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Personal Statement
I believe in health care that is based on a personal commitment to meet patient needs with compassion and care....more
I believe in health care that is based on a personal commitment to meet patient needs with compassion and care.
More about Dr. Yogish
Dr. Yogish is a popular General Physician in Koramangala, Bangalore. He is currently practising at Health First Clinic in Koramangala, Bangalore. Save your time and book an appointment online with Dr. Yogish on Lybrate.com.
Lybrate.com has a number of highly qualified General Physicians in India. You will find General Physicians with more than 28 years of experience on Lybrate.com. You can find General Physicians online in Bangalore and from across India. View the profile of medical specialists and their reviews from other patients to make an informed decision.
Lybrate.com has a number of highly qualified General Physicians in India. You will find General Physicians with more than 28 years of experience on Lybrate.com. You can find General Physicians online in Bangalore and from across India. View the profile of medical specialists and their reviews from other patients to make an informed decision.
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Health First ClinicGanapathi Temple Rd, Koramangala 6 Block, Koramangala,Landmark: Near Panchmukhi Ajarya Temple, Bangalore Get Directions
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20 years old male from Jhansi asked
I am 21 bachelor and a virgin teenager. Can I take Zandu Vigorex tablets. Are there any side effects. If so suggest any other herbal booster. I just wanna take it to aid my stamina. Should I?
General Physician, Jalgaon
To increase your stamina and to strengthen penis... Wake up early go for morning walk in greenery daily Do yogasanas and pranayam daily Do perineal exercises daily Can take Musali pak by Patanjali 2 tsf twice a day for 3 months
32 years old female from Jammu asked
I am 32 yrs female muje pichhle 2 mahine se har 2week mein fever ho jata hai sardi bhut lgti hai fever mein body feel very much tired 4yrs se married hun but no child yet kabhi concieve hi nahi hua.
General Physician, Mumbai
Get your blood checked for cbc, mp , widal , sgpt and urine r/m and revert back to us with reports and For cold take tablet cetrizine at night and For fever take tablet paracetamol 650 mg
32 years old female from Gonda asked
Sir I want to ask about my friend. She is 32 year old. I think she is suffering from chronic infection and symptoms is ulcer, nodules deformities of fingers and toes and wasting of body parts. In this condition what can we do.
DHMS (Hons.)
Homeopath, Patna
Hi, your presentation is not much, clear to reach a diagnosis, unless the affectrd parts are viewed, properly. However, tk, pleny of water take, protein rich diet, on time. Clean the affected part to check infection. Following, assosiated symptoms are required: is, there any sort of burning sensestion.-any sort of discharge.-itching sensestion. Take, care.
31 years old male asked
I am a 30year old male and have suffer back pain from last 2/3 month as well as some cramps in inside of chest and below chest and also lightly pain in back side of head.
Homeopath, Faridabad
Hello. Maintain a right standing and sitting posture, keep your back always straight, avoid carrying weight on back. Hold or carry weight divided equally in both the hands or shoulders (to avoid over straining on one particular hand or shoulder). Lie down on bed on your back without pillow. Exercise: Practice Shavasana or Corpse Pose as it looks like a dead body. If practiced correctly and with full efforts, can relax every body part, relaxing not only the body but also the mind: Spread the legs one to two feet apart, the toes are turned outwards, the heels facing each other, a comfortable distance apart. Bring the arms a little away from the body, palms turned upward. Relax the neck and allow it to turn to the side if it is more comfortable. Close the eyes and focus the attention on the body, breathing normally. Medication: Take homoeopathic medicines - Arnica 200 and Rhus Tox. 200 - take both of them thrice daily for 1 week. Also, apply Schwabe's Topi-Arnica and Topi MP Gel on the affected area in circular motions gently thrice a day. Get back to me thereafter.
year old male from Guwahati asked
I am a 65 years old male. Recently I was diagnosed to be suffering from prostate gland (frequent urination) and the doctor advised me to go for an operation. Kindly tell me a treatment without operation is possible or not.
MBBS, MS- GENERAL Surgery
General Surgeon,
Yes treatment is possible without surgery but you need to provide me with investigations like psa report and ultrasound of lower abdomen.
22 years old male from Chennai asked
Bachelor of Ayurveda, Medicine and Surgery (BAMS)
Sexologist, Jodhpur
Excessive of anything is bad same applies with masterbation. 2 things happen when one masterbate excessively 1. Premature ejaculation you can correlate when you started mastrrbation what was time and check current timing. You will find time to ejaculate might hv reduced significantly. Some people even ejaculate while rubbing in bed 2. Second problem is weak erection. Over a period you will feel lesser or weaker erections. One may feel shrinking penis due to this. Consult to cure side effects of excessive masterbation. Results within 10 days once you start treatment.
30 years old female from Hyderabad asked
Hello doctor. My mom is diabetic patient her age is 59. She is using tables when we consult to doctor they said blood count is low so you should take injection for this and they wrote renocel 4000 I you injection and we are giving that injection weekly 2 times but. Skin getting rashes like red red doubts so kindly help me in this matter. Waiting for your reply.
Dietitian/Nutritionist, Chandigarh
There are several means to gain control over your diabetes. Few of them as below: 1. Eating at regular intervals 2. Avoiding foods high in processed and invisible sugar 3. Good sleep hygiene 4. Consistent workouts to be able to give you specific guidelines, we would need to gain a deeper insight into your lifestyle and work with you in greater proximity. Do let me know if you're keen and we could take this prospect forward. Meanwhile, feel free to consult me privately.
19 years old male from Titwala asked
I am 18 years old and having a acidity problem and my face is so oily what I should to prevent from both?
Bachelor of Ayurveda, Medicine and Surgery (BAMS)
Ayurveda, Faridabad
Tips to cure acidity:: 1. Drink 100-500 ml of tender coconut water twice a day for relief from hyperacidity Mix equal parts of powdered fennel, licorice root, basil leaves and coriander seeds. Have ½ teaspoon of this mixture with ½ teaspoon of powdered rock candy (or unrefined sugar) 15 minutes before lunch and dinner. 2.Make a powdered mixture of equal parts of rock candy (or unrefined sugar), fennel, and green cardamom. Whenever you feel heartburn, mix 1 teaspoon of the mixture in a glass of cold milk and drink. 3.Avoid tea, coffee, and carbonated or alcoholic beverages.Especially stop drinking bed tea. 4.Avoid using garlic, ginger, onions, tomatoes, and vinegar in cooking. 5.Curd should be strictly avoided at night. 6. Practice Yoga and Pranayama such as Vajrasana, Bhujangasana, Salabhasana, Bhastrika Pranayama,Shitali Pranayama and Shitkari Pranayama. 7.Drink plenty of fluids and water. 8. Avoid constipation. 9. Revert back after 10 days. Take Care.
41 years old male from Mumbai asked
Hi, I am having problem to sleep, some time unable to sleep full night (Sleepless). Resulting now has heart problem, My Blood Test Report was very bad like Lipid Profile.
