pet, any animal kept by human beings as a source of companionship and pleasure.

While a pet is generally kept for the pleasure that it can give to its owner, often, especially with horses, dogs, and cats, as well as with some other domesticated animals, this pleasure appears to be mutual. Thus, pet keeping can be described as a symbiotic relationship, one that benefits both animals and human beings. As the keeping of pets has been practiced from prehistoric times to the present and as pets are found in nearly every culture and society, pet keeping apparently satisfies a deep, universal human need.

The history of pets is intertwined with the process of animal domestication, and it is likely that the dog, as the first domesticated species, was also the first pet. Perhaps the initial steps toward domestication were taken largely through the widespread human practice of making pets of captured young wild animals. Eventually, a working relationship developed between the dogs and their human captors. The dog was swifter, had stronger jaws, and was better at tracking prey; therefore, it could be of great use in hunting and guarding duties. From human beings, on the other hand, the dogs were assured of a constant supply of food as well as warmth from the fire. There is indirect evidence that the dog may have been domesticated and kept as a pet since Paleolithic times, as can be surmised from the paintings and carvings that archaeologists have found in ancient campsites and tombs. In Mesopotamia, dogs that look remarkably like the present-day mastiff were shown participating in a lion hunt. Domestic pets were often depicted in the scenes of family life in ancient Egypt; hunting dogs of the greyhound or saluki type accompany their master to the chase, and lap dogs frequently sit under the chair of their master or mistress.

Next to the dog, horses and cats are the animals most intimately associated with human beings. Surprisingly, both these animal groups were domesticated rather late in human history. There is no evidence that horses were domesticated in Paleolithic or Mesolithic times, but by about 2000 bce horses used in chariot battles were an established phenomenon throughout the Middle East. It seems that riding astride horses was a practice developed a few centuries later (see horsemanship). The cat too does not seem to have been domesticated as a pet until the New Kingdom period (about the 16th century bce) in Egypt. This is all the more strange as the ancient Egyptians had tamed many types of animals, such as lions, hyenas, monkeys, the Nile goose, and dogs, since the Old Kingdom period. But once cats were finally domesticated, their popularity was enormous. Gradually, the cat became one of the most universally worshiped animals.

As has been noted, the primary bond distinguishing a pet-and-owner relationship is affection. As useful as many of these animals are, what differentiates a pet from other economically useful livestock is the degree of contact between the animals and human beings. Often, this relationship has been unabashedly sentimentalized in myth, art, and literature. The affection between Alexander the Great and his favourite horse, Bucephalus, has become legendary, while in the modern age the popularity of such canine motion-picture stars as Rin Tin Tin and Lassie is further evidence of the importance placed on the relationship between owner and pet.

The pet-and-owner relationship, however, is not only founded on companionship; since the earliest period of domestication, pets have fulfilled practical, economic ends. Catching other animals to feed their human masters is one of the most fundamental uses of pets, and not only dogs have served in this capacity but cats, hyenas, and lions have also been used for hunting. The aristocratic, rather arcane sport of falconry made use of the natural talent of hawks to aid in hunting game birds. Pets have also been used for the purpose of guardingeither other livestock, the home or territory of their owners, or the owners themselves. Any pet that has a sharp sense of smell or hearing and that makes a loud noise when aroused can be used as a guard, although dogs are the best-known examples. It is thought that the Nile goose, a favourite household pet of the ancient Egyptians, may have served such a purpose. The herding and guarding of livestock is another practical use of pets, in particular the dog. Over the centuries, many specialized breeds of dog have been developed to suit this purpose.

Often, pets have been used as a source of food when other sources become scarce. This has been the case with dogs throughout their history of domestication in both the Old World and the New World. Guinea pigs, domesticated as pets in the New World, also assured a stable food supply.

Pets have also been used to eliminate animal pests. The rat-catching ability of cats is celebrated in fairy tales such as Puss n Boots and Dick Whittington, as is the snake-catching talent of the mongoose in Rudyard Kiplings Rikki-tikki-tavi.

Finally, pets themselves have become a self-perpetuating industry, bred for a variety of purposes, including their value as breeding animals. Pets that are bred for aesthetic purposes may have full-fledged show careers. Other pets may be bred for racing or other competitive sports, around which sizable industries have been built.

Animals kept as pets can be classified according to the type of premises or habitat they usually occupy. Dogs, cats, and birds such as canaries and parakeets are kept as household pets. Other birds, such as jays, magpies, and members of the crow family, are kept in aviaries. When kept as pets, reptiles and amphibians frequently require special conditions of heat and moisture. For this reason, they are best kept in glassed enclosures called vivaria. The most common vivarium pets are snakes, lizards, turtles, frogs, and toads. Many people keep fish as aquarium pets. Fishes constitute a completely separate section of the pet world, and an international industry exists for catching, breeding, transporting, and supplying stock. Hutch, or cage, pets can be kept indoors or outdoors under protected conditions. These pets include rabbits, guinea pigs, rats, mice, hamsters, gerbils, and, recently, chinchillas. Paddock pets are those that must be stabled outdoors and include such animals as horses, ponies, donkeys, and mules. Several kinds of insects are also kept as pets. These include walking-stick insects (kept in simple containers at room temperature) and ants (kept in artificial nests).

Of increasing concern is the sale of exotic pets (e.g., jaguars, alligators, ocelots, monkeys, apes, kinkajous, etc.). Rarely are the owners of such pets able to provide the basic nutritional or habitat needs of these animals; most of the animals soon die or are sent to a zoo. Furthermore, in order to obtain the young, which are considered most desirable as pets, many adults of the wild species are killed, seriously depleting populations already endangered. Several countries have passed laws to prohibit the importation of endangered species as pets, but an active black market flourishes.

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Pet | Definition, Types, History, & Facts | Britannica

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2016 psychological thriller film directed by Carles Torrens

Pet is a 2016 psychological thriller film directed by Carles Torrens, written by Jeremy Slater, and starring Dominic Monaghan, Ksenia Solo, Jennette McCurdy, and Nathan Parsons.[4][5][6] The film premiered at South by Southwest on March 11, 2016 [7][8] and was released to theaters on December 2, 2016 by Orion Pictures and Samuel Goldwyn Films.

Seth is a severely introverted man who works at an animal control center. He has developed an obsession with Holly, a waitress and former high school classmate of his, after he saw her writing in her journal on the bus. Seth seeks advice from security guard Nate, who tells him to be confident and approach her. Seth extensively researches Holly's online profiles and tries to ask her out, but she rebuffs him. At home, Holly tells her friend Claire about Seth, and takes a drunken phone call from her ex-boyfriend Eric.

Holly receives a bouquet at work the next day, and, assuming they are from Eric, goes to the bar where he works. Seth, who has been following her, confronts her, leading to a physical confrontation when he refuses to accept her rejection. Eric attacks Seth, but Seth is able to steal Holly's journal, which he spends the next several days reading. At work, Seth discovers a trapdoor to a room in an abandoned wing of the animal shelter, where he sets about constructing a steel cage. He follows Holly home, breaks into her apartment, and kidnaps her.

After awakening in the cage, Holly vows to survive, speaking to an image of Claire. Seth informs Holly that he loves her and has imprisoned her to "save" her. Through reading Holly's journal, Seth has learned that Eric slept with Claire, which Holly confronted the latter about during a car ride. In her anger, Holly continued accelerating the car until they were hit by a truck. Although injured, Claire had survived, but Holly fatally stabbed her with a glass shard; Claire's death was attributed to the crash. Since then, Holly has committed a series of gruesome murders and written about them in her journal, leading to Seth finding a "purpose" in life by preventing Holly from hurting anyone else.

Over the next several days, they engage in psychological mind games against each other as Holly begins to slip details to draw Seth in. Seth maintains that Holly committed the other murders out of guilt for not being caught over Claire, but Holly counters that she kills simply for pleasure. A suspicious Nate follows Seth and discovers Holly. She deliberately distracts Nate so that Seth has time to overpower him. At Holly's urging, he smashes Nate's skull with a cinder block, then follows her instructions to dispose of the body.

