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1. We sought to determine if catecholamine biosynthetic enzymes of spontaneously hypertensive rats (SHR) differed from those of normotensive Wistar—Kyoto (WKY) and Sprague—Dawley (SD) control rats before birth.
2. By immunocytochemical and biochemical methods we compared strains for the time of appearance and maturation of the enzymes tyrosine hydroylase (TH), dopamine-β-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT) in sympathetic ganglia and adrenals.
3. The time of appearance of enzymes was identical in all three strains: TH and DBH first appeared in sympathetic ganglia on embryonic day 11 (E11) and in adrenal medulla on E16. PNMT, restricted to adrenal medulla, appeared later on E18.
4. The activity of adrenal TH prenatally on E18 and E21 and at day of birth (P1) in SHR was approximately two fold that in WKY or SD rats. In contrast PNMT was lower in SHR but only on E18.
5. Thus, although the timing of the first expression of adrenergic phenotypes is similar in SHR and normotensive controls, the differences in TH activity in adrenals suggest an enhanced biosynthetic capacity for catecholamines in this strain before birth.
6. We conclude that SHR differ from normotensive rats from the first expression of some of the genes controlling catecholamine biosynthesis.
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Choking - unconscious adult or child over 1 year
Choking is when someone cannot breathe because food, a toy, or other object is blocking the throat or windpipe (airway).
This article discusses choking in adults or children over age 1 who have lost alertness (are unconscious).
Causes
Choking may be caused by:
• Eating too fast, not chewing food well, or eating with dentures that do not fit well
• Drinking alcohol (even a small amount of alcohol affects awareness)
• Being unconscious and breathing in vomit
• Breathing in or swallowing small objects (young children)
• Injury to the head and face (for example, swelling, bleeding, or a deformity can cause choking)
• Swallowing problems caused by a stroke or other brain disorders
• Enlarging tonsils or tumors of the neck and throat
• Problems with the esophagus (food pipe or swallowing tube)
Symptoms
Symptoms of choking when a person is unconscious include:
• Bluish color to the lips and nails
• Inability to breathe
First Aid
Tell someone to call 911 or the local emergency number while you begin first aid and CPR.
If you are alone, shout for help and begin first aid and CPR.
1. Roll the person onto their back on a hard surface, keeping the back in a straight line while firmly supporting the head and neck. Expose the person's chest.
2. Open the person's mouth with your thumb and index finger, placing your thumb over the tongue and your index finger under the chin. If you can see an object and it is loose, remove it.
3. If you do not see an object, open the person’s airway by lifting the chin while tilting the head back.
4. Place your ear close to the person's mouth and watch for chest movement. Look, listen, and feel for breathing for 5 seconds.
5. If the person is breathing, give first aid for unconsciousness.
6. If the person is not breathing, begin rescue breathing. Maintain the head position, close the person's nostrils by pinching them with your thumb and index finger, and cover the person's mouth tightly with your mouth. Give two slow, full breaths with a pause in between.
7. If the person's chest does not rise, reposition the head and give two more breaths.
8. If the chest still does not rise, the airway is likely blocked, and you need to start CPR with chest compressions. The compressions may help relieve the blockage.
9. Do 30 chest compressions, open the person's mouth to look for an object. If you see the object and it is loose, remove it.
10. If the object is removed, but the person has no pulse, begin CPR with chest compressions.
11. If you do not see an object, give two more rescue breaths. If the person's chest still does not rise, keep going with cycles of chest compressions, checking for an object, and rescue breaths until medical help arrives or the person starts breathing on their own.
If the person starts having convulsions or seizures, give first aid for this problem.
After removing the object that caused the choking, keep the person still and get medical help. Anyone who is choking should have a medical examination. This is because the person can have complications not only from the choking, but also from the first aid measures that were taken.
DO NOT
Do NOT try to grasp an object that is lodged in the person's throat. This may push it farther down the airway. If you can see the object in the mouth, it may be removed.
When to Contact a Medical Professional
Seek medical help right away if someone is found unconscious.
In the days following a choking episode, contact a doctor right away if the person develops:
• A cough that does not go away
• Pneumonia and fever
• Shortness of breath
• Wheezing
These could be signs that the object entered the lung instead of being expelled
Prevention
To prevent choking:
• Eat slowly and chew food completely.
• Do not drink too much alcohol before or during eating.
• Keep small objects away from young children.
• Make sure dentures fit properly.
Alternative Names
Choking - unconscious adult or child over 1 year; First aid – choking - unconscious adult or child over 1 year; CPR - choking - unconscious adult or child over 1 year
References
American Red Cross.First Aid/CPR/AED Participant's Manual.
Berg RA, Hemphill R, Abella BS, et al. Part 5: Adult basic life support: 2010 American Heart Association Guidelines for cardiopulmonary resuscitation and emergency cardiovascular care.Circulation.http://www.ncbi.nlm.nih.gov/pubmed/20956221
Berg MD, Schexnayder SM, Chameides L, et al. Part 13: Pediatric basic life support: 2010 American Heart Association guidelines for cardiopulmonary resuscitation and emergency cardiovascular care.Circulation.http://www.ncbi.nlm.nih.gov/pubmed/20956229
Cukor J, Manno M. Pediatric respiratory emergencies. In: Marx J, Hockberger RS, Walls RM, eds.Rosen's Emergency Medicine: Concepts and Clinical Practice.
Thomas SH, Goodloe JM. Foreign bodies. In: Marx JA, Hockberger RS, Walls RM, eds.Rosen's Emergency Medicine: Concepts and Clinical Practice.
Update Date 4/12/2015
Related MedlinePlus Health Topics
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Thick fog will also be seen on west coast and the mountainous region. The frame can be extruded as a one-piece construction. commande-contrôle d’une salle de commande ou des pupitres et panneaux de commande des appareils, pour débiter des billes en pièces de bois brutes et scier, fendre, tailler et raboter ce premier débit en pièces de différentes dimensions et scier ou fendre des bardeaux et des bardeaux de fente; Operate automated lumbermill equipment from control rooms or equipment consoles to saw logs into rough lumber; saw, trim and plane rough lumber into dressed lumber of various sizes; and saw or split shingles and shakes Mais tu vas t’irriter, à force de te raboter. ls it greedy to want to feed your family? J’irais jusqu’au jardin de tuiles pour voir si ils n’ont pas ces tuiles d’adobe. It is said to be a way of kick-starting the metabolism. Include a letter from your doctor explaining your need for all medications you are carrying, including any over-the-counter medications, in English and the language of your destination(s).
Cancer was named after the crab, because it describes the way that the disease kills – like a crab eating you. House employees of a French or foreign citizen, travelling with their employer to France for a short visit (less than 3 months) As any foreign citizen coming to France to work, on full or part time basis, domestic and private employee of either French or foreign citizen, coming to France with his employer in order to work, has to hold a work authorisation, stamped by French authorities. There are several different types of glaucoma, including open-angle glaucoma and acute angle-closure glaucoma. As of the year 2000, results were not yet available from controlled double-blind studies so the value of IDET remained to be proven. deterrent, stop. During attacks they may use topical treatments, such as steroid creams. Bathing frequently, especially with soaps, can contribute to dry skin.
There are two additional funds for the self-employed and agricultural workers. Microbiology made advances, a science that started with Antonie Philips van Leeuwenhoek (1632 – 1723), who first observed microorganisms with a microscope. Early stages of recolonization were studied from 1995 to 1997 after cessation of dredging. Paid Online Questionnaires, Content Writing, Search Marketing are all examples of Wirk. Your visa application must contain, aside from the other required supporting documents, two passport photos meeting the ICAO standards. Aromatherapy is a widely used term for a range of traditional therapies that use essential oils. To immunize against viral diseases, the virus used in the vaccine has been weakened or killed.
PHN is a nerve pain (neuralgia) that persists after a shingles rash has cleared. While in the general dictionary you will find usual words and expressions from the famous publisher Collins, in the Collaborative Dictionary you will discover slang terms, technical translations, familiar words and expressions, regionalisms that are difficult to find in the traditional online dictionaries. However, until more is known, pycnogenol should be used cautiously or avoided by women who are pregnant. PHN is a nerve pain (neuralgia) that persists after a shingles rash has cleared. Conine taught there in 1891, later serving on the board of directors. Ruth, Yelm; Attorney J.R. Traditional painkillers such as paracetamol, anti-inflammatories and codeine usually do not help very much.
Two young children have died, several others are afflicted and the school has been closed. The following depicts part of there life there. Traditional painkillers such as paracetamol, anti-inflammatories and codeine usually do not help very much. Today’s cough drops and peppermint sticks descend from this tradition. Though it’s worth mentioning if you plan to stay in Europe for more than 2 months, I’d recommend a quick look in case you’re not asking these Schengen questions (yet). This leaflet only deals with the most common. There is general agreement that the term “Creole” derives from the Portuguese word crioulo, which means a slave born in the master’s household.
Note: In calculating the moving wall, the current year is not counted. NBC have flatly denied the claim that botox played any part in the sportscasters’ condition. It’s waterproof, it regulates our body temperature and it helps make vitamin D – essential for strong, healthy bones. Geological history of Britain and Ireland. Nonetheless, resistance to biomedicine through recourse to alternative therapies is mixed with ongoing dependence on the biomedical system, since patients seek strategic combinations of both systems to maximize health and other benefits. Also, was it my imagination or was the shingle pain similar, although not in the same location, to CHs? There are many causes of chest pain.
Life on the home front during World War II was a significant part of the war effort for all participants and had a major impact on the outcome of the war.
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Nutrition Therapy for Cancer Patients
By , Onlymyhealth editorial team
May 30, 2011
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Nutrition is important for good health and implies a healthy diet that includes eating a combination of vitamins, proteins, carbohydrates, fat and water, which the body needs, to stay healthy and active.
Nutrition therapy is important for cancer patients to enable them to have the nutrients needed to keep up their body weight, strength; stay healthy and combat infection. Well-nourished cancer patients get the calories and nutrients they need to combat the ill effects of cancer treatment. They have a better prognosis and enjoy a better quality of life.
The effects of cancer treatments make it difficult for many to eat well. Cancer treatments that affect nutrition are surgery, chemotherapy, radiation therapy, immunotherapy and stem cell transplants. When the head, neck, oesophagus, stomach and intestines are affected by the aggressiveness of these treatments, the patient faces the unintended side effects of these treatments such as -
• Anorexia (loss of appetite)
• Mouth sores
• Dry mouth
• Trouble swallowing
• Nausea
• Vomiting
• Diarrhoea
• Constipation
• Depression
• Anxiety
• Pain
Further, cancer treatments affect taste, smell and appetite and hence such people are unable to eat enough to absorb the key nutrients from food and become malnourished, diminishing their ability to fight the disease. It is therefore important that cancer symptoms and side effects that affect eating are detected early. Both nutrition therapy and medicines could be used for increasing appetite, improving digestion, preventing nausea and vomiting and reducing pain.
Nutrition therapy for cancer patients who are under active treatment and those who are in the process of recovery has three main goals -
1. Treating nutrition problems that prevent muscle and bone loss
2. Help the patient’s immune system fight infection and help the body recover and heal
3. Keep up and improve the patient’s quality of life.
Methods of giving nutrition therapy for cancer patients unable to eat may be invasive in some cases and involves what is medically known as enteral feeding, In this case, a feeding tube is inserted directly into the stomach or intestines. The other method is called the parenteral method where the nutrients are infused into the bloodstream through the veins. The liquids in both casesare formulas that contain water, proteins, fats, carbohydrates, vitamins and minerals in accordance to the patient’s specific needs.
These methods of feeding have to be continued at home by a care giver after the patient has recovered and discharged from the hospital. Removal of these items when the patient has recovered at home should be done under medical supervision.
Treatment of cancer uses various methods depending on the specific type or location and is generally surgery, chemotherapy, radiotherapy, immuno therapy and stem cell transplants. While these may ultimately lead to the remission of the tumours, the patient has to endure the ill effects of such aggressive treatments. The chief of these is lack of appetite (anorexia), compounded by mouth sores, dryness of mouth, diarrhoea, continuing pain, depression and anxiety. As a result the patient cannot take the vitamins and minerals, proteins and carbohydrates that the body needs to sustain the treatment and fight infection.