PG Diploma in Emergency Medicine Services (PGDEMS), Bachelor of Ayurveda, Medicine and Surgery (BAMS), MD - Alternate Medicine
Ayurveda, Ghaziabad
Hi One of the simplest and most effective ways to induce sleep is to rub some oil on the scalp and the soles of the feet before going to bed. Use sesame oil, brahmi oil, jasmine oil, or coconut oil and massage gently for a few minutes. Slightly warming the oil before applying is helpful. 1-1 drop of cow ghee in both nostrils at night time before sleep. Take sarpgandha vati 1-1 tab twice a day. Take pranacharya no tens capsule 1-1 twice a day. Avoid spicy food. Junk food. Fermented food. Tea.coffee. Do pranayama early in the morning.
25 years old female from Ahmedabad asked
My mom is suffering from diabetes. Some time Her sugar level increase or decrease. How to maintain her sugar level with out any medicine?
Diploma in Naturopathy & Yogic Science (DNYS), M.Sc - Dietitics / Nutrition
Dietitian/Nutritionist, Vadodara
Yoga and diet will definitely help in diabetes. There are some asana and pranayam which will help you to control your blood sugar level, like 1 ardh matsyendrasana 2 purna matsyendrasana 3 vakrasana 4 kapalbhati and many more diet has key role in diabetes. Diabetic diet is different for person to person. It is depend of person lifestyle, his blood sugar level. Medication and many more factor. But here are some common tips 1 eat high fiber food like oats, bran, sprouts, beans 2 avoid eating refine flour and its product 3 eat fresh fruits instead of fruit juice. 4 after taking medicines take your meal within half hour.
21 years old female from Delhi asked
My brother have pain in his testicle balls because of continuous running and exercise. Is it a serious problem or can be solved with some home remedy.
BASM, MD, MS (Counseling & Psychotherapy), MSc - Psychology, Certificate in Clinical psychology of children and Young People, Certificate in Psychological First Aid, Certificate in Positive Psychology
Psychologist, Palakkad
Dear user. Taking rest for a week and drinking lots of water should be helpful. Do consult a doctor if the pain persist for more than 3 days. Take care.
36 years old male from Purulia asked
I am 38 years old suffering type 2 diabetic from last 8 years my blood sugar is frequently up anddown gap of two or 3 month. I have to use 4 mg glimepiride tab 1000 mg metformin daily. Please give me a food suggestion and life style.
MBBS, CCEBDM, Diploma in Diabetology
Endocrinologist, Hubli-Dharwad
Mr. lybrate-user , You have mentioned about the drugs being taken for diabetes, but no dosage. There is no mention about fasting, PP glucose level and HbA1c%, so it is not clear whether diabetes is well controlled at all. At your age it is very essential to achieve a very strict blood glucose control with fasting closer to 100 mg, PP 150 to 160 mg & HbA1c%<6.8. If not then you are inviting long term complications of diabetes much early. As regards dietary advice, unless all the details are known, it is difficult to suggest a specific diet. However, please avoid sweets, deep fried foods, junk foods, pastries, bakery items, sweetened aerated drinks, sweet corn syrup containing products and food stuff made up of maida. Avoid alcohol and smoking completely. As regards lifestyle regular one hour exercise a day is most important. Brisk walking is good enough. But if you can jog or run please do. Thanks.
18 years old male from Mumbai asked
Actually I am having cold from a month. I had gone to the dispensary and the doctors gave me medicine but after 3-4 days I got affected again. Can I get to know what would be the best treatment.
MBBS, cc USG
General Physician, Gurgaon
Frequent/ continuous cough cold may be because of many reasons like: 1. Any ent problem like dns polyp sinusitis 2. Low immunity 3. Allergic condition you need to avoid dust you can take vitamin c tablet, one tablet with food for 15 days there are some supplements name nutrilite daily and protein available that can be start after personal consultation and these supplement can reduce severity and no. Of episodes of cough/cold.
34 years old female from Chennai asked
My wife often suffers from headache and she takes Saridon whenever she gets headache. Is there any side effects by taking Saridon. Kindly advice on this.
General Physician, Cuttack
1.Take paracetamol 500mg,1 tablet sos after food up to a maximum of 3 tablets daily, 2.Drink plenty of water 3.Take rest If you have recurrent attack of headache 1.It could be a tension headache due to stress and strain or migraine. 2.Avoid stress, physical and mental exertion 3.Go for regular exercise 4.Practice yoga, meditation and deep breathing exercise to calm your mind, control your emotion and relieve stress 5.Check for refractive error, sinusitis Hemoglobin, BP 6. Consult Neurologist to rule out other causes of headache 7. consult for further advice Avoid taking saridon regularly. It will have side effects like gastritis etc
21 years old male asked
Hello sir, I have red eyes from two days, some fever is there, Please tell me solution and medicine.
Homeopath, Faridabad
Hi, Take belladonna 200, 5 drops , thrice a day. BC no. 11, 5 tabs every 4 hourly. Revert me after 2 days with feedback.
18 years old male from Rudrapur asked
From puberty till now whenever I masturbate sperm never come out. I am so worried about this but when I use sex doll then it ejaculates and its too quick.
General Physician,
You have no problem in ejacualtion and therfore you are safe and in masturbation your mind may be anxious or get diverted
23 years old male from Hyderabad asked
I am suffering from gas trouble for so many day could you give a suggestion how to stop a gas formation in my stomach.
Bachelor of Ayurveda, Medicine and Surgery (BAMS)
Ayurveda, Bangalore
Please follow tips 1. Eat when you feel hungry, chew the food 2. Do not drink water just before and after food 3. Chew funnel seeds or lavang, elaichi after food 4. Sit in vjrasana after each meal 5. Drink buttermilk with or after meals 6. Avoid over eating, eating in between meals 7. Exercise or practice yoga, sooryanamskra regularly alovera juice, hinguvachadi pills, chitrakadi vati, jeeraka rasayana help to reduce gas formation and increase digestion.
23 years old male from Saharanpur asked
My name is Abdul wahid. I am 23 years old. I suffered chest problem 1 to 2 years ago. I feel fullness of chest and ache around sternum and backache upper part. I have ask its problem from a allopathic doctor at lybra e. He told me that you have esophagus problem so what should I do for it.
BPTh/BPT, MPTh/MPT
Physiotherapist, Noida
Do the cat/cow stretch. Get on all fours, with your arms straight and your hands directly under your shoulders; your knees hip-width apart.
23 years old male from Nadia asked
I'm a student of engineering, I could not read in book at midnight. My body very weak and not healthy. Maximum time of the day I am sleeping. What can I do.
MD-Ayurveda, Bachelor of Ayurveda, Medicine and Surgery (BAMS)
Sexologist, Haldwani
Hello lybrate-user- night is meant for sleeping. Studying at night will disturb your sleep cycle and hence will make your brain weak, it will also ruin your eye power. So go to bed before 10 pm, wake up as early as possible if you want to study because in morning time the brain tissue is almost 95% active and has a higher capacity to memorize as compared to that of night hours. Moreover waking up early morning will also benefit your health and you can give your 100% in other fields in future too.
24 years old male asked
My wife's body temperature drops - her lips turn pale like when you were to long in cold water - that happens from one moment to the other. She uses a heating-pad and then she spikes a fever - her entire body is overly hot. There are some weeks where she has it almost every day and other times she feels good for days. She also has heart racing / high blood pressure. The doctors hear in West Jefferson, NC don't really help.
