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Phoenix HospitalPadma Nagar,Chikuwadi-Borivali West,Landmark: Near Vrindavan Tower, Mumbai, Mumbai Get Directions
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Endometrial Ablation Procedure
Treatment of Treatment of Breast Cancer
Management of Abortion
Hormonal Replacement Therapy Treatment
Caesarean Section Procedure
Contraceptive Advice
Delivery Procedure
Treatment of Gynae Problems
Gynecology Laparoscopy Procedures
Hpv Vaccination
Treatment Of Female Sexual Problems
Treatment Of Menopause Related Issues
Treatment Of Menstrual Problems
Treatment of Mirena (Hormonal Iud)
Pap Smear Procedure
Polycystic Ovary Syndrome Treatment
Ureteroscopy Procedure
Treatment of Uterine Bleeding
Antenatal And Postnatal Exercise
Antenatal Checkup
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At some point of time we all go through a rough patch and these are the times when we suffer from certain disorders. In slightly more medical terms, sexual disorder is known as sexual dysfunction. Sexual disorders are health conditions that could surface in both males and females. This not only acts as a hurdle in the path of an alternative understanding of sexuality, but also deters spontaneous reporting of sexual problems.
Patients largely feel the need to redress such problems; the fear of being ridiculed or being mocked deters them continually. Another way of approaching the problem and its subsequent treatment is to identify the factors leading to its development. Your way of life plays an eminent role in deciding the quality of your health. You may seek help from a professional to bring about fruitful changes in your lifestyle.
Sexual disorders can be categorized as follows:
- Desire disorders: It often happens that a male or a female repeatedly feels laidback about indulging in sexual activity. The problem could be persistent in general or it could be directed towards a single partner. You may or may not develop this condition all of a sudden. Low levels of estrogen in females and low levels of testosterone in males are known to lead to such sexual irregularities.
- Sexual arousal disorders: Females at an older age or after childbirth often suffer from a sexual arousal disorder. The condition could also be found in young adults. It is characterized by a dry vagina and the lack of pleasure during foreplay or intercourse. Amid males this problem is prevalently known as erectile dysfunction. A man might not be able to enjoy physical contact with his partner. Men usually face difficulties in having a proper erection. The male organ in such a case is only partly erected.
- Sexual orgasm disorders: If you are on certain medications or have been experiencing frequent bouts of anxiety, attaining climax can be a headache. Patients of this disorder either fail to have an orgasm or it is extremely delayed.
- Sexual pain disorders: Pain disorders are more common in women. In most cases, painful intercourse is the result of lack of arousal. If the vagina is not sufficiently lubricated, a woman might experience severe pain. Pain can also be the symptom of a serious underlying disease like vaginismus. A medical condition where a man faces painful erection due to excess accumulation of blood in the penis is referred to as priapism.
1. Get a goodnight's sleep -" poor sleep takes a toll on everything, from your job performance to your daily chores, and especially your memory, says Dr. Adarsh kumar, internal medicine, national heart institute. Sleep is a key time for the brain to solidify the connections between neurons, thus helping us remember more of our tasks. Rule of thumb: get 7-8 hours of sleep everyday. And yes, naps count. Experts are of the opinion that if you do only one thing to improve your memory, getting more sleep should be it. If you have trouble sleeping fast, drink a warm cup of milk a half hour before hitting the sheets. Yes, this age-old remedy actually works. Now stop staring at the ceiling, and slip into slumber.
2. Jog your memory - literally. Exercise increases your heart rate which gets blood flowing to your brain, thus keeping your memory sharp. Running, swimming, biking - any form of exercise - for at least 30 minutes helps enlarge the hippocampus, which is regarded as the'memory center of the brain. In fact, physical activities that require hand-eye coordination or complex motor skills are particularly beneficial for brain building. If you don't have time for a full workout, squeeze in a 10-minute walk around the block in your schedule or a few jumping jacks. It's enough to reboot your brain.
3. Quit multitasking - can't find your keys? it's probably because you weren't paying attention when you put them down. When you're juggling too many things, you're bound to forget. As it turns out, the brain doesn't actually multitask. Instead, it switches focus from one thing to the other, which is why it is difficult to read a book and hold a conversation at the same time. Multitasking will slow you down, so make it a point to concentrate on the task at hand. It's crucial. Studies suggest to say it out loud" I left my keys on the dresser, etc. So the brain can process it. Your brain actually need about eight seconds to commit a piece of information to your memory, so if you're talking on the phone and carrying groceries when you put down your car keys, you're unlikely to remember where you left them.
4. Go back to your roots - ashwagandha, a go-to choice in ayurvedic medicine, is known for promoting memory since it helps prevent nerve cell damage. Dr. Ashutosh gautam, clinical operations and coordination manager at baidyanath says" ashwagandha can be taken in the form of powder or tablet. It also improves the brain's memory functions like attention and concentration, hence helping with the symptoms of parkinson's, alzheimer's and other neuro degenerative diseases. Fish oil, has also been associated with encouraging the growth of neurons and lowering the risk of dementia. Fish oil contains dha, an omega-3 fatty acid which helps improve your memory power.
5. Use mnemonic devices - mnemonic devices are tools which help you memorize in an easier format - words, lists, concepts.
Acronyms: they are basically abbreviations used a word to help you jog your memory. For example: cart can be carrots, apples, raspberries and tomatoes, which can be used to remember your grocery list.
Rhymes: if you need to remember a name, get creative" mary loves cherry" or" simon is a fireman.
Acrostics: these are life savers during exams, especially. Whenever you need to mug up a sentence, combine the initial letters and use as a memory cue. For example: how we all remembered the 9 planets during childhood" my very educated mother just showed us nine planets.
6. Get organized - if your house is in a mess, you're more likely to forget things. Jot down tasks, declutter your home and note down appointments. Set aside a particular place at home to keep your keys, and limit distractions. Live by to-do lists, keep them upto date and check off the items you've completed. Physically writing down new information actually helps reinforce it.
7. Meditation is key - according to a 2015 study from the ucla brain mapping center, the brain starts to decline in your 20s and continues to decrease both in size and volume. Meditating regularly delays cognitive decline and prevents neurodegenerative diseases like dementia, alzheimer's and parkinson's. Meditation produces a positive charge in the brain's gray matter overtime, which is important for memory, learning and self-awareness. In addition, meditation has been shown to reduce stress, which can do a number on memory.
8. Stay mentally active - note: crossword puzzles and sudoku are your new best friend. Challenge your brain, take a different route to work, learn a new language, read a section of the newspaper you usually skip, do things out of the ordinary. Stay engaged, because mentally stimulating activities help keep your brain in shape - and might even keep memory loss at bay. People who are cognitively active have better memory as they age, it's true. So quiz yourself, flex your brain and improve your memory power.
9. Balance your stress - chronic stress and depression, both contribute to memory loss and the destruction of brain cells. One of the best things you can do is to laugh. Yes, it's that simple. Laughter engaged multiple regions of the brain and simultaneously reduces stress. Social interaction also helps ward off depression and stress, so look for opportunities to get together with family, loved ones, friends and work colleagues. When you're invited to share a meal or attend an event, go!
10. Food for thought - did you know that the brain is an energy hungry organ? despite comprising only 2% of the body's weight, the brain gobbles up more than 20% of daily energy intake. So a healthy diet might be as good for your brain as it is for your overall health, and eating right may in fact be more important than you think. After all, you are what you eat. The brain demands a constant supply of glucose which is obtained from recently eaten carbohydrates like whole grains, fruits and greens. Because when the glucose level drops, it results in confused thinking. No, this does not give you the license to slurp on sugary drinks. Instead eat throughout the day to optimize brain power- not too much, not too little. Memory superfoods include antioxidant-rich, colorful fruits, green leafy vegetables and whole grains which protect your brain from harmful free radicals. Choose low-fat protein sources such as fish and drink at least 8 glasses of water daily since dehydration can lead to memory loss and confusion.