The police become suspicious of Seth's role in Nate's disappearance. Holly convinces him that he can save her if he proves his love to her by cutting off his finger. He does, but this leads to Holly grabbing his knife and threatening to kill herself if he doesn't release her. She says she finally believes that he loves her before slitting his throat.

Some time later, Holly is back together with Eric, and the "fictional" events from her journal are being published by a vanity press. Holly finds evidence that Eric has been cheating again, but declines to hurt him. Instead, she travels to a warehouse, where it is revealed that Seth is being kept in a cage, still alive but horribly mutilated and tortured; she thanks him for "saving" her by allowing her to take out all of her murderous impulses on him instead.

It was announced in August 2015 that filming had begun with actors Dominic Monaghan, Ksenia Solo, Jennette McCurdy, and Nathan Parsons.[4][5][6]

The film premiered at the March 2016 South by Southwest Film festival.[7][8][9]

Pet was released to nine theatres on December 2, 2016 with total gross of $8,004.[10][11] It was set for a June 16, 2017 theatrical opening in Spain.[12]

Reviews for Pet have been mixed, with critics alternatively praising the subversive themes and strong leading performances, or lamenting the implausible twists. The review aggregator website Rotten Tomatoes reported that 56% of critics have given the film a positive review based on 18 reviews, with an average rating of 5.16/10.[13] On Metacritic, the film has a weighted average score of 48 out of 100 based on 8 critics, indicating "mixed or average reviews".[14] Reviews that have criticized the twists include Screen Anarchy, which wrote "Without spoiling anything more, the twists push past the merely unlikely into a strange minefield of 'what in the world?'",[15] and RogerEbert.com, which noted "The rank, idiotic implausibilities continue to mount..."[16]

The Hollywood Reporter gave the bottom line of "This graphically violent horror thriller features too many plot twists for its own good", but the review has also words of praise for the direction and cast: "Still, the film is engrossing, thanks to the directors skill at delivering sustained tension, and the excellent performances."[17]

An entirely positive review came from The A.V. Club's Alex McCown, who stated: "Part of the wicked fun of Pet, a dark little exercise in sadism and black humor, is how it upends the traditional conventions of the 'wronged woman turns the tables on her abuser' narrative. (...) The films zigs where you expect a depraved zag, resulting in a smart and unsettling tale."[18]

Another overall positive review has been given by Katie Walsh of Los Angeles Times, who wrote: "The constant power flipping allows for some interesting explorations of both the misogyny and misandry demonstrated by the main characters, and the way they justify their actions through the philosophical lens of love and sacrifice. 'Pet' is a modern-day fable of unchecked desire that descends quickly into a bloody, morbid cautionary tale."[19]

Francisco Marinero of El Mundo rated Pet 2 out 5 stars, writing that the film, which as usual in American independent pictures stars a "nondescript guy sadly alone in domestic routines", "is conceived as a function of the bombshells in its writing and characters", favouring gruesomeness over suspense.[20]

Link:
Pet (film) - Wikipedia

OverviewWhat is DiabetesWhat is diabetes?

Diabetes happens when your body isn't able to take up sugar (glucose) into its cells and use it for energy. This results in a build up of extra sugar in your bloodstream.

Mismanagement of diabetes can lead to serious consequences, causing damage to a wide range of your body's organs and tissues including your heart, kidneys, eyes and nerves.

The process of digestion includes breaking down the food you eat into various different nutrient sources. When you eat carbohydrates (for example, bread, rice, pasta), your body breaks this down into sugar (glucose). When glucose is in your bloodstream, it needs help a "key" to get into its final destination where it's used, which is inside your body's cells (cells make up your body's tissues and organs). This help or "key" is insulin.

Insulin is a hormone made by your pancreas, an organ located behind your stomach. Your pancreas releases insulin into your bloodstream. Insulin acts as the key that unlocks the cell wall door, which allows glucose to enter your bodys cells. Glucose provides the fuel or energy tissues and organs need to properly function.

If you have diabetes:

Or

If glucose cant get into your bodys cells, it stays in your bloodstream and your blood glucose level rises.

The types of diabetes are:

Less common types of diabetes include:

Diabetes insipidus is a distinct rare condition that causes your kidneys to produce a large amount of urine.

Some 34.2 million people of all ages about 1 in 10 have diabetes in the U.S. Some 7.3 million adults aged 18 and older (about 1 in 5) are unaware that they have diabetes (just under 3% of all U.S. adults). The number of people who are diagnosed with diabetes increases with age. More than 26% of adults age 65 and older (about 1 in 4) have diabetes.

Factors that increase your risk differ depending on the type of diabetes you ultimately develop.

Risk factors for Type 1 diabetes include:

Risk factors for prediabetes and Type 2 diabetes include:

Risk factors for gestational diabetes include:

The cause of diabetes, regardless of the type, is having too much glucose circulating in your bloodstream. However, the reason why your blood glucose levels are high differs depending on the type of diabetes.

Symptoms of diabetes include:

Other symptoms

Type 1 diabetes symptoms: Symptoms can develop quickly over a few weeks or months. Symptoms begin when youre young as a child, teen or young adult. Additional symptoms include nausea, vomiting or stomach pains and yeast infections or urinary tract infections.

Type 2 diabetes and prediabetes symptoms: You may not have any symptoms at all or may not notice them since they develop slowly over several years. Symptoms usually begin to develop when youre an adult, but prediabetes and Type 2 diabetes is on the rise in all age groups.

Gestational diabetes: You typically will not notice symptoms. Your obstetrician will test you for gestational diabetes between 24 and 28 weeks of your pregnancy.

If your blood glucose level remains high over a long period of time, your bodys tissues and organs can be seriously damaged. Some complications can be life-threatening over time.

Complications include:

Complications of gestational diabetes:

In the mother: Preeclampsia (high blood pressure, excess protein in urine, leg/feet swelling), risk of gestational diabetes during future pregnancies and risk of diabetes later in life.

In the newborn: Higher-than-normal birth weight, low blood sugar (hypoglycemia), higher risk of developing Type 2 diabetes over time and death shortly after birth.

Diabetes is diagnosed and managed by checking your glucose level in a blood test. There are three tests that can measure your blood glucose level: fasting glucose test, random glucose test and A1c test.

Less than 100

Less than 140

Less than 5.7%

Gestational diabetes tests: There are two blood glucose tests if you are pregnant. With a glucose challenge test, you drink a sugary liquid and your glucose level is checked one hour later. You dont need to fast before this test. If this test shows a higher than normal level of glucose (over 140 ml/dL), an oral glucose tolerance test will follow (as described above).

Type 1 diabetes: If your healthcare provider suspects Type 1 diabetes, blood and urine samples will be collected and tested. The blood is checked for autoantibodies (an autoimmune sign that your body is attacking itself). The urine is checked for the presence of ketones (a sign your body is burning fat as its energy supply). These signs indicate Type 1 diabetes.

If you have symptoms or risk factors for diabetes, you should get tested. The earlier diabetes is found, the earlier management can begin and complications can be lessened or prevented. If a blood test determines you have prediabetes, you and your healthcare professional can work together to make lifestyle changes (e.g. weight loss, exercise, healthy diet) to prevent or delay developing Type 2 diabetes.

Additional specific testing advice based on risk factors:

Diabetes affects your whole body. To best manage diabetes, youll need to take steps to manage your risk factors, including:

You hold the keys to managing your diabetes by:

Checking your blood glucose level is important because the results help guide decisions about what to eat, your physical activity and any needed medication and insulin adjustments or additions.

The most common way to check your blood glucose level is with a blood glucose meter. With this test, you prick the side of your finger, apply the drop of blood to a test strip, insert the strip into the meter and the meter will show your glucose level at that moment in time. Your healthcare provider will tell you how often youll need to check your glucose level.