PG Diploma in Emergency Medicine Services (PGDEMS), Bachelor of Ayurveda, Medicine and Surgery (BAMS), MD - Alternate Medicine
Ayurveda, Ghaziabad
it happens to most of the girls in cold water. Her lips become pale. So if this is happening she should go away from the cold water and use warm blanket. Dont give any heating pad to her. It happens because of weakness. She should take ayurvedic medicines for this.
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There are two main types of sand filters that are used for water treatment: rapid sand filters (also known as Rapid Gravity Filters or RGFs) and slow sand filters. Sand filters should not be confused with Roughing Filters, which tend to be horizontal-flow gravel filters used as a roughing treatment for turbid raw waters prior to sand filtration.
The two sand filtration processes differ from each other in several ways. In general however, rapid sand filters are usually fully automated, complex and costly, forming part of a wider treatment process in industrialized countries. They are not known to be suitable for household-level use, as is the case with versions of the slow sand filter, for example the intermittently operated slow sand filter (or IOSFF). For the sake of clarity though, a brief comparison is laid out below. For a fuller explanation of rapid sand filtration, please refer to Schulz and Okun (1984 [ref_01]).
Note that the unit metres/hour can be confusing, and is in fact a compaction of m3/m2/hour; that is, how much water volume can go through a certain area in one hour. Also sometimes this unit is expressed per day rather than per hour. For a fuller explanation of the relationship of water traveling through sand, check out Darcys Law explained.
Ref 01: Schulz, C. R.; Okun, D. A. (1984). Surface water treatment for communities in Developing Countries. IT, London
Ref 02: Unpublished information supplied by WEDC, 1999.
Ref 03: Huisman, L; Wood, W.E. (1974). Slow Sand Filtration. WHO, Geneva, Switzerland. p.44. Available from IRC


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Get Ready for Guests
Friends and relatives visiting this holiday season? Check out these easy ways to get your spare bedroom in tip-top shape.
Easy Early Mornings
A large pile of pillows on the bed looks inviting, but it also presents a challenge for visitors when it comes time to make the bed. Dress your guest bed simply with pairs of pillows neatly arranged. If you want to offer extra toppings, such as quilts, stack them in the closet for easy access.
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5 names.
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|David Dunston Actor, Star David Dunston was born in West Philadelphia changed his life & moved away from the tough streets to pursued his career as an actor / music producer / artist manager he is featured in TV Shows & Movies such as: 'Green Leaf' Oprah Whinfrey Network / 'The Quad' / 'Star' / 'Saints & Sinners' / Love & HipHop Momma Dee Music Video 'Hurt You're / T.I...
|Glenn Cannon Actor, Hawaii Five-O From 1955 - 1960, Glenn Cannon was in New York City. He appeared on Broadway in A Moon for the Misbegotten and The Good Woman of Setzuan, and Off Broadway in 20 plays, among which were the famed productions of The Three Penny Opera at the Theatre DeLys and The Iceman Cometh at Circle in the Square. His tours included leading roles in West Side Story...
|Albert Linville Actor, Damn Yankees!
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5 names.


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This easy-to-use refrence from gardening expert Barbara Pleasant covers the pros and cons of weeds, recommends proven organic methods for controlling unwanted weeds, and presents an illustrated encyclopedia of dozens of the most common weeds.
Master gardener Barbara Pleasant takes the guesswork and anxiety out of growing food, explaining in simple language exactly how to start, maintain and eventually expand an organic vegetable garden, even in the smallest of spaces. Choose one of 24 no-fail, small-scale garden plans and find out how easy it is to enjoy your own fresh food all season long!
Jeff Hertzberg and Zoë François have proved that people want to bake fresh, homemade bread with their groundbreaking, hugely popular first cookbook, Artisan Bread in Five Minutes a Day. Now, the Artisan revolution continues for those who are changing the way they eat and bake because of health concerns or lifestyle choices. Healthy Bread in Five Minutes a Day takes Hertzberg and François' established methods and applies them to breads rich in nutrition and flavor.
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Season 1, Episode 16 El Toro Grande
En route to California to purchase a bull, Joe and Hoss are ambushed---and robbed of $15,000. Joe: Michael Landon. Hoss: Dan Blocker. Ben: Lorne Greene. Xavier: Ricardo Cortez. Cayetena: Barbara Luna.
Guest Cast
Ricardo Cortez Xavier
Armand Alzamora Eduardo El Montalban
Rodd Redwing Indian Brave
Alma Beltran Maria Theresa Losaro
Barbara Luna Cayetena
Alan Roberts Epifanio Lopez
Ralph Moody Indian Chief
Penny Stanton Esmerelda Lopez
Tina Menard Floriana
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I can totally hear Van Halen in your music.
your music reminds me of pink Floyd...... that's a great thing I love pink floyd
I really dig your sound! Rock on! And what a nice language.
~ Excellent Work ~ Best Regards ~
Maks, I have decided on my own to make you #1 in Russia. If there is a committee I need to talk to you let me know. GREAT MUSIC and PRESENCE! Charlie
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Listening to " Line Of Lot " cool song man!
Check out our Songs when you get a chance. Thanks. Keep up the good work. Peace!
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Explore Garden Mosaics, Mosaic Pathway, and more!
Explore related topics
Not sure I like the stone path or bench, but the loose multicolored stones as edgers are just enough whimsy
Heart quilting design?
Garden path made of pebbles
pebble mosaics...what a beautiful idea!
What a way to Garden?: Garden mosaic detail
Alternative Gardning: How to Make Garden Stepping Stones
what to do with that old stump by vagabond.traveller.wanderer
DIY Concrete Ladybug DIY Concrete Ladybug by diyforever
Diseño empedrado


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About six years ago I stumbled upon the Competing Values Framework and have been a big fan ever since. As a leadership consultant, I immediately integrated this tool and my first client was a major, worldwide hotel where I worked with one of its busiest properties in mid-town New York City. What became quickly apparent was how easy the assessment is to conduct, how easy the results are to understand, and how interesting the whole approach can be.
Calling customer service, you immediately hear whether they really care or whether they feel indifferent. Intuitively we all know the importance of employee engagement. And research confirms it. In a study of professional service firms for instance, it was found that offices with engaged employees were up to 43% more productive. I'm a much happier customer when the service employee cares about my problem and puts in maximum effort to solve it...
The engaged employee is engaged in the work, their co-workers and the workplace. This is what employers and executives are looking for: People who are committed and participate, who think about improving, changing and so on. They have this magic "ownership" that is crucial for successful change. They have intrinsic motivation and feel responsible. They supply energy and ideas and will go the extra mile. At least, if you don't frustrate them unnecessarily and give them a chance to participate.
"The OCAI assessment made the need for market culture apparent. Team opinion shifted."
How do you take advantage of the results of your organizational culture assessment? It's essential to work these out during an OCAI workshop. Berrie Stam, regional manager at Philadelphia Care (care for disabled people), shares their story with us. "We achieved a great outcome. The team suddenly understood that they need more than just getting along well. Resistance to targets and results disappeared when the team realized that they are necessary for a successful future."
Philadelphia's management team, consisting of location managers, health care counselors, and executives participated in the useful OCAI workshop. People usually have an overall picture of their organizational culture. So does the Philadelphia team, as we saw during a meeting on a sunny day during springtime in Zwolle. They describe their culture with the key word "solidarity".