Vitamin c and b: vitamin c, found in citrus fruits, is associated with mental agility, whereas vitamin b is known to guard against age-related brain shrinkage and cognitive impairment. Load up on blackcurrants, fish, green leafy vegetables, mushrooms, peanuts, sesame seeds and eggs to rev up your brain power.
Nuts and seeds: a handful of seeds and nuts may help improve your memory power significantly. Pumpkin seeds are loaded with zinc that play a great role in sharpening your memory. Brain-shaped walnuts are a good source of omega-3 and other essential nutrients crucial for brain functioning and memory skills. Sunflower seeds are good sources of vitamin e, just sprinkle them on top of your salad to give your brain a boost. Even peanuts are packed with vitamin e, a potent antioxidant. Almonds and hazelnuts also help boost memory.
Berries: blueberries are the top source of substances called anthocyanins which are brain-boosting antioxidants. Eating blueberries daily can help combat against the onset of short term memory loss. Even strawberries, when consumed regularly can help delay age-related memory decline.
Green vegetables: broccoli, kale, collard greens, spinach - all green vegetables are full of iron, vitamin e, k and b9 (folate), and phytonutrients like vitamin c which are extremely important for brain cell development. Vitamin k is known to be helpful in cognitive enhancement and increasing mental alertness.
Avocados: enriched with vitamin e, avocados are loaded with antioxidants which help in keeping the brain healthy and alert. Creamy avocados are also associated with lowering the risk of alzheimer's.
Tomatoes: tomatoes are a good source of lycopene which acts against brain cell degeneration and aids in the maintenance and production of new brain cells.
Whole grains: whole grains are considered to be the powerhouse of energy and help you focus better. On consuming fiber-rich whole grains, energy is released in the body in the form of sugar which assists the brain in functioning properly, keeping us alert.
Fish: omega-3 fatty acids as well as dha and epa are essential for healthy brain functioning as well as for healthy brain neurons. Include salmon, mackerel, tuna and other fish in your diet. Vegetarian? you can substitute with soybean oil and flaxseed oil.
Red wine: resveratrol, a powerful antioxidant, predominantly present in red wine protects against cell damage and alzheimer's disease. So, consuming a glass of red wine a day may actually keep memory loss at bay.
migraine-: ayurvedic view
Migraine headaches are commonly defined as episodic or recurrent severe headaches, typically of a throbbing or stabbing nature, usually on one side of the head, the neck or behind the eye.
~~migraine pain commonly lasts for about 6 to 48hrs.
~~migraine is triggered by some factors which result in excessive stimulation (constriction and sudden dilatation) of blood vessels in the head.
~~migraine may or may not be preceded by an aura; aura may cause visual disturbances such as flashing lights, zig-zags, sparkles or blurring of a visual field.
~~migraine is associated with nausea, vomiting, dizziness and or cognitive disturbances.
~~patient may also experience chills, sweating, fatigue and weakness etc.
A. Causes-:
Improper metabolism, suppression of natural urges, anger, jealousy, grief, poor elimination, intake of dry, pungent and salty food, intake of polluted food, mental and physical stress, viewing of tv for long periods, reading with insufficient light, sleeplessness etc.
B. Origin of migraine headaches:
Due to the causative factors, pitta dosha obstructs the flow of vata dosha in the brain, causing throbbing pain.
C. Pain management-:
1. Avoiding the cause and trigger factors.
2. Keeping a diary record of specific foods were eaten, environmental exposure, sleep patterns may help to identify personal trigger factors.
3. Sneha nasya therapy at home-: sneha nasya, instillation of medicated oils to the nostrils by shadbindu tailam or anu tailam yield significant benefits.
4. Apply a paste of freshly ground clay or freshly ground sandalwood to the forehead, let dry, rub off by hand and wash.
5. Mix a quarter teaspoon of clove powder with 1 teaspoon of cinnamon oil. Apply this paste on the affected area for 20-30 minutes.
6. Add chamomile powder to 1 cup of boiling water. Transfer the liquid to a cup. Drink when hot. It gives fast relief from migraine.
7. Take half glass of carrot juice. Add half glass of spinach juice. Mix well. Drink this juice twice daily.
8. Grind 10-12 grains of black pepper and 10-12 grains of rice with water to make it a paste. Apply this paste on the affected area of the head for 15-20 minutes.
9. Sugar powder 16 parts, vamshalochana 8 parts, pippali (long pepper) 4 parts, ela (cardamom) 2 parts, cinnamon - 1 part. Make fine powder and mix it, keep in tight moisture-free container. Take half teaspoon thrice daily with honey.
10. Aromatherapy- pitta calming aromas typically flower fragrances like lily, cloves, lotus, lavender, peppermint, rose, sandalwood are beneficial.
11. According to Ayurveda virechan (laxative therapy) is an effective way to reduce excess pitta. Castor oil or warm milk with ghee or triphla at bedtime is helpful.
12. Practicing meditation is highly recommended.
13. Morning and evening walk followed by brisk exercise is recommended.
14. Avoid hard physical exertional exercises and aggressive sports competitions.
15. Avoid working for long hours continuously; take short breaks. Have a good night's sleep (8hrs) in a dark room.
D. Diet:
~~apple, mango, papaya, cheeku, guava.
~~salad 1hr before lunch, 1hr before dinner.
~~buttermilk, boiled rice sauted with cinnamon, cumin seeds and garlic or asfoetida are good in the daytime.
~~have warm and easily digestible foods, boiled and steamed vegetables, soups, vegetable juices, porridge, brown rice and whole-wheat flour.
~~avoid yogurt, especially at night.
~~avoid refined, oily, spicy, cold, sour foods (fermented, pickled) and stale food, aerated drinks.
~~avoid coffee, tea, alcohol, smoking.
~~avoid cheese, chocolate and wine.
E. Yogasanas and pranayama:
Hastapadasana, setu bandhasana, marjariasana, paschimottanasana, bhastrika and anulom-vilom pranayama.
Note: when suffering from a migraine attack:
~lie down in a dark, quiet place and get some sleep.
~~avoid eating.
~~drink only water.
~~apply hot or cold compresses.
~~ consult doctor
She has diabetes from childhood. If she gets pregnant is it safe to do Caesarean? And what are the chances that the born baby will also be affected with diabetes?
Hello Doctor, Last month, I started doing more exercise. Lost around 1 kg and around abdomen. But now I am delayed on my period by 10 days. Did 5 home pregnancy tests and all are negative. Not involved in unsafe sex last month. So, attributed my delay in period to rigorous exercising. But pimples like never before. And some light white vaginal discharge. What is this discharge about, should I be worried for pregnancy. Also, I was diagnosed with mild pcod. Also, there was a small spotting last month around 14th day.
What if the sonography reports show twins but the nutrition bag is just one even aftr 11weeks of pregnancy?
I am 22 years old female, I missed the periods 12 days before today is 13th day and I checked through pregnancy test kit as it came positive. As I'm very young I don't need pregnancy. Will you please prescribe any medicine for my abortion.
I had 2 miscarriages. My al reports are finenow m planning for my pregnancy again n n very scared is there any tips so that I can hv successful pregnancy this time.
We are planning for a baby by next year, but we both are having hypo thyroid n I get always severe period cramp, so I want to know that any test should I do or any suggestion regarding this?
~ Ayurvedic Name: Dandak Jwara
~ Other Names: Dandy fever,Three-day fever, Breakbone fever.
~What's is a Dengue Fever?
1-Dengue fever is a disease caused by one of a number of viruses.
2-These viruses are spread by the bite of mosquitoes and are transmitted to the human body.
3-The disease is very common in tropical and subtropical areas of the world (including some islands in the Caribbean, Mexico, most countries of South and Central America, the Pacific, Asia and parts of tropical Africa). In these locations, the dengue fever arbovirus is endemic, meaning that the dengue fever virus naturally and consistently lives in that location and the virus are not carried by mosquitoes.
