New MRI method fingerprints tissues and diseases

A new method of magnetic resonance imaging (MRI) could routinely spot specific cancers, multiple sclerosis, heart disease and other maladies early, when they’re most treatable, researchers at Case Western Reserve University and University Hospitals (UH) Case Medical Center suggest in the journal Nature.

Each body tissue and disease has a unique fingerprint that can be used to quickly diagnose problems, the scientists say.

By using new MRI technologies to scan for different physical properties simultaneously, the team differentiated white matter from gray matter from cerebrospinal fluid in the brain in about 12 seconds, with the promise of doing this much faster in the near future.

The technology has the potential to make an MRI scan standard procedure in annual check-ups, the authors believe. A full-body scan lasting just minutes would provide far more information and require no radiologist to interpret the data, making diagnostics cheap, compared to today’s scans, they contend.

"The overall goal is to specifically identify individual tissues and diseases, to hopefully see things and quantify things before they become a problem," said Mark Griswold, a radiology professor at Case Western Reserve School of Medicine and UH Case Medical Center. "But to try to get there, we’ve had to give up everything we knew about the MRI and start over."

Griswold has been working on this goal with Case Western Reserve’s Vikas Gulani, MD, an assistant professor of radiology, and Nicole Seiberlich, assistant professor of biomedical engineering, for a decade. During the last three years, they developed the technology and proved the concept with graduate student Dan Ma; Kecheng Liu, PhD, collaborations manager from Siemens Medical Solutions Inc.; Jeffrey L. Sunshine, MD, professor of radiology and a radiologist at UH Case Medical Center; and Jeffrey L. Duerk, dean of Case School of Engineering and professor of biomedical engineering.

(Image: Rex Features)

New Hope for Reversing the Effects of Spinal Cord Injury

Walking is the obvious goal for individuals who have a chronic spinal cord injury, but it is not the only one. Regaining sensation and continence control also are important goals that can positively impact an individual’s quality of life. New hope for reversing the effects of spinal cord injury may be found in a combination of stem cell therapy and physical therapy as reported in Cell Transplantation by scientists at the University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School.

“Our phase one/two clinical trial had one goal: to give patients who have no other treatment options some hope,” said Hatem E. Sabaawy, MD, PhD, an assistant professor of medicine in the molecular and regenerative medicine program at Robert Wood Johnson Medical School. “Early findings have concluded that we have met our goal and can improve the quality of life for individuals with spinal cord injuries by providing a safe treatment that restores some neurological function.”

Dr. Sabaawy led a clinical trial that included 70 patients who had cervical or thoracic spinal cord injuries and were previously treated for at least six months without response. The patients were randomized into two groups, both of which were given physical therapy treatment. One of the groups also received stem cells derived from their own bone marrow injected near the injury site. Using the American Spinal Injury Association Impairment (AIS) Scale, patients received neurological and physical evaluations monthly for 18 months to determine if sensory and motor functions improved.

“Of primary importance, there was a notable absence of side effects in patients treated with stem cells during the course of our investigation,” added Dr. Sabaawy, who also is a resident member of The Cancer Institute of New Jersey at Robert Wood Johnson Medical School.

None of the patients in the control group who received only physical therapy showed any improvement in sensory or motor function during the same time frame. Although the scale of injuries differed, all patients who were treated with a combination of bone-marrow derived stem cells and physical therapy responded to tactile and sensory stimuli as early as 4 weeks into the study. After 12 weeks, they experienced improvements in sensation and muscle strength, which was associated with enhanced potency and improved bladder and bowel control that eventually allowed patients to live catheter-free. Patients who showed improvement based on the AIS scale also were able to sit up and turn in their beds.

“Since the emergence of stem cells as a potential therapy for spinal cord injury, scientists have diligently sought the best application for using their regenerating properties to improve a patient’s mobility,” said Joseph R. Bertino, MD, University Professor of medicine and pharmacology, interim director, Stem Cell Institute of New Jersey and chief scientific officer at The Cancer Institute of New Jersey. “Dr. Sabaawy’s discovery that treatment is more successful when stem cell therapy is combined with physical therapy could provide a remarkable, and hopefully sustainable, improvement in the overall quality of life for patients with spinal cord injury.”

At the end of 18 months, 23 of the 50 patients who received both physical therapy and stem cell therapy showed a significant improvement of at least 10 points on the AIS scale. Several were able to walk with assistance. In addition, more gains were made in motor skill control by patients with thoracic spinal cord injuries, suggesting that patients with thoracic spinal cord injuries may respond better to the combined treatment.

Dr. Sabaawy however cautioned that more studies are needed with a larger number of patients to test different cell dose levels and intervals at which stem cell therapy should be delivered.

“Although a cure for spinal cord injury does not yet exist, it is clear that the regenerative and secretory properties of bone-marrow derived stem cells can improve symptoms of paralysis in some patients when coupled with the current standard of care that physical therapy provides,” said Dr. Sabaawy. “We will continue monitoring our patients for long-term safety effects of stem cell therapy and work to expand our research through a phase two clinical trial that can be conducted at multiple centers nationwide and internationally.”

(Image courtesy: University of Alberta, Faculty of Rehabilitation Medicine)

Some brain cells are better virus fighters

Viruses often spread through the brain in patchwork patterns, infecting some cells but missing others. New research at Washington University School of Medicine in St. Louis helps explain why. The scientists showed that natural immune defenses that resist viral infection are turned on in some brain cells but switched off in others.

“The cells that a pathogen infects can be a major determinant of the seriousness of brain infections,” says senior author Michael Diamond, MD, PhD, professor of medicine. “To understand the basis of disease, it is important to understand which brain regions are more susceptible and why.”

While some brain infections are caused by bacteria, fungi or parasites, often the cause is a virus, such as West Nile virus, herpesvirus or enteroviruses.

For their study, now available online in Nature Medicine, the researchers focused on granule cell neurons, a cell type that rarely becomes infected. They compared gene profiles in granule cells from the cerebellum with the activity in cortical neurons in the cerebral cortex, which are more vulnerable to infection.

The comparison revealed many differences, including a number of genes in cortical neurons that were less well-expressed—meaning that for those specific genes there were fewer copies of mRNA, the molecules that relay genetic information from DNA to the cell’s protein-making mechanisms.

Next, the researchers transferred individually 40 of those genes into cortical neurons and screened the cells for susceptibility to viral