Advancements in technology have given us another way to monitor glucose levels. Continuous glucose monitoring uses a tiny sensor inserted under your skin. You don't need to prick your finger. Instead, the sensor measures your glucose and can display results anytime during the day or night. Ask your healthcare provider about continuous glucose monitors to see if this is an option for you.

Ask your healthcare team what your blood glucose level should be. They may have a specific target range for you. In general, though, most people try to keep their blood glucose levels at these targets:

Having a blood glucose level that is lower than the normal range (usually below 70 mg/dL) is called hypoglycemia. This is a sign that your body gives out that you need sugar.

Symptoms you might experience if you have hypoglycemia include:

You might pass out if your hypoglycemia is not managed.

If you have too much glucose in your blood, you have a condition called hyperglycemia. Hyperglycemia is defined as:

or

Treatments for diabetes depend on your type of diabetes, how well managed your blood glucose level is and your other existing health conditions.

Oral medications and insulin work in one of these ways to treat your diabetes:

Over 40 medications have been approved by the Food and Drug Administration for the treatment of diabetes. Its beyond the scope of this article to review all of these drugs. Instead, well briefly review the main drug classes available, how they work and present the names of a few drugs in each class. Your healthcare team will decide if medication is right for you. If so, theyll decide which specific drug(s) are best to treat your diabetes.

Diabetes medication drug classes include:

Many oral diabetes medications may be used in combination or with insulin to achieve the best blood glucose management. Some of the above medications are available as a combination of two medicines in a single pill. Others are available as injectable medications, for example, the GLP-1 agonist semaglutide (Ozempic) and lixisenatide (Adlyxin).

Always take your medicine exactly as your healthcare prescribes it. Discuss your specific questions and concerns with them.

There are many types of insulins for diabetes. If you need insulin, you healthcare team will discuss the different types and if they are to be combined with oral medications. To follow is a brief review of insulin types.

There are insulins that are a combination of different insulins. There are also insulins that are combined with a GLP-1 receptor agonist medication (e.g. Xultophy, Soliqua).

Insulin is available in several different formats. You and your healthcare provider will decide which delivery method is right for you based on your preference, lifestyle, insulin needs and insurance plan. Heres a quick review of available types.

Yes. There are two types of transplantations that might be an option for a select number of patients who have Type 1 diabetes. A pancreas transplant is possible. However, getting an organ transplant requires taking immune-suppressing drugs for the rest of your life and dealing with the side effects of these drugs. However, if the transplant is successful, youll likely be able to stop taking insulin.

Another type of transplant is a pancreatic islet transplant. In this transplant, clusters of islet cells (the cells that make insulin) are transplanted from an organ donor into your pancreas to replace those that have been destroyed.

Another treatment under research for Type 1 diabetes is immunotherapy. Since Type 1 is an immune system disease, immunotherapy holds promise as a way to use medication to turn off the parts of the immune system that cause Type 1 disease.

Bariatric surgery is another treatment option thats an indirect treatment for diabetes. Bariatric surgery is an option if you have Type 2 diabetes, have obesity (body mass index over 35) and considered a good candidate for this type of surgery. Much improved blood glucose levels are seen in people who have lost a significant amount of weight.

Of course other medications are prescribed to treat any existing health problems that contribute to increasing your risk of developing diabetes. These conditions include high blood pressure, high cholesterol and other heart-related diseases.

Although diabetes risk factors like family history and race cant be changed, there are other risk factors that you do have some control over. Adopting some of the healthy lifestyle habits listed below can improve these modifiable risk factors and help to decrease your chances of getting diabetes:

No. Type 1 diabetes is an autoimmune disease, meaning your body attacks itself. Scientists arent sure why someones body would attack itself. Other factors may be involved too, such as genetic changes.

Chronic complications are responsible for most illness and death associated with diabetes. Chronic complications usually appear after several years of elevated blood sugars (hyperglycemia). Since patients with Type 2 diabetes may have elevated blood sugars for several years before being diagnosed, these patients may have signs of complications at the time of diagnosis.

The complications of diabetes have been described earlier in this article. Although the complications can be wide ranging and affect many organ systems, there are many basic principles of prevention that are shared in common. These include:

If you have diabetes, the most important thing you can do is keep your blood glucose level within the target range recommended by your healthcare provider. In general, these targets are:

You will need to closely follow a treatment plan, which will likely include following a customized diet plan, exercising 30 minutes five times a week, quitting smoking, limiting alcohol and getting seven to nine hours of sleep a night. Always take your medications and insulin as instructed by your provider.

If you havent been diagnosed with diabetes, you should see your healthcare provider if you have any symptoms of diabetes. If you already have been diagnosed with diabetes, you should contact your provider if your blood glucose levels are outside of your target range, if current symptoms worsen or if you develop any new symptoms.

Sugar itself doesn't directly cause diabetes. Eating foods high in sugar content can lead to weight gain, which is a risk factor for developing diabetes. Eating more sugar than recommended American Heart Association recommends no more than six teaspoons a day (25 grams) for women and nine teaspoons (36 grams) for men leads to all kinds of health harms in addition to weight gain.

These health harms are all risk factors for the development of diabetes or can worsen complications. Weight gain can:

Most people with diabetes see their primary healthcare provider first. Your provider might refer you to an endocrinologist/pediatric endocrinologist, a physician who specializes in diabetes care. Other members of your healthcare team may include an ophthalmologist (eye doctor), nephrologist (kidney doctor), cardiologist (heart doctor), podiatrist (foot doctor), neurologist (nerve and brain doctor), gastroenterologist (digestive tract doctor), registered dietician, nurse practitioners/physician assistants, diabetes educator, pharmacist, personal trainer, social worker, mental health professional, transplant team and others.

In general, if you are being treated with insulin shots, you should see your doctor at least every three to four months. If you are treated with pills or are managing diabetes through diet, you should be seen at least every four to six months. More frequent visits may be needed if your blood sugar isn't managed or if complications of diabetes are worsening.

Although these seem like simple questions, the answers are not so simple. Depending on the type of your diabetes and its specific cause, it may or may not be possible to reverse your diabetes. Successfully reversing diabetes is more commonly called achieving remission.

Type 1 diabetes is an immune system disease with some genetic component. This type of diabetes cant be reversed with traditional treatments. You need lifelong insulin to survive. Providing insulin through an artificial pancreas (insulin pump plus continuous glucose monitor and computer program) is the most advanced way of keeping glucose within a tight range at all times most closely mimicking the body. The closest thing toward a cure for Type 1 is a pancreas transplant or a pancreas islet transplant. Transplant candidates must meet strict criteria to be eligible. Its not an option for everyone and it requires taking immunosuppressant medications for life and dealing with the side effects of these drugs.

Its possible to reverse prediabetes and Type 2 diabetes with a lot of effort and motivation. Youd have to reverse all your risk factors for disease. To do this means a combination of losing weight, exercising regularly and eating healthy (for example, a plant-based, low carb, low sugar, healthy fat diet). These efforts should also lower your cholesterol numbers and blood pressure to within their normal range. Bariatric surgery (surgery that makes your stomach smaller) has been shown to achieve remission in some people with Type 2 diabetes. This is a significant surgery that has its own risks and complications.

If you have gestational diabetes, this type of diabetes ends with the birth of your child. However, having gestational diabetes is a risk factor for developing Type 2 diabetes.

The good news is that diabetes can be effectively managed. The extent to which your Type 1 or Type 2 diabetes can be managed is a discussion to have with your healthcare provider.

Yes, its possible that if diabetes remains undiagnosed and unmanaged (severely high or severely low glucose levels) it can cause devastating harm to your body. Diabetes can cause heart attack, heart failure, stroke, kidney failure and coma. These complications can lead to your death. Cardiovascular disease in particular is the leading cause of death in adults with diabetes.

Although having diabetes may not necessarily increase your risk of contracting COVID-19, if you do get the virus, you are more likely to have more severe complications. If you contract COVID-19, your blood sugars are likely to increase as your body is working to clear the infection. If you contract COVID-19, contact your healthcare team early to let them know.