~Who gets Dengue Fever?
Dengue fever can occur to people of all ages. Children usually have a milder disease than adults.
~What's are the causes of Dengue Fever ?
When a mosquito carrying dengue fever virus bites a healthy human, the virus travels in the blood stream and then starts multiplying itself in the whole body. Dengue fever is not contagious, which means one person cannot pass it directly to another.
Dengue fever is also a type of arbovirus, which is short for arthropod-borne virus. The main virus causing dengue fever is carried by a mosquito named "Aedes aegypti" which transmits the virus to humans through its bite. An organism that carries a disease, without actually developing the disease itself, is called a vector. Aedes aegypti works as a vector for transmitting the disease.
An another cause for dengue fever is a virus name flaviviruses. More than 100 million cases of dengue fever occur every year in a whole world. A little bit percent of these develop into dengue hemorrhagic fever. It is possible for a traveler who has returned to the United States to pass the infection to someone who has not traveled. Risk factors for dengue hemorrhagic fever include having antibodies to dengue virus from prior infection and being younger than 12, female, or Caucasian.
~What's are the symptoms of Dengue Fever?
The symptoms of dengue fever are generally seen in around three days after transition of virus to humans through mosquitoes bite and after that patient starts feeling very weak. Dengue fever, however, is not a fatal disease.
Dengue fever may vary on an individual basis for each patient. One should always remember before looking for treatment that over-medication in dengue fever is likely to lead to further complications and some times it may also turn slight serious. A mild attack of the fever may pass off without having to administer any medicine.
~Some of the main symptoms of dengue fever are listed below:
Dengue fever usually starts suddenly with a rapidly climbing high fever, that's why the temperature in dengue fever is called a 'saddleback' type temperature.
severe headaches.
Retro-orbital pain behind the eye.
nausea & vomiting.
loss of appetite.
rashes develop on the feet or legs 3 to 4 days after the beginning of the fever.
Swelling and pain in muscles and joints.
The joint pain in the body has given dengue fever the name that is "breakbone fever".The common symptoms of dengue fever may go in around 10 days, but complete recovery from dengue fever can take more than a month.
~Herbal Treatments for Dengue Fever
There is no specific treatment for the treatment of classical dengue fever, and like most people you will also recover completely within 2 weeks. For severe dengue symptoms, including shock and coma, early and aggressive emergency treatment with fluid and electrolyte replacement can be lifesaving.
Some treatment options that may enhance the curing of classical dengue fever are listed:
1) Mix 360 mg of Shunthi Churna with 125 mg of Hinguleshwar and now take this mixture at least 4 times daily, you may take with tea or hot water. If case the temperature of the patient rises beyond 104°F (40°C), then you may apply cold pad, on the forehead till the temperature subsides.
2) Another ayurvedic medicine used for treatment of dengue fever is tulsi (holy basil). Medicinal properties of tulsi are very useful in ayurvedic therapy. The leaves of tulsi are used to produce soothing effect on the nervous system. Tulsi leaves strengthen the stomach and induce copious perspiration.
3) Amrutharistam,Amruthottaram kashayam,sudarsanam gulika,Pachanamrutham kashayam can be given for treatment.
Adenoids refer to the mass of lymphatic tissue situated at the intersection of the back of the nose and throat. They are a part of the body's immune system and play a noteworthy part in battling infections in kids. Adenoids fight off the microbes and infection that enter the body through the nose. They typically occur in children of five to seven years of age and from that point, they begin to decrease. They vanish when the child gets older. This is the physiological development and relapse of the adenoids. However, permanent adenoids are an issue in view of a number of annoying symptoms they show in a child. The major among these are breathing through the mouth and wheezing because of nasal blockage.
Homeopathic medicines are a very safe option than surgery in kids with extended adenoids. It has been noted that adenoids, when surgically removed, tend to regrow. Homeopathic medicines focus on strengthening the child's immune system. The cures are as follows:
- Baryta Carb: Baryta Carb is one of the top recorded homeopathic medications for swollen adenoids. It is likewise the medicine to endorse where both tonsils and adenoids are broadened. The child feels sensitive to ice in extremes. The tiniest exposure to icy air causes cough. Cough also shows up at the slightest change in climate. Sneezing and thick yellow nasal release may be seen. Pain in the throat increases on gulping.
- Calcarea Carb: Calcarea Carb is another brilliant homeopathic medicine for amplified adenoids with the tendency to get infected. Calcarea Carb will be recommended where the youngster is exceptionally helpless to cold and falls sick after each such introduction. This weak immune system for diseases brings about broadened adenoids or tonsils.
- Agraphis Nutans and Merc Sol: Agraphis Nutans and Merc Sol are extremely helpful homeopathic medicines for treating ear grumblings because of expanded adenoids. Agraphis Nutans is the best treatment for deafness because of expanded adenoids. Homeopathic prescription Merc Sol is highly used in case of an ear infection connected with swollen adenoids.
- Kali Sulphuricum: Kali Sulphuricum is among one of the top recorded homeopathic solutions for adenoids that regrow after surgery. The symptoms controlling utilisation of Kali Sulphuricum are a blocked nose, mouth breathing and wheezing. Yellow coloured nasal release is also common.
- Ammonium Carb and Sambucus Nigra: Ammonium Carb and Sambucus Nigra are incredibly helpful homeopathic medicines for developed adenoids. A youngster that needs these drugs will have an increased nasal blockage. The nasal hindrance is most set apart around evening time. Breathing through the mouth because of a blocked nose is also very common. Ammonium Carb is likewise used for a seeping nose, particularly in the morning. If you wish to discuss about any specific problem, you can consult a homeopath.
I sex with my husband in period 3rd day on 5th September it start itching till it is not stopped what can I do please help me is it a serious problem help me.
Hi my sis her age is 30 years she recently got married 3months back so due to her job she & her hubby stays in different cities, now they are planning for kids so her Dr. Has advised them as they couldn't not meet regularly each time, to have injections to form eggs to my sister & have sexs on those days so that she gets pregnant, but my sister is not sure to have injection as it. Is safe or not, or in comings days will it creates any problems. So please advice her.
Im checked my urine using with the pregnancy test kit. The result is positive. Then im checked from the laboratory then also the result is positive. But my period date is not over. Before my period date I get the result. My last month period started from 19 th january and ended on 26 th january. Is it ture result.
For a married couple, having a child is the next step towards completing the family picture. However, for various reasons, some attributed to the male and some to the female, this picture remains incomplete.
Talking to a doctor is one of the best starting points. There could be deep rooted causes for this issue and homeopathy aims at treating the root cause and not just infertility. A good homeopath will ask you numerous questions to find out associated symptoms, family history, etc. and then arrive at a remedy that would work best for you. This is a highly customised therapy and so self-medication based on a friend or family member's recommendation is best avoided.
Infertility in females is often caused by the following reasons:
- Irregular menstruation
- Hormonal imbalance
- Advancing age
- Emotional stress
- Obesity
- Excessive smoking
- Alcohol consumption
- Sexually transmitted diseases (Chlamydia, gonorrhoea)
- Structural abnormalities in the pelvic area including fibroids, pelvic adhesions, blocked fallopian tubes, etc.
- Endometriosis
- Polycystic Ovarian Syndrome (PCOS)
- Pelvic Inflammatory disease
- Thyroid disorders
- Diabetes
With more and more women coming out in the open to discuss, researchers have started working towards identifying the problem and working towards the treatment. The success rate of homeopathy in treating infertility is also on the rise. As with any medical condition, homeopathy treats not just the problem or symptom at hand, but the person holistically. Read on to know some of the common homeopathic remedies, but make sure you have a detailed discussion with your doctor to identify what would work best for you. Self-medication is best avoided.
- In women with reduced sexual drive, Agnus and Sepia are widely used. The vaginal is extremely dry which could be painful during sex. There is also a bearing down sensation of the uterus in these women.