"The OCAI assessment improves understanding and incites reflection," according to Ytje Jensma. She is an operational manager at the UMCG Rehabilitation Center. "It turned out that we have a dominant clan culture—more than I expected. I have seen how important a friendly working environment is. A private hospital was understaffed in spite of good salaries, because doctors and patients were bossing the nurses around."
'Come to the cliff', he said. They said, 'We are afraid.' 'Come to the cliff', he said. They came. He pushed them. And they flew.
-Guillaume Appolinaire
Many people and organizations are still recovering from the credit crunch. The wrongdoers: banks and other financial organizations or banksters. Their image is cold, competitive, hungry for power, and even greedy. President Obama announced strong measures to control banks.
So what would the overall organizational culture be in Dutch financial organizations? We dived into our database and calculated the profiles for the financial sector: a results-oriented business indeed, but there's more focus on people than expected…. Striking is their strong wish for less focus on performance and more emphasis on innovation. But should we support that?
What's the diagnosis for the health care sector? The National Survey on Organizational Culture in Health Care in the Netherlands found that managers and professionals are dissatisfied with their organizational culture. This study, based on the Organizational Culture Assessment Instrument, was carried out among 1,613 doctors, nurses, caretakers, managers, and boards of directors. It turns out that more focus on people, market orientation, and innovation is urgently needed.
Vrouwenlaan 1068017 HS ZwolleNetherlandsPhone: +31382301503Time zone: GMT +1Chamber of commerce: 280156545VAT: NL807512813B01


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Chauvet EVE E-100Z
Chauvet EVE E-100ZDetails
Product Ref: 73598View Full Description
Buy on finance from £16.35 per month Finance from £16.35 per month
Please note: This is only available where all items in the order are dispatched in a single delivery.
To make it as easy as possible for you to get the gear you want, we offer a range of instant finance options. If you spend £250 or more online, you can choose to pay over 48, 36 or 24 months. The majority of decisions are given instantly, so you could get your new gear sooner than you thought!
Representative Example
To help you understand how our finance options work, the examples below are based on a total order value of £250:
|Deposit to Pay
|Total Payable
|Classic Credit 48 Months 14.9%
|No Deposit
|48 months: £6.83
Final payment: £6.83
No settlement fee
|Classic Credit 36 Months 15.9%
|No Deposit
|36 months: £8.65
Final payment: £8.65
No settlement fee
|Classic Credit 24 Months 18.9%
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|24 months: £12.41
Final payment: £12.41
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How to Apply
Applying for finance couldn't be easier - if your order totals £250 or more, simply proceed through the checkout as normal, then select Pay by Finance on the payment page, then click the orange APPLY FOR INSTANT FINANCE button. This will start your application with V12 Retail Finance, our finance partner, and completing this form should take approximately 10 minutes.
You should get an instant answer on whether your application has been successful, though in some rare cases V12 may need to look at the application in more detail.
Please note: This is only available on orders placed via the website consisting of a single delivery, and is not available in store or via phone currently.
Full DescriptionView Full Description
- LED Ellipsoidal shines a hard-edged, warm white spot in any direction
- Easily highlight areas by altering the projected beam with framing shutters
- Flexibility to project from any distance using the manual zoom
- Includes holders for glass and steel gobos and a gel frame
- Save time running extension cords by power linking multiple units
- Eliminate the need for DMX control with standalone operation
- DMX Channels: 1 or 3
- DMX Connectors: 3-pin XLR
- Light Source: 1 LED (warm white) 100 W (2.3 A), 50,000 hours life expectancy
- Strobe Rate: 0 to 30 Hz
- Illuminance: (14°) 15,100 lux @ 2 m, (30°) 3,800 lux @ 2 m
- Power Linking: 9 units @ 120 V; 16 units @ 230 V
- Input Voltage: 100 to 240 VAC, 50/60 Hz (auto-ranging)
- Power and Current: 106 W, 0.9 A @ 120 V, 60 Hz 103 W, 0.5 A @ 230 V, 50 Hz
- Gobo Size: M (66 mm outside, 48 mm image, 2 mm max thickness)
- Zoom Angle (Manual): 14° to 30°
- Weight: 12.6 lb (5.7 kg)
- Size: 24 x 7.2 x 16.9 in (611 x 183 x 430 mm)
- Approvals: CE, MET
What's Included?
- Power cord
- Gel Frame
- Framing Shutters
- Gobo Holders for Glass and Steel Gobos
- Warranty card
The product price includes 2 years' warranty and 30 day money back guarantee.
However, the Gear4music extended warranty scheme provides an enhanced service, and is designed to offer a higher level of protection than many 'free' warranties offered by some retailers and manufacturers.
The highlights of this enhanced warranty service are:
- Warranty is extended to 4 years from the date of delivery.
- Our Money Back guarantee is extended from 30 days to 60 days (terms apply after 30 days)
- Collection and delivery costs are included for the first 2 years
- Claim 50% cash back on the cost of the policy after 4 years if it has not been used!
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Please note that this extended warranty is currently only available on products sold within the EU or EEA, and is subject to the terms and conditions as detailed on the extended warranty certificate provided.
For customers in outlying areas of the country, the price of the Extended Warranty may be marginally higher, due to the potential for higher collection and redelivery transport costs.
This enhanced warranty is in addition to the standard Gear4music and manufacturer warranty policies, and the Gear4music terms and conditions of trade. No statement or condition of sale contained within these terms affects or restricts your statutory rights as defined under the Consumer Contracts Regulation 2014.
All Products are new unless stated, and include a minimum of one year warranty.
This offer excludes computer software, in-ear monitors that have been opened, and items supplied to your personal specification.
If you also purchase items that do not qualify for free delivery, some delivery charges may be applied to your order which will be shown in your shopping basket.


text: Home Gallery Video Ford Motor Company The Last New Fiesta in the World Ford Motor Company: The Last New Fiesta in the World Published on Apr 25, 2014 Rate this Ad You must be registered to rate the ad. Please Login or Register Now About Description In a post-apocalyptic world, a man finds something that may change his life: the Ford Fiesta. CreditsDateApr 02, 2014Agency:JWT - Brazil Client:Ford Motor CompanyVP Creative Director:Vico BenevidesCreative Director:Marcelo Jesus MarianoCopywriter:Pedro FurtadoDirector:Felipe VellascoCreative Director:Rafael FreireArt Director:Daniel Massih Tagscars Industryautomobile Latest Creativity News on Ad Age Witchy Woman: A Creative Director Reveals Her Wicca Ways Creativity Top 5: The Best Brand Ideas of the Week BBDO New York Is Named Agency of the Year at the 2017 Webby Awards Related work Ford Motor Company: Ford F-150 Raptor Global Team Blue Ford Motor Company: The Wrestler Blue Hive Ford Motor Company: Ford By Design Billboard Creator Team Detroit


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Emilie Lewis
Friday, March 04, 2016
Looking for a job in Denver while you're living in D.C.? Moving down the street can be difficult, but moving more than a few hours away brings its own set of challenges and complications. One of the most important things to think about when moving is your career. Unless you're moving for a job, you'll probably need to transition to a new company and finding something can be tricky. To help you make the switch smooth, we talked with Dan Gomez-Palacio, Director of Career Services at Columbia College to get some of his top job search tips.