Nutrient therapy for cancer patients steps in at this point by devising means to introduce these items into the body by introducing these vital foods directly into the stomach or intestines, or through the veins into the bloodstream.
Read more articles on Cancer Treatment.
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Can a Photo Facial Cause Swollen Gland?
I am experiencing swollen glands in my neck after having IPL. Could this be a side effect?
Doctor Answers 1
Photofacial
It is very unlikely to be a side effect from IPL. Thank you for your question and good luck with everything.
Danville Plastic Surgeon
4.9 out of 5 stars 25 reviews
These answers are for educational purposes and should not be relied upon as a substitute for medical advice you may receive from your physician. If you have a medical emergency, please call 911. These answers do not constitute or initiate a patient/doctor relationship.
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Main Line: 817-346-7333
EXCEPTIONAL EYE CARE IN FORT WORTH FOR THIRTY YEARS
Styes Explained
A stye, also known as a hordeolum, is a small yet painful lump that develops outside or inside a person’s eyelid. Just like a pimple, it is an abscess that contains pus. It is usually caused by poor eye hygiene, which can lead to a staphylococcus infection.
This is a common problem that any eye specialist has encountered. At some point in our lives, we have all experienced this condition. Keep in mind that the stye should not be confused with the chalazion, which is a painless round bump found in the middle portion of an eyelid.
Different Types of Styes
There are two different types of styes. The first is the external stye. It is a small spot that develops next to a solitary eyelash. This type will feel tender and patients will observe a noticeable swelling. However, the external stye will resolve in a few days after it bursts.
A stye found on the underside of a lid can cause the same amount of pain and discomfort. The internal stye is quite uncomfortable as in most cases it can be felt by the eyeball itself. Since it cannot be seen, the structure may heal and leave a fluid-filled cyst or a node. This will usually require excision or draining by a skilled eye specialist.
What Can Cause Stye Formation?
As mentioned previously, this problem is caused by staphylococci. These bacteria live on the skin surface, also called the epithelium, with other types of bacteria. When we fail to clean our skin properly, the staphylococci engorge themselves with debris and dead skin cells, which leads to pimple formation.
Symptoms of Stye Formation
In some instances the area may feel itchy a day or two before. The discomfort begins when inflammation sets in. The abscess formation will cause a whitish bump to form, while the surrounding area is usually reddish in color.
Stye Prevention
If you suffer from recurrent styes, you may need to improve your eyelid hygiene regimen. Any eye specialist will tell you that it would be best to use warm water and no-tears baby shampoo to rid the eyelids of dirt and debris. Use gentle rubbing movements to prevent irritating this delicate area.
Stye Treatment
A stye will usually resolve on its own. You can use a warm compress to relieve the discomfort. However, if it interferes with your vision or is extremely uncomfortable, it would be best to see an eye specialist. The doctor may prescribe oral or topical antimicrobial solutions depending on the case.
Styes are uncomfortable and unsightly, so most people who suffer from this condition avoid public appearances. If you have recurrent styes and practice proper eye hygiene, make sure to see an eye specialist, because you may have an undiagnosed skin condition that needs to be treated.
At Marvel Eye Center, we strive to help our patients in any way we can. If you have a stye condition that does not resolve itself, contact us today and we’ll work with you to treat this common eye problem. Our office number is (817) 346-7333.
Marvel Eye Center
About The Marvel Eye Center
EXCEPTIONAL EYE CARE IN THE DALLAS-FORT WORTH AREA FOR THIRTY YEARS
© Copyright 2017 Marvel Eye Center | Web Marketing Powered by Ceatus Media Group LLC | Sitemap | Privacy Policy
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An inhibitory septum to lateral hypothalamus circuit that suppresses feeding
Patrick Sweeney, Yunlei Yang
Research output: Contribution to journalArticlepeer-review
50 Scopus citations
Abstract
Feeding behavior is orchestrated by neural circuits primarily residing in the hypothalamus and hindbrain. However, the relative influence of cognitive and emotional brain circuits to the feeding circuitry in the hypothalamus and hindbrain remains unclear. Here, using the cell-type selectivity of genetic methods, circuit mapping, and behavior assays, we sought to decipher neural circuits emanating from the septal nucleus to the lateral hypothalamus (LH) that contribute to neural regulation of food intake in mice. We found that chemogenetic and optogenetic activation of septal vesicular GABA transporter (vGAT)-containing neurons or their projections in the LH reduced food intake in mice. Consistently, chemogenetic inhibition of septal vGAT neurons increased food intake. Furthermore, we investigated a previously unknown neural circuit originating from septal vGAT neurons to a subset of vGAT neurons in the LH, an area involved in homeostatic and hedonic control of energy states. Collectively, our data reveal an inhibitory septohypothalamic feeding circuit that might serve as a therapeutic target for the treatment of eating disorders such as anorexia nervosa.
Original languageEnglish (US)
Pages (from-to)11185-11195
Number of pages11
JournalJournal of Neuroscience
Volume36
Issue number44
DOIs
StatePublished - Nov 2 2016
Externally publishedYes
Keywords
• Chemogenetics
• Food intake
• Lateral hypothalamus
• Optogenetics
• Septum
ASJC Scopus subject areas
• Neuroscience(all)
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Headaches: Causes And Treatments
Causes and Treatments of headaches You may experience headaches once in a while or maybe even every day. They come in a variety of levels and ways but what are the causes? Are you getting them more often than you’d like? Turns out, you may be able to prevent their onset if you can pinpoint the source. Below, we outline 5 different types of headaches and what triggers them.
Tension headaches- This is the most common type. These headaches feel like constant pressure or tension around your head, particularly around your temples and your neck. It is believed that these may be caused by the contraction of the scalp and neck muscles. They can be offset if you’re not getting enough sleep at night, dealing with a lot of stress or anxiety, from alcohol or even from straining your eyes. You can treat them with over the counter acetaminophen or ibuprofen. These types of headaches shouldn’t cause too much concern unless you’re dealing with them excessively.
Treatment: Over-the-counter products can alleviate pain in a prompt and effective manner. Do not become too dependent on these medications or take them too often, however, or you may fall victim to rebound headaches.
Cluster headache- Have you ever had a burning or piercing sensation in or around one eye? If so, then you’ve probably experienced a cluster headache, which typically occur in cycles or groups throughout the course of the day. Often times, they appear on one side of the head and can cause excruciating pain; lying down can often worsen the condition. In fact, during an attack, most sufferers will pace as they find that they cannot sit still. While the cause may be traced back to genetics, the true cause is still unknown. They’re also pretty rare, affecting up to 1% of the population.
Treatment: Due to the nature of these headaches, including its quick onset lasting only a brief amount of time, there is no cure; you can minimize the amount of pain you’re in, however. For quick relief, inhaling 100% oxygen at a rate of 12 liters a minute through a mask can make a dramatically alleviate your pain.
Rebound headaches- Ironically, too much pain medicine could actually be attributing to your pain. Rebound headaches occur due to the overuse of pain relieving drugs such as aspirin, acetaminophen, or prescription drugs. It often begins with a patient taking lots of medicine for their headaches. This increase can have a counterintuitive effect on the body, as they begin developing headaches more frequently. They then take more medicine, which results in even more headaches and eventually it gets to the point where they are taking the medicine daily. These medicines can place the mind in an excited state, which can cause the headache.Additionally, rebound headaches may be offset from dips of the medicine levels in the bloodstream.
Treatment: To stop these types of headaches, one must stop using short-acting pain relief medication. It may be hard to stop the perpetual cycle at first, but a medical professional can help guide you with long-term treatment options.
Sinus headaches- Your sinuses are the air-filled cavities which can be found behind your forehead, nose, and cheekbones. If you have inflamed sinuses, the pressure in this area can trigger your sinus headache. These types of headaches are may feel worse as you bend forward or lie down. Sinus headaches are hard to self-diagnose; most people think they have a sinus headache but 80-90% of the time, they’re really suffering from a migraine.
Treatment: To treat a sinus headache, one must take care of the underlying cause. The root of your pain is due to these inflamed sinus cavities; therefore treatment options should include a saline nasal spray, humidifier, or prescription antibiotics. Using a humidifier should help as well, as it will add moisture to the air, allowing your sinuses to function properly.
Migraines- Often caused by blood vessel enlargement and the release of chemicals from nerve fibers surrounding enlarged blood vessels, this is one of the most debilitating types of headaches. If you’re suffering from a migraine, you may also suffer from symptoms such as nausea; sensitivity to light, odor, and sound; stomach pain and even appetite loss. Migraines can last a few hours or even a few days and affect more women than men. Often misdiagnosed as sinus or tension headaches, migraines can be triggered by a number of factors: bright lights, allergies, alcohol, smoking, stress, or even by certain foods, such as cheese and red wine.
Treatment: Small amounts of caffeine can help alleviate migraine pain. Over-the-counter medicine can help relieve pain as well. Opt for an ice pack, which provides anti-inflammatory relief, for an easy and effective remedy.
While most headaches will go away on their own, if they’re occurring frequently or causing you excruciating pain, it’s always best to seek medical attention as it may be the sign of something more serious, such as a stroke. If your headache does not respond to treatment, contact your medical professional to ensure that the root of your pain isn’t caused by a more serious condition.
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Skip Nav
When Is Best to Join the Gym
Why Winter's the Best Time to Join the Gym
For many people joining the gym symbolises the beginning of the next step of their health and fitness journey. While you can get amazing results by working out from the comfort of your home, a gym membership comes with the advantages of trainers with years of knowledge and expertise, fitness classes, equipment, and machines that are way too pricey and bulky for home use. If you're deliberating over joining the gym, read on for three reasons that might convince you to take the plunge ahead of the New Year.
It Can Be Cheaper
By the time it gets to the end of the year, gym attendance and memberships start to tail off because most people are busy preparing for the festive season. Many gyms offer joining deals that involve some form of incentive to entice new customers, which can sometimes mean you're paying significantly less than you would by joining at any other time.
It's Quieter
January and May-June are generally the busiest months at the gym, the former because many people are getting into the swing of their New Year's health and fitness resolutions and the latter because some people workout in preparation for Summer holidays. The Winter months are a great time to join the gym because the quieter atmosphere allows you to familiarise yourself with all the equipment and try out classes that tend to have mile-long waiting lists at other times of the year.
You Get a Head Start
As the saying goes, it's easier to stay fit than get fit. If you usually quit your fitness goals to take the Winter off, then struggle to get back into it come Spring, getting a gym membership and signing up for a few weekly classes might help you remain consistent throughout Winter. You'll also be able to find your training stride ahead of the January rush and ease yourself into a consistent routine.
Latest Health & Fitness
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JoVE Visualize What is visualize?
Related JoVE Video
Pubmed Article
Quantitative susceptibility mapping differentiates between blood depositions and calcifications in patients with glioblastoma.
PLoS ONE
PUBLISHED: 01-28-2013
The application of susceptibility weighted imaging (SWI) in brain tumor imaging is mainly used to assess tumor-related "susceptibility based signals" (SBS). The origin of SBS in glioblastoma is still unknown, potentially representing calcifications or blood depositions. Reliable differentiation between both entities may be important to evaluate treatment response and to identify glioblastoma with oligodendroglial components that are supposed to present calcifications. Since calcifications and blood deposits are difficult to differentiate using conventional MRI, we investigated whether a new post-processing approach, quantitative susceptibility mapping (QSM), is able to distinguish between both entities reliably.
Authors: Ardian Hana, Andreas Husch, Vimal Raj Nitish Gunness, Christophe Berthold, Anisa Hana, Georges Dooms, Hans Boecher Schwarz, Frank Hertel.