- In women with reduced menstruation, Pulsatilla and Sepia are widely used. The periods are never on the expected date, and when they occur, the flow is quite scanty and suppressed. Pulsatilla is also used when ovarian cysts are present. Sepia is useful in women that are prone to miscarriages.
- In women with excessive menstruation, Calcarea and Aletris are widely used. The periods happen before time, is too long with profuse bleeding. This excessive bleeding also causes anaemia, weakness, and fatigue. These women might also have frequent abortions.
- In women who have experienced a miscarriage in their third month earlier, with uterine and ovarian inflammation, Sabina is used.
- In women who are not able to retain sperms, Natrum carb is useful. There could be an offensive smelling vaginal discharge which is also very irritating and itching.
These are just some of the common remedies, but there are more, and each patient would require different therapy based on associated symptoms. If you wish to discuss about any specific problem, you can consult a doctor and ask a free question.


text:
Building an l shaped shed photos
Posted By Cira H. on 27.04.2013
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Building an l shaped shed photos - shading
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Family Law Titles
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These Regulations revoke and replace the Fostering Services Regulations 2002. They make provision about the way in which fostering agencies and local authority fostering services are managed and run, and in particular about the process for approving foster carers and safeguarding children placed with foster carers.
These Regulations come into force on 1 April 2011.
"the principal (monthly) periodical dealing with contemporary issues" Sir Mark Potter P
This ready reference guide for all family court practitioners and judges provides a portable...


text: Nov 18, 2011 GG002 by Henry Leave a comment → Nov 18, 2011 by Henry Leave a comment → /?php if (isset($src)){ ?> /?php } ?> Related Posts Kicking Off ESP PhotoWeek 2012 UPDATE: Live images from the road – click here for the gallery I am sure you guys already know that we now have five photographers in our team. Read more → Samuel + Diana Wedding Never had a whole after party room dedicated to a football club before :D But Samuel is one heck of a hooligan. Too bad Liverpool lost that nite, hehehe… Read more → My First Kill It was a first for me, to kill an animal myself in order to eat it. I found myself shouting "Any last words?" to the crab three times because I was so nervous about to do this horrible-horrible thing.But in the end, it was an enlightening experience. I learned that:1. Crabs don't want to die […] Read more →


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Keith Hanna at Edinburgh Fringe 2015 in 'Eejit of Love'
August 27th, 2015 by admin
Keith Hanna takes to the stage in this year's Edinburgh Fringe
Festival as Cowboy in the satirical musical comedy Eejit of Love.
Eejit of Love is written and directed by Jody Trehy and was originally developed at Greenwich Theatre and produced as part of the Dublin Fringe Festival where it attracted fans from tweenies to grannies as audiences returned two or three times to sing along with the extravagant characters.
The show follows Billy and Eileen, two Irish country bumpkins who find themselves swept up in the allure of reality TV, testing their relationship and their own willpower!
For more information and to book tickets click: https://edinburghfestival.list.co.uk/event/508411-eejit-of-love/


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The Upper East Side continues to increase in prestige.
Yet, even with its affluent and dignified aesthetic, The Upper East Side maintains an outstanding sense of community and warmth. Families choose this neighborhood for its five-star schools, quiet atmosphere, and marvelously close proximity to Central Park.learn more about this neighborhood
#6Neighborhood popularity:based on Naked Apartments searches
Building Mix
- 27% Pre War
- 21% Luxury Highrise
- 20% Walk-Up
- 12% Elevator/Laundry Building
- 5% Brownstone
Legal Disclosure: Neighborhood descriptions and data are created exclusively by Naked Apartments without contributions or approval from the agents listed on this page.
East 69th st by Gryffindor is licensed under CC BY-SA 3.0
Elegant Two Bedroom (convertible 3 bedroom) with outstanding Central park views. Marble entry and gallery take you to a 26' Living room. On the other side Formal Dining Room, Library or 3rd bedroom, separate Breakfast Room and Gourmet granite counter topped kitchen. Two full beautiful marble baths (Master bath with separate glass shower and tub, powder room and generous closets throughout including 2 walk-ins. Full-Service Doorman and Concierge Building with Attended Garage & Valet Service, Half acre resident's garden, Health club and Common storage room. Contact MAJA today for an appointment.
Apartment Details
$16,600, Two Bedrooms, 2.0 Bathrooms
Last Updated Over 4 Years
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- Web ID: 209799
Doorman, Elevator, Garage, Gym, Outdoor Space, Storage Room
Nearby Subway Stations
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N Q R 4 5 6Lexington Av-59 St (0.37 mi)
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Firefighters were called to a car fire in Belmont this morning.
It took them five minutes to tackle the blaze in a Chrysler Voyager in Hulverston Close which happened at about 12.10am.
They then waited for the police to arrive as the fire is suspected to have been started deliberately.
- Water stations, urinal bays and MP runners: The 2017 London Marathon in numbers
- Record number of runners to take on today's London Marathon
- Fly-tipping getting worse since new bin collections introduced, residents claim
- Five arrested and drugs seized in series of raids across Surrey
- Man with 'distinctive Jesus tattoo' wanted in connection with assault


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Deadline extended for Webby Awards, recognising "excellence on the internet"
The deadline for entries to The Webby Awards has been extended until 27 January, meaning you have a couple of extra weeks to be considered for the globally recognised prize.
The annual awards, now in its 21st year, recognises excellence across six types: websites; advertising, media and PR; film and video; mobile sites and apps; social; and podcasts and digital audio. Winners are selected by members of the International Academy of Digital Arts and Sciences, based on criteria including creativity, engagement, reach and functionality. There are over 100 categories, in which each entrant has two chances to win: by judging panel vote and public vote.
Last year's awards honoured 378 winners including a star-studded line-up for the Special Achievement prizes, such as Lena Dunham and Jenni Konner, who were recognised for their work on Lenny Letter.
Alicia Garza, Opal Tometi and Patrisse Cullors, founders of the #BlackLivesMatter organisation, were awarded The Webby Social Movement of the Year for their roles in the Black Lives Matter movement.
R/GA was named Agency of the Year, and The Onion was given the Lifetime Achievement award.
Shane Smith, founder and CEO of Vice Media, was handed the Webby for Best Cultural Website by rapper Action Bronson (pictured above).
Entries close 27 January 2017; find out more about the competition and criteria here.
- Best of the Web is here, and so is the weekend!
- Animator Saiman Chow's trippy idents for Adult Swim's Rick and Morty
- Friday Mixtape: Legendary record label, 4AD
- Risograph photograph journal, This is the Same Ocean, returns with a sixth issue
- Illustrator Gizem Vural impresses us with attention-grabbing personal work and commissions
- Colophon Foundry re-releases its road-sign inspired typeface, Montefiore, with new specimen
- Jon Burgerman on his utterly brilliant Instagram experiments
- The photos Juergen Teller took while waiting for Rihanna
- Animator and director James Curran's amusing 30-day Gifathon project in Tokyo
- "Before I was a graphic designer I had nearly no idea what one was": meet Austin Redman
- Meet Berlin-based studio Büro Bum Bum
- Matthew Raw: the east London artist making clay great again


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A few hours after a gunman walked up to a young TV reporter and her cameraman and shot them dead on live TV, a message appeared on Twitter. Short and unpunctuated, it read: "I filmed the shooting see Facebook"
The grim narrative that unfolded Wednesday was shocking to watch. Part gun-related workplace dispute and part racial hate crime — the gunman was African-American, the victims white — the horrifying shootings played out on live local TV, then on social media, then again on TV everywhere, as police pi...