One of the first things Gomez-Palacio suggested was to keep a list of potential employers and organizations as you go about your job search. Be sure to highlight any bonuses such as help relocating or great benefits. If you are looking to change professions, look into potential education requirements and, if needed, local or online universities near your relocation area.
To get started, check out websites such as Indeed.com, SimplyHired.com and Glassdoor.com. These sites "can provide a lot of information in specific geographic areas and help you not only look for open positions, but also potential employers," Gomez-Palacio said. Even if a company doesn't have your perfect position, they might still be worth looking into should a job open up. You can also use these sites as a jumping-off point to go to company specific sites.
Gomez-Palacio also suggested looking into the local Chamber of Commerce website. This will give you a good idea of the local businesses as well as helping you narrow down large and small companies depending on what you're looking for. The Commerce website can also help you see lists of the largest employers, relocation information, a list of businesses/organizations in your industry and networking opportunities.
Another way to go about the job search is to start within your industry. Many professional organizations will have local chapters which actively host career sites and networking opportunities. Most will have a free job bank and information. Gomez-Palacio pulled out a few suggestions for our Denver example:
For some cities, universities and colleges are the largest employer, and looking into their career services could also help. Many host career fairs and advertise their career partners on the website, giving you an idea of the local businesses and organizations currently hiring. You can also peruse sites such as Craigslist, but Gomez-Palacio warned that "you have to be wary of scams." Using LinkedIn lets you connect with friends and employees and do preliminary searches for specific jobs or locations. You can even use Twitter and "follow local job hashtags and job sources" for updates and potential open positions.
There are a number of ways to go about the job search, but doing it from a different city can be tricky. Be open to Skype or FaceTime interviews and try to have a firm grasp on your timeline for moving and when you can actually start your new job. Remember to start broad and narrow it down from there and don't forget about good, old-fashioned networking - good luck!
Job Search, Change, Family, Career Change, Columbia College
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Emilie Lewis
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After earning a degree in English, Emilie joined the Marketing Department at Columbia College as the Communications Coordinator. While at Columbia College her daily work included writing copy for brochures, emails, the college's website and anything else that requires words. Emilie is a voracious reader, a Ravenclaw and the proud owner of a horse named Shadowfax. She is now pursuing her master's degree in Boston.
Head shots: no longer just for performers and models. Today, more than ever, head shots are an important part of being a professional. Before you get yours taken, scroll through our guide of professional head shot best practices.
You know what red flags you need to avoid on your resume, so now let's get into the details. These three parts of your resume are essential, and it's important that you get them right.
You've decided you want a career in nursing, but now what? Our handy infographic should help you decide which path is right for you based on certifications, education needed, salary, duties and projected employment.
We think it's time to stop picturing college students only as teenagers looking for the next party. Instead, we'd like to expand that view to include what students actually look like: you.
Whether you're taking them in-seat or online, classes are a time commitment. Putting together a weekly schedule can help you manage the time commitment, and keep you on track for success!


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Pre-K applications will be accepted through May 1, 2017. You may contact our School Registrar, Linda Easter at the School Board Office with questions.
April 26th- School Board Self-Evaluation/Supt. Midyear Evaluation Work Session. 6 p.m. in the Government & School Board Administration Building
May 26th- Early Release for Students
May 29th- Memorial Day- All schools and offices closed.
June 10th- CCHS Class of 2017 Commencement Ceremony
June 16th- Last day of school for students
Charles City County Public Schools is now a nationally published school division in recognition for our efforts at data, community and stakeholder input and strategic planning and engagement. We are humbled and immensely proud of the work that this school division continues to perform and achieve. Each member of our team contributes richly to its end and we are honored to be a part of this effort and organization.
You can read the article by clicking here.
JROTC Cadets of the Charles City High Panther Honor Guard, commanded by Miles Ragin, prepare to receive officials from Cadet Command. The Panther Battalion underwent a Program for Accreditation review. This triennial review is used to evaluate Secondary Educational Institutions and JROTC Instructors, as defined by public laws, contracts, Army Regulations, and USACC policies to ensure compliance with minimum program criteria. As a result, Charles City High School JROTC program is deemed fully accredited.
© Copyright 2015+ Charles City County Public Schools


text: Went to Finns on a Friday evening. Although it was busy, we were quickly seated and waited on. The food was good and the prices reasonable. Most entrees were between 12 and 20 dollars. Bottled wine was mostly between 20 and 25 dollars. Desserts were all $6.50. Like most places they don't put prices on the beer, which annoys me,... More


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What you need to create that right-at-home feel.
Your coupon will be applied to the highest regular-price, eligible item in your cart.
Offer good for one item at regular price only. Limit one coupon per customer per day. Must present coupon at time of purchase. Offer is not valid with any other coupon, discount or previous purchase. One cut or one bolt of fabric or trim "by the yard" equals one item. Online fabric & trim discount is limited to 10 yards, single cut. Excludes Heidi Swapp™ Minc.™ Foil Applicator, CRICUT® products, Gemini Machine™ Crafters Companion, Silhouette CAMEO® Machine, candy & snack products, gum & mints, helium tanks, gift cards, custom orders, special orders, labor, rentals or class fees. Cash Value 1/10¢.
What you need to create that right-at-home feel.
While away the DIY hours, starting here.
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Dress up cupcakes or other tasty desserts with Glitter Baking Cup Wraps. These thick paper wraps feature a shiny, glittery surface, a scalloped top border, and three slots so you can adjust the circumference.
These glamorous baking cup covers easily fold and fasten around any standard cupcake treat. Use them at parties and events to make even simple sweets look positively pretty!
Package contains 24 baking cup wraps.
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Last Minute Christmas Gifts Starting from $5 to $50
December 24, 2016 No comments Article
Yes it's Christmas Eve, and if you're still looking for gifts for friends and families at the last minute (tonight or tomorrow), here are a few recommendations! More information about the brands and gifts in the links below! 😉 Shop QueDuBon bags Quote "Ena" for 30% off, min. $20 and online purchase only. Until March


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English english
Explore related topics
du vocabulaire bien utile lors des séjours en immersion Anglais in France!
English More Más
The difference between A, AN, SOME and ANY. - English Grammar Rules.
The difference between Do, Does, Did and Done in English - Grammar Chart
Learning what action verbs are - Repinned by Chesapeake College Adult Ed. We offer free classes on the Eastern Shore of MD to help you earn your GED - H.S. Diploma or Learn English (ESL) . For GED classes contact Danielle Thomas 410-829-6043 firstname.lastname@example.com For ESL classes contact Karen Luceti - 410-443-1163 firstname.lastname@example.com . www.chesapeake.edu
Dakota 2/11/16 English tenses illustration: past, present, future (simple, progressive, perfect)
Different shop names in English. Print out this cute graphic chart and turn it into flash cards!