Published: 08-26-2014
ABSTRACT
DTI is a technique that identifies white matter tracts (WMT) non-invasively in healthy and non-healthy patients using diffusion measurements. Similar to visual pathways (VP), WMT are not visible with classical MRI or intra-operatively with microscope. DTI will help neurosurgeons to prevent destruction of the VP while removing lesions adjacent to this WMT. We have performed DTI on fifty patients before and after surgery between March 2012 to January 2014. To navigate we used a 3DT1-weighted sequence. Additionally, we performed a T2-weighted and DTI-sequences. The parameters used were, FOV: 200 x 200 mm, slice thickness: 2 mm, and acquisition matrix: 96 x 96 yielding nearly isotropic voxels of 2 x 2 x 2 mm. Axial MRI was carried out using a 32 gradient direction and one b0-image. We used Echo-Planar-Imaging (EPI) and ASSET parallel imaging with an acceleration factor of 2 and b-value of 800 s/mm². The scanning time was less than 9 min. The DTI-data obtained were processed using a FDA approved surgical navigation system program which uses a straightforward fiber-tracking approach known as fiber assignment by continuous tracking (FACT). This is based on the propagation of lines between regions of interest (ROI) which is defined by a physician. A maximum angle of 50, FA start value of 0.10 and ADC stop value of 0.20 mm²/s were the parameters used for tractography. There are some limitations to this technique. The limited acquisition time frame enforces trade-offs in the image quality. Another important point not to be neglected is the brain shift during surgery. As for the latter intra-operative MRI might be helpful. Furthermore the risk of false positive or false negative tracts needs to be taken into account which might compromise the final results.
23 Related JoVE Articles!
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Tumor Treating Field Therapy in Combination with Bevacizumab for the Treatment of Recurrent Glioblastoma
Authors: Ayman I. Omar.
Institutions: Southern Illinois University School of Medicine.
A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA approved in April 2011 for the treatment of patients 22 years or older with rGBM. The device delivers alternating electric fields and is programmed to ensure maximal tumor cell kill1. Glioblastoma is the most common type of glioma and has an estimated incidence of approximately 10,000 new cases per year in the United States alone2. This tumor is particularly resistant to treatment and is uniformly fatal especially in the recurrent setting3-5. Prior to the approval of the TTF System, the only FDA approved treatment for rGBM was bevacizumab6. Bevacizumab is a humanized monoclonal antibody targeted against the vascular endothelial growth factor (VEGF) protein that drives tumor angiogenesis7. By blocking the VEGF pathway, bevacizumab can result in a significant radiographic response (pseudoresponse), improve progression free survival and reduce corticosteroid requirements in rGBM patients8,9. Bevacizumab however failed to prolong overall survival in a recent phase III trial26. A pivotal phase III trial (EF-11) demonstrated comparable overall survival between physicians’ choice chemotherapy and TTF Therapy but better quality of life were observed in the TTF arm10. There is currently an unmet need to develop novel approaches designed to prolong overall survival and/or improve quality of life in this unfortunate patient population. One appealing approach would be to combine the two currently approved treatment modalities namely bevacizumab and TTF Therapy. These two treatments are currently approved as monotherapy11,12, but their combination has never been evaluated in a clinical trial. We have developed an approach for combining those two treatment modalities and treated 2 rGBM patients. Here we describe a detailed methodology outlining this novel treatment protocol and present representative data from one of the treated patients.
Medicine, Issue 92, Tumor Treating Fields, TTF System, TTF Therapy, Recurrent Glioblastoma, Bevacizumab, Brain Tumor
51638
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A Matrigel-Based Tube Formation Assay to Assess the Vasculogenic Activity of Tumor Cells
Authors: Ralph A. Francescone III, Michael Faibish, Rong Shao.
Institutions: University of Massachusetts, University of Massachusetts, University of Massachusetts.
Over the past several decades, a tube formation assay using growth factor-reduced Matrigel has been typically employed to demonstrate the angiogenic activity of vascular endothelial cells in vitro1-5. However, recently growing evidence has shown that this assay is not limited to test vascular behavior for endothelial cells. Instead, it also has been used to test the ability of a number of tumor cells to develop a vascular phenotype6-8. This capability was consistent with their vasculogenic behavior identified in xenotransplanted animals, a process known as vasculogenic mimicry (VM)9. There is a multitude of evidence demonstrating that tumor cell-mediated VM plays a vital role in the tumor development, independent of endothelial cell angiogenesis6, 10-13. For example, tumor cells were found to participate in the blood perfused, vascular channel formation in tissue samples from melanoma and glioblastoma patients8, 10, 11. Here, we described this tubular network assay as a useful tool in evaluation of vasculogenic activity of tumor cells. We found that some tumor cell lines such as melanoma B16F1 cells, glioblastoma U87 cells, and breast cancer MDA-MB-435 cells are able to form vascular tubules; but some do not such as colon cancer HCT116 cells. Furthermore, this vascular phenotype is dependent on cell numbers plated on the Matrigel. Therefore, this assay may serve as powerful utility to screen the vascular potential of a variety of cell types including vascular cells, tumor cells as well as other cells.
Cancer Biology, Issue 55, tumor, vascular, endothelial, tube formation, Matrigel, in vitro
3040
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Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells Using the Neurosphere Assay
Authors: Hassan Azari, Sebastien Millette, Saeed Ansari, Maryam Rahman, Loic P. Deleyrolle, Brent A. Reynolds.
Institutions: University of Florida , Shiraz University of Medical Sciences.
Stem-like cells have been isolated in tumors such as breast, lung, colon, prostate and brain. A critical issue in all these tumors, especially in glioblastoma mutliforme (GBM), is to identify and isolate tumor initiating cell population(s) to investigate their role in tumor formation, progression, and recurrence. Understanding tumor initiating cell populations will provide clues to finding effective therapeutic approaches for these tumors. The neurosphere assay (NSA) due to its simplicity and reproducibility has been used as the method of choice for isolation and propagation of many of this tumor cells. This protocol demonstrates the neurosphere culture method to isolate and expand stem-like cells in surgically resected human GBM tumor tissue. The procedures include an initial chemical digestion and mechanical dissociation of tumor tissue, and subsequently plating the resulting single cell suspension in NSA culture. After 7-10 days, primary neurospheres of 150-200 μm in diameter can be observed and are ready for further passaging and expansion.
Neuroscience, Issue 56, Glioblastoma Multiforme, Tumor Cell, Neurosphere Assay, Isolation, Expansion
3633
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Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI
Authors: Heling Zhou, Dawen Zhao.
Institutions: University of Texas Southwestern Medical Center.
Breast cancer brain metastasis, occurring in 30% of breast cancer patients at stage IV, is associated with high mortality. The median survival is only 6 months. It is critical to have suitable animal models to mimic the hemodynamic spread of the metastatic cells in the clinical scenario. Here, we are introducing the use of small animal ultrasound imaging to guide an accurate injection of brain tropical breast cancer cells into the left ventricle of athymic nude mice. Longitudinal MRI is used to assessing intracranial initiation and growth of brain metastases. Ultrasound-guided intracardiac injection ensures not only an accurate injection and hereby a higher successful rate but also significantly decreased mortality rate, as compared to our previous manual procedure. In vivo high resolution MRI allows the visualization of hyperintense multifocal lesions, as small as 310 µm in diameter on T2-weighted images at 3 weeks post injection. Follow-up MRI reveals intracranial tumor growth and increased number of metastases that distribute throughout the whole brain.
Medicine, Issue 85, breast cancer brain metastasis, intracardiac injection, ultrasound imaging, MRI, MDA-MB231/Br-GFP cells
51146
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Preparation of a Blood Culture Pellet for Rapid Bacterial Identification and Antibiotic Susceptibility Testing
Authors: Antony Croxatto, Guy Prod'hom, Christian Durussel, Gilbert Greub.
Institutions: University Hospital Center and University of Lausanne.
Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. The analysis of Gram stains from positive blood culture can be rapidly conducted and already significantly impact the antibiotic regimen. However, the accurate identification of the infectious agent is still required to establish the optimal targeted treatment. We present here a simple and fast bacterial pellet preparation from a positive blood culture that can be used as a sample for several essential downstream applications such as identification by MALDI-TOF MS, antibiotic susceptibility testing (AST) by disc diffusion assay or automated AST systems and by automated PCR-based diagnostic testing. The performance of these different identification and AST systems applied directly on the blood culture bacterial pellets is very similar to the performance normally obtained from isolated colonies grown on agar plates. Compared to conventional approaches, the rapid acquisition of a bacterial pellet significantly reduces the time to report both identification and AST. Thus, following blood culture positivity, identification by MALDI-TOF can be reported within less than 1 hr whereas results of AST by automated AST systems or disc diffusion assays within 8 to 18 hr, respectively. Similarly, the results of a rapid PCR-based assay can be communicated to the clinicians less than 2 hr following the report of a bacteremia. Together, these results demonstrate that the rapid preparation of a blood culture bacterial pellet has a significant impact on the identification and AST turnaround time and thus on the successful outcome of patients suffering from bloodstream infections.
Immunology, Issue 92, blood culture, bacteriology, identification, antibiotic susceptibility testing, MALDI-TOF MS.
51985
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Optimization of High Grade Glioma Cell Culture from Surgical Specimens for Use in Clinically Relevant Animal Models and 3D Immunochemistry
Authors: Laura A. Hasselbach, Susan M. Irtenkauf, Nancy W. Lemke, Kevin K. Nelson, Artem D. Berezovsky, Enoch T. Carlton, Andrea D. Transou, Tom Mikkelsen, Ana C. deCarvalho.
Institutions: Henry Ford Hospital.
Glioblastomas, the most common and aggressive form of astrocytoma, are refractory to therapy, and molecularly heterogeneous. The ability to establish cell cultures that preserve the genomic profile of the parental tumors, for use in patient specific in vitro and in vivo models, has the potential to revolutionize the preclinical development of new treatments for glioblastoma tailored to the molecular characteristics of each tumor. Starting with fresh high grade astrocytoma tumors dissociated into single cells, we use the neurosphere assay as an enrichment method for cells presenting cancer stem cell phenotype, including expression of neural stem cell markers, long term self-renewal in vitro, and the ability to form orthotopic xenograft tumors. This method has been previously proposed, and is now in use by several investigators. Based on our experience of dissociating and culturing 125 glioblastoma specimens, we arrived at the detailed protocol we present here, suitable for routine neurosphere culturing of high grade astrocytomas and large scale expansion of tumorigenic cells for preclinical studies. We report on the efficiency of successful long term cultures using this protocol and suggest affordable alternatives for culturing dissociated glioblastoma cells that fail to grow as neurospheres. We also describe in detail a protocol for preserving the neurospheres 3D architecture for immunohistochemistry. Cell cultures enriched in CSCs, capable of generating orthotopic xenograft models that preserve the molecular signatures and heterogeneity of GBMs, are becoming increasingly popular for the study of the biology of GBMs and for the improved design of preclinical testing of potential therapies.
Medicine, Issue 83, Primary Cell Culture, animal models, Nervous System Diseases, Neoplasms, glioblastoma, neurosphere, surgical specimens, long-term self-renewal
51088
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gDNA Enrichment by a Transposase-based Technology for NGS Analysis of the Whole Sequence of BRCA1, BRCA2, and 9 Genes Involved in DNA Damage Repair
Authors: Sandy Chevrier, Romain Boidot.
Institutions: Centre Georges-François Leclerc.
The widespread use of Next Generation Sequencing has opened up new avenues for cancer research and diagnosis. NGS will bring huge amounts of new data on cancer, and especially cancer genetics. Current knowledge and future discoveries will make it necessary to study a huge number of genes that could be involved in a genetic predisposition to cancer. In this regard, we developed a Nextera design to study 11 complete genes involved in DNA damage repair. This protocol was developed to safely study 11 genes (ATM, BARD1, BRCA1, BRCA2, BRIP1, CHEK2, PALB2, RAD50, RAD51C, RAD80, and TP53) from promoter to 3'-UTR in 24 patients simultaneously. This protocol, based on transposase technology and gDNA enrichment, gives a great advantage in terms of time for the genetic diagnosis thanks to sample multiplexing. This protocol can be safely used with blood gDNA.
Genetics, Issue 92, gDNA enrichment, Nextera, NGS, DNA damage, BRCA1, BRCA2
51902
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Use of Artificial Sputum Medium to Test Antibiotic Efficacy Against Pseudomonas aeruginosa in Conditions More Relevant to the Cystic Fibrosis Lung
Authors: Sebastian Kirchner, Joanne L Fothergill, Elli A. Wright, Chloe E. James, Eilidh Mowat, Craig Winstanley.
Institutions: University of Liverpool , University of Liverpool .
