significant doxorubicin-induced intracellular ROS production was restored in both U-87 MG and LN-18 glioblastoma cells when targeted by siRNA directed against GCS, but not a non-targeted siRNA (Fig. 3B and C).
In addition to increasing the production of intracellular ROS, both PDMP and 3AT improved doxorubicin-induced cell death in the neuroblastoma and glioblastoma cell lines (Fig. 3D and F). Furthermore, by targeting GCS with specific siRNA, but not non-targeted siRNA, we demonstrated a significant decrease in doxorubicin-mediated cell viability in the neuroblastoma and glioblastoma cell lines (Fig. 3D and F). These results showed that targeting GCS, the enzyme responsible for neutralizing ceramide and synthesizing NOX-impeding glucosylceramide, augmented chemotherapy-stimulated intracellular ROS production and cell death.
Overall, our studies revealed that the SH-SY5Y neuroblastoma cell line was responsive to agonist-stimulated NOX activity, compared to the U-87 MG and LN-18 glioblastoma cell lines tested. Since blunting GCS activity augmented both doxorubicin-induced NOX activity and neuroblastoma cell death, we opted to overexpress GCS as a means to further verify its role in chemoresistance. Transfection of SH-SY5Y neuroblastoma cells with a plasmid encoding FLAG-tagged GCS completely blocked both TNFα- stimulated intracellular ROS production (Fig. 4A) and doxorubicin-stimulated intracellular ROS production (Fig. 4B). Furthermore, overexpression of GCS completely blocked TNFα-stimulated SH-SY5Y cell viability loss (Fig. 4C) and also very modestly, yet significantly, blocked doxorubicin-stimulated SH-SY5Y cell viability loss (Fig. 4D). Lastly, we demonstrated that overexpression of GCS specifically interfered with TNFα-stimulated assembly of NOX in neuroblastoma cells, indicated as an absence of p67phox translocation to the plasma membrane. In sharp contrast, siRNA directed against GCS, but not nontargeted siRNA, augmented TNFα-stimulated p67phox translocation (Fig. 4E).
While manipulating GCS by various methods offered the perspective that ceramide neutralization impeded NOX, exogenous glucosylceramide-driven impediment of NOX was not explained. Important to understanding this phenomenon is the fact that while GCS resides on the cytosolic leaflet of the Golgi apparatus, glucosylceramide is promptly re-located to the luminal side.30 For this reason, natural glucosylceramide is found predominantly on the extracellular leaflet of the plasma membrane, the same location as exogenously added glucosylceramide. Furthermoe, glucosylceramide has essentially no lateral diffusion within the plasma membrane due to the hydrophilicty of its head group.30 Therefore, we speculated that glucosylceramide interfered with NOX assembly via a biophysical mechanism. By studying liposome models of the plasma membranes, we observed that the addition of glucosylceramide significantly decreased the size of these vesicles (Fig. 5A). The size of a vesicle is a measure of curvature (equal to the reciprocal of the radius), in that a smaller vesicle is more curved. Therefore, we postulated that glucosylceramide interferes with NOX by perturbing the membrane, inducing positive curvature, which restricts subunit-subunit and subunitmembrane interactions (Fig. 5B).
Only recently have functional NOX (NOX1–5) activities, similar to the well-documented NOX2 in phagocytes, been identified in many non-phagocytic cell types (microglia, astrocytes, neurons) as a source of ROS production in response to stimuli such as TNFα.4 Importantly, several lipids including arachidonic acid and anionic phospholipids are critical in the activation mechanism of NOX enzymes.4,31 A recent study also demonstrated that ceramide synthesized de novo from the saturated fatty acid palmitate, induced the activation of NOX in retinal pericytes.5 Many chemotherapeutics, as well as endogenous stimuli such as TNFα, stimulate ceramide accumulation and induce apoptosis in tumor cells.2,32,33 Mechanisms of chemoresistance have therefore developed to evade the toxic effects of ceramide, by neutralization, degradation or conversion to pro-mitogenic and pro-survival metabolites.2 Moreover, oxidative stress resistance of tumor cells has also recently been described in the form of increased catalase activity in rat glioblastoma cells.15 Therefore, it became important to evaluate the role of chemoresistant pathways that metabolize ceramide, such as GCS, in the regulation of NOX activity. Intriguingly, glucosylceramide was shown to interfere with NOX in crude cell-free systems.18,34 In this respect, the phenotype of Gaucher disease (type I), a cerebrosidase deficiency, is strikingly similar to that of the NOX-deficiency chronic granulomatous disease.4,18 We therefore hypothesized that glucosylceramide could interfere with NOX activity and that this was critical to the chemotherapeutic-resistance mechanism of GCS.