Also read: Georgia officer seen in video hitting man asks for job back
Read more
Also read:
19:04 Funeral services held for Ohio victim of Facebook killer16
10:13 Paris shooting: Election campaign halts as investigation gathers pace27
20:09 SEE IT: Teenage boy gets stomped on during Coney Island brawl16
23:37 Three people killed in 'unprovoked' Fresno shooting rampage 'motivated by hate,' police say20


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This event is an exciting development of the 'On Your Doorstep' themes, this time specifically focussing on the experience of rural communities and rural churches. It will resource churches to workContinue reading
Tag: Rural
Meeting Gloucestershire's farming families
Bishop Rachel has spent time this week visiting local farming families
Brexit and the Rural Future – Implications for churches and communities
A day conference exploring the issues of Brexit and likely effects on rural communities, churches, agriculture and the environment. This is not a day to debate the pros and consContinue reading
Growing the Rural Church…
I've just returned from my first ever General Synod of the Church of England (Suffragan Bishops like myself are not automatically members of Synod, unlike Diocesan Bishops). It was anContinue reading


text: Home > Focus Agri-tourism in Aquino's SONA? Posted at Jul 20 2013 09:05 PM | Updated as of Jul 21 2013 05:05 AM Share Save Facebook Twitter GPlus LinkedIn Watch also in iWantv or TFC.tv Some members of the private sector hope President Aquino will put the spotlight on the so-called "agri-tourism" in the State of the Nation Address on Monday. Abner Mercado explains agri-tourism in this report. -- Prime Time, ANC, July 20, 2013 Share Save Facebook Share on Twitter GPlus LinkedIn Read More: SONA pnoy agri-tourism /news/04/26/17/afp-bodies-of-high-value-terror-suspects-found-in-lanao-clash-site/sports/04/26/17/thai-imports-power-smashers-ready-for-pvl/video/news/04/26/17/in-the-loop-pnp-officer-and-suspected-abu-sayyaf-member-to-face-charges/news/04/26/17/police-official-tagged-in-jee-slay-back-in-camp-crame-as-detainee/news/04/26/17/ph-rises-in-press-freedom-index-but-worth-watching-due-to-new-strongman-duterte-rsf


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These Daily Kos posts are just too fascinating to ignore. Electing an African American President, if it happens, will be a big deal....it's not like racism is dead or anything. Check this out.
Friday, October 31, 2008
Black People are Voting!
Posted by Jim Crocker at 5:43 PM
Labels: black vote, how to vote, Obama, US Election
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72, Lansdale, PAWho's Who of Professionals, 2 yrs. in a row,' 03 and ' 04...... Who's Who Executives in Telecommunications '98 .......my marriage was annulled many years ago but I have found no marriage partner per se...have had a few long term relationships and proposals but marriage never happened. My career has kept me busy through the long hard years working in advertising (print, radio, t.v.) in Pa., Ct., Ca., and Fla. and telecom/data in PA. but nothing long term, either the company closed or I left due to medical problems after 2-3 yrs. I enjoy my children and grandchildren and see them quite often considering all of them live on the west coast. My hobbies include travel, gourmet cooking and antiques. I am a typical 60's type of person, a first child, somewhat of a perfectionist : I worked on the Republican National Convention in Phila. as a media specialist, love Europe the last place visited was Prague, good food and wine,a good book,beautiful music and God.( I am a Christian). I enjoy geneology and am a member of "D.O.L.L.U.S., my great-grandfather was a commissioned officer during the civil war. I am semi-retired and currently consult from a home office to businesses for the banking industry.
49, Downingtown, PAI am passionate about my Catholic faith and my children. I feel strongly that I am responsible in helping my children and my future spouse to get to heaven. I hope to find someone who shares a love of our beautiful Catholic faith and sees the importance of a strong and loving family with Jesus at the center. My parents and family have been the most influential in my life. I am a school teacher of 1st grade. Love teaching children and seeing them growing educationally and developmentally. During my leisure time I love to go out to eat, go to movies, spend time outdoors as well as spend time with my children at their activities or at home together.
54, Warrington, PAI am a kind, caring, and compassionate person. I am a dedicated Catholic who believes that Jesus is the center of our lives. I am a hard working professional immersed in an occupation that provides a service to others to reach their fulliest potential in education. I have been in many long term relationships throughout my life; I am now seeking a partner who is willing to fully commit themeselves to true companionship. I believe that I am a very simplistic person who is well rounded in life; I see the goodness in all people. My life has been very full with my career, family, and friends. I am now at a point in my life where I would like to share my life with that special someone.
45, Warminster, PAHi, my name is Steph. I am devout, and am looking forward to finding a man who is devout, as well. I look forward to knowing someone who lives the acronym JOY: Jesus first, others second, yourself third. I believe in serving others, and I try to lead by example when it comes to this. My passion is my Faith, but I desire to find someone who can lead me & not always look to me for leadership. That may not make sense, but that's because sometimes I have a hard time putting into words what I think in my head & feel in my heart. I have a dry sense of humor & can be sarcastic, but I have toned that down somewhat, as I found that it can be a turnoff. I can also be silly, which sometimes can be perceived as immature, but I see it as young at heart. I'm currently reading Matthew Kelly's REDISCOVER CATHOLICISM. So far it's an excellent book. I also like motorcycles & cars. My older son is learning to be a mechanic & loves cars, and he & I will sit and watch some of the shows on Velocity. I like to think of myself as a closet gearhead!
62, Oxford, PAWidowed Mother of 3 wonderful college students. Family focused, eclectic in listening to music, enjoy easy hikes, reading and watching movies. I grew up on Long Island in a large Irish Catholic family and went to college upstate NY. I work in the medical device industry and I am just starting to look around, wondering God has another spouse in mind for me.
25, Ridley Park, PAHi, I'm Stephanie, but most people call me Steph. My one true passion is working with children, teenagers especially. My goal in life is to become a high school math teacher and a girls softball coach. I am currently on the road to making my dreams a reality by substitute teaching during the school year, and working at a summer day camp throughout the summer. I absolutely love my summer camp job. It combines my love of children of all ages, away from the technology, and active outdoors! My Mommom (mom's mom) is my biggest insperation. My parents and my brother are also influential in my life, without them, I would not be the person I am today. I am a girl of varied interests always looking for a new adventure. I am shy, but once I open up you get nothing but the real me! I am an optimistic and like to think that God has a plan for everything.
36, Philadelphia, PARecently moved back to Philly, very happy to be back near my family. I lived near DC for 10 years. I'm a pediatrician, so of course I love kids :) I'm looking for a long-term relationship with someone I can explore the city and have great conversations with.
68, Chalfont, PAIf you like homemade pies I'm your girl. One of my hobbies is trying to perfect the chocolate chip cookie and lemon meringue pie. Ok now that I have you attention (not kidding about the baking) I'll tell you about me. I'm a formal fashion model and still look great if I do say so, even though that was awhile ago. I went into a full time career in sales and marketing in the corporate world and now semi-retired. My job does not not have the pressure or insane commute so I can still enjoy a great deal of down time. I love to walk, run, bike, sit on the beach at the jersey shore, dance, day trip to NY, theater, concerts, cooking, historical sites, art galleries and museums. Love my family and my cat and dog. I like working on indoor and outdoor projects on my lovely home. Will be planting a flower and herb garden soon and also looking forward to firing up the grill. Looking for a grill master, dance partner and cookie taster.
28, Hatboro, PAHi~ I'm a graphic designer and illustrator. I hope to one day make my own children's books. I'm currently working on improving many aspects of my life, and my faith is a major part of it. I enjoy going on walks, especially in nature. Playing video games (Minecraft, Persona 3, 4 and 5, and Earthbound Just to name a few). Playing board games (I don't get to play these much, but when I do I thoroughly enjoy them). Playing Dungeons and Dragons (I miss playing this so much!!). Going to plays, musicals, movies, concerts, etc. Needle felting, Drawing, Coming up with fun ideas/plans, traveling (especially road trips!).