There is growing concern about the relevance of in vitro antimicrobial susceptibility tests when applied to isolates of P. aeruginosa from cystic fibrosis (CF) patients. Existing methods rely on single or a few isolates grown aerobically and planktonically. Predetermined cut-offs are used to define whether the bacteria are sensitive or resistant to any given antibiotic1. However, during chronic lung infections in CF, P. aeruginosa populations exist in biofilms and there is evidence that the environment is largely microaerophilic2. The stark difference in conditions between bacteria in the lung and those during diagnostic testing has called into question the reliability and even relevance of these tests3. Artificial sputum medium (ASM) is a culture medium containing the components of CF patient sputum, including amino acids, mucin and free DNA. P. aeruginosa growth in ASM mimics growth during CF infections, with the formation of self-aggregating biofilm structures and population divergence4,5,6. The aim of this study was to develop a microtitre-plate assay to study antimicrobial susceptibility of P. aeruginosa based on growth in ASM, which is applicable to both microaerophilic and aerobic conditions. An ASM assay was developed in a microtitre plate format. P. aeruginosa biofilms were allowed to develop for 3 days prior to incubation with antimicrobial agents at different concentrations for 24 hours. After biofilm disruption, cell viability was measured by staining with resazurin. This assay was used to ascertain the sessile cell minimum inhibitory concentration (SMIC) of tobramycin for 15 different P. aeruginosa isolates under aerobic and microaerophilic conditions and SMIC values were compared to those obtained with standard broth growth. Whilst there was some evidence for increased MIC values for isolates grown in ASM when compared to their planktonic counterparts, the biggest differences were found with bacteria tested in microaerophilic conditions, which showed a much increased resistance up to a >128 fold, towards tobramycin in the ASM system when compared to assays carried out in aerobic conditions. The lack of association between current susceptibility testing methods and clinical outcome has questioned the validity of current methods3. Several in vitro models have been used previously to study P. aeruginosa biofilms7, 8. However, these methods rely on surface attached biofilms, whereas the ASM biofilms resemble those observed in the CF lung9 . In addition, reduced oxygen concentration in the mucus has been shown to alter the behavior of P. aeruginosa2 and affect antibiotic susceptibility10. Therefore using ASM under microaerophilic conditions may provide a more realistic environment in which to study antimicrobial susceptibility.
Immunology, Issue 64, Microbiology, Pseudomonas aeruginosa, antimicrobial susceptibility, artificial sputum media, lung infection, cystic fibrosis, diagnostics, plankton
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One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures
Authors: Wendy L.J. Hansen, Judith Beuving, Annelies Verbon, Petra. F.G. Wolffs.
Institutions: Maastricht University Medical Center, Erasmus Medical Center.
Bloodstream infections are associated with high mortality rates because of the probable manifestation of sepsis, severe sepsis and septic shock1. Therefore, rapid administration of adequate antibiotic therapy is of foremost importance in the treatment of bloodstream infections. The critical element in this process is timing, heavily dependent on the results of bacterial identification and antibiotic susceptibility testing. Both of these parameters are routinely obtained by culture-based testing, which is time-consuming and takes on average 24-48 hours2, 4. The aim of the study was to develop DNA-based assays for rapid identification of bloodstream infections, as well as rapid antimicrobial susceptibility testing. The first assay is a eubacterial 16S rDNA-based real-time PCR assay complemented with species- or genus-specific probes5. Using these probes, Gram-negative bacteria including Pseudomonas spp., Pseudomonas aeruginosa and Escherichia coli as well as Gram-positive bacteria including Staphylococcus spp., Staphylococcus aureus, Enterococcus spp., Streptococcus spp., and Streptococcus pneumoniae could be distinguished. Using this multiprobe assay, a first identification of the causative micro-organism was given after 2 h. Secondly, we developed a semi-molecular assay for antibiotic susceptibility testing of S. aureus, Enterococcus spp. and (facultative) aerobe Gram-negative rods6. This assay was based on a study in which PCR was used to measure the growth of bacteria7. Bacteria harvested directly from blood cultures are incubated for 6 h with a selection of antibiotics, and following a Sybr Green-based real-time PCR assay determines inhibition of growth. The combination of these two methods could direct the choice of a suitable antibiotic therapy on the same day (Figure 1). In conclusion, molecular analysis of both identification and antibiotic susceptibility offers a faster alternative for pathogen detection and could improve the diagnosis of bloodstream infections.
Immunology, Issue 65, Infection, Medicine, Microbiology, Bacteria, real-time PCR, probes, pathogen detection, blood culture, 16S rDNA gene, antibiotic resistance, antibiotic susceptibility testing
3254
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Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme
Authors: Brian C. Baumann, Jay F. Dorsey, Joseph L. Benci, Daniel Y. Joh, Gary D. Kao.
Institutions: University of Pennsylvania .
Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality treatment consisting of surgical resection, post-operative radiation therapy and temozolomide chemotherapy 1. New therapeutic approaches are clearly needed to improve patient survival and quality of life. The development of more effective treatment strategies would be aided by animal models of GBM that recapitulate human disease yet allow serial imaging to monitor tumor growth and treatment response. In this paper, we describe our technique for the precise stereotactic implantation of bio-imageable GBM cancer cells into the brains of nude mice resulting in tumor xenografts that recapitulate key clinical features of GBM 2. This method yields tumors that are reproducible and are located in precise anatomic locations while allowing in vivo bioluminescent imaging to serially monitor intracranial xenograft growth and response to treatments 3-5. This method is also well-tolerated by the animals with low perioperative morbidity and mortality.
Cancer Biology, Issue 67, Medicine, Molecular Biology, glioblastoma multiforme, mouse, brain tumor, bioluminescent imaging, stereotactic rodent surgery
4089
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Method for Novel Anti-Cancer Drug Development using Tumor Explants of Surgical Specimens
Authors: Kaushal Joshi, Habibe Demir, Ryosuke Yamada, Takeshi Miyazaki, Abhik Ray-Chaudhury, Ichiro Nakano.
Institutions: The Ohio State University Medical Center, The Ohio State University Medical Center.
The current therapies for malignant glioma have only palliative effect. For therapeutic development, one hurdle is the discrepancy of efficacy determined by current drug efficacy tests and the efficacy on patients. Thus, novel and reliable methods for evaluating drug efficacy are warranted in pre-clinical phase. In vitro culture of tumor tissues, including cell lines, has substantial phenotypic, genetic, and epigenetic alterations of cancer cells caused by artificial environment of cell culture, which may not reflect the biology of original tumors in situ. Xenograft models with the immunodeficient mice also have limitations, i.e., the lack of immune system and interspecies genetic and epigenetic discrepancies in microenvironment. Here, we demonstrate a novel method using the surgical specimens of malignant glioma as undissociated tumor blocks to evaluate treatment effects. To validate this method, data with the current first-line chemotherapeutic agent, temozolomide (TMZ), are described. We used the freshly-removed surgical specimen of malignant glioma for our experiments. We performed intratumoral injection of TMZ or other drug candidates, followed by incubation and analysis on surgical specimens. Here, we sought to establish a tumor tissue explant method as a platform to determine the efficacy of novel anti-cancer therapies so that we may be able to overcome, at least, some of the current limitations and fill the existing gap between the current experimental data and the efficacy on an actual patient's tumor. This method may have the potential to accelerate identifying novel chemotherapeutic agents for solid cancer treatment.
Medicine, Issue 53, Glioblastoma multiforme, glioma, temozolomide, therapeutics, drug design
2846
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Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model
Authors: Tamalee R. Kramp, Kevin Camphausen.
Institutions: National Cancer Institute.
Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the understanding of the mechanics of tumor invasion, and direct intracerebral inoculation of tumor provides the opportunity of observing the invasive process in a physiologically appropriate environment2. As far as human brain tumors are concerned, the orthotopic models currently available are established either by stereotaxic injection of cell suspensions or implantation of a solid piece of tumor through a complicated craniotomy procedure3. In our technique we harvest cells from tissue culture to create a cell suspension used to implant directly into the brain. The duration of the surgery is approximately 30 minutes, and as the mouse needs to be in a constant surgical plane, an injectable anesthetic is used. The mouse is placed in a stereotaxic jig made by Stoetling (figure 1). After the surgical area is cleaned and prepared, an incision is made; and the bregma is located to determine the location of the craniotomy. The location of the craniotomy is 2 mm to the right and 1 mm rostral to the bregma. The depth is 3 mm from the surface of the skull, and cells are injected at a rate of 2 μl every 2 minutes. The skin is sutured with 5-0 PDS, and the mouse is allowed to wake up on a heating pad. From our experience, depending on the cell line, treatment can take place from 7-10 days after surgery. Drug delivery is dependent on the drug composition. For radiation treatment the mice are anesthetized, and put into a custom made jig. Lead covers the mouse's body and exposes only the brain of the mouse. The study of tumorigenesis and the evaluation of new therapies for GBM require accurate and reproducible brain tumor animal models. Thus we use this orthotopic brain model to study the interaction of the microenvironment of the brain and the tumor, to test the effectiveness of different therapeutic agents with and without radiation.
Medicine, Issue 61, Neuroscience, mouse, intracranial, orthotopic, radiation, glioblastoma
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Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice
Authors: Robert S. McNeill, Ralf S. Schmid, Ryan E. Bash, Mark Vitucci, Kristen K. White, Andrea M. Werneke, Brian H. Constance, Byron Huff, C. Ryan Miller.
Institutions: University of North Carolina School of Medicine, University of North Carolina School of Medicine, University of North Carolina School of Medicine, University of North Carolina School of Medicine, University of North Carolina School of Medicine, Emory University School of Medicine, University of North Carolina School of Medicine.
Current astrocytoma models are limited in their ability to define the roles of oncogenic mutations in specific brain cell types during disease pathogenesis and their utility for preclinical drug development. In order to design a better model system for these applications, phenotypically wild-type cortical astrocytes and neural stem cells (NSC) from conditional, genetically engineered mice (GEM) that harbor various combinations of floxed oncogenic alleles were harvested and grown in culture. Genetic recombination was induced in vitro using adenoviral Cre-mediated recombination, resulting in expression of mutated oncogenes and deletion of tumor suppressor genes. The phenotypic consequences of these mutations were defined by measuring proliferation, transformation, and drug response in vitro. Orthotopic allograft models, whereby transformed cells are stereotactically injected into the brains of immune-competent, syngeneic littermates, were developed to define the role of oncogenic mutations and cell type on tumorigenesis in vivo. Unlike most established human glioblastoma cell line xenografts, injection of transformed GEM-derived cortical astrocytes into the brains of immune-competent littermates produced astrocytomas, including the most aggressive subtype, glioblastoma, that recapitulated the histopathological hallmarks of human astrocytomas, including diffuse invasion of normal brain parenchyma. Bioluminescence imaging of orthotopic allografts from transformed astrocytes engineered to express luciferase was utilized to monitor in vivo tumor growth over time. Thus, astrocytoma models using astrocytes and NSC harvested from GEM with conditional oncogenic alleles provide an integrated system to study the genetics and cell biology of astrocytoma pathogenesis in vitro and in vivo and may be useful in preclinical drug development for these devastating diseases.
Neuroscience, Issue 90, astrocytoma, cortical astrocytes, genetically engineered mice, glioblastoma, neural stem cells, orthotopic allograft
51763
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Diffusion Tensor Magnetic Resonance Imaging in the Analysis of Neurodegenerative Diseases
Authors: Hans-Peter Müller, Jan Kassubek.
Institutions: University of Ulm.
Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo. The present applications are designed to investigate differences of WM involvement patterns in different brain diseases, especially neurodegenerative disorders, by use of different DTI analyses in comparison with matched controls. DTI data analysis is performed in a variate fashion, i.e. voxelwise comparison of regional diffusion direction-based metrics such as fractional anisotropy (FA), together with fiber tracking (FT) accompanied by tractwise fractional anisotropy statistics (TFAS) at the group level in order to identify differences in FA along WM structures, aiming at the definition of regional patterns of WM alterations at the group level. Transformation into a stereotaxic standard space is a prerequisite for group studies and requires thorough data processing to preserve directional inter-dependencies. The present applications show optimized technical approaches for this preservation of quantitative and directional information during spatial normalization in data analyses at the group level. On this basis, FT techniques can be applied to group averaged data in order to quantify metrics information as defined by FT. Additionally, application of DTI methods, i.e. differences in FA-maps after stereotaxic alignment, in a longitudinal analysis at an individual subject basis reveal information about the progression of neurological disorders. Further quality improvement of DTI based results can be obtained during preprocessing by application of a controlled elimination of gradient directions with high noise levels. In summary, DTI is used to define a distinct WM pathoanatomy of different brain diseases by the combination of whole brain-based and tract-based DTI analysis.