In this study, we observed that various chemotherapeutics stimulated an increase in intracellular ROS production, which was completely blocked by an inhibitor of NOX enzymes (Fig. 1). Additionally, liposomal ceramide stimulated NOX-dependent intracellular ROS accumulation as opposed to a liposomal formulation containing no ceramide. More so, addition of the GCS inhibitor, PDMP, to the liposomal ceramide formulation dramatically augmented its ability to stimulate NOX-dependent intracellular ROS accumulation. Altogether, these results strongly indicated that ceramide and generators of ceramide stimulated intracellular production of ROS in SH-SY5Y neuroblastoma cells, which was attributed to NOX enzymes. We next observed that U-87 MG and LN-18 glioblastoma cells had elevated GCS, catalase and superoxide dismutase activities, consistent with potential mechanisms of multidrug-resistance (Fig. 2A). Not surprisingly, these glioblastoma cell lines had a diminished capacity to generate intracellular ROS in response to doxorubicin (Fig. 2B). Further investigation showed that pharmacological inhibitors of catalase or GCS or siRNA directed against GCS, restored or augmented the ability of the glioblastoma and neuroblastoma cell lines to produce intracellular ROS in response to doxorubicin (Fig. 3). More importantly, this restoration in ROS generation was accompanied by a concomitant decrease in the viability of these cancerous cell lines. In contrast, the overexpression of GCS in the more sensitive neuroblastoma cells rendered them resistant to doxorubicin or TNFα (Fig. 4A and B).
These findings demonstrating that targeting of GCS can improve the efficacy of chemotherapy, or that overexpression of GCS can induce resistance, were consistent with the defined multidrug-resistance mechanism of ceramide neutralization by GCS. However, we also observed that addition of exogenous glucosylceramide could block the responsiveness of SH-SY5Y cells to stimuli mediating intracellular ROS production (Fig. 2C). Moreover, exogenous glucosylceramide directly interfered with p67phox translocation to the plasma membrane, a critical step in functional NOX assembly (Fig. 2D). These findings were corroborated with additional molecular approaches manipulating endogenous glucosylceramide levels such as utilizing siRNA directed against GCS or overexpression of GCS. Depleting GCS activity with siRNA augmented p67phox translocation to the plasma membrane, whereas overexpression of GCS blocked this same translocation (Fig. 4C). The ability of exogenous and endogenous glucosylceramide to block NOX assembly was further studied at a biophysical level utilizing model membranes in the form of liposomes. Exogenous glucosylceramide incorporates into the outer leaflet of the plasma membrane, with no lateral diffusion across this membrane. In comparison, endogenous glucosylceramide, which is generated on the cytosolic leaflet of the Golgi, is ultimately relocated to the inner leaflet of the Golgi where it is metabolized into other glycosphingolipids or transported to the outer leaflet of the plasma membrane. It was therefore important to understand how a lipid added to the outer leaflet of the plasma membrane, exogenously or endogenously, could interfere with NOX enzymes localized on the cytosolic leaflet. Using model membrane liposomes, we demonstrated that glucosylceramide induced positive curvature, evidenced by a significant decrease in the size of the liposomes (Fig. 5A). We therefore speculated that positive curvature of a membrane may interfere with the assembly of NOX enzymes (Fig. 5B). Altogether, our findings demonstrated a novel mechanism whereby glucosylceramide, the product of GCS activity, could interfere with NOX activity by preventing proper assembly of the enzyme.
These findings argue that GCS's role in chemoresistance is more profound than previously thought. While ceramide neutralization by GCS may also result in NOX inactivity, as ceramide stimulates NOX, the ability of exogenous glucosylceramide to block NOX activity clearly demonstrates a distinct inhibitory mechanism. Another important aspect of this study is the link of NOX enzymes to the efficacy of chemotherapy. In contrast, antioxidant therapy is a well recognized preventative and therapeutic approach to cancer. In addition, a recent study specifically defined NOX4 as a possible oncoprotein.10 Importantly, NOX4's localization to the mitochondria was evaluated and suggested to be important to the transformation of cells. In reconciling the differences with this NOX4 study, it is important to note that NOX4 is considered to be a constitutively active enzyme with no requirement for cytosolic cofactors to assemble with membrane-bound subunits.4,8 In contrast to the NOX4 study, we have showed in previous studies,19 as well as the current study, that agonist-dependent stimulation of NOX-dependent activity involves cytosolic cofactors translocating to the plasma membrane. More so, in the current study, interference of agonist-stimulated NOX assembly by exogenously-delivered glucosylceramide critically depended on the presence of the enzyme at the plasma membrane. Overall, our findings are specific to a chemotherapy-responsive NOX enzyme that is distinct from the potential cancer-promoting NOX4.