Blood vessels are located throughout our bodys tissues and organs. They surround our bodys cells, providing a transfer of oxygen, nutrients and other substances, using blood as the exchange vehicle. In simple terms, diabetes doesnt allow glucose (the bodys fuel) to get into cells and it damages blood vessels in/near these organs and those that nourish nerves. If organs, nerves and tissues cant get the essentials they need to properly function, they can begin to fail. Proper function means that your hearts blood vessels, including arteries, are not damaged (narrowed or blocked). In your kidneys, this means that waste products can be filtered out of your blood. In your eyes, this means that the blood vessels in your retina (area of your eye that provides your vision) remain intact. In your feet and nerves, this means that nerves are nourished and that theres blood flow to your feet. Diabetes causes damage that prevents proper function.

Unmanaged diabetes can lead to poor blood flow (poor circulation). Without oxygen and nutrients (delivered in blood), you are more prone to the development of cuts and sores that can lead to infections that cant fully heal. Areas of your body that are farthest away from your heart (the blood pump) are more likely to experience the effects of poor blood flow. So areas of your body like your toes, feet, legs and fingers are more likely to be amputated if an infection develops and healing is poor.

Yes. Because unmanaged diabetes can damage the blood vessels of the retina, blindness is possible. If you havent been diagnosed with diabetes yet but are experiencing a change in your vision, see primary healthcare provider or ophthalmologist as soon as you can.

Scientists dont have firm answers yet but there appears to be a correlation between hearing loss and diabetes. According to the American Diabetes Association, a recent study found that hearing loss was twice as common in people with diabetes versus those who didnt have diabetes. Also, the rate of hearing loss in people with prediabetes was 30% higher compared with those who had normal blood glucose levels. Scientists think diabetes damages the blood vessels in the inner ear, but more research is needed.

Yes, its possible to develop headaches or dizziness if your blood glucose level is too low usually below 70 mg/dL. This condition is called hypoglycemia. You can read about the other symptoms hypoglycemia causes in this article. Hypoglycemia is common in people with Type 1 diabetes and can happen in some people with Type 2 diabetes who take insulin (insulin helps glucose move out of the blood and into your bodys cells) or medications such as sulfonylureas.

Yes, its possible for diabetes to cause hair loss. Unmanaged diabetes can lead to persistently high blood glucose levels. This, in turn, leads to blood vessel damage and restricted flow, and oxygen and nutrients cant get to the cells that need it including hair follicles. Stress can cause hormone level changes that affect hair growth. If you have Type 1 diabetes, your immune system attacks itself and can also cause a hair loss condition called alopecia areata.

People with Type 1 diabetes need insulin to live. If you have Type 1 diabetes, your body has attacked your pancreas, destroying the cells that make insulin. If you have Type 2 diabetes, your pancreas makes insulin, but it doesnt work as it should. In some people with Type 2 diabetes, insulin may be needed to help glucose move from your bloodstream to your bodys cells where its needed for energy. You may or may not need insulin if you have gestational diabetes. If you are pregnant or have Type 2 diabetes, your healthcare provider will check your blood glucose level, assess other risk factors and determine a treatment approach which may include a combination of lifestyle changes, oral medications and insulin. Each person is unique and so is your treatment plan.

You arent born with diabetes, but Type 1 diabetes usually appears in childhood. Prediabetes and diabetes develop slowly over time. Gestational diabetes occurs during pregnancy. Scientists do believe that genetics may play a role or contribute to the development of Type 1 diabetes. Something in the environment or a virus may trigger its development. If you have a family history of Type 1 diabetes, you are at higher risk of developing Type 1 diabetes. If you have a family history of prediabetes, Type 2 diabetes or gestational diabetes, youre at increased risk of developing prediabetes, Type 2 diabetes or gestational diabetes.

Diabetes-related ketoacidosis is a life-threatening condition. It happens when your liver breaks down fat to use as energy because theres not enough insulin and therefore glucose isnt being used as an energy source. Fat is broken down by the liver into a fuel called ketones. The formation and use of ketones is a normal process if it has been a long time since your last meal and your body needs fuel. Ketones are a problem when your fat is broken down too fast for your body to process and they build up in your blood. This makes your blood acidic, which is a condition called ketoacidosis. Diabetes-related ketoacidosis can be the result of unmanaged Type 1 diabetes and less commonly, Type 2 diabetes. Diabetes-related ketoacidosis is diagnosed by the presence of ketones in your urine or blood and a basic metabolic panel. The condition develops over several hours and can cause coma and possibly even death.

Hyperglycemic hyperosmolar nonketotic syndrome (HHNS) develops more slowly (over days to weeks) than diabetes-related ketoacidosis. It occurs in patients with Type 2 diabetes, especially the elderly and usually occurs when patients are ill or stressed. If you have HHNS, you blood glucose level is typically greater than 600 mg/dL. Symptoms include frequent urination, drowsiness, lack of energy and dehydration. HHNS is not associated with ketones in the blood. It can cause coma or death. Youll need to be treated in the hospital.

This means your kidneys are allowing protein to be filtered through and now appear in your urine. This condition is called proteinuria. The continued presence of protein in your urine is a sign of kidney damage.

A note from Cleveland Clinic

Theres much you can do to prevent the development of diabetes (except Type 1 diabetes). However, if you or your child or adolescent develop symptoms of diabetes, see your healthcare provider. The earlier diabetes is diagnosed, the sooner steps can be taken to treat and manage it. The better you are able to manage your blood sugar level, the more likely you are to live a long, healthy life.

Originally posted here:
Diabetes: An Overview - Cleveland Clinic

The use of stem cell therapy in treating patients with autism spectrum disorder (ASD) is not recommended and its promotion and advertisement will be considered professional misconduct, the National Medical Commission (NMC) has said.

The NMCs ethics and medical registration board set up a committee to look into the prescription, recommendation, or administration of such treatment after doctors and parents complained about the mushrooming stem cell therapy centres and their advertisements promising cure for autism. The committee submitted its report on December 6 and it was uploaded on December 14.

The medical fraternity has welcomed the order, saying, there is not scientific basis to this line of treatment for which patients have been shelling out for years.

Dr Samir Dalwai, developmental-behavioural paediatrician, Nanavati Max Hospital, Vile Parle, said, Autism is a chronic condition and the results are slow. And there is a stigma attached to it. However, desperate parents fall for such promotional gimmicks as the therapy is marketed well not just in the city but across the country.

Every week, the hospital gets at least one patient in the 7-8 age group whose parents have tried stem cell therapy to treat autism, but in vain. Because of the stigma or being told by doctors that there wont be much, or slow improvement in their child, parents fall for the stem cell advertisements. Many a time, patients stopped the treatment midway and opted for the therapy hoping to see the miracle cure it promises, he said, adding each therapy reportedly costs 3- 4 lakh.

The neuro-developmental paediatric chapter of Indian Academy of Paediatrics had also written to all government authorities to take action in this regard.

Parul Kumtha, trustee, Forum for Autism, said the therapy neither had enough proof of efficacy nor did it have the Food and Drug Administrations approval.

Many parents have mortgaged their jewellery, property to bear the cost of treatment. If any medical centres are doing research on stem cell therapy, then they should not charge the patients. Also, as the NMC order says, there is no cure for autism and it is medically unethical to promise it as a cure, she said.

Dr Milan Balakrishnan, psychiatrist and member of Bombay Psychiatric Society, said it has been observed that after the therapy, there is a rise in irritability and aggression in the patient.

Recently, a 15-year-old boy with autism was brought to us. He had a lot of anger, and he was beating up his parents too. We had to manage the outbursts with medication. It was during the course of treatment that the parents told us about stem cell therapy and the aggression their son developed as a new symptom after the procedure, he said.

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The Federal Reserve just raised rates. Heres what it could mean for inflation and interest rates in the new year.