Medicine, Issue 77, Neuroscience, Neurobiology, Molecular Biology, Biomedical Engineering, Anatomy, Physiology, Neurodegenerative Diseases, nuclear magnetic resonance, NMR, MR, MRI, diffusion tensor imaging, fiber tracking, group level comparison, neurodegenerative diseases, brain, imaging, clinical techniques
50427
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Detection of Architectural Distortion in Prior Mammograms via Analysis of Oriented Patterns
Authors: Rangaraj M. Rangayyan, Shantanu Banik, J.E. Leo Desautels.
Institutions: University of Calgary , University of Calgary .
We demonstrate methods for the detection of architectural distortion in prior mammograms of interval-cancer cases based on analysis of the orientation of breast tissue patterns in mammograms. We hypothesize that architectural distortion modifies the normal orientation of breast tissue patterns in mammographic images before the formation of masses or tumors. In the initial steps of our methods, the oriented structures in a given mammogram are analyzed using Gabor filters and phase portraits to detect node-like sites of radiating or intersecting tissue patterns. Each detected site is then characterized using the node value, fractal dimension, and a measure of angular dispersion specifically designed to represent spiculating patterns associated with architectural distortion. Our methods were tested with a database of 106 prior mammograms of 56 interval-cancer cases and 52 mammograms of 13 normal cases using the features developed for the characterization of architectural distortion, pattern classification via quadratic discriminant analysis, and validation with the leave-one-patient out procedure. According to the results of free-response receiver operating characteristic analysis, our methods have demonstrated the capability to detect architectural distortion in prior mammograms, taken 15 months (on the average) before clinical diagnosis of breast cancer, with a sensitivity of 80% at about five false positives per patient.
Medicine, Issue 78, Anatomy, Physiology, Cancer Biology, angular spread, architectural distortion, breast cancer, Computer-Assisted Diagnosis, computer-aided diagnosis (CAD), entropy, fractional Brownian motion, fractal dimension, Gabor filters, Image Processing, Medical Informatics, node map, oriented texture, Pattern Recognition, phase portraits, prior mammograms, spectral analysis
50341
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Flexible Colonoscopy in Mice to Evaluate the Severity of Colitis and Colorectal Tumors Using a Validated Endoscopic Scoring System
Authors: Tomohiro Kodani, Alex Rodriguez-Palacios, Daniele Corridoni, Loris Lopetuso, Luca Di Martino, Brian Marks, James Pizarro, Theresa Pizarro, Amitabh Chak, Fabio Cominelli.
Institutions: Case Western Reserve University School of Medicine, Cleveland, Case Western Reserve University School of Medicine, Cleveland, Case Western Reserve University School of Medicine, Cleveland.
The use of modern endoscopy for research purposes has greatly facilitated our understanding of gastrointestinal pathologies. In particular, experimental endoscopy has been highly useful for studies that require repeated assessments in a single laboratory animal, such as those evaluating mechanisms of chronic inflammatory bowel disease and the progression of colorectal cancer. However, the methods used across studies are highly variable. At least three endoscopic scoring systems have been published for murine colitis and published protocols for the assessment of colorectal tumors fail to address the presence of concomitant colonic inflammation. This study develops and validates a reproducible endoscopic scoring system that integrates evaluation of both inflammation and tumors simultaneously. This novel scoring system has three major components: 1) assessment of the extent and severity of colorectal inflammation (based on perianal findings, transparency of the wall, mucosal bleeding, and focal lesions), 2) quantitative recording of tumor lesions (grid map and bar graph), and 3) numerical sorting of clinical cases by their pathological and research relevance based on decimal units with assigned categories of observed lesions and endoscopic complications (decimal identifiers). The video and manuscript presented herein were prepared, following IACUC-approved protocols, to allow investigators to score their own experimental mice using a well-validated and highly reproducible endoscopic methodology, with the system option to differentiate distal from proximal endoscopic colitis (D-PECS).
Medicine, Issue 80, Crohn's disease, ulcerative colitis, colon cancer, Clostridium difficile, SAMP mice, DSS/AOM-colitis, decimal scoring identifier
50843
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Identification of Disease-related Spatial Covariance Patterns using Neuroimaging Data
Authors: Phoebe Spetsieris, Yilong Ma, Shichun Peng, Ji Hyun Ko, Vijay Dhawan, Chris C. Tang, David Eidelberg.
Institutions: The Feinstein Institute for Medical Research.
The scaled subprofile model (SSM)1-4 is a multivariate PCA-based algorithm that identifies major sources of variation in patient and control group brain image data while rejecting lesser components (Figure 1). Applied directly to voxel-by-voxel covariance data of steady-state multimodality images, an entire group image set can be reduced to a few significant linearly independent covariance patterns and corresponding subject scores. Each pattern, termed a group invariant subprofile (GIS), is an orthogonal principal component that represents a spatially distributed network of functionally interrelated brain regions. Large global mean scalar effects that can obscure smaller network-specific contributions are removed by the inherent logarithmic conversion and mean centering of the data2,5,6. Subjects express each of these patterns to a variable degree represented by a simple scalar score that can correlate with independent clinical or psychometric descriptors7,8. Using logistic regression analysis of subject scores (i.e. pattern expression values), linear coefficients can be derived to combine multiple principal components into single disease-related spatial covariance patterns, i.e. composite networks with improved discrimination of patients from healthy control subjects5,6. Cross-validation within the derivation set can be performed using bootstrap resampling techniques9. Forward validation is easily confirmed by direct score evaluation of the derived patterns in prospective datasets10. Once validated, disease-related patterns can be used to score individual patients with respect to a fixed reference sample, often the set of healthy subjects that was used (with the disease group) in the original pattern derivation11. These standardized values can in turn be used to assist in differential diagnosis12,13 and to assess disease progression and treatment effects at the network level7,14-16. We present an example of the application of this methodology to FDG PET data of Parkinson's Disease patients and normal controls using our in-house software to derive a characteristic covariance pattern biomarker of disease.
Medicine, Issue 76, Neurobiology, Neuroscience, Anatomy, Physiology, Molecular Biology, Basal Ganglia Diseases, Parkinsonian Disorders, Parkinson Disease, Movement Disorders, Neurodegenerative Diseases, PCA, SSM, PET, imaging biomarkers, functional brain imaging, multivariate spatial covariance analysis, global normalization, differential diagnosis, PD, brain, imaging, clinical techniques
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Setting-up an In Vitro Model of Rat Blood-brain Barrier (BBB): A Focus on BBB Impermeability and Receptor-mediated Transport
Authors: Yves Molino, Françoise Jabès, Emmanuelle Lacassagne, Nicolas Gaudin, Michel Khrestchatisky.
Institutions: VECT-HORUS SAS, CNRS, NICN UMR 7259.
The blood brain barrier (BBB) specifically regulates molecular and cellular flux between the blood and the nervous tissue. Our aim was to develop and characterize a highly reproducible rat syngeneic in vitro model of the BBB using co-cultures of primary rat brain endothelial cells (RBEC) and astrocytes to study receptors involved in transcytosis across the endothelial cell monolayer. Astrocytes were isolated by mechanical dissection following trypsin digestion and were frozen for later co-culture. RBEC were isolated from 5-week-old rat cortices. The brains were cleaned of meninges and white matter, and mechanically dissociated following enzymatic digestion. Thereafter, the tissue homogenate was centrifuged in bovine serum albumin to separate vessel fragments from nervous tissue. The vessel fragments underwent a second enzymatic digestion to free endothelial cells from their extracellular matrix. The remaining contaminating cells such as pericytes were further eliminated by plating the microvessel fragments in puromycin-containing medium. They were then passaged onto filters for co-culture with astrocytes grown on the bottom of the wells. RBEC expressed high levels of tight junction (TJ) proteins such as occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. The transendothelial electrical resistance (TEER) of brain endothelial monolayers, indicating the tightness of TJs reached 300 ohm·cm2 on average. The endothelial permeability coefficients (Pe) for lucifer yellow (LY) was highly reproducible with an average of 0.26 ± 0.11 x 10-3 cm/min. Brain endothelial cells organized in monolayers expressed the efflux transporter P-glycoprotein (P-gp), showed a polarized transport of rhodamine 123, a ligand for P-gp, and showed specific transport of transferrin-Cy3 and DiILDL across the endothelial cell monolayer. In conclusion, we provide a protocol for setting up an in vitro BBB model that is highly reproducible due to the quality assurance methods, and that is suitable for research on BBB transporters and receptors.
Medicine, Issue 88, rat brain endothelial cells (RBEC), mouse, spinal cord, tight junction (TJ), receptor-mediated transport (RMT), low density lipoprotein (LDL), LDLR, transferrin, TfR, P-glycoprotein (P-gp), transendothelial electrical resistance (TEER),
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Creating Anatomically Accurate and Reproducible Intracranial Xenografts of Human Brain Tumors
Authors: Angela M. Pierce, Amy K. Keating.
Institutions: University of Colorado School of Medicine.
Orthotopic tumor models are currently the best way to study the characteristics of a tumor type, with and without intervention, in the context of a live animal – particularly in sites with unique physiological and architectural qualities such as the brain. In vitro and ectopic models cannot account for features such as vasculature, blood brain barrier, metabolism, drug delivery and toxicity, and a host of other relevant factors. Orthotopic models have their limitations too, but with proper technique tumor cells of interest can be accurately engrafted into tissue that most closely mimics conditions in the human brain. By employing methods that deliver precisely measured volumes to accurately defined locations at a consistent rate and pressure, mouse models of human brain tumors with predictable growth rates can be reproducibly created and are suitable for reliable analysis of various interventions. The protocol described here focuses on the technical details of designing and preparing for an intracranial injection, performing the surgery, and ensuring successful and reproducible tumor growth and provides starting points for a variety of conditions that can be customized for a range of different brain tumor models.
Medicine, Issue 91, intracranial, glioblastoma, mouse, orthotopic, brain tumor, stereotaxic, micropump, brain injection
52017
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Cortical Source Analysis of High-Density EEG Recordings in Children
Authors: Joe Bathelt, Helen O'Reilly, Michelle de Haan.
Institutions: UCL Institute of Child Health, University College London.
EEG is traditionally described as a neuroimaging technique with high temporal and low spatial resolution. Recent advances in biophysical modelling and signal processing make it possible to exploit information from other imaging modalities like structural MRI that provide high spatial resolution to overcome this constraint1. This is especially useful for investigations that require high resolution in the temporal as well as spatial domain. In addition, due to the easy application and low cost of EEG recordings, EEG is often the method of choice when working with populations, such as young children, that do not tolerate functional MRI scans well. However, in order to investigate which neural substrates are involved, anatomical information from structural MRI is still needed. Most EEG analysis packages work with standard head models that are based on adult anatomy. The accuracy of these models when used for children is limited2, because the composition and spatial configuration of head tissues changes dramatically over development3. In the present paper, we provide an overview of our recent work in utilizing head models based on individual structural MRI scans or age specific head models to reconstruct the cortical generators of high density EEG. This article describes how EEG recordings are acquired, processed, and analyzed with pediatric populations at the London Baby Lab, including laboratory setup, task design, EEG preprocessing, MRI processing, and EEG channel level and source analysis.
Behavior, Issue 88, EEG, electroencephalogram, development, source analysis, pediatric, minimum-norm estimation, cognitive neuroscience, event-related potentials
51705
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Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors
Authors: Steven J. Smith, Stephen H. Hughes.
Institutions: National Cancer Institute.