In summary, ceramide neutralization through the formation of glucosylceramide not only decreases the proapoptotic stimulus ceramide but also abolishes NOX assembly and activity (Fig. 5C) and in conjunction with increased antioxidant enzyme activities, greatly alleviates cell death-inducing oxidative stress associated with chemotherapy or physiological agonists. Consequently, pharmacological inhibition of antioxidant enzymes, such as catalase or pharmacological or molecular inhibition of GCS restored doxorubicin-toxicity, offering promise as a useful therapeutic avenue for the treatment of advanced cancers of the CNS. While treatment of patients with siRNA is currently not possible due to toxicity associated with delivery systems, our non-toxic cationic nanoliposomal formulation is designed specifically to overcome this hurdle. Additionally, our nanoliposomal formulation combining C6-ceramide and PDMP overcomes the current hurdle to clinical delivery of the GCS inhibitor PDMP associated with its low solubility. Nanoscale delivery systems further offer considerable clinical potential in the delivery of lower, yet more concentrated, therapeutic doses.26,35 Many advanced CNS tumors, such as glioblastomas, respond poorly to chemotherapeutic and radiation therapies, while in comparison, resistance in neuroblastomas is less common.14,36 Complicating matters and driving the demand for new and better therapeutics, is the fact that glioblastoma multiform, the most common primary brain tumor, has one of the worst survival rates among all cancers.14 This research offers an important advance to the understanding of mechanisms of multidrug-resistance in glioblastoma and the design of better therapeutics. On the other hand, it is worth noting that degenerative pathologies, including acute and chronic CNS disorders as well as psychiatric disorders, exhibit oxidative stress as a major component.37,38 This is important because in contrast to the goals of cancer therapy, the goals in the treatment of degenerative disorders are to improve the survival and function of normal cells. While ceramide neutralization and degradation are hallmarks of chemoresistance, ceramide accumulation is characteristic of degenerative disorders.6,32,33,38 Altogether, nano-encapsulated pharmacological or molecular agents targeting GCS offer exceptional promise in the treatment of chemoresistant CNS cancers, based on our findings that downregulating GCS activity not only can override ceramide neutralization by tumor cells, but also reengage stimuli-dependent NOX activity, restoring an ability to produce ROS and ultimately inducing tumor cell death.
DMEM and penicillin/streptomycin solution were obtained from Mediatech (Herndon, VA). Solutions of GlutaMAX-1 and trypsin/EDTA, Hank's Balanced Salt Solution (HBSS) and Amplex Red reagent, as well as fluorescent siRNA, were from Invitrogen (Carlsbad, CA). Fetal bovine serum was from Atlanta Biologicals (Atlanta, GA). The ON-TARGET plus SMART pool siRNA directed against GCS and non-targeting siRNA was from Dharmacon (Lafayette, CO). GCS cDNA (Gene: UGCG; Accession: BC038711), was purchased from Thermo Fisher Scientific (Huntsville, AL). Streptavidin-agarose beads, sulfo-NHS-biotin, protease inhibitor cocktail (PIC), a Pico Super Signal chemoluminescent kit and a BCA protein assay kit were obtained from Pierce (Rockland, IL). Polyclonal rabbit anti-GCS, rabbit anti-FLAG, rabbit anti-GAPDH and p67phox antibodies, and corresponding horseradish peroxidase-conjugated secondary antibodies were from Santa Cruz (Santa Cruz, CA). An XTT cell proliferation assay kit was obtained from Trevigen (Gaithersburg, MD). VECTASHIELD hard set mounting medium with 4,6-diamidino-2-phenylindole (DAPI) was purchased from Vector Laboratories (Burlingame, CA). Recombinant human tumor necrosis factor alpha (TNFα) was purchased from Millipore (Temecula, CA). 2′, 7′-dihydrodichlorofluorescein diacetate, doxorubicin, paclitaxel, methotrexate, fenretinide and 4-hydroxy tamoxifen were from EMD Biosciences (San Diego, CA). Gemcitabine was a gift from Dr. Yixing Jiang. D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) was purchased from Biomol (Plymouth Meeting, PA). All other lipids were from Avanti Polar Lipids (Alabaster, AL). All other reagents were purchased from Sigma (St. Louis, MO).