We have some new material for you about learning how to tell the time in English. First we have this video Here is a summary chart for your reference: You can find more details about telling the ti…
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ESL, ESL vocabulary, different kinds of bread, bread, ESL conversation, ESL food


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Gateway to the dramatic Rockies, Calgary is the perfect destination for anyone looking to get away for a weekend or a few days. Calgary is filled with many activities, from winter wonderland adventures to walks that transport you through history. Don't forget to add these to your Calgary bucket list when planning your next Canada […]
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Top 5 Sights to Visit in Calgary
I For One, Welcome Our Icy Overlords: Ski Resorts To Check Out This Winter
As September grinds on, it's undeniable: winter is on its way. The light is diminishing at an accelerating rate each and every day, the first major cool spell of the upcoming fall is starting to sweep its way across North America, making people reach for their hoodies and sweaters and in the higher places in […]
Exploring The Urban Center Of Canada: Top Attractions In Toronto
At a population of just over six million, the Greater Toronto Area is the urban hub of Canada, America's neighbor to the north. As one of North America's safest cities, locals and visitors can feel at ease as they they explore its unique and bustling central districts, be it day or night. With the worst […]
Take Me To The Lake: Canada's Best Getaways By the Water
Ah, summer! With school out and work vacations well underway across much of Canada and the United States, the freedom of these long, hazy, and carefree days are the stuff of dreams for most. However, the inescapable truth of this time of year, especially north of the border, is that days like these are fleeting, […]
Remote Places On The BC Coast To Escape The World
With the latest outrage on the nightly news weighing on your psyche, combined with the drama that most people have in their lives at one point or another, the time for a break from the modern world has arrived. However, the typical city break or getaway to a popular national park won't cut the mustard […]
Five Places Where it is Impossible to Take A Bad Photo
As somebody who is interested in photography, you are always looking to get away to places across the globe where truly amazing scenes are just ripe for the shooting. Despite this, many destinations have a limited amount of subjects that make for interesting photos, or lighting conditions and other considerations can drastically affect the quality […]
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Aberrant stalk development and breakdown of tip dominance in Dictyosteliumcell lines with RNAi-silenced expression of calcineurin B
© Boeckeler et al; licensee BioMed Central Ltd. 2006
Received: 28 October 2005
Accepted: 02 March 2006
Published: 02 March 2006
Calcineurin, the Ca2+/calmodulin-dependent protein phosphatase, plays important roles in various cellular processes in lower and higher eukaryotes. Here we analyze the role of calcineurin in the development of Dictyostelium discoideum by RNAi-mediated manipulation of its expression.
The cnbA gene of Dictyostelium discoideum which encodes the regulatory B subunit (CNB) of calcineurin was silenced by RNAi. We found a variety of silencing levels of CNB in different recombinant cell lines. Reduction of CNB expression in a given cell line was correlated with developmental aberrations. Cell lines with strongly reduced protein levels developed slower than wild type cells and formed short stalks and spore heads with additional tips. Formation of short stalks results from incomplete vacuolization of prestalk cells during terminal differentiation. Expression of the stalk-specific gene ecmB was reduced in mutant cells. Aberrant stalk development is a cell autonomous defect, whereas the breakdown of tip dominance can be prevented by the presence of as low as 10% wild type cells in chimeras.
Silencing of calcineurin B in Dictyostelium by expression of RNAi reveals an unexpected link between increased intracellular calcium levels, possibly triggered by the morphogen DIF, activation of calcineurin, and the terminal stage of morphogenesis.
Calcineurin (CN), the Ca2+/calmodulin-dependent protein phosphatase, is highly conserved from yeast to mammalian cells. It consists of a catalytic (CNA) and a regulatory subunit (CNB) which form a heterodimer upon Ca2+ binding to CNB. The enzyme has been shown to play important roles in various cellular processes in lower and higher eukaryotes. In mammals these include T cell activation via dephosphorylation of the cytosolic component of nuclear factor of activated T cells (NFAT) [1], cardiac development and hypertrophy, learning and memory, and axonal pathfinding [2]. In yeast calcineurin is involved in the regulation of ion homeostasis and cell cycle control [3]. Whereas most organisms have at least two genes for the calcineurin subunits, the Dictyostelium discoideum genome contains single copy genes for CNA and CNB, whose expression changes during development of the organism. As in higher organisms CNB is required for high affinity binding of protein substrates by the holoenzyme [4]. CNA is highest in vegetative cells and after aggregation [4, 5]. The coding mRNA for CNB is processed by an unorthodox mechanism starting during early development to give rise to a shorter isoform encoding a CNB protein with a truncated N terminus that does not contain the N myristoylation consensus site found in the full-length protein [4]. Several attempts to delete the single gene for CNA (canA) in D. discoideum by insertion of selectable markers or to reduce its expression by antisense-mRNA failed (U. Kessen and R.M., unpublished results). 30-fold overexpression of the single cnbA gene which encodes the regulatory B subunit resulted in moderately accelerated multicellular development with recombinant populations completing morphogenesis about 2–3 hours earlier than wild type cells (F. Fouladi and B.W., unpublished observation). Pharmacological inhibition of D. discoideum CN using gossypol impaired growth and cellular signaling [6]. Development of wild type cells treated with 25 μM gossypol was totally blocked for more than 24 hours. Studies using the classical inhibitors, Cyclosporin A (CsA) and FK506, indicated that CN regulates Ca2+ stress-induced transcription activation of the Ca2+-ATPase patA gene [7] as well as expression of the "calcium upregulated" (Cup) class of Ca2+-binding proteins [8]. In an in vitro differentiation assay CsA and FK506 strongly inhibited stalk cell formation in the wild-type and spore formation in a sporogenous Dictyostelium strain [9]. These agents also reduced the expression of prestalk and prespore-specific transcripts, assigning a function for CN as a general activator of differentiation
During development D. discoideum cells aggregate in response to cAMP, form slugs and differentiate into two major cell types, prespore and prestalk cells. These cells organize in a spatial pattern with the prespore region localizing to the posterior and the organizer, prestalk region to the anterior of the slug. Development is completed by the formation of fruiting bodies consisting of a stalk of dead cells and the spore head. Several lines of evidence link the level of cytosolic Ca2+ (Ca2+i) to cell type differentiation. Cells in S and early G2 cell cycle phases, which show a tendency to become prestalk cells, have high intracellular calcium [10]. At the slug stage, a gradient of Ca2+i along the anterior-posterior axis has been found which is inversely correlated with the amount of sequestered Ca2+ and the calcium-sequestering activity in high-capacity Ca2+ stores [11]. High Ca2+i has been shown to be necessary for induction of the prestalk-specific gene ecmB by the morphogen, differentiation-inducing factor (DIF), and a sustained increase in Ca2+i was proposed to be responsible for prestalk-specific gene expression [12]. This work also showed that the increase in Ca2+i is brought about by DIF. It is unclear how DIF increases Ca2+i, and the mechanism by which Ca2+i ultimately leads to activation of ecmB expression has remained elusive.
RNA interference-mediated gene silencing was recently established in Dictyostelium [13]. RNAi-mediated silencing of gene expression is induced by double-stranded RNA and its processing to 23 mers which cause the degradation of endogenous target mRNAs [14, 15]. The technique appears to be well suited for the analysis of potentially essential genes in the haploid Dictyostelium genome since it can lead to different levels of silencing from almost wild type to maximally reduced levels in individual cell lines. This would allow to determine the repression level of a given gene that is just tolerable for cell survival.
We show here that a hairpin RNA construct targeted against cnbA effectively reduces the cellular CNB concentration. Recombinant cell lines showed retardation of multicellular development, culminating in fruiting bodies with short stalks and the formation of additional tips on the spore mass of developing fruiting bodies.