Although a number of anti HIV drugs have been approved, there are still problems with toxicity and drug resistance. This demonstrates a need to identify new compounds that can inhibit infection by the common drug resistant HIV-1 strains with minimal toxicity. Here we describe an efficient assay that can be used to rapidly determine the cellular cytotoxicity and efficacy of a compound against WT and mutant viral strains. The desired target cell line is seeded in a 96-well plate and, after a 24 hr incubation, serially dilutions of the compounds to be tested are added. No further manipulations are necessary for cellular cytotoxicity assays; for anti HIV assays a predetermined amount of either a WT or drug resistant HIV-1 vector that expresses luciferase is added to the cells. Cytotoxicity is measured by using an ATP dependent luminescence assay and the impact of the compounds on infectivity is measured by determining the amount of luciferase in the presence or the absence of the putative inhibitors. This screening assay takes 4 days to complete and multiple compounds can be screened in parallel. Compounds are screened in triplicate and the data are normalized to the infectivity/ATP levels in absence of target compounds. This technique provides a quick and accurate measurement of the efficacy and toxicity of potential anti HIV compounds.
Immunology, Issue 86, HIV, cytotoxicity, infectivity, luciferase, drug resistance, integrase, reverse transcriptase
51400
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Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment
Authors: Bob S. Carter, Christopher Farrell, Christopher Owen.
Institutions: Havard Medical School, Massachusetts General Hospital.
Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel. Surgical clipping is an important treatment for aneurysms which attempts to exclude blood from flowing into the aneurysmal segment of the vessel while preserving blood flow in a normal fashion. Improper clip placement may result in residual aneurysm with the potential for subsequent aneurysm rupture or partial or full occlusion of distal arteries resulting in cerebral infarction. Here we describe the use of an ultrasonic flow probe to provide quantitative evaluation of arterial flow before and after microsurgical clip placement at the base of a middle cerebral artery aneurysm. This information helps ensure adequate aneurysm reconstruction with preservation of normal distal blood flow.
Medicine, Issue 31, Aneurysm, intraoperative, brain, surgery, surgical clipping, blood flow, aneurysmal segment, ultrasonic flow probe
1294
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Simultaneous fMRI and Electrophysiology in the Rodent Brain
Authors: Wen-ju Pan, Garth Thompson, Matthew Magnuson, Waqas Majeed, Dieter Jaeger, Shella Keilholz.
Institutions: Emory University, Georgia Institute of Technology, Emory University.
To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in which functional MRI data and in vivo intracortical recording can be performed simultaneously. The combination of MRI and electrical recording is technically challenging because the electrodes used for recording distort the MRI images and the MRI acquisition induces noise in the electrical recording. To minimize the mutual interference of the two modalities, glass microelectrodes were used rather than metal and a noise removal algorithm was implemented for the electrophysiology data. In our studies, two microelectrodes were separately implanted in bilateral primary somatosensory cortices (SI) of the rat and fixed in place. One coronal slice covering the electrode tips was selected for functional MRI. Electrode shafts and fixation positions were not included in the image slice to avoid imaging artifacts. The removed scalp was replaced with toothpaste to reduce susceptibility mismatch and prevent Gibbs ringing artifacts in the images. The artifact structure induced in the electrical recordings by the rapidly-switching magnetic fields during image acquisition was characterized by averaging all cycles of scans for each run. The noise structure during imaging was then subtracted from original recordings. The denoised time courses were then used for further analysis in combination with the fMRI data. As an example, the simultaneous acquisition was used to determine the relationship between spontaneous fMRI BOLD signals and band-limited intracortical electrical activity. Simultaneous fMRI and electrophysiological recording in the rodent will provide a platform for many exciting applications in neuroscience in addition to elucidating the relationship between the fMRI BOLD signal and neuronal activity.
Neuroscience, Issue 42, fMRI, electrophysiology, rat, BOLD, brain, resting state
1901
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What is Visualize?
JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.
How does it work?
We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.
Video X seems to be unrelated to Abstract Y...
In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.
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Austin
Tea, Chocolate Chemical May Boost Memory
An aging hippy asks his friend, “Did you know that smoking marijuana affects your short-term memory?”
His friend responses, “No, I did not know that”.
Long pause in conversation at this point and then the aging hippy asks his friend again, “Did you know that smoking marijuana affects your short-term memory?” - end of joke.
What Is Exercise Intensity?
Q: What is exercise intensity?
A: In simplest terms, it's how hard an exercise is at a point in time. The technical definition is the level of momentary exertion during exercise.
Q: Why is it important?
A: Exercise of sufficient intensity is necessary to stimulate the body to make a change. When the body is worked beyond what it is equipped to handle, the body adapts as a form of self-protection.
Syndicate content
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Documentation: Herrenhausen Nature Symposium "Genome Editing for Gene and Cell Therapy"
The symposium focussed on genome editing, cell therapy, gene therapy, delivery in general, and biomedical applications. (Photo: freshidea - Fotolia.com)
The symposium focussed on genome editing, cell therapy, gene therapy, delivery in general, and biomedical applications. (Photo: freshidea - Fotolia.com)
November 3-4, 2016
Herrenhausen Palace, Hanover
Organizers: Volkswagen Foundation in collaboration with Nature Medicine
Christine Borowski (Nature Medicine, USA); Hannah Stower (Nature Medicine, USA); Markus Elsner (Nature Biotechnology, Germany); Alison Farrell (Nature Medicine, USA); Oliver Grewe (Volkswagen Foundation, Germany)
Summary Report
Genome editing revolutionizes biomedical research and inspires new therapeutic approaches for diseases from HIV/AIDS to cancer to immune deficiencies. The 13th Herrenhausen symposium discussed current applications, developments in research and the challenges to translate these into the clinic.
Genome editing uses meganucleases, Zinc Finger (ZNF), so-called TALEN nucleases or the bacterial CRISPR/Cas system to insert, delete or replace DNA sequences in a living organism. These nucleases create double-strand breaks at precisely defined locations in the genome, which are then repaired by the cell's DNA repair mechanism. Depending on the conditions, it either stitches the loose ends together via non-homologous end joining (NHEJ) or replaces the damaged part through homologous directed repair (HDR) using the intact copy on the other chromosome as template. HDR can be exploited for genome editing by providing an appropriate donor DNA instead. Whereas ZNF and TALEN nucleases must be engineered to recognize a specific target sequence, CRISPR/Cas can be 'programmed' via a specific guide RNA (gRNA).
In practice, donor DNA, gRNA and the nuclease must be delivered to the cell; the nuclease can be delivered either as protein, DNA or messenger RNA. The nuclease then continues to cut their target DNA sequence until the cell's repair mechanism has removed the sequence through deletion, insertion or via HDR. Clinical applications involve either ex vivo manipulation of patients' own stem cells outside the body and re-injecting the corrected cells – which is standard practice in classical gene therapy– or in vivo treatment by delivering nuclease and donor DNA to target cells, usually via Adeno-associated viruses (AAV) or lentiviruses.
Toni Cathomen, in his keynote talk, gave an overview on the current state of gene and cell therapies and the future impact of genome editing: whereas gene therapy provides an additional intact copy of the mutated gene, genome editing can fix the error itself. Cathomen described various pre-clinical developments to find cures for hemophilia, HIV or immune disorders and explained the major challenges for genome editing in clinical use. These are in particular specificity to avoid off-target editing elsewhere in the host genome, efficiency in terms of the number of cells that are correctly edited, and delivery of the editing machinery to the correct cells.
Luigi Naldini described the success of gene therapy for SCID, which is now a safe and efficient cure for this fatal immune deficiency. It uses ex vivo treatment of the patients' blood stem cells to add an intact copy of the mutated gene and re-injecting the corrected cells. Naldini's group is now exploring the use of genome editing to correct the gene, which still yields a lower percentage of 'repaired' cells compared to gene therapy. Optimized procedures and stimulating blood stem cells prior to editing to activate their DNA repair mechanism could improve efficiency.
Matthew Porteus and Daniel Bauer discussed the use of genome editing to cure sickle cell anemia. Porteus' group uses AAV to transport CRISPR/Cas, chemically modified gRNA and donor DNA into blood stem cells to repair mutations in the ß-hemoglobin gene that cause the disease. Bauer takes a different route by reactivating the γ-hemoglobin gene instead, which is usually shut down after birth. Instead of the more challenging HDR to repair mutations, this requires only NHEJ to cleave a specific genome sequences that keeps the gene silent.
Katherine High presented results of clinical trials to test the safety and efficacy of investigational treatments for hemophilia and progressive vision loss. The latter uses AAV-mediated in vivo gene transfer to the retinal pigment epithelial cells in affected individuals. This treatment for vision loss involves injecting the vector with the repair machinery directly into the eye where it targets and repairs the cells that support the light-sensing cells.
Carl June and Isabelle Rivière demonstrated the use of gene editing for generating optimized CAR T cells – immune cells that are engineered to recognize and attack tumor cells. Rivière showed how gene editing could improve these cells' efficiency against leukemia while June discussed generating CAR T cells that destroy solid tumors.
James Wilson's experiments in mice to cure metabolic liver diseases by correcting gene defects with CRISPR/Cas showed that repairing only a small percentage of cells is sufficient to restore normal function. His work also unveiled that the efficiency of gene editing differs between adults and newborns, potentially owing to the fact that the newborn's cells are rapidly proliferating and therefore more amenable to HDR.
Frank Buchholz explores gene editing to cure AIDS by destroying the quiescent HIV genomes in the human genome. His group has been exploring using recombinases instead of nucleases as these can excise longer DNA sequences from the genome. They engineered recombinases against HIV DNA and are testing these with HIV-infected blood stem cells.
Carlos Moraes explained how genome editing of mitochondrial DNA could be used to cure inherited mitochondrial diseases. Mitchochondria, the cell's energy-producing organelles, contain many copies of their own genome; it is therefore not necessary to repair gene defects but just destroy erroneous copies to restore proper function.
Preclinical developments and clinical trials of genome editing are not just pushing the boundaries of medical research but also challenging the regulation of drug development. Natalie Mount commented that regular and open interaction with regulatory agencies is therefore key for a successful transition of these Advanced Therapy Medicinal Products into the clinic.
A major issue for genome editing is to avoid off-target effects in other areas of the genome. One solution is to increase the nuclease's specificity for the target sequence. Benjamin Kleinstiver presented his work on engineering Cas9 and the related Cpf1 nuclease to increase both target specificity and cutting efficiency.
Cutting is only the first step in gene editing, followed by NHEJ or HDR. Eric Hendrickson demonstrated that it generates quite different results depending on how the cell repairs DNA breaks. Understanding these mechanisms could help to further improve genome editing and gene therapy through the choice of appropriate gRNA and donor DNA.
Delivering the nuclease and donor DNA to the target cell is another major challenge for gene therapy. Thierry VandenDriessche uses tissue-specific promoter sequences to ensure that Cas is expressed only in specific cell types. Proof of principle experiments demonstrate a potential cure for myotonic dystrophy, a muscle-wasting disease, by repairing the gene defect in muscle cells.
Mark Kay's group has engineered AAV that infect only specific cell types such as liver or nerve cells. As AAV integrates its DNA into the host cell genome, Kay uses a nuclease-free approach called gene ride to transfer a corrective DNA sequence into the target cell. One application is fixing a point mutation in liver cells that causes Crigler Najjar Syndrome, which leads to brain damage in infants.
Niren Murthy has developed a delivery system for the CRISP/Cas system based on gold nanoparticles. Experiments in mice models of Duchenne Muscular Dystrophy show some efficiency in correcting the dystrophin gene in muscle tissue with little off-target effects. Aditi Mehta presented her research on inhalable nanoparticles to deliver Cas9 and donor DNA in an attempt to target and destroy tumor cells in lung cancer.
As quiescent cells are more likely to use NHEJ whereas growing and dividing cells prefer HDR, Tony Gutschner has linked Cas activity to the cell cycle to improve the efficiency of genome engineering.
The longer CRISPR/Cas remains active in the cell, the higher the risk of off-target effects. Gianluca Petris' group has thus developed a CRSPR/Cas9 system that self-destructs after it completed its mission.
Lukas Dow demonstrated the use of gene editing to model human disease in mice. They use CRISPR/Cas to cause mutations similar to those found in human colorectal cancer. Growing mutated stem cells into organoids – a three-dimensional organ bud with realistic micro-anatomy – allows them to study the effect of these mutations on cancer growth and metastasis.