Human SH-SY5Y neuroblastoma cells, human U-87 MG glioblastoma cells and human LN-18 glioblastoma cells were grown in DMEM medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 U/ml streptomycin (humidified atmosphere, 5% CO2, 37°C) in 100 mm tissue culture dishes (Falcon). For subculture, cells were washed with PBS (phosphate buffer saline) then treated with 0.5 mg/ml trypsin and 0.2 mg/ml EDTA (5–15 min), resuspended and centrifuged (200x gmax, 2 min) and plated in 100 mm tissue culture dishes (2 × 107 cells per dish), 6-well tissue culture plates (5 × 106 cells per well) or 96-well tissue culture plates (5 × 103 to 5 × 104 cells per well).
Aliquots of lipids were made at appropriate ratios (Table 1), and dried to a film under a stream of nitrogen, then hydrated by addition of 0.9% NaCl to a final lipid concentration of 25 mg/ml. Solutions were sealed, heated at 60°C (60 min) and subjected to vortex mixing and sonicated until light no longer diffracted through the suspension. The lipid vesicle-containing solution was quickly extruded at 60°C by passing the solution 10 times through 100 nm polycarbonate filters in an Avanti Mini-Extruder (Avanti Polar Lipids, Alabaster, AL). Nanoliposome solutions were stored at 4°C until use, protected from light when necessary. To prepare siRNA- or plasmid-loaded cationic nanoliposomes, siRNA or plasmid DNA, was aliquoted into 0.9% NaCl and nanoliposomes were added in a 10:1 weight ratio to nucleic acid. The solution was allowed to incubate overnight at room temperature prior to use. The RNA interference sequences utilized (Table 2), were from the ON-TARGET plus SMART pool product line of Dharmacon (Lafayette, CO).
Liposome suspensions were diluted 1:10 in 0.9% NaCl prior to size determination. Light scattering was used to quantify the size of liposomes using a Malvern Instruments Zetasizer Nano (Malvern, Worcestershire, United Kingdom).
Glucosylceramide synthase cDNA (Gene, UGCG; Accession, BC038711) was transferred by PCR sub-cloning into the mammalian expression vector pcDNA3.1 (Invitrogen, Carlsbad, CA). The construct was sequence-verified by the Molecular Genetics Core Facility at the Pennsylvania State University College of Medicine.
Plasma membrane proteins were prepared by biotinylation and streptavidin affinity purification as previously described.19 SH-SY5Y cells grown in 6-well plates for transfected with siRNA or plasmid (200 nM or 3 µg/ml, respectively, 48 h) or preincubated with arachidonyl trifluoromethyl ketone (ATK, 10 µM, 60 min) or C8-glucosylceramide (10 µM, 60 min), prior to TNFα stimulation (100 ng/ml, 15 min). Next, cultures were washed with HBSS pH 7.5 containing 0.1 g/l CaCl2 and 0.1 g/l MgCl2 (HBSS-CM), incubated with 0.5 mg/ml sulfo-NHS-biotin (20 mM HEPES in HBSS-CM pH 8.0, 40 min, on ice) and excess sulfo-NHS-biotin was neutralized with 50 mM glycine (HBSS-CM, on ice, 15 min). Cells were scraped into HBSS-CM, centrifuged (200x gmax, 2 min), lysed in 500 µL loading buffer (20 mM HEPES in HBSS pH 7.5, 1% Triton X-100, 0.2 mg/ml saponin, 1% PIC) and after brief sonication, incubated with streptavidin-agarose beads (25 µl, 2 h, 4°C). Beads were collected by centrifugation (2,500x gmax, 2 min) and re-suspended in 150 µl of loading buffer (5 min, boiling water bath) to release bound protein. Supernatants were obtained by centrifugation (2,500x gmax, 2 min) and analyzed by western blotting.