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Live Cell Imaging Market accounted for US$ 1.8 billion in 2022 and is estimated to be US$ 5.0 billion by 2032 and is anticipated to register a CAGR of 9.2% - By PMI  India Shorts

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Is your child suffering from severe headaches and dysfunctional motor skills, it can be a sign of Childhood Leukaemia  APN News

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GD2 enriches for breast CSCs. We recently reported that, following the induction of EMT, human mammary epithelial cells show functional properties similar to those of human bone marrowderived MSCs (13). Therefore, we hypothesized that the cell markers expressed on the surface of MSCs could also be expressed on the surface of breast CSCs. To test this hypothesis, we analyzed for the expression of several known MSC cell surface markers (i.e., CD105, CD90, CD106, CD166, CD73, CD271, MSCA-1, and GD2) on HMECs that had been experimentally transformed to become tumorigenic using oncogenic V12-H-Ras (HMLER cells) (21). Absolute expression of most of the markers analyzed could not divide HMLER cells into two distinct subpopulations (Supplemental Figure 1; supplemental material available online with this article; doi:10.1172/JCI59735DS1), similar to CD44hiCD24lo cells (12). However, ganglioside GD2, one of the cell surface markers for MSCs, was able to separate HMLER cells into GD2+ (4.5% 2.4%) and GD2 (92.7% 3.8%) populations (Figure 1A and Supplemental Figure 1). Strikingly, GD2+ HMLER cells isolated using FACS appeared spindle-shaped, with limited cell-cell contacts; conversely, the GD2 cells displayed cobblestone epithelial morphology (Figure 1B). Moreover, the GD2+ HMLER cells proliferated approximately 5-fold slower than the GD2 HMLER cells (Figure 1C).

GD2 identifies CSCs in breast cancer. (A) HMLER cells were stained with anti-GD2 antibody by indirect staining and analyzed on an LSR II flow cytometer. GD2+/ gates were drawn based on IgG2a isotype control. FSC, forward scatter. (B) GD2+/ HMLER cells were cell sorted and cultured in cell culture dishes for 4 days. Scale bars: 50 m. (C) 2 104 GD2+/ HMLER cells were cultured in 6-well cell culture dishes in triplicate. Total cells were counted on days 2, 4, and 6 using a Vi-CELL (Beckman Coulter) cell counter. (D) HMLER or MDA-MB-231 cells (1 103) were sorted into each well of 24-well ultra-low attachment dishes containing mammosphere growth medium using the FACSAria II cell sorter. Cells were cultured for 12 days, and the photos were taken using a light microscope. Scale bars: 100 m. (E and F) Number of mammospheres formed from GD2+/ HMLER (E) and MDA-MB-231 (F) cells. The experiment was performed in triplicate. P < 0.01 (G) GD2+/ MDA-MB-231 cells were sorted (1 cell or 5 cells/well) into 96-well ultra-low-attachment dishes containing mammosphere growth medium. Cells were cultured for 12 days, and mammospheres were counted using a light microscope. Scale bars: 200 m. (H) Number of mammospheres formed from single GD2+/ MDA-MB-231 cells. *P < 0.002. (I) Size of mammospheres measured using a hemocytometer. *P < 0.0001.

To further investigate the functional properties of GD2+ and GD2 cells, we sorted HMLER and MDA-MB-231 cells based on GD2 expression and examined them by mammosphere assay. Interestingly, the GD2+ cells from HMLER and MDA-MB-231 cells formed 2-fold more mammospheres compared with GD2 cells (Figure 1, DF, P < 0.01). Direct sorting of GD2+ and GD2 MDA-MB-231 cells into low-attachment 96-well plates at either 1 or 5 cells per well also resulted in a 2-fold increase in sphere formation by GD2+ cells regardless of the number of cells per well compared with GD2 cells (Figure 1, G and H). In addition, the mammospheres generated by GD2+ cells were 3 times larger than those generated by GD2 cells (Figure 1, G and I), indicating that the GD2+ cells are capable of growing better in suspension cultures.

CSCs are known to be more migratory and invasive (1, 3). To examine the migration and invasion potential of GD2+/ cells, we fractionated HMLER cells into GD2+ and GD2 cells and analyzed them for migration and invasion using Boyden chamber Matrigel invasion assays. After 24 hours of incubation, GD2+ HMLER cells migrated to a more than 4-fold greater extent compared with GD2 cells, indicating that GD2+ cells are highly migratory (Supplemental Figure 2). The hallmark of CSCs is their ability to initiate tumor better than their bulk tumor counterparts (1, 2). To determine the tumor-initiating potential of GD2+ cells, we sorted GD2+ and GD2 MDA-MB-231 cells and transplanted them subcutaneously into the flank of NOD/SCID mice at limiting dilutions. At lower cell numbers including 100 or 10 cells/site, the GD2+ cells generated 2- and 5-fold more tumors, respectively, compared with the GD2 fraction (Table 1). However, at higher cell numbers (10,000 or 1,000 cells/site), there were no significant differences in tumor initiation between GD2+ and GD2 cells. These data firmly established that GD2 is a marker of cells capable of initiating tumors at a higher frequency than cells without GD2.

Generation of tumors by GD2+/ cells in vivo

Percentage of GD2+ cells is highest in cell lines with a basal molecular signature. On the basis of gene expression profile (22), breast cancer cell lines have been classified into 3 groups: luminal, basal A, and basal B. We randomly selected 12 breast cancer cell lines representing these 3 subgroups and analyzed them for GD2 expression. Interestingly, the majority of these lines, independent of the subgroup, contained a subpopulation of GD2+ cells at variable levels (Table 2). However, basal cell lines contained a much greater number (mean 9%, range 1.2%17%, n = 6) of GD2+ cells compared with luminal cell lines (median 0.2%, range 03%, n = 6, Table 1, P = 0.00237). Since basal-derived cell lines show greater tumor initiation potential and contain more CSCs based on the previously reported CD44hiCD24lo profiles (23), this finding once again confirms GD2 as a stem cell marker.

Expression of GD2 in breast cancer cell lines

GD2 identifies the CD44hiCD24lo population in breast cancer cell lines and patient samples. Since we found that GD2, similar to previously reported CD44 and CD24 cell surface markers, is capable of separating cancer cells into two populations with differing tumor-initiating potential (7), we hypothesized that GD2 would be mostly expressed in the CD44hiCD24lo cancer cell fraction. To test this, we initially analyzed the expression of CD44hiCD24lo cells in GD2+ HMLER cells and found that more than 85% (85% 3.5%) of GD2+ HMLER cells also displayed a CD44hiCD24lo CSC profile, whereas less than 1% (0.7% 0.2%) of GD2 HMLER cells were CD44hiCD24lo (Figure 2A). In addition, through reverse gating analysis of CD44hiCD24lo HMLER cells, we noted that more than 84% (84% 2.5%) of CD44hiCD24lo HMLER cells were also positive for GD2 (Figure 2B), whereas less than 5% of CD44loCD24hi HMLER cells were GD2+ (4.3% 1.2%). To further determine the correlation between the expression of GD2 and the CD44/CD24 profiles, we sequentially gated HMLER cells into GD2hi, GD2lo, and GD2neg cells. This analysis revealed that GD2 expression levels correlated strongly with the CD44hiCD24lo phenotype (Supplemental Figure 3A). Moreover, by determining the MFI, we found that GD2 expression levels correlated positively with CD44 expression (correlation index, r2 = 0.85; P < 0.0003; Supplemental Figure 3B). To validate the coexpression of GD2 on CD44hiCD24lo cells, we used anti-GD2 antibody from a different source (Abcam, clone 2Q549) to stain HMLER cells in a 4-step staining procedure as explained before (24), along with anti-CD44 and anti-CD24 antibodies. Analysis of GD2+ cells revealed that these cells coexpress CD44hiCD24lo, confirming our initial findings with the 14G2a clone (Supplemental Figure 4).