Kosuke Yusa presented the application of gene editing for drug screening. His group uses CRISPR/Cas with a library of gRNAs to mutate cancer cells. Mutations that cause cell death could hint to genes essential for the cancer cell's survival and thereby potential drug targets. His research has already yielded two such candidate genes for treating leukemia.
The discussion showed that many challenges remain to harness the full potential of gene editing. The choice of nuclease, target cells and DNA sequence, delivery vehicle and the mode of DNA repair determine efficiency and specificity, making it a personalized therapy. "Every patient gets a unique product," Porteus remarked.
All academic titles have been omitted.
Author
Holger Breithaupt, embl, Science reporter
Speakers and Chairs
Daniel Anderson, Massachusetts Institute of Technology, USA, Daniel Bauer, Harvard Medical School, USA, Frank Buchholz, Technische Universitäten Dresden, Germany, Toni Cathomen, Universitätsklinikum Freiburg, Germany, Lukas Dow, Weill Cornell Medical College, USA, Tony Gutschner, The University of Texas MD Anderson Cancer Center, USA, Eric Hendrickson, University of Minnesota, USA, Katherine High, University of Pennsylvania, USA, Carl June, University of Pennsylvania, USA, Mark Kay, Stanford School of Medicine, USA, Benjamin Kleinstiver, Harvard Medical School, USA, Carlos Moraes, University of Miami, USA, Natalie Mount, Cell and Gene Therapy Catapult, UK, Niren Murthy, University of California, Berkeley, USA, Luigi Naldini, Ospedale San Raffaele, Italy, Matt Porteus, Stanford University School of Medicine, USA, Isabelle Rivière, Memorial Sloan Kettering Cancer Center, USA, Thierry VandenDriessche, Free University of Brussels, Belgium, James Wilson, Perelman School of Medicine University of Pennsylvania, USA, Kosuke Yusa, Wellcome Trust Sanger Institute, UK
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Clinical UM Guideline
Subject: Hospital Beds and Accessories
Guideline #: CG-DME-15Current Effective Date: 10/08/2013
Status:ReviewedLast Review Date: 08/08/2013
Description
Hospital beds are specialized beds used primarily in the treatment of individuals with an illness or injury. Hospital bed accessories are durable medical equipment items used in conjunction with a hospital bed.
Clinical Indications
Hospital Beds
Medically Necessary:
A fixed height hospital bed is considered medically necessary if one or more of the following criteria are met:
1. The individual has a medical condition that requires positioning of the body in ways not feasible with an ordinary bed to alleviate pain, prevent contractures, promote good body alignment or avoid respiratory infections.
2. The individual requires the head of the bed to be elevated more than 30 degrees most of the time due to congestive heart failure, chronic pulmonary disease, or problems with aspiration. Pillows or wedges must have been considered and ruled out. Elevation of the head/upper body less than 30 degrees does not usually require the use of a hospital bed.
3. The individual requires special attachments, such as traction equipment, that can only be attached to a hospital bed.
A variable height hospital bed is considered medically necessary if the individual meets one or more of the criteria for a fixed height hospital bed and requires a bed height different than a fixed height hospital bed to permit transfers to chair, wheelchair, or standing position. This includes, but is not limited to:
A semi-electric hospital bed is considered medically necessary if the individual meets one or more of the criteria for a fixed height bed and requires frequent changes in body position or has an immediate need for a change in body position.
A heavy-duty, extra-wide hospital bed is considered medically necessary if the individual meets one or more of the criteria for a fixed height hospital bed and the individual's weight is more than 350 pounds, but does not exceed 600 pounds.
An extra heavy-duty hospital bed is considered medically necessary if the individual meets one or more of the criteria for a hospital bed and the individual's weight exceeds 600 pounds.
An enclosed crib or enclosed bed is considered medically necessary for individuals with seizures, disorientation, vertigo, and neurological disorders, where the individual needs to be restrained to bed. Clinical documentation must be provided that states less invasive strategies (i.e., bed rails, bed rail protectors, or environmental modifications) have been tried and have not been successful.
A request for a hospital grade, pediatric crib will be reviewed for medical necessity on an individual basis.
Not Medically Necessary:
If the above criteria are not met, the hospital bed will be considered not medically necessary.
A total electric hospital bed is considered not medically necessary. The height adjustment feature is considered to be a convenience feature.
Ordinary (Non-Hospital) beds are considered not medically necessary. An ordinary bed does not meet the definition of durable medical equipment as it is not primarily medical in nature and is not primarily used in the treatment of a disease or injury.
Power or manual lounge beds are considered not medically necessary since they are not primarily medical in nature and are considered to be a comfort or convenience item.
Bed Accessories
Medically Necessary:
Trapeze equipment is considered medically necessary if the individual is confined to bed and needs this device to sit up because of a respiratory condition, to change body position for other medical reasons, or to get in or out of bed. Heavy duty trapeze equipment is considered medically necessary if the individual meets the criteria for regular trapeze equipment and weighs more than 250 pounds.
A bed cradle is considered medically necessary when it is necessary to prevent contact with the bed coverings. This includes, but is not limited to individuals with burns, decubitus or diabetic ulcers, or gouty arthritis.
Side rails are considered medically necessary when they are required by the individual's condition and they are an integral part of, or an accessory to, a hospital bed.
If an individual's condition requires a replacement innerspring mattress or foam rubber mattress it will be considered medically necessary for an individual-owned hospital bed.
Not Medically Necessary:
The following bed accessories are considered not medically necessary since they are not primarily medical in nature, are not mainly used in the treatment of a disease or injury and are normally of use to people who do not have a disease or injury.
A frame/canopy for use with a hospital bed and limb restraints is considered not medically necessary since these items are not primarily medical in nature.
Coding
The following codes for treatments and procedures applicable to this document are included below for informational purposes. A draft of future ICD-10 Coding (effective 10/01/2014) related to this document, as it might look today, is included below for your reference. Inclusion or exclusion of a procedure, diagnosis or device code(s) does not constitute or imply member coverage or provider reimbursement policy. Please refer to the member's contract benefits in effect at the time of service to determine coverage or non-coverage of these services as it applies to an individual member.
HCPCS
Beds
E0250-E0251Hospital bed, fixed height, with any type side rails
E0255-E0256Hospital bed, variable height, hi-lo, with any type side rails
E0260-E0261Hospital bed, semi-electric (head and foot adjustment), with any type side rails
E0265-E0266Hospital bed, total electric (head, foot, and height adjustments), with any type side rails
E0290-E0291Hospital bed, fixed height, without side rails
E0292-E0293Hospital bed, variable height, hi-lo, without side rails
E0294-E0295Hospital bed, semi-electric (head and foot adjustment), without side rails
E0296-E0297Hospital bed, total electric, (head, foot and height adjustments), without side rails
E0300Pediatric crib, hospital grade, fully enclosed, with or without top enclosure
E0301-E0304Hospital bed, heavy duty/extra heavy duty (includes codes E0301, E0302, E0303, E0304)
E0328Hospital bed, pediatric, manual, 360 degree side enclosures, top of head board, foot board and side rails up to 24 inches above the spring, includes mattress
E0329Hospital bed, pediatric, electric or semi-electric, 360 degree side enclosures, top of head board, foot board and side rails up to 24 inches above spring, includes mattress
Accessories
E0271-E0272Mattress
E0273Bed board
E0274Over-bed table
E0280Bed cradle, any type
E0305Bed side rails, half-length
E0310Bed side rails, full-length
E0315Bed accessory: board, table or support device, any type
E0316Safety enclosure frame/canopy for use with hospital bed, any type
E0910Trapeze bars, also known as Patient Helper, attached to bed, with grab bar
E0911Trapeze bar, heavy duty, for patient weight capacity greater than 250 pounds, attached to bed, with grab bar
ICD-9 Diagnosis
All diagnoses
ICD-10 DiagnosisICD-10-CM draft codes; effective 10/01/2014
All diagnoses
Discussion/General Information
Descriptions
A fixed height hospital bed is one with manual head and leg elevation adjustments but no height adjustment.
A variable height hospital bed is one with manual height adjustment and with manual head and leg elevation adjustments.
A semi-electric bed is one with manual height adjustment and with electric head and leg elevation adjustments.
A total electric bed is one with electric height adjustment and with electric head and leg elevation adjustments.
An ordinary bed is one that is typically sold as furniture. It consists of a frame, box springs and mattress. It is a fixed height and has no head or leg elevation adjustments. It is normally for use in the absence of illness or injury.
Power or manual lounge beds, like other ordinary beds, are typically sold as furniture and are not considered durable medical equipment as they are used in the absence of illness or injury. The following are examples of lounge beds:
The U.S. Food and Drug Administration (FDA) in 2005 determined that the Vail Enclosure Bed poses a significant public health risk because individuals can become entrapped and suffocate, resulting in severe neurological damage or death. Vail Products, Inc of Toledo, Ohio, has permanently ceased manufacture, sale and distribution of all Vail enclosed bed systems.
This Clinical UM Guideline is based on Centers for Medicare and Medicaid Services (CMS) criteria.
References
Peer Reviewed Publications:
1. Hampton S. Can electric beds aid pressure sore prevention in hospitals? Br J Nurs. 1998; 7(17):1010-1017.
Government Agency, Medical Society, and Other Authoritative Publications:
1. Centers for Medicare and Medicaid Services. National Coverage Determination. Available at: http://www.cms.gov/mcd/indexes.asp?clickon=index. Accessed on June 14, 2013.
• Durable Medical Equipment Reference List. NCD #280.1. Effective May 5, 2005.
• Hospital Beds. NCD #280.7. This is a longstanding national coverage determination. The effective date of this version has not been posted.
2. Electronic Data Systems Corp. Jurisdiction D. Local Coverage Determination for Hospital Beds and Accessories (L11572). Revised 2/04/2011. Available at: http://www.cms.gov/mcd/index_local_alpha.asp?from=alphalmrp&letter=A. Accessed on June 14, 2013.
3. NHIC. Jurisdiction A. Local Coverage Determination for Hospital Beds and Accessories (L5049). Revised 2/04/2011. Available at: http://www.cms.gov/mcd/index_local_alpha.asp?from=alphalmrp&letter=A. Accessed on June 14, 2013.
4. U.S. Food and Drug Administration (FDA), Center for Devices and Radiological Health (CDRH). Medical Device Recalls - Class I Recall: Vail Products, Inc. Enclosed Bed Systems. Rockville, MD: FDA. June 24, 2005. Available at: http://www.fda.gov/MedicalDevices/Safety/ListofRecalls/ucm063853.htm. Accessed on June 14, 2013.
Index
Hospital Beds and Accessories
History
StatusDateAction
Reviewed08/08/2013Medical Policy & Technology Assessment Committee (MPTAC) review. Websites and References updated.
01/01/2013Updated Coding section with 01/01/2013 HCPCS descriptor change.
Reviewed08/09/2012MPTAC review. Websites and References updated.
Reviewed08/18/2011MPTAC review. Websites and References updated.
Reviewed08/19/2010MPTAC review. Websites and References updated.
Revised08/27/2009MPTAC review. Removed not medically necessary statement addressing the Vail enclosure bed. Removed place of service. References updated.
Reviewed08/28/2008MPTAC review. References updated.
01/01/2008Updated coding section with 01/01/2008 HCPCS changes.
Revised08/23/2007MPTAC review. Addition of medically necessary statement for enclosure beds. References and coding updated.
Revised12/07/2006MPTAC review. Enclosure beds moved from medically necessary to not medically necessary. Added medically necessary language addressing heavy duty trapeze equipment. References and coding updated.
New12/01/2005MPTAC initial guideline development.
Pre-Merger Organizations
Last Review Date
Document Number
Title
Anthem, Inc.
No Document
Anthem CO/NV
DME.211Hospital Beds and Accessories
Anthem MW
04/08/2005
DME.004Hospital Beds & Other Bed Accessories
Anthem ME
Benefit DetailHospital Bed
Anthem CT
10/01/2004
DME Coverage Criteria Guideline, Section DHospital Beds and Accessories
WellPoint Health Networks, Inc. No Document
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For a long time now caffeine has had a bad rap. And while drinking bottomless cups of joe may make you jittery or keep you awake at night, moderate amounts of caffeine can give you a boost in more ways than one. Recent studies show that it may actually have some powerful health benefits. Just stick to two or three cups of coffee a day—that's been shown to be a safe intake.