21, North Wales, PAHey! My name is Jackie. I am a pre-school teacher and I LOVE kids. I spend a lot of time with my cats and my dog, or with my friends. I love to explore and travel! That is super important to me. I am Catholic and I hold my faith very close to my heart. I try my hardest to abide by Catholic values and beliefs. I am very committed to the things I set my mind to, especially a relationship. I am not looking for a fling, or just someone to talk to. I am serious about what I am looking for, and I expect that in return. I am loyal and respectful, and if you spend some time on me, I hope to be well worth that. I look forward to getting to know you and having some fun together!
50, Philadelphia, PALooking for someone to share fun, laughter, and life. I have a variety of interests music, movies, museums, and books. Capable of having fun enjoying a night on the town or home on the couch.
60, Bensalem, PA
Catholic Dating By Major City:
- Philadelphia,
- Pittsburgh,
- Harrisburg,
- Erie,
- California,
- Lancaster,
- Reading,
- West Chester,
- Scranton,
- Allentown,
- Bethlehem,
- York,
- Mechanicsburg,
- State College,
- Greensburg,
- Easton,
- Norristown,
- Altoona,
- Upper Darby,
- Wayne
Catholic Dating By Diocese:
- Diocese of Allentown,
- Diocese of Altoona-Johnstown,
- Diocese of Erie,
- Diocese of Greensburg,
- Diocese of Harrisburg,
- Archdiocese of Philadelphia,
- Diocese of Pittsburgh,
- Diocese of Scranton


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- Internships /
- Sales /
- Washington-DC-Remote Position-Sales and Marketing Intern
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The Impact Consultants
Posted: May 04 2015
Company Location: Sterling, VA
Application Deadline: Available Year-round
Position: 3 Part-time, Paid
Always be on the alert for potentially fraudulent job postings online - never send money to a potential employer.
DescriptionThe Impact Consultants, LLC is a DC-based firm that offers a variety of services related to business intelligence and data visualization to it's clients. The Sales and Marketing intern will play an integral role in the business goals and mission of the firm. You will gain coveted skills as you assist in acquiring clients and developing business marketing strategies. One benefit of working with The Impact Consultants, LLC is that you may work remotely from within the Washington, DC metro area and create your own schedule. The internship is paid based on commission. Candidates must have a reliable computer, access to high speed internet, knowledge of how to conduct meetings, product demonstrations and presentations both digitally and in person, and have excellent verbal and written communication skills
Responsibilities- Provide research on potential and existing clients and the competitive landscape
- Managing and follow up on digital marketing campaigns
- Assist in identifying potential business opportunities
- Monitor news and developments relating to, or impacting, key clients/prospects
- Prepare periodic reports on screening, tracking, and monitoring clients and prospects
Requirements- Pursuing a Bachelors Degree in Business (concentrating in Sales, Entrepreneurship, Marketing)
- Ability to work in a fast-paced environment and collaborate effectively as a team member
- Ability to communicate at all levels with internal/external clients by phone, email and face to face
- Proficiency in Microsoft Office (e.g. Word, Excel, and Power Point)
- A positive attitude with good organizational, presentation and writing skills


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1. To dream that you try to enter a place in which is guarded with security, signifies that you go over obstacles to solve trouble.
2. Dreaming that you are in security, means that you are uncertain about intimate relations.
3. Dreaming that you see security guards on site, suggests that you threaten something at home and at work.
4. To dream that you guard something or someone, means you should take care of your financial situation and budget.
[Other similar dream interpretations]
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α1-antitrypsin and its C-terminal fragment attenuate effects of degranulated neutrophil-conditioned medium on lung cancer HCC cells, in vitro
- Inga Zelvyte1,
- Tim Stevens1,
- Ulla Westin1 and
- Sabina Janciauskiene1Email author
© Zelvyte et al; licensee BioMed Central Ltd. 2004
Received: 05 March 2004
Accepted: 21 November 2004
Published: 21 November 2004
Tumor microenvironment, which is largely affected by inflammatory cells, is a crucial participant in the neoplastic process through promotion of cell proliferation, survival and migration. We measured the effects of polymorphonuclear neutrophil (PMN) conditioned medium alone, and supplemented with serine proteinase inhibitor α-1 antitrypsin (AAT) or its C-terminal fragment (C-36 peptide), on cultured lung cancer cells.
Lung cancer HCC cells were grown in a regular medium or in a PMN-conditioned medium in the presence or absence of AAT (0.5 mg/ml) or its C-36 peptide (0.06 mg/ml) for 24 h. Cell proliferation, invasiveness and release of IL-8 and VEGF were analyzed by [3H]-thymidine incorporation, Matrigel invasion and ELISA methods, respectively.
Cells exposed to PMN-conditioned medium show decreased proliferation and IL-8 release by 3.9-fold, p < 0.001 and 1.3-fold, p < 0.05, respectively, and increased invasiveness by 2-fold (p < 0.001) compared to non-treated controls. In the presence of AAT, PMN-conditioned medium loses its effects on cell proliferation, invasiveness and IL-8 release, whereas VEGF is up-regulated by 3.7-fold (p < 0.001) compared to controls. Similarly, C-36 peptide abolishes the effects of PMN-conditioned medium on cell invasiveness, but does not alter its effects on cell proliferation, IL-8 and VEGF release. Direct HCC cell exposure to AAT enhances VEGF, but inhibits IL-8 release by 1.7-fold (p < 0.001) and 1.4-fold (p < 0.01) respectively, and reduces proliferation 2.5-fold (p < 0.01). In contrast, C-36 peptide alone did not affect these parameters, but inhibited cell invasiveness by 51.4% (p < 0.001), when compared with non-treated controls.
Our data provide evidence that neutrophil derived factors decrease lung cancer HCC cell proliferation and IL-8 release, but increase cell invasiveness. These effects were found to be modulated by exogenously present serine proteinase inhibitor, AAT, and its C-terminal fragment, which points to a complexity of the relationships between tumor cell biological activities and local microenvironment.
The relationship between inflammation, innate immunity and cancer is widely accepted. Early in the neoplastic process, inflammatory cells and their released molecular species influence the growth, migration and differentiation of all cell types in the tumor microenvironment, whereas later in the tumorigenic process neoplastic cells also divert inflammatory mechanisms, such as metalloproteinase production, and chemokine/cytokine functions to favour tumor spread and metastasis [1–3]. Human polymorphonuclear neutrophils (PMN) comprise 50–70% of circulating leukocytes and induce inflammatory reactions that can be either cytotoxic for tumour cells or promote tumour growth and metastasis [4, 5]. For example, animal studies show that circulating neutrophils isolated from tumor-bearing animals reduce the number of metastatic foci in the lung. Other studies reveal that neutrophils stimulate tumour cell attachment to endothelial monolayers [6, 7] and neutrophil-released cytokines, growth and angiogenic factors enhance tumour growth and spread [8]. Most of the neutrophil-induced tumour-promoting effects are attributed to their abilities to release proteases. Neutrophil degranulation results in the release of serine proteases, such as elastase, cathepsin G and protease-3, which may contribute to the activation of matrix metalloproteinases (MMPs) that mediate tumor cell invasiveness [9–12]. Recently, Schwartz and co-workers found that neutrophil-secreted factors can activate pro-MMP-2, which is important in ECM degradation and tumor cell invasion [14]. It is now also apparent that tumor cells can be sources of proteolytic enzymes themselves. For example, some cells possess immunoreactive neutrophil elastase [15], which is suggested to play a role in increasing tumor cell invasiveness of surrounding tissue [16]. Levels of immunoreactive neutrophil elastase in tumour extracts are reported to be an independent prognostic factor for patients with breast cancer and non-small cell lung cancer [17–19].