SH-SY5Y, U-87 MG or LN-18 cells were seeded in 96-well tissue culture dishes (5 × 104 cells per well) and grown for 48 hours. Cultures were incubated with 50 µM of the oxidation-sensitive fluorescent indicator 2′, 7′-dihydrodichlorofluorescein diacetate (H2DCFDA) for one hour in the presence or absence of pharmacological inhibitors, N-acetyl-L-cysteine (5 mM) or C8-glucosylceramide (10 µM). Alternatively, cells were transfected with siRNA (200 nM) or plasmid (3 µg/ml), 48 h prior to exposure, the last hour of which included H2DCFDA loading. H2DCFDA is de-acetylated in the cytosol to dihydrodichlorofluorescein and increases in fluorescence upon oxidation by H2O2 to dichlorofluorescein (DCF). Following exposure to stimuli, cultures were washed with PBS, agitated in lysis buffer (2 M Tris-Cl pH 8.0, 2% w/v SDS, 1 mM Na3VO4) and total DCF fluorescence intensity was quantified in 100 µl of cell lysates using a Beckman Coulter Multimode DTX 880 microplate reader (Fullerton, CA) with a 495 nm excitation filter and a 525 emission filter. All DCF fluorescence data were normalized to the average DCF fluorescence under control conditions (relative DCF-fluorescence values).
Generation of fluorescent NBD-glucosylceramide was quantified as follows. Samples of whole cell lysates containing 5 µg of total protein (BCA protein assay) were reacted with 100 µM NBD-ceramide in 5 mM MnCl2 pH 7.2, 30 µg/ml BSA and 1 mM UDP-glucose (100 µl reaction volume, 60 min, 37°C). After termination of the reaction (100 µl dichloromethane), lipids were extracted (Bligh-Dyer) and separated by thin layer chromatography (chloroform/acetone/methanol/acetic acid/water at 10:4:3:2:1). NBD-glucosylceramide was quantified using a GE Healthcare Typhoon Imager (Piscataway, NJ) running ImageQuant software.
Interference of ferrictyochdrome reduction by superoxide was measured as an indication of superoxide dismutase (SOD) activity. Samples of whole cell lysates containing 5 µg of total protein (BCA protein assay) were added to a solution of 50 µM hypoxanthine, 6 nM xanthine oxidase, 100 µM EDTA and 10 µM ferricytochrome c in a 50 mM potassium phosphate pH 7.8 (100 µl reaction volume, 30 min, 25°C) and ferricytochrome reduction was quantified using a Beckman Coulter Multimode DTX 880 microplate reader (Fullerton, CA), reading absorbance at 550 nm.
Interference of horseradish peroxidasemediated formation of the fluorescent resorufin from Amplex Red was quantified as an indication of catalase activity. Samples of whole cell lysates containing 5 µg of total protein (BCA protein assay) were mixed with 0.1 M Tris-Cl pH 7.4 and 10 mM MgCl2 (volume of 75 µl) prior to incubation with 10 µM H0O2 (25 µl) for 30 min at 37°C. Next, an equal volume (100 µl) of horseradish peroxidase (2 U/ml) and Amplex Red (100 µM) was added to the reaction mixture and resorufin fluorescence was quantified using a Beckman Coulter Multimode DTX 880 microplate reader (Fullerton, CA), with a 530 nm excitation filter and a 590 emission filter.
SH-SY5Y, U-87 MG or LN-18 cells were seeded in 96-well tissue culture dishes (5 × 103 cells per well) and grown for 48 hours. Cultures were co-incubated with pharmacological inhibitors and either 100 ng/ml TNFα or 5 µg/ml doxorubicin, for 48 hours. Alternatively, cells were transfected with siRNA (200 nM) or plasmid (3 µg/ml), 48 hours prior to TNFα or doxorubicin exposure. XTT reagent prepared according to the manufacturer (Trevigen, Gaithersburg, MD) was added and allowed to incubate for 4 hours. Viability was determined by absorbance at 490 nm (650 nm ref.), using a Beckman Coulter Multimode DTX 880 microplate reader (Fullerton, CA). All values were normalized to the average values under control conditions.
One-way or two-way, analysis of variance (ANOVA), were used to determine statistically significant differences between treatments (p < 0.05). At least three independent experiments were performed for each condition. Post hoc comparisons of specific treatments were performed using a Bonferroni test. Unpaired t-tests were used to directly compare mean diameters of respective liposome formulations. All error bars represent standard error from the mean (SEM). All statistical analyses were carried out using GraphPad Prism 4 software (La Jolla, CA).
We would like to thank Keystone Nano, Inc., (State College, PA) and in particular Amy Knupp and Dr. Mylisa Parette, for the use of and assistance with, the Malvern Zetasizer Nano instrument. We graciously thank Dr. Yixing Jiang, of the Milton S. Hershey Penn State Medical Center, for kindly providing gemcitabine. Discussions with Dr. Marvin Schulte, Dr. Barbara Taylor and Dr. Kristin O'Brien, of the University of Alaska-Fairbanks, were helpful during the preparation of this manuscript. This research was funded in part by USDA grant 2005-34495-16519 (T.B.K.), NIH grant U54 NS41069 (T.B.K.) and NIGMS grant GM77391 (M.C.C.). Additional financial support was made possible by the College of Natural Sciences and Mathematics, University of Alaska-Fairbanks.