GD2 identifies CD44hiCD24lo stem cell phenotype in breast cancer cells. (A) HMLER cells were stained with anti-GD2 antibody and with CD44-APC and CD24-FITC using the 4-step staining protocol described in Methods. Cells were electrically gated on GD2+/ cells and displayed in a pseudocolor dot plot with CD44 on the y axis and CD24 on the x axis using FlowJo data analysis software. (B) In an identical experiment, CD44hi/loCD24lo/hi cells were displayed on a pseudocolor dot plot with GD2 on the y axis and FSC on the x axis. (C) Primary breast tumor samples were processed as described in Methods, and the single cells in suspension were stained with anti-GD2, CD44-APC, CD24-FITC, CD45-FITC, and DAPI using the 4-step staining protocol. Cells were initially gated on DAPI-negative cells to exclude dead cells, and the cells were then gated on CD45 cells to exclude hematopoietic cells. GD2+CD45 cells were displayed on a dot plot, with CD44 on the y axis and CD24 on the x axis. Analysis was perfumed using an LSR II flow cytometer. Data were analyzed using FlowJo software.

To further investigate the correlation between GD2 expression and the CD44hiCD24lo profile, we also analyzed primary breast tumor samples (n = 12, Table 3). Using multi-parameter flow cytometry, we excluded CD45+ inflammatory and other hematopoietic cells from dissociated tumor samples. The non-hematopoietic CD45 fraction was then analyzed for the expression of GD2, CD44, and CD24. This analysis of the CD45 fraction revealed that GD2 was expressed, at variable levels from 0.5% to 35% (median 4.35%, range 0.5%35.8%), in tumor samples (Table 3). Importantly, similar to what we observed in cell lines, more than 95.5% (95.5% 2.7%) of GD2+CD45 tumor cells also co-segregated with the CD44hiCD24lo phenotype (Figure 2C). In contrast, only 2.4% (2.4% 0.4%) of GD2CD45 cells exhibited the CD44hiCD24lo phenotype (Figure 2C). Together these findings clearly indicated that GD2 is a marker of a subset of cancer cells with stem cell properties. To validate that the identified GD2+ cells are in fact tumor and not MSCs, we stained human breast tumor tissues with anti-GD2 and epithelial-specific antipan-cytokeratin antibodies. We found coexpression of GD2 and cytokeratin in some of the breast cancer cells, suggesting that GD2 identifies breast tumors cells (Supplemental Figure 5, A and B).

Breast cancer patient samples analyzed: patient number, tumor type, percentage of GD2+ cells

GD2+ and CD44hiCD24lo cells have similar gene signatures. Since we found that GD2 is capable of independently enriching for CSCs as a single marker compared with the previously known double marker CD44hiCD24lo, we compared the global gene expression profiles in these two populations isolated from HMLER cells using microarray analysis. We initially compared the GD2+ fraction with the GD2 fraction of cells (GD2 set) and the CD44hiCD24lo with the CD4loCD24hi fraction (CD44 set) and identified gene signatures specific to the GD2+ and CD44hiCD24lo fractions (Figure 3A). Comparison of the top 600 differentially expressed genes in the GD2 set (GEO GSE36643) and the CD44 set (GSE36643) identified 231 genes as being identical in the two sets (Supplemental Table 1). In addition, we applied Pearsons 2 test with a Yates continuity correction to assess the association between these two cell types and found that the identified 231 genes correlated (100%) between the two groups described above (Figure 3B). This gene expression analysis along with the cell surface protein analysis shown in Figure 2 indicated that GD2+ cells share not only functional properties but also a gene signature with CD44hiCD24lo cells.

GD2+ and CD44hiCD24lo cells have a similar gene signature. (A) Heat maps derived from microarray analysis of CD44hi/loCD24lo/hi and GD2+/ populations of HMLER cells. (B) Two hundred thirty-one genes of the top 600 differentially expressed genes were identical in GD2+ versus GD and CD44hiCD24lo versus CD44loCD24hi groups. These genes were cross-classified in a 2-by-2 table by GD2+ up-/downregulation and CD44hiCD24lo up-/downregulation. Pearsons 2 test with a Yates continuity correction was applied to assess the association. Statistical significance was assessed at the 0.05 level. (C) Biosynthesis reaction of GD2. (D and E) To measure the expression of GD2S/GD3S mRNA, CD44hiCD24lo or CD44loCD24hi and GD2+/ cells from HMLER (D) or MDA-MB-231 cells (E) were FACS sorted, and mRNA was analyzed using qRT-PCR. *P < 0.001.

Among the genes differentially expressed between GD2+ and GD2 populations, GD3S, a key enzyme involved in the biosynthesis of GD3 (an intermediate for GD2, Figure 3C), was found to be upregulated approximately 9-fold in GD2+ compared to GD2 cells (Supplemental Table 2). The microarray data were validated by qRT-PCR (Figure 3D). However, expression of the gene encoding GD2S, which is involved in conversion of GD3 to GD2, was not altered (Figure 3D). Expression of a number of genes involved in migration and invasion, including MMPs (MMP2, MMP7, and MMP19), and EMT-associated markers, including N-cadherin and vimentin, were expressed at higher levels, whereas E-cadherin was expressed at low levels in GD2+ cells (Supplemental Table 2). We confirmed these findings by qRT-PCR (Supplemental Figure 6). In addition, CD44 mRNA was upregulated and CD24 mRNA downregulated in GD2+ relative to GD2 cells, which was confirmed by FACS analysis (Figure 2A). In addition, the stem cell marker nestin was also found to be upregulated in GD2+ cells compared with GD2 cells (Supplemental Figure 7). These and other genes that were differentially expressed in GD2+ versus GD2 cells are listed in Supplemental Table 2. Conversely, as in GD2+ versus GD2 cells, GD3S was overexpressed more than 10-fold in CD44hiCD24lo compared with CD4loCD24hi cells (Supplemental Figure 8), but no significant difference was found in the expression of GD2S between CD44hiCD24lo and CD4loCD24hi cells. This again demonstrates that the expression of GD3S and GD2 strongly correlates with the CD44hiCD24lo phenotype. Similar to HMLER cells, GD2+ cells from MDA-MB-231 cells expressed GD3S at a more than 5-fold-higher level than GD cells, and consistent with our earlier finding, no significant differences in GD2S expression were observed (Figure 3E).

GD2 cells can spontaneously generate GD2+ cells. Since we observed only a 2-fold difference in mammosphere formation and a 2- to 5-fold difference in tumor initiation between GD2+ and GD2 populations, we investigated whether this was due to the generation of GD2+ cells from GD2 cells. In fact, GD2+ and GD2 cells were sorted from HMLER (Figure 4A) and MDA-MB-231 cells (Figure 4B) and cultured in vitro for 12 days in their respective growth media. Surprisingly, approximately 10% of GD2+ HMLER cells had become GD2, and 15% of GD2 cells had spontaneously generated GD2+ cells, and this proportion was almost identical to that in the unfractionated original HMLER cells (Figure 4A). Similarly, the GD2+ and GD2 cells from MDA-MB-231 cells also generated 81% (81% 2.5%) of GD2 and 12% of GD2+ cells, respectively, again reflecting the percentage of GD2+ cells within the parental MDA-MB-231 cell composition. To investigate the generation of GD2+ cells from GD2 cells and vice versa in vivo, GFP-labeled MDA-MB-231 cells were sorted into GD2+ and GD2 fractions, and 1 106 GD2+ and GD2 cells (GD2+/ cells) were subcutaneously transplanted into NOD/SCID mice. Four weeks later the tumors were dissected, and single-cell suspensions were prepared as described in Methods. The cells were then stained with anti-GD2 antibody and analyzed by flow cytometry. Tumors generated by GD2+ cells consisted of nearly 91% 4.5% GD2 cells, whereas 2.4% 1.1% of cells in GD2 derived tumors were positive for GD2. These findings indicate that GD2+ cells can spontaneously achieve a GD2 phenotype and vice versa in vivo (Supplemental Figure 9, A and B).