A large number of studies support the notion that proteases play an important role in the progression of malignant tumors. Therefore, the expression of proteinase inhibitors is considered to be an antimalignant event [20]. Alpha-1 antitrypsin (AAT), a major inhibitor of human serine proteases in serum, is produced mainly by the liver, but also by extrahepatic cells, including neutrophils and certain cancer cells [21, 22]. AAT is an acute phase protein and its concentration rises up to 3–4-fold above normal during acute inflammation [23]. Several types of cancer, including non-small cell lung adenocarcinoma, have been associated with increased serum levels of AAT [20, 24]. Clinical studies have shown that high circulating levels of AAT directly correlate with tumour progression [24–26]. Recently we also found that plasma levels of AAT are significantly elevated in lung cancer patients and, particularly in cases with metastases [27]. The role of AAT is poorly understood in tumor diseases although it is suggested that it can inhibit tumor cell growth and invasion, in vitro [28]. Moreover, not only the native, inhibitory form of AAT, but also conformationally modified, non-inhibitory forms are suggested to play a role in modulating tumour growth and invasiveness. For example, Kataoka and co-workers have shown that the C-terminal fragment of AAT can enhance the growth and invasiveness of human pancreas adenocarcinoma cells, in vivo [29]. Our in vitro studies revealed that the C-terminal fragment of AAT, corresponding to amino acid sequence 358–396 (C-36 peptide), induces breast tumour cell proliferation and invasiveness [30]. Recently we found that C-36 peptide induces neutrophil chemotaxis, adhesion, degranulation and superoxide generation, in vitro [31], which we propose may indirectly affect tumor cell biological activities, in vivo. Together these findings prompted us to design an experimental model in vitro that would allow us to evaluate how neutrophil-derived products themselves and in combination with exogenously added native AAT or its C-36 peptide influence lung cancer HCC cell growth, invasion and metastatic properties.
HCC cell proliferation
HCC cell invasiveness
Quantitative evaluation of HCC cell invasiveness (migrated cells/per view, in ten randomly selected fields).
HCC cells in a regular medium
HCC cells in PMN-conditioned medium
Control (medium)
AAT (0.5 mg/ml)
C-36 (0.06 mg/ml)
Release of Vascular Endothelial Growth Factor (VEGF) and interleukin-8 (IL-8)
Enzymatic activity in PMN-conditioned medium
Molecular profile of AAT in HCC cell culture supernatants
Local and systemic inflammatory mediators derived by tumor-associated PMN and tumor cells are suggested to play a role in the development of lung tumors [36, 37]. Different studies have reported an increase of proteolytic activity in cancer and have suggested a role of proteases in tumor progression and metastasis [38–40]. For example, Guner and co-workers have shown that the plasma concentration of elastase in patients with malignant lung carcinoma is 10-fold higher compared to benign lesions of the lungs [41]. An increased concentration of neutrophil elastase is found to be closely associated with progression of non-small cell lung cancer (NSCL). In parallel, an increase in levels of proteinase inhibitors, such as AAT, has also been reported in tumor cases, including lung tumors. These elevated levels of proteinase inhibitors are attributed to the inflammatory reactions accompanying the tumorigenesis [42, 43].
PMN, the most abundant circulating blood leukocytes, are potent effectors of inflammation and release a wide profile of serine and metalloproteinases [44]. Infiltration of tumors with PMN is associated with a favourable prognosis in some studies in humans, however for individual patients there is no predictable relationship between PMN infiltration and cancer prognosis [45, 46]. Thus, in the present study we aimed to investigate how degranulated neutrophil-conditioned medium alone and supplemented with the serine proteinase inhibitor, AAT, or its C-terminal peptide (C-36), affects functional activities of a non-small cell lung adenocarcinoma cell line (HCC), in vitro.
We found that PMN-conditioned medium expresses multiple effects on lung cancer HCC cell functional activities (decreases in cell proliferation and IL-8 release and stimulation of cell invasion through the ECM membrane), but has no significant effects on VEGF release. These dual neutrophil activities toward cancer cells have been described by other investigators. It has been suggested that through the release of cytokines, chlorinated oxidants and defensins, neutrophils may cause direct tumor killing [47, 48] while in parallel, through release of serine proteinases and MMPs, breakdown of basement membranes and ECM and increase in tumor invasiveness are promoted [49]. Our findings that PMN-conditioned medium decreases HCC cancer cell proliferation and IL-8 release, but increases invasiveness, in part support the hypothesis that neutrophils can both, inhibit and promote tumor cell growth and invasiveness. In parallel experiments, we found that PMN-conditioned medium supplemented with AAT loses its effects on HCC cell invasion, proliferation and IL-8 release, but enhances VEGF. In contrast, C-36 peptide added to PMN-conditioned medium abolished the effects of medium on cell invasiveness, but had no significant influence on medium effects on cell proliferation, IL-8 and VEGF release.
The major function of AAT is to inhibit neutrophil derived serine proteases, particularly neutrophil elastase [50]. A local increase of proteinases and PMN elastase-AAT complexes has been demonstrated in bronchoalveolar lavage from patients with lung cancer [51]. AAT is also known to interact with other components of degranulated-PMN. For example, AAT binds to defensins and neutralizes their effects on cell cytotoxicity and migration [52]. In our in vitro model, AAT added to degranulated PMN-conditioned medium occurs in a complexed form. Thus, the modulating effects of AAT on PMN-medium activities toward HCC cancer cells can be attributed to AAT alone, but also to its interaction(s) with components of the medium. Verification of the effects of AAT on separate components released by degranulated PMN needs further investigations.
It is important to point out that AAT alone as well as in combination with PMN-conditioned medium induced VEGF release from HCC cells. VEGF is one of the most potent angiogenic molecules, regulating both angiogenesis and vascular permeability, and hence promotes tumor progression and development in NSCLC [53]. Many compounds, including anti-VEGF antibody and anti-VEGF receptor antibody have been developed as VEGF inhibitors and these compounds were reported to inhibit growth of a wide variety of tumor cell lines in vitro and in animal models, in vivo [54]. Therefore, our in vitro findings that AAT expresses potent effects on VEGF release allow classify AAT as a tumor promoter. Contrary data exist for a relationship between the levels of AAT and cancer advancement. Some investigators propose that AAT plays a protective role in tumorogenesis. For example, Finlay and co-workers showed that AAT might be directly involved in breast cancer progression by acting as a tumor suppressor [55]. Yavelow and co-workers have shown that AAT inhibits human breast cancer cells MCF-7 growth [56] and AAT was also shown to block the activation of pro MMP-2 and tumor cell invasion [11]. However, immunohistochemical studies revealed that patients with AAT-positive colon, gastric and lung adenocarcinomas had a worse prognosis than AAT-negative ones [57–59]. Together these findings suggest that AAT may play multiple roles in cancerogenesis in vivo, in addition to its role as proteinase inhibitor.
During tumor progression, inflammatory cells produce large amounts of IL-8, an autocrine growth factor [53, 60]. Levels of IL-8 have also been shown to correlate with lung cancer angiogenesis. Current knowledge of the degranulation process of neutrophils in vitro suggests that the chemokine IL-8 promotes rapid release of all neutrophil granular stores after cell exposure to cytoskeleton-disrupting agents [61]. In the present study, we found that the amount of IL-8 released by HCC can be reduced by AAT or PMN-conditioned medium alone, whereas these inhibitory effects are abolished when AAT and PMN-conditioned media are added together. Based on our findings, we hypothesize that IL-8 and other angiogenic factors released from HCC cells are regulated by PMN components, which may potentially initiate tumor cell activities or induce anti-tumor responses dependent on the tumor cell microenvironment.