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.


text:
Joan S Serchuck
New York, NY
Also Known As
Get Background Check on Joan
Joan S Serchuck
Jerome S Serchuck
Joan Qpr Serchuck
Joan S Serchuck is in her 80's
Joan Has Lived In
New York, NY
Palm Beach, FL
Manhattan, NY
Joan's Relatives
Joan S Serchuck
Zodiac SignPisces
Additional Infomation on Joan S Serchuck
Joan S Serchuck's Contact Information
Known Cities Lived In
Find out where Joan S Serchuck has lived as well as Joan S Serchuck's phone numbers and email addresses.
Joan S Serchuck Has Lived in 2 States
New York Address for Joan S Serchuck
30 E 7xxx Sx
New York NY
New York NY
Has Lived InGet Full Address Report
New York, NY (2 Addresses)
Palm Beach, FL (3 Addresses)
Phone Numbers Associated with Joan S Serchuck
Get Full Phone Report
(212) ***-**** - New York, NY
(561) ***-**** - West Palm Beach, FL
(561) ***-**** - West Palm Beach, FL
All trademarks, product names, company names or logos on this page are the property of their respective owners.
Related People Searches


text:
Old Dubliner Bed and Breakfast
No prices found through our partners. Please contact the business directly or check some of our recommended alternatives.
TripAdvisor Reviews Old Dubliner Bed and Breakfast
Travel Blogs from Dublin
Two Super Bowls in One Day! Fun is Fun.
... the Browns. (which I was fine with) Lets face it, no one wants to watch NFL when you have to Super Bowls playing on the same day. (that is the only way to describe this situation). Since it was Sunday....everyone was out to watch the games.
I got to the bar around 4PM knowing full well it was going to be pretty dicey and I was not wrong. This place was packed. Standing room only and ...
Grrreat Day
... and jelly. We were saved!!! Chewed them down, stood up and raced to the next bus. Aunt Peggy, this would be a good time for you to grab some lunch, quick trip to the bathroom and come back and fasten your seat belt. As we pulled away from the dock once again, I wondered if it could get any better. This morning our guide had been a delightful retired school teacher named Annie Gallagher!! We boarded the bus and there she was, to be with us all ...
... Dublin.... Andrew enjoyed the motorway at 120 on cruise control and Blake slept soundly. That is until we arrived into busy Dublin city and needed to concentrate on navigating to our accommodation when Blake decided to wake up ravenous and scream! Fortunately I managed to feed him and Andrew could focus enough to deliver us to our third flash accommodation of the trip - The Croke Park Hotel. We have another lovely room and Mum, Dad and Lucy are a couple of floors above ...
Luck o the Irish
... Dublin to explore a bit. We started off at the Guiness Storehouse which is amazing. It is a seven story brewery tour that explains all the history of guiness. Very interesting. It ends with a pint of guiness in the Gravity Bar which has 360 degree views of Dublin.
After that we went for a wander through town. Sam was very excited to find a Timmies!! So had to have a token donut! ...
On the Road Again, for the Last Time in Eire
... minutes later, she was made naked before our eyes and only about 3 kilos of wool were left behind. Their wool, while so incredibly soft, is just a by-product of the industry. If farmers hire out a professional shearer it can cost more to have the coats clipped than the wool will bring at the market. In fact, Fenton was wearing an authentic counterfeit Tommy Hilfiger sweater, made of real acrylic somewhere in southeast Asia. As he explained, Irish farmers wear ...
Other places to stay in Dublin
Jurys Inn Dublin Parnell StreetNorthside, Dublin | 3 star hotelfrom $101
- Perfect -- simple, clean, affordable. Great location
- Great Hotel
- Very good. location perfect
Hilton Garden Inn Dublin Custom HouseNorthside, Dublin | 4 star hotelfrom $116
- A good choice for it's price range
- Previous Harsh Reviews not quite true
- Avoid
Maldron Hotel Cardiff LaneTrinity College, Dublin | 4 star hotelfrom $162
- Fantastic hotel
- Great weekend!!
- Fantastic value
Bewley's Hotel BallsbridgeBallsbridge, Dublin | 3 star hotelfrom $89
- One price is great, even during concerts!