Isolation of cnbARNAi transgenic cell lines
Downregulation of calcineurin B
Characterization of cnbA-RNAi mutant 01–5
Cell lines with slight reduction of CNB expression displayed no or only mild phenotypic aberrations which were mainly manifest in the erection of fruiting bodies with somewhat shorter stalks than observed with wild type organisms (not shown). However, phenotypic alterations were much more prominent in cell lines with pronounced silencing of CNB. To characterize the phenotypic consequences of silencing of cnbA mRNA 10 independent transformants with drastically reduced expression of CNB were further analyzed. The phenotypes of all these transformants were very similar to cell line 01–5 which is described in more detail below.
We first compared expression of cnbA mRNA in cell line 01–5 and parental AX2 cells by Northern blotting. Figure 4 shows that the recombinant cell line had drastically reduced amounts of endogenous cnbA mRNA (0.7 kb). A strong signal corresponding to the size of the RNAi construct (1.2 kb) indicates massive expression of the transfected gene. Degradation of both RNAi and endogenous cnbA mRNA is indicated by the smear detected in the mRNA isolated from 01–5 cells.
Cell-autonomous and non cell-autonomous effects of RNAi expression
Analysis of aberrant stalks by light and electron microscopy
The defect in stalk formation which was observed in cell lines with strongly reduced CNB expression occurs due to smaller and incompletely vacuolated stalk cells and this result points to an important role for CNB or the CN holoenzyme in the regulation of stalk development in Dictyostelium. DIF-1 induces the expression of prestalk-specific genes via an increase in the intracellular free Ca2+ [12]. Based on our results we speculate that the effects of increased Ca2+i are mediated by CN. In analogy to the well-characterized role of CN in dephosphorylation of transcription factor complexes of the NFAT type we propose that calcineurin, in response to an increase in cytosolic Ca2+, dephosphorylates a cytosolic component of a transcription complex, which is then translocated to the nucleus and activates – in concert with nuclear components of the active complex – the transcription of prestalk genes. This hypothesis is directly supported by our observation that RNAi mutant 01–5 failed to properly induce expression of the prestalk-specific gene ecmB during development.
The phenotypic defects which we report here complement previous studies on the effects of mutations in the Dictyostelium glycogen synthase kinase-3 homolog GskA [18] which has been shown to promote posterior cell patterning and inhibit anterior cell differentiation [reviewed in [19]]. GskA is regulated through an intracellular protein tyrosine kinase/phosphatase pathway mediated by the cAMP receptors CAR1 and/or CAR3 and CAR4. GskA is activated by CAR1- and/or CAR3-mediated activation of the tyrosine kinase ZAK1 [20], and inactivated by CAR4-mediated activation of a protein tyrosine phosphatase [21]. GskA null cells show accelerated early development and form only stalk cells during morphogenesis [18]. Both the gskA null and the cnbA RNAi phenotypes are cell autonomous [18]; see above, fig. 8A. The direct link between GskA and CN is supported by recent results that come from analysis of the NFAT pathway. In T cells phosphorylation of NFAT by GSK3 represses NFAT-dependent gene expression by inhibition of NFAT binding to DNA [22]. Inhibition of re-export of NFAT to the cytosol when GSK3 is inactivated, leads to increased NFAT-dependent activation of gene expression [23].
Deletion of the Dictyostelium aarA gene, a homolog of mammalian β-catenin, leads to a closely similar breakdown of tip dominance and to formation of ectopic tips during late development as silencing of CNB. In both cases, the defect is non-cell autonomous [24]; see above, fig. 8B. It has been shown that AarA plays a necessary role in the formation of adherence junctions at the neck of the rising stalk [25]. β-catenin is also a transcriptional co-activator in the metazoan canonical Wnt pathway. In the control of dorsoventral axis formation in Xenopus by extracellular signals, a CN-independent NFAT mutant inhibited anterior development of the primary axis, whereas a dominant negative NFAT mutation induced ectopic dorsal axis formation and the expression of target genes of the canonical Wnt pathway, suggesting that CN and NFAT are part of the noncanonical Wnt/Ca2+ pathway which leads to inhibition of the canonical Wnt pathway upstream of β-catenin [26] (for a Wnt/Ca2+ pathway overview see [27]). It is therefore possible to propose a model for Dictyostelium tip dominance where a pathway similar to the canonical metazoan Wnt pathway is acting through CAR1/CAR3 to activate GskA, and a noncanonical "Wnt/Ca2+" pathway through CAR4 to activate CN via an intracellular Ca2+ signal. The postulated transcription factor target for CN in Dictyostelium is so far unknown.
Calcineurin B is a homolog of members of the 4 EF hand calcium sensor protein family which include, for example, recoverin, the neuronal calcium sensor-1 (NCS-1), and the plant calcineurin B-like proteins [(CBLs [28]]. It has recently been demonstrated that N myristoylation, a common feature of CNB and the Ca2+ sensor protein family, increases the cooperativity of Ca2+ binding to NCS-1 and leads to larger conformational changes upon Ca2+ binding than in the nonmyristoylated protein [29]. If Ca2+ sensing is indeed a major function of calcineurin B, the phenotypic defects observed in our cnbA RNAi mutants may be due to the failure to correctly sense the slow, sustained increase in Ca2+i from ca. 50 to ca. 150 nM during a period of about 8 hours [12], which was proposed to induce prestalk gene expression. Starting during early development D. discoideum cells express two CNB isoforms, one of them lacking the myristoylation consensus site. Further work has to show whether it is the myristoylated CNB isoform which plays a crucial role in the mediation of DIF effects on prestalk gene expression.
Dictyostelium calcineurin B knock down mutants produced smaller stalk cells and as a consequence smaller stalks due to incomplete vacuolization of stalk cells. We propose that calcineurin mediates the effects of a differentiation-inducing factor (DIF)-elicited increase in free intracellular calcium in prestalk cells via the dephosphorylation of a cytosolic transcription complex component which activates the expression of prestalk genes. Culminating fruiting bodies formed ectopic tips on the spore heads which points to a link between calcineurin and the wnt pathway in the social amoebae.
Growth and development of Dictyostelium discoideum
D. discoideum AX2 cells were grown at 22°C in AX medium [30]. Development was induced by washing cells twice with ice-cold Sørensen phosphate (SP) buffer [17 mM (KH2/Na2H)PO4], pH 6.0 and spreading 2 × 107 cells on HABP nitrocellulose filters (0.45 μm, diameter 47 mm, Millipore, Eschborn, Germany) supported by two paper filters (diameter 47 mm) soaked with buffer.
Transformation of Dictyostelium discoideum
D. discoideum AX2 cells were grown in AX medium to a density of 5 × 106 cells/ml, washed twice with ice-cold H-50 buffer, resuspended at 2 × 107 cells/ml, and 100 μl of this suspension was electroporated with 10 μg of plasmid DNA [31, 32]. Transformed cells were grown on suitable selective media and clonal populations were obtained by serial dilution in microtiter plates. In initial experiments we observed that the phenotypic aberrations which correlated with RNAi expression became gradually weaker upon serial passages of the cells. Therefore, immediately after isolation, clones of recombinant cells were grown to final numbers of approx. 1.5 × 109 (ca. 31 to 32 generations) and allowed to form spores on SP agar plates which were harvested and kept in SP buffer at -70°C until used to inoculate experimental cultures. These underwent additional 13 to 15 generations.