AAT is the main inhibitor of neutrophil elastase and proteinase 3 [62] and it has been reported that an imbalance between AAT and neutrophil elastase may predispose to lung cancer development [63]. Patients who carry the deficiency allele of AAT have a significantly higher risk of developing squamous cell or bronchoalveolar carcinoma of the lungs [64]. On the other hand it was reported that strong expression of AAT in lung adenocarcinoma correlates with poor prognosis [65], although it was not shown whether the inhibitory activity of AAT was normal and whether elastase levels were high among this group of patients. Moreover, AAT is a good substrate for MMPs: neutrophil collagenase, gelatinase-B, stromelysin-1 and -3, and matrilysin can effectively cleave AAT [66–69]. The cleavages by these MMPs occurs at peptide bonds within AAT active site loop, resulting in a generation of cleaved forms of AAT. Comparative proteome analysis performed to identify protein alterations in plasma of prostate, lung and breast-cancer patients showed significant elevation of AAT and its N-terminal fragment [70], which points to a role for different molecular forms of AAT in cancer progression. Recent studies provide evidence that the C-terminal fragment of AAT may enhance tumor growth and invasiveness in vitro and in vivo [29, 30]. We found that the C-36 peptide displays striking concentration-dependent pro-inflammatory effects on human neutrophils, including induction of neutrophil chemotaxis, adhesion, degranulation and superoxide generation [31]. Interestingly, in our HCC cancer cell model, C-36 peptide reduced HCC cell invasiveness and also abolished PMN-conditioned medium-induced cell invasiveness, whereas there were no significant effects on other parameters measured, such as cell proliferation, VEGF and IL-8 release.
Our data show that AAT in various molecular forms expresses differential effects on lung tumour cell responses and provide an experimental evidence for complexity in the interactions of neutrophil-released molecular species and lung cancer cells.
Materials and methods
AAT and its C-terminal peptide
Native, purified human plasma AAT was purchased from the Clinical Chemistry Department (UMAS, Malmö). The quality of AAT preparations was confirmed by 7.5% SDS-PAGE and determination of anti-elastase activity as described by Gaillard et al. [71]. Synthetic C-terminal fragment of AAT (C-36, corresponding to residues 359–394) was obtained from Saveen, Biotech AB (Denmark) and was greater than 98% purity. The C-36 peptide was prepared in sterile, endotoxin-free Tris buffered saline (0.015 M Tris, pH 7.4 containing 0.15 M NaCl) just before use. The endotoxin content in AAT was tested by quantitative E-TOXATE Assay (Sigma, USA). According to international standards no more than 0.08 enzyme U/ml of endotoxin is allowed to be present in AAT solutions [72]. In our experimental model we used AAT preparation with endotoxin levels below 0.05 enzyme U/ml. If necessary, AAT samples were purified using Detoxi-Gel™ endotoxin removing gel (Pierce, Rockford, IL, USA) according to the manufacturers recommendations.
Neutrophil isolation
Neutrophils were isolated from the peripheral blood of healthy donors using Polymorphprep™ (Axis-Shield PoC AS, Oslo, Norway) according to the manufacturers recommendations. Neutrophils were harvested as the lower cellular band above the red cell pellet. Residual erythrocytes were removed by a hypotonic lysis using ice cold 0.2% NaCl (w/v) for 30 s, followed addition of an equal volume of 1.6% NaCl to restore isotonicity. The neutrophil purity was typically 90% as determined by an AC900EO AutoCounter (Swelab Instruments, Sweden) and cell viability exceeded 95% according to trypan blue staining.
Preparation of PMN-conditioned medium
PMN-conditioned medium was prepared by degranulating neutrophils (5 × 106 cells/ml) according to Videm and Strand [72]. Briefly, isolated neutrophils were incubated on a shaker for 1 h at 37°C and immediately after on ice for 3 min, and centrifuged at 240 × g for 10 min at 4°C. Supernatants were collected and analyzed for the cytokine release and enzyme activity.
Cell culture
Human non-small cell lung carcinoma (HCC) cells were established from the lung of 54-year old women with non-small cell lung the adenocarcinoma type (DSMC No. ACC 534). The cells were routinely grown at 37°C in a humid air containing 5% CO2, in 125-cm2 flasks in RPMI-1640 medium, supplemented with 10% FBS, 100 IU/ml penicillin, and 100 μg/ml streptomycin. Medium was changed every 2–3 days. Prior to experiments HCC cells were seeded into 12-wells plates at a density of 3 × 105 cells/ml, and cultured until confluent. After, cells were washed and a serum-free medium containing test substances was added for 24 h. Two sets of experiments were designed: the first, in which HCC cells were exposed to native AAT (0.5 mg/ml) or C-36 peptide (0.06 mg/ml) in a RPMI-1640 and the second, in which the cells were cultured in PMN-conditioned medium alone or supplemented with native AAT or C-36 peptide.
[3H] Thymidine incorporation assay
HCC cells were incubated with test substances for 20 h. [3H] Thymidine was then added (0.2 μCi/ml) for a further 4 h at 37°C. The medium was then aspirated, the cells were washed twice with 0.5 M NaCl and incubated for 5 min with 5% trichloroacetic acid. The cells were then washed with distilled water, dissolved in 1 ml 0.5 M NaOH, neutralised with 200 μl HCl and radioactivity determined in a β-counter (Packard 300CD liquid scintillation spectrometer; Packard Instruments). Protein concentration in cell lysates was determined by the Lowry method using HSA as a standard.
Matrigel invasion assay
Cell invasion assay was performed in an invasion chamber, a 24-well tissue culture plate with 12 inserts. The inserts contain an 8 μm pore size polycarbonate filters over which is placed a thin layer of ECMatrix™. HCC cells were suspended in a serum free medium containing 1 × 106 cells/ml alone or together with test substances. Each suspension was added to the upper chamber of an ECM Invasion system. Medium containing 10% of fetal bovine serum (FBS), serving as a chemoattractant, was added to the lower chamber. The chambers were incubated at 37°C in 5% CO2 for 24 h. Invasive cells were stained and counted using microscope (Olympus BX41, PC program Olympus MicroImage) at a 100 × magnification. The number of cells per field, from 10 randomly selected fields, are presented.
Enzyme activity assay
HCC cells were cultured alone or stimulated with test substances for 24 h. Cell free supernatants were then analysed by zymography using 10% and 12% Tris-Glycine gels containing 0.1% gelatin or β-casein, respectively (NOVEX, Invitrogen Life Technologies, UK). Briefly, supernatants were diluted in a sample buffer and separated by electrophoresis at 125 V for 90 min. The gels were then renatured and developed over night in a developing buffer (NOVEX, Invitrogen Life Technologies, UK) at 37°C. After washing, the gels were stained with Coomassie Blue R-250. Proteases that can utilize casein or gelatine as a substrate showed up as clear zones in the gels.
Endothelial growth factor (VEGF) and IL-8 analysis
HCC cell supernatants were analysed for the VEGF and IL-8 levels by using commercially available quantitative ELISA kits (R&D systems, Minneapolis, USA) according to manufacturers instructions. The lowest detectable concentration for VEGF and IL-8 was 15.6 pg/ml and 10 pg/ml, respectively.
HCC cells supernatants were analysed by 10% SDS-PAGE gels and when transferred to a polivinylidene fluoride (PVDF) membrane (Millipore, Millipore Corporation, Bedford, MA 01730) using a semi-dry immunoblot transfer system. The blot was visualised using polyclonal rabbit antibody to human AAT (1:500) (DAKO, A/S, Denmark), secondary horseradish peroxidase-conjugated swine anti-rabbit antibody (1:800) (DAKO, A/S, Denmark) and peroxidase substrate, DAB (3,3-diaminobenzidine tetrahydrochloride) (Sigma, USA).
The differences in the means of experimental results were analysed for their statistical significance with the one-way ANOVA combined with a multiple-comparisons procedure (Scheffe multiple range test), with an overall significance level of α = 0.05. Statistical Package (SPSS for Windows, release 11.0) was used for the statistical calculations.
Authors contributions
Inga Zelvyte – AB, Tim Stevens – JY, Ulla Westin – JY, Sabina Janciauskiene ES, FG
List of abbreviations
C-terminal fragment of AAT
fetal bovine serum
non small-cell lung cancer
- MMPs:
matrix metalloproteinases
membrane type matrix metalloproteinase
phosphate buffered saline
polymorphonuclear neutrophil
vascular endothelial growth factor.
This work was supported by grants from Tore Nilsson Foundation, Swedish Research Council, Astra Zeneca R&D Lund, and Medical Faculty Lund University.
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