GD2-depleted cells are able to generate GD2+ cells in culture: a possible role of EMT. (A and B) GD2+/ cells from HMLER (A) MDA-MB-231 (B) cells were FACS sorted and cultured in MEGM medium for 10 days. After incubation, the cells were stained with GD2 antibody (BD) and analyzed on an LSR II flow cytometer. Note the regeneration of GD+ cells in a GD2-depleted population. (C) To determine the possible role of EMT, HMLER cells transduced with two known EMT inducers (Twist and Snail) were stained with anti-GD2 antibody and analyzed on an LSR II flow cytometer. Expression of GD2 is shown on the y axis and FSC on the x axis. Lower panels in A and C represent antibody staining controls without primary antibody for each cell line. (D) Graphic representation of percentage of GD2+ vector control or Twist- or Snail-transduced HMLER cells. (E and F) mRNA expression analysis of GD3S (E) and GD2S (F) in vector- or Twist- or Snail-transduced cells was performed by real-time TaqMan qRT-PCR. *P < 0.001.

Induction of EMT generates GD2+ cells. Since we recently reported that the induction of EMT in HMLER cells results in the acquisition of stem cell properties (12), we also examined the expression of GD2 on HMLER cells induced to undergo EMT by the ectopic expression of either Twist or Snail. Strikingly, we found that the induction of EMT by Snail or Twist resulted in a significant increase in the percentage of GD2+ populations from the initial 18% (control) to 40% in HMLER-Snail cells and 100% in HMLER-Twist cells (Figure 4, C and D). Corroborating our previous data suggesting a correlation between GD3S and CSCs, we also found that the expression of GD3S mRNA increased in the EMT-derived HMLER cells following induction of EMT by 2.5-fold in Snail cells and 8-fold in Twist cells (Figure 4E), which correlates with the total percentage of GD2+ cells in their respective population (40% in Snail and 100% in Twist cells). In contrast, we found no significant difference in the expression of GD2S (Figure 4F), supporting the hypothesis that GD3S is the key regulator in the biosynthesis of GD2.

GD3S is necessary for CSC properties. To investigate the functional role of GD2 in CSCs, we suppressed the expression of GD3S, the critical enzyme involved in the biosynthesis of GD2, in MDA-MB-231 cells using a lentiviral-based shRNA expression vector and achieved more than 80% knockdown (Figure 5A). As expected, GD3S knockdown reduced the percentage of GD2+ cells from 12.3% (12.3% 1.7%) to 5.5% (5.5% 0.8%) in MDA-MB-231 cells (Figure 5, B and C). Since GD3S is known to regulate a-series gangliosides including GM3, we tested whether knockdown of GD3S could induce the expression of GM3 in MDA-MB-231 cells. Flow cytometric analysis revealed that expression of GM3 was increased from 0.4% 0.3% (control cells) to 15% 1.4% (in GD3S knockdown [GD3S-KD] cells), suggesting that knockdown of GD3S was efficient in these cells (Supplemental Figure 10, A and B). In addition, functional analysis revealed that GD3S-KD-MDA-MB-231 (GD3S-KD-MDA231) cells migrated approximately 3-fold less in Transwell Matrigel invasion assays (Figure 5D) and formed 3-fold fewer mammospheres compared with controls (Figure 5E). To further investigate the effects of suppression of GD3S on tumor formation, we subcutaneously injected MDA-MB-231cells expressing either control shRNA or the GD3S shRNA into the flank of NOD/SCID mice. Strikingly, even after 8 weeks, 1 106 GD3S shRNA cells had not formed tumors, whereas the control shRNA cells had formed tumors in 4 of 4 mice (Figure 5F). The growth rate (tumor size) was also dramatically altered, as plotted in Figure 5G.

Knockdown of GD3S reduces cell proliferation, mammosphere formation, and tumor initiation in MDA-MB-231 cells. (A) To measure knockdown of GD3S, vector control, or GD3S-KD-MDA231 cells were analyzed for mRNA expression of GD3S by real-time TaqMan RT-PCR. Relative expression of GD3S is shown. *P < 0.0001. (B) To measure levels of GD2 on the cell surface, vector control or GD3S-KD-MDA231 cells were stained with anti-GD2 antibody and analyzed on an LSR II flow cytometer (BD). GD2 expression is shown on the y axis and FSC on the x axis. (C) Percentage of GD2+ cells in vector control and GD3S-KD-MDA231 cells. *P < 0.01. (D) To measure cell migration, vector control and GD3S-KD-MDA231 cells were cultured in the presence or absence of 30% serum in a Transwell migration chamber. The average number of cells per microscopic field is shown. *P < 0.001. (E) Mammosphere formation assay using either vector control or GD3S-KD-MDA231 cells was performed by seeding 1,000 cells per well in 24-well cell culture dishes containing mammosphere growth medium. After 10 days, the mammospheres were counted under a light microscope. Scale bar: 200 m. Numbers of mammospheres formed from either vector control or GD3S-KD-MDA231 cells are shown. *P < 0.0001. (F) To examine tumor initiation potential, 1 106 vector control or GD3S-KD-MDA231 cells were transplanted subcutaneously into flanks of NOD/SCID mice. At the end of the ninth week, mice were shaved to remove excess hair on the tumors, and photographs were taken. (G) The tumors size was measured between 4 and 9 weeks. P < 0.000001.

Triptolide, a small molecule inhibitor, inhibits GD3S expression and CSC properties. Triptolide, a small molecule anti-inflammatory drug, has been shown to inhibit GD3S in a melanoma cancer cell line (25). Therefore, we investigated whether triptolide could inhibit GD3S in breast cancer cell lines as well. MDA-MB-231 and SUM-159 cells were treated with different concentrations of triptolide for 24 hours. Triptolide inhibited GD3S mRNA expression in both cell types in a dose-dependent manner, with greater than 95% inhibition at 125 nM (Figure 6, A and B). To test whether inhibition of GD3S by triptolide also inhibited GD2 expression, we treated MDA-MB-231 cells with different concentrations of triptolide for either 24 or 48 hours. Absolute cell counts were measured using flow cytometry. A dose- and time-dependent decrease in GD2+ cells was observed after triptolide treatment, indicating the successful inhibition of GD3S by triptolide (Figure 6C). Of note, a decrease in GD2+ cells was seen under conditions that induced apoptosis in less than 5% of cells.

Triptolide inhibits the expression of GD3S, induces apoptosis in MDA-MB-231 cells, and blocks tumor growth in NOD/SCID mice. MDA-MB-231 cells (A) or SUM-159 cells (B) (5 105/well) of 6-well cell culture dishes were treated with 25, 50, 75, 100, or 125 nM triptolide for 24 hours. /, no treatment. Total RNA was extracted, and GD3S expression was measured by qRT-PCR. (C and D) To measure GD2+ cell growth inhibition, 5 105 MDA-MB-231 (C) and SUM-159 (D) cells were plated in each well of 6-well cell culture dishes and treated with 25, 50, 75, 100, or 125 nM triptolide for 24 or 48 hours. After incubation, the cells were detached with trypsin and stained with anti-GD2 antibody and Sytox Red (for dead cells; Invitrogen). The stained cells were analyzed on an LSR II flow cytometer. Absolute numbers of live cells were calculated by measuring 1,000 events for Trucount beads as explained in Methods. (E) To determine the inhibition of tumor growth, 1 106 MDA-MB-231 cells were subcutaneously transplanted into NOD/SCID mice (n = 8; 4 mice/group). A group of the mice were treated with 0.15 mg/kg/d triptolide, and the control group was treated with PBS every day by i.p. injection. At the end of 8 weeks, mice were sacrificed, and tumors were dissected out and photographed. (F) Tumor sizes from the mice in experiment in E were measured every week after tumor engraftment, and the measurements are shown. P < 0.001, week 3. (G) The survival analysis was based on Kaplan-Meier estimation, and groups were compared by the log-rank test. Control (n = 4, black line) and triptolide (n = 4, blue line) were analyzed for cumulative survival. Survival was defined as the time (in weeks) from transplantation until death. P = 0.015.

To further examine whether triptolide could also inhibit tumor growth in vivo, we introduced 1 106 MDA-MB-231 cells subcutaneously into NOD/SCID mice (2 injections per mouse and 4 mice per group). After the tumors reached 50 mm3, we randomly divided the mice into two groups and treated half of the mice with triptolide (0.15 mg/kg/d) and the other half with PBS (control mice) every day by i.p. injection. Interestingly, after 4 weeks, triptolide-treated animals showed a dramatic decrease in tumor growth compared with control mice. Fifty percent of triptolide treated breast tumors were completely tumor free, and there was a more than 8-fold reduction in tumor volume in 25% of mice (Figure 6D). In addition, tumors in triptolide-treated mice were 3-fold smaller in size and 4-fold lighter by weight (Figure 6E and Supplemental Figure 11). Moreover, in a repeat, identical experiment, triptolide significantly prolonged survival of the treated mice (log-rank, control vs. triptolide, P = 0.0015) (Figure 6F). These findings indicate that GD3S plays a major role in regulating GD3S expression and the resulting GD2+ population. Specifically, it affects cell proliferation and tumor initiation of GD2+ breast cancer cells and when inhibited, greatly diminishes tumor growth and increases metastasis-free survival of breast cancerbearing mice.

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I. Background:

A. Information And Statistics About Diabetes

Diabetes mellitus is a disease of metabolism presenting as a complex group of syndromes that have in common elevated blood glucose levels. It occurs because the insulin produced by the beta cells of the pancreas is either absent, insufficient, or not used properly by target tissues. As a result, the body is unable to metabolize macronutrients in food in the normal way. Since the body cannot convert glucose into energy, high levels of glucose remain in the blood and spill into the urine, eventually resulting in micro-vascular complications (for example, kidney disease and eye disease) and macro-vascular complications (for example, stroke and ischemic heart disease).

There are two major types of diabetes that affect the Medicare population, Type 1 diabetes, previously called insulin dependent diabetes mellitus, and Type 2 diabetes, previously called non-insulin dependent diabetes mellitus.

The goals in the management of diabetes are to achieve normal metabolic control and reduce the risk of micro and macro-vascular complications. Numerous epidemiologic and interventional studies point to the necessity of maintaining good glycemic control to reduce the risk of the complications of diabetes.

Despite this knowledge, diabetes remains the leading cause of blindness, lower extremity amputations, and kidney disease requiring dialysis. Diabetes and its complications are primary or secondary factors in an estimated 9 percent of hospitalizations (Aubert, RE, et al., Diabetes-related hospitalizations and hospital utilization. In: Diabetes in America. 2nd ed. National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Disease, NIH, Pub. No. 9514681995: 553570).

Overall, beneficiaries with diabetes are hospitalized 1.5 times more often than beneficiaries without diabetes. Ten percent of these hospitalizations are a direct result of uncontrolled diabetes, and more than half of these admissions occur in beneficiaries 65 and older (National Hospital Discharge Survey, U.S. National Center for Health Statistics, U.S. Department of Health and Human Services, 1990). In expanding the Medicare program to include outpatient diabetes self-management training services, the Congress intended to empower Medicare beneficiaries with diabetes to better manage and control their conditions. The Conference Report indicates that the conferees believed that this provision will provide significant Medicare savings over time due to reduced hospitalizations and complications arising from diabetes. (H.R. Conf. Rep. No. 105217, at 701 (1997)).

According to the National Health and Nutrition Examination Survey (NHANES):

(Diabetes Fact Sheet, The Centers for Disease Control & Prevention https://www.cdc.gov/diabetes/pubs/pdf/ndfs_2011.pdf)

According to the Department of Health and Human Services Centers for Disease Control and Prevention (CDC):

B. Statutory Authority for Diabetic Self-Management Training (DSMT)

Section 4105(a) of the Balanced Budget Act of 1997 (BBA) (pub. L. 105-33), enacted on August 5, 1997, provides for Medicare coverage for DSMT services provided by a certified provider. Section 4105 of the BBA amended section 1861 of the Social Security Act (The Act) by adding a new section (q)(q).

Section 1861(qq) of the Social Security Act (the Act) provides CMS with the statutory authority to regulate Medicare outpatient coverage of DSMT services.

Section 1861(q)(q)(2) provides that the Secretary may recognize a physician, individual, or entity that is recognized by an organization as meeting standards for furnishing these services as a certified DSMT provider. This statute also provides that a physician or other individual or entity shall be deemed to have met such standards if they meet applicable standards originally established by the National Diabetes Advisory Board.

Section 1861(q)(q)(2)(B) of the Act states that a physician, or such other individual or entity, meets the quality standards if the physician, or individual or entity, meets quality standards established by the Secretary, except that the physician or other individual or entity shall be deemed to have met such standards if the physician or other individual or entity meets applicable standards originally established by the National Diabetes Advisory Board and subsequently revised by organizations who participated in the establishment of standards by such Board, or is recognized by an organization that represents individuals (including individuals under this title) with diabetes as meeting standards for furnishing the services.''

Additionally, section 4105(c)(1) of the BBA requires the Secretary to establish outcome measurements for purposes of evaluating the improvement of the health status of Medicare beneficiaries with diabetes.

A final rule (65 FR 83130) was published in the Federal Register on December 29, 2000 which implemented the BBA provisions addressing the coverage, payment, quality standards, and accreditation requirements for DSMT. This final rule also implemented the DSMT regulations which are codified at Title 42 of the Code of Federal Regulation (CFR) sections 410.140 to 410.146.

The CMS regulations at 42 CFR 410.144 provide the authority for the CMS to require the DSMT AOs to use one of the following types of accreditation standards: (1) the accreditation standards set forth at 410.144(a); (2) the accreditation standards issued by the National Standards for Diabetes Self-Management Education Support (NSDSMES) (410.144(b)); or (3) other accreditation standards, so long as they have been submitted to CMS and approved as meeting or exceeding the CMS quality standards described at 410.144(a).

The American Diabetes Association (ADA) and the American Association of Diabetic Educators (AADE) are the two national DSMT AOs approved by CMS to accredit entities that furnish DSMT services. These DSMT AOs are approved by CMS for six-year terms. Section 410.143(a) sets forth the ongoing responsibilities of the DSMT AOs. The requirement at section 410.143(b) sets forth the oversight activities that CMS, or its agent, will perform to ensure that a CMS approved DSMT AO and the entities the organization accredits continue to meet a set of quality standards described at 410.144.

Section 410.145 of the regulations specifies requirements that DSMT entities must meet. Section 410.146 requires that approved entity must collect and record in an organized systematic manner, patient assessment information at least on a quarterly basis for a beneficiary who receives DSMT training.

II. Information about the Diabetic Self-Management Training Accrediting Organizations

A. General Information

B. Information About The ADA Education Recognition Program (ERP)

C. Information About the AADE

III. Contact Information for CMS-designated DSMT Accrediting Organizations

There are two accrediting organizations approved by CMS to accredit DSMT entities. The contact information for these DSMT AO is listed below.

American Diabetes Association (ADA)

2451 Crystal City Drive

Suite 800

Arlington, VA 22202

703-549-1500

Phone: 800-342-2383

Website: http://www.diabetes.org

American Association of Diabetic Educators (AADE)

200 W. Madison

Suite 800

Chicago, IL 60606

Phone: 800-338-3633

Website: https://www.diabeteseducator.org

IV. Oversight and Validation Process for DSMT Accrediting Organizations Accreditation Processes

V. Where to Submit Questions Related to the DSMT Accreditation Program

Questions about the DSMT Accreditation Program may be submitted to the DSMT Accreditation email box at DSMTAccreditation@cms.hhs.gov.

We monitor this email box on a frequent basis and will respond to your email as soon as possible.

Read more here:
Diabetic Self-Management Training (DSMT) Accreditation Program