Gallstones
Researchers at the Harvard School of Public Health speculate that caffeine causes the gallbladder to contract, reducing the chance for gallstones to build up. A study of men between the ages of 40 and 75 found that coffee drinkers have fewer gallstones than those who don't drink it.
Parkinson's Disease
When Harvard researchers studied the effects of caffeine on Parkinson's disease, they discovered that women who drank one to three cups of coffee a day cut their risk of developing Parkinson's in half. Drinking more than three cups, however, seems to reduce the positive effect.
Breast Cancer
If you're worried about caffeine increasing your risk of breast cancer, relax. A Toronto study found that women who carry the BRCA I gene that puts them at risk for developing breast cancer can actually decrease that risk by drinking coffee. The study showed that those who drank six or more cups reduced their risk even more than those who drank between one and three cups.
Type 2 Diabetes
Drinking coffee has been linked with better glucose tolerance and a lower risk of type 2 diabetes. A recent study from the University of Minnesota in Minneapolis showed that women who drank more than six cups of coffee a day were 22 percent less likely to develop type 2 diabetes—but those who preferred decaf reduced their risk even more, by 33 percent. Therefore, the researchers speculate that the antioxidants in coffee, not the caffeine, may provide the benefit.
Did You Know?
-Non–plant-based beverages such as energy drinks are not likely to have the same benefits as coffee or tea.
-Drinking caffeinated beverages before a workout may help you keep exercising longer without feeling tired.
-In the past, it was thought that drinking coffee increased your chance of developing cardiovascular disease, but recent research shows that the effects of caffeine on the heart seem to be neutral.
What Do You Think?
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Sports and fitness nutrition wildman
Report was sports and fitness nutrition wildman vitamin means
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Vitamin consists of eating and ingesting appropriately to benefit from health of your body and improve your complete wellness stage. Between several sections are smaller sections with motivational statements. Ideally, you need to eat fish because it's low in fat and energy and nonetheless has a excessive quantity of protein. As Asian ladies undertake a western diet their breast most cancers rates climb. You may carry this app when you are going out on camping and study more on the place you are. If a toddler is consuming a properly-balanced weight-reduction plan, they should not require vitamin dietary supplements. Normally the evening meal sports and fitness nutrition wildman planned around a primary dish. With a view to cover the sickly appearance of farmed salmon meat, the fish are fed a pink pigment to change their tissue sports and fitness nutrition wildman. 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What you could have written could also be seen, disclosed to, or collected by third parties and could also be utilized by others in sports and fitness nutrition wildman we are unable to control or predict, including to contact you or sports and fitness nutrition wildman be used for unauthorized or illegal functions. Headquartered in Alameda, CA, Chef-Ok (an acronym for Culinary Health Schooling for Youngsters) gives culinary education schemes geared to youngsters and teenagers. For the reason that skill of your body to make Sports and fitness nutrition wildman declines with age and since your cells haven't any technique to substitue for Q10 in mobile power production it could readily be understood that a lack of Q10 will increase the amount of fatigue and the speed of cell malfunction - options that are elementary to every aspect of a person's health (since mobile power is needed for any cell to do anything). Please notice that these diet values are estimated based on our commonplace serving portions. 39 Subsequently, the proposed modifications improper synthesis. The puppies of huge breed canines could develop skeletal abnormalities, if overfed on vitality meals, as a result of this kind of meals accelerates the pace at sports and fitness nutrition wildman they grow. Testosterone is a male hormone that helps with muscle growth too. Calories per serving: one hundred fifty. Different snack concepts for developing muscle consist of obtaining a banana in addition to a glass great value creamed corn nutrition milk. Researchers are looking at a lot of illnesses including breast cancer, autism and schizophrenia using genomic information to single out threat elements and preventions. It additionally makes it simpler to check comparable meals to see which is more healthy. For example, decreasing easy sugars (glucose, sucrose, fructose and lactose) can forestall diabetes, and excessive fiber diets (especially soluble fiber) can help management diabetes. Once the immunity of the physique jobs for nutritionist in san diego declining and getting strained, it is not simple to get it again to its earlier good health. In case you are a runner who's going to make use of the BMI calculator to determine whether or not or not you might be fit, wholesome, or having success along with your running or fitness program, you want to be aware of some shortcomings of the BMI calculation. That is to recreate the nutrition in the meals after they remove or wreck all of it by way of processing. The risk elevated to 26 p. Insulin and glucose overload results in hypertension, polycystic ovarian syndrome, heart disease, diabetes, and cancer. Here is what any good private coach will at all times let you know: No amount sports and fitness nutrition wildman calorie restriction or tablet-popping will ever offer you FITNESS. If you're from one other Australian tertiary establishment you could be permitted to underake cross-institutional study in one or more models of examine on the University of Sydney. If they're allowed to overeat, puppies can eat too many calories, develop too quickly and develop well being problems. Extracting those seeds is a labor of love, though one you shouldn't attempt when carrying mild clothes. Round 50 of the selenium in a man is within the testes and seminal ducts; males lose selenium in semen, subsequently it's good to preserve ranges topped up.
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15.02.2013 at 21:34 Akinolar:
Yes cannot be!
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The Addiction Series: The Highjacker
Addiction Policy Forum has created a series of videos that can help us understand the disease of addiction. Starting with THE HIGHJACKER, we learn how substance use disorders (SUDs) affect tissue function in two main parts of the brain: the limbic system (responsible for basic survival instincts) and the prefrontal cortex (where decision-making and impulse control live).
The animated series aims to expand public understanding about addiction and replace the myths and misinformation that keep substance use disorders (SUDs) from being treated like any other medical condition.
Substance use activates the dopamine process in the survival center much more powerfully than natural rewards like food or sex. When repeated it can hijack the brain, making it think that the substance is the most important thing for survival.
Over time, more and more of the substance is needed to activate the same level of reward, causing the brain’s circuits to become increasingly imbalanced--eroding a person’s self-control and ability to make sound decisions, while producing intense impulses to seek and use the substance.
This is what it means when scientists say that addiction is a brain disease.
The good news – SUDs are preventable and treatable, and brain scans show that once an individual is in recovery, brain tissue can get better.
A special thanks to the National Institute on Drug Abuse (NIDA) for the science highlighted in the series.
More episodes to come...
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@article {Tisminetzky:2012-01-01T00:00:00:1087-3244:20, author = "Tisminetzky, Mayra and Bray, Bethany C. and Miozzo, Ruben and Aupont, Onesky and McLaughlin, Thomas", title = "Classes of Depression, Anxiety, and Functioning in Acute Coronary Syndrome Patients", journal = "American Journal of Health Behavior", volume = "36", number = "1", year = "2012-01-01T00:00:00", abstract = " Objectives: To describe change in subgroups characterized by patterns of depression, anxiety, and functional impairment; examine treatment effects on subgroup membership; examine effects of sex and age on subgroup membership. Methods: Latent class models were used to meet the first 2 objectives using 79 patients with depression/anxiety. Generalized estimating equations were used to meet the third objective. Results: Three subgroups characterized by different combinations of psychiatric disorders and functioning were identified. Patients who received treatment were more likely to transition to a less impaired subgroup. Conclusions: Unique information about holistic treatment effects can be gained when multiple outcomes are considered simultaneously.", pages = "20-30", url = "http://www.ingentaconnect.com/content/png/ajhb/2012/00000036/00000001/art00003", doi = "doi:10.5993/AJHB.36.1.3", keyword = "ANXIETY, DEPRESSION, ACUTE CORONARY SYNDROME, LATENT CLASS ANALYSIS" }
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SMAS imbrication vs SMAS plication
Many of the docs at RS have said that facelifts using SMAS plication do not involve true mobilization. Does this mean that if my surgeon is cutting an elliptical section out of the SMAS that I am getting greater skin flap mobilization and the probability longer lasting result than I would get from a more simple plication facelift? Please don't tell me to just look at before and after photos.
Doctor Answers 8
SMAS plication works very well
I stopped doing SMAS dissection, excision and imbrication 20 years ago because the SMAS is so thin and most patients it simply tears after excision and imbrication. In my experience the plication of the SMAS works very well and the benefit is that the SMAS correction holds. The skin flap mobilization works perfectly well after this technique and because the distal attachments are still in place and male better than totally mobilizing the skin flap in my opinion.
Should I get an Imbrication or a plication
There are many different techniques for performing a facelift. The most important factor in choosing a surgeon is finding one who has tried and true results with whichever technique he or she is the most experienced with. In my experience, I get the best results using an imbrication technique.
Todd C. Miller, MD
Newport Beach Facial Plastic Surgeon
5.0 out of 5 stars 34 reviews
SMAS imbrication versus plication
In our practice, we perform imbrication in the majority of facelifts. On a rare occasion on a multiple revisional facelift, we may use plication sutures instead. Imbrication tends to give a better and longer lasting result than plication. Tightening the SMAS is only one component of the face lift. Also look to see how the fatty deposits are are removed in the neck and the neck muscles are tightened. For many examples of imbricating facelifts, please see the link below.
William Portuese, MD
Seattle Facial Plastic Surgeon
4.7 out of 5 stars 125 reviews
SMAS facelifts
SMAS mobilization is a result of a technique that loosens and frees up this layer to achieve repositioning of the muscles of the face. The amount of mobilization depends on how much your surgeon frees up this layer. Just because you have part of the SMAS remove does not mean you get more mobilization and a longer-lasting result.
Earl Stephenson, Jr, MD, DDS, FACS
SMAS treatment
There are many different ways to lift the soft tissues of the face and SMAS imbrication, plication, and elevation all can do it a bit differently.
Steven Wallach, MD
New York Plastic Surgeon
4.1 out of 5 stars 23 reviews
SMAS
There are many derivations of the use/treatment of the SMAS during a facelift. SMAS elevation with mobilization, imbrication or plication. All of these approaches have their place in facelifts, both in first time patients and secondaries. There are a number of factors which will allow choosing one approach over another including, overlying skin quality, soft tissue fullness/deficiency, primary or secondary cases with/without prior SMAS lift, SMAS mobility etc. Each case much be evaluated individually.
David L. J. Wardle, MD
Ottawa Plastic Surgeon
SMAS Procedures
The SMAS is a sheet of tissue that envelops the face. It is attached to the deep tissues in many places around the face and attaches to the superficial tissues wherever the skin folds during facial animation. In my experience, the best way to get a great, long-lasting result is by lifting the SMAS and separating it from the underlying attachments. If you do not release these attachments, then the SMAS moves very little and the results are not as satisfying.
Plication does not allow for the release of the underlying attachments.
Robert M. Freund, MD
New York Plastic Surgeon
4.8 out of 5 stars 29 reviews
SMAS Imbication vs SMAS plication for a Long Lasting Result?
I personally would answer neither, because I think those approaches do not include a significant SMAS elevation and mobilization, and therefore are inherently limited in their ability to achieve skin flap mobilization. In general these approaches are used because they take less time as they require less dissection and potentially have less risk. I believe that a SMAS approach which involves thorough elevation, mobilization, and repositioning of the SMAS results in better correction of the mid-face/lid cheek junction with more complete and longer lasting results. However, there are no good long term studies which definitively show that a more extended SMAS elevation and mobilization technique results in better long term results.
One complaint that I have with any technique the involves cutting out a section of the SMAS is that you are throwing away precious tissue, mostly fat, which is extremely important in maintaining a natural youthful appearance. We all know now that facial aging involves a significant amount of loss of facial volume over time in addition to sagging or descent of the facial tissues. This why SMAS mobilization alone is not sufficient to achieve a comprehensive facial rejuvenation. I incorporate autologous fat grafting into every facelift procedure done today because it results in a more complete, softer, natural rejuvenation. Use of autologous fat has not been in widespread use for long enough to draw any conclusions about whether it helps provide a longer lasting result, but I think it is an essential component to almost any facial rejuvenation procedure done today.
These answers are for educational purposes and should not be relied upon as a substitute for medical advice you may receive from your physician. If you have a medical emergency, please call 911. These answers do not constitute or initiate a patient/doctor relationship.
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