- Excellent Hotel-Great Value
- Nice Hotel in a Quiet Neighborhood
BEST WESTERN PLUS Academy Plaza HotelRotunda, Dublin | 3 star hotelfrom $97
- Would stay there again!
- It's only OK
DoubleTree By Hilton Hotel Dublin - Burlington RoadBallsbridge, Dublin | 4 star hotelfrom $118
- Nice overall
- Loved the Burlington
- Top class hotel for great prices
Mespil HotelPembroke West, Dublin | 3 star hotelfrom $114
- Good value for money
- Good for the price, short on charm
- Good service but really needs a refurb
Gresham HotelNorthside, Dublin | 4 star hotelfrom $145
- Don't even consider it....
- Gresham Hotel
Jurys Inn Dublin ChristchurchLiberties, Dublin | 3 star hotelfrom $125
- Great time was had by all
- Good Location/Average Hotel
- Not Great
Bewleys At Newlands CrossExit 1A Naas Road, Dublin | 3 star hotelfrom $68
The Croke Park HotelNorth Inner City, Dublin | 4 star hotelfrom $130
- Impressive
- Nice Stay
- Thumbs Up.
The Westbury HotelSouth-East Inner City, Dublin | 5 star hotelfrom $277
- Nice touches at the Wesbury
- Very friendly staff
- Overpriced and Overrated
Clarion Hotel Liffey ValleyNorth Inner City, Dublin | 4 star hotelfrom $77
- Great rooms, cold reception
- Great 'new' hotel
- Not impressed.
Camden Court HotelTrinity College, Dublin | 3 star hotelfrom $129
- Great value hotel
- OK- but nothing special
- Great weekend spot
Radisson Blu Hotel, Dublin AirportDublin Airport, Dublin | 4 star hotelfrom $127
- Poor
- Great airport hotel
- Good Airport Hotel
Green Isle HotelNaas Road, Dublin | 3 star hotelfrom $65
- Not what was expected
- Great refurb
- Awful hotel
Ashling HotelLiberties, Dublin | 4 star hotelfrom $118
- Location and room ok, but service rather poor
- Happy With the Ashling
- Not the Best West
Radisson Blu Royal Hotel, DublinLiberties, Dublin | 4 star hotelfrom $196
Crowne Plaza Hotel Dublin-NorthwoodNorthwood Park, Dublin 9, Dublin | 4 star hotelfrom $105
- Spot-on, well-above average Airport Hotel
- Top Class Airport Hotel
- Good hotel but......
Four Seasons Hotel DublinPembroke East, Dublin | 5 star hotelfrom $248
- Excellent, world-class service
- Almost perfect
- Corporate ambiance and spotty service
The Regency Hotel DublinArtane - Whitehall, Dublin | 3 star hotelfrom $80
- Pleasantly Supprised
- Great choice
- Stay Away
Maldron Hotel Dublin AirportDublin Airport, Dublin | 4 star hotelfrom $105
- Poor reviews unjustified
- Woeful
- Definitely not a Four Star accomodation
The Shelbourne Dublin, A Renaissance HotelMansion House, Dublin | 5 star hotelfrom $273
- Superior Shelbourne
- Fabulous Shelbourne
- Excellence in Dublin
Trinity City HotelSouth-East Inner City, Dublin | 3 star hotelfrom $166
- Great location, good restaurant
- Trinity Capital Hotel, Dublin, Republic of Ireland
- Central in Dublin
Hilton Dublin Airport HotelNorth Inner City, Dublin | 4 star hotelfrom $92
- A home away from home in Dublin!
- Excellent hotel
- Very nice room


text:
i first saw this plant on toptropicals.com and i fell in love! i ordered some seeds and planted a few. none of them came up. i did not scarify them. is that why?
dichrostachys cinerea (kalahari christmas tree) question.
mekos Fair Play, SC(Zone 7b) Jan 06, 2010
Did you soak them first? Mine said emerse seeds in HOT water and let cool, then soak seeds in water for 24 hours. Sow in sandy soil. should sprout 2-10 days. Did you try that? If not maybe try again this way.
Post a Reply to this Thread
More Tropical Zone Gardening Threads
queen palm died -what caused it?!started by uglymotherpluck
last post by uglymotherpluckAug 28, 20155Aug 28, 2015
Thailandstarted by mistifarang
last post by mistifarangAug 26, 20154Aug 26, 2015
Destroyed leavesstarted by mistifarang
last post by mistifarangSep 04, 20153Sep 04, 2015
what the problemstarted by sheed
last post by sheedAug 27, 20151Aug 27, 2015
Thailandstarted by mistifarang
last post by mistifarangSep 01, 20156Sep 01, 2015


text:
Keira Knightley and James Righton have survived the first year of raising their baby girl together when Keira was hit by unwarranted criticism from Begin Again director John Carney. Good thing that she has such a dependable husband in James Righton to get through the rough waters together. Show More Summary
From Kristen to Cara and Kendall to Lily-Rose, Karl Lagerfeld has certainly adopted an "If it ain't broke, don't fix it" attitude when it comes to his Chanel muses. His wide-ranging roster of go-to models, actresses and general girls-about-town have fronted campaigns for many different categories... Continue reading
Keira Knightley might seem dainty, but she has no problem dropping an F-bomb when the occasion calls for it—like when chastising the young British citizens who plan to skip the country's upcoming referendum on European Union membership. read more
John Carney, who worked with Keira Knightley on 2013's Begin Again, has recently criticized the actress for being too pretentious and not possessing the depth required for serious film actresses. In being asked what he would do differently, Carney said that he would not work with supermodels again, referring to Ms. Show More Summary
"I'll never make a film with supermodels again." This was director John Carney's passionate declaration, made in an interview with The Independent on May 28. The "supermodel" in question was Keira Knightley, star of his well-received...Show More Summary
In an interview with The Independent Saturday, director John Carney, known for his films Once, Begin Again, and Sing Street, had a surprising amount to say about his Begin Again star Keira Knightley—none of it flattering. What was particularly odd was that Carney seemed more preoccupied with insulting Knightley than promoting his latest film, Sing Street.
It's been quite a week for airing cinematic grievances. John Crawley has now backtracked on his comments disparaging working with Keira Knightley on Begin Again, while Kate Beckinsale also made some rather unsavoury comments about Pearl Harbor director Michael Bay, too. Show More Summary
John Carney also offered a private apology to Knightley
LONDON — Irish film director John Carney has taken to Twitter to share a public apology to Keira Knightley after he criticised her acting ability in a recent interview. Carney directed Knightley in the 2013 drama Begin Again. In a recent...Show More Summary
At least he's admitting he was wrong! Director John Carney recently BASHED Keira Knightley's acting abilities from his 2013 flick Begin Again, and it really wasn't pretty. But now, the Irish filmmaker has revealed that he's personally...Show More Summary
Begin Again director John Carney has offered up a wide-ranging mea culpa to star Keira Knightley, just a few days after he talked some pretty extensive and far-flung shit about her while promoting his latest film. Carney—who broke into...Show More Summary
A couple of days ago, John Carney, the director of Sing Street and Begin Again, happened to make some choice remarks about how he basically didn't want to work with Keira Knightly ever again. We were as surprised as you were, frankly,...Show More Summary
After dismissing Keira Knightley as a supermodel with a giant entourage and telling The Independent that "acting requires a certain level of honesty and self-analysis that I don't think she's ready for yet," Begin Again director John...Show More Summary
John Carney would like to right a wrong. Over the weekend, the director got a little too candid in an interview and criticized Keira Knightley, who starred in his 2014 film Begin Again. There, he said that acting "requires a certain level of honesty and self-analysis that I don't think she's ready ... More »
Begin Again director John Carney has issued an apology for his statements about Keira Knightley, insulting her acting talents. "From a director who feels like a complete idiot," John began in his lengthy statement, posted to his Twitter account. "In trying to pick holes in my own work, I ended up blaming someone else. That's [...]
NEW YORK (AP) — "Begin Again" director John Carney has apologized for slamming actress Keira Knightley in a recent interview in which he negatively compared her to "a supermodel." Carney is the Irish filmmaker of the Oscar-winning "Once." He posted a statement Wednesday on Twitter from "a director who feels like a complete idiot." He […]
This weekend, Begin Again and Song Street director John Carney made the mistake of discussing his true feelings about Keira Knightley in an interview with the Independent. Carney said of Knightley: "I don't want to rubbish Keira, but you know it's hard being a film actor and it requires a certain level of honesty and self-analysis that I don't […]
Never forget that Hollywood is a small town: not long after Begin Again and Sing Street's director John Carney trash-talked the former film's star actress, Keira Knightley, in an interview with the Independent, her former colleagues rushed to her defense on Twitter. Show More Summary
Days after director John Carney lambasted certified British gem Keira Knightley for not being a "proper" actress, her former colleagues took to the press and social media to defend Knightley's character and professionalism. Read mor...