Construction of plasmids
cnbA cDNA was amplified by reverse transcription of total RNA from vegetative D. discoideum using the primer pair 5'GGGCCATGGGGAATCAACATTCATTATTA3' and 5'TTATTCTGACCAATTTACGCTAAG3'. The product was cloned into pDrive (Qiagen, Hilden, Germany) to obtain plasmid pBW104. Construction of the RNAi-encoding plasmid for cnbA was done in three steps. First, a fragment of 437 bp amplified with primers 5'CCGGATCCAAGCTTAAAGAACAATTAGAACAAATGA3' and 5'CCGTCGACTCACCTTCAATAATAGTTTTATCAACA3' was cloned in sense orientation into BamHI/SalI-digested pUC18 [33], yielding pKB05. Second, pKB06 was generated by ligating a longer cnbA fragment of 538 bp, amplified with primers 5'CCCTGCAGAAAGAACAATTAGAACAAATGA3' and 5'CCGTCGACTTCTGACCAATTTACGCTAAGTTTT3', in reverse orientation into pKB05 after digestion with PstI and SalI. Finally the fused fragments were subcloned into PstI/BamHI-digested pDNeoII [34] to obtain pKB07 (see below, fig. 1).
Northern blotting
Total RNA was isolated using the RNeasy Mini Kit (Qiagen, Hilden, Germany), chromatographed on 0.9% agarose gels containing 6.6% formaldehyde and blotted onto nylon membranes which were hybridized with DIG-labelled cDNA probes and stained with CSPstar as recommended by the manufacturer (all reagents from Roche Molecular Diagnostics, Mannheim, Germany).
SDS/polyacrylamide gel electrophoresis and Western blotting
Proteins were chromatographed on 15% polyacrylamide/0.1% SDS gels [35], transferred to nitrocellulose membranes and probed with 1:5000 diluted rabbit antiserum against recombinant D. discoideum calcineurin B as described [4].
Analysis of developmental chimeras
AX2 wild type and mutant strain 01–5 were cultured in AX medium. Cells were washed twice in SP buffer and resuspended at 2 × 107 cells/ml. The cells were mixed in different ratios and 2 × 107 cells spread on HABP nitrocellulose filters. Filters were incubated in the dark at 22°C for 30 h.
Analysis of fruiting body size and spore viability
Wild type and mutant strains were cultured in AX medium. Cells were washed twice in SP buffer and resuspended to 4 × 106 cells/ml. 100 μl each were spread into individual wells of a 96 well microtiter plate filled with 200 μl SP agar each. Organisms were allowed to develop for 48 h, fruiting bodies were collected, resuspended in SP buffer and frozen at -70°C. After one day spores were spread on nutrient agar with Klebsiella planticola, incubated for three days at 22°C and the numbers of clones were counted.
Embedding of fruiting bodies
Dictyostelium fruiting bodies developed on HABP filters were fixed for 3 days in a humid chamber with glutaraldehyde (2.5%) in 0.1 M Na/K phosphate buffer (pH 7.0), using both fixative solution (completely absorbed by the HABP filter) and the arising fixative vapour. The fruiting bodies were then embedded in 1.5% low melting agarose (LGT Agarose, Marine Colloids Inc., US) at 20°C. After hardening the agarose by cooling down to 0°C, blocks including fruiting bodies were prepared. The specimens were then fixed a second time for 2 h at RT with glutaraldehyde in 0.1 M Na/K phosphate buffer (pH 7.0) containing paraformaldehyde (2%) and tannic acid (0.2%). After washing 3 times with the same buffer for 20 min each a third fixation with osmium tetroxyde (1% in 50 mM Na/K phosphate buffer, pH 7.0) for 12 h at 20°C was performed, followed by repeated washing with the same buffer. The specimens were dehydrated in a graded series of ethanol. They were incubated in propylene oxide, followed by a mixture of propylene oxide/epoxide resin (v/v) and pure epoxide resin ERL [36]. The resin was polymerized for 2 days at 60°C.
Light microscopy
Transmission electron microscopy
Ultrathin sections of ca. 50–80 nm were prepared and mounted on 400 mesh cupper grids. The samples were contrasted with 2% w/v uranyl acetate for 10 min and with 0.2% w/v lead citrate in 0.2 M NaOH for 2 min. Electron micrographs were obtained with a Zeiss EM 10 microscope using Scientia negative films. The negatives were scanned with an Epson 1680 Pro scanner at a resolution of 1200 dpi.
List of abbreviations used
differentiation-inducing factor
neuronal calcium sensor-1
nuclear factor of activated T cells
glycogen synthase kinase.
We are grateful to Angela Lahrz for skilled technical assistance and to Mirko Trilling for help in the construction of pBW104. We thank Freya Kaulbars for help in microscopical analyses, Petra Dersch for critically reading the manuscript and Wolfgang Nellen for most helpful suggestions and for communicating unpublished results. This work was supported by the Deutsche Forschungsgemeinschaft (grant MU 1286/3).
Authors' Affiliations
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Excitement builds for Centenary celebrations
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Canadian Eric Walberg is known worldwide as a journalist specializing in the Middle East, Central Asia and Russia. A graduate of University of Toronto and Cambridge in economics, he has been writing on East-West relations since the 1980s.
He has lived in both the Soviet Union and Russia, and then Uzbekistan, as a UN adviser, writer, translator and lecturer. Presently a writer for the foremost Cairo newspaper, Al Ahram, he is also a regular contributor to Counterpunch, Dissident Voice, Global Research, Al-Jazeerah and Turkish Weekly, and is a commentator on Voice of the Cape radio.
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Very Urgenlty Branch Managers 35 Sales / Operations /mis / Marketing ( 1 - 20 yrs)
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RS 1,00,000 To 50,00,000 Ahmedabad, Ajmer, Alwar, Jaipur, Jodhpur, Kota-Kota, Udaipur, Delhi
Managers 35 sales / operations /MIS / marketing Experience 2 to 20 years Any company sales and operation exp. Salary pkg 3 lacs to 15 lacs Call for immediate telephonic interview.
Key Skills : sales,marketing,operations,sales manager,marketing manager,business development manager
Area Sales manager (ASM) ( 3 - 4 yrs)
Jobingo Hr Solution
RS 3,00,000 To 3,25,000 Ajmer, Jaipur, Kota-Kota
Greetings from Jobingo HR Solution…!!! This is to inform you Currently We Have Urgent Openings for Our FMCG Clients. Job Title/Designation: Area Sales manager (ASM) No. of Vacancies: 2 Education/Qualification: Must be graduate & MBA Experienc...
Key Skills : area sales manager,area sales executive,area sales incharge,area sales head
Shapers Management Services Private Limited
RS 1,00,000 To 5,00,000 Ajmer, Bikaner, Chittaurgarh, Sri Ganganagar, Hanumangarh, Jaipur, Jaisalmer, Jodhpur, Kota-Kota, Udaipur
We have Openings for Sales Managers for Leading Mnc Life Insurance Companies. please Find the Job Description Below / Overall Purpose of the Job: - to Recruit, Train & Create High Performing Fc's (financial Consultants) to Achieve Sales Targets...
Key Skills : insurance sales,banking,retail sales,sales manager,business development,retail operation,field marketing,counter sales,retail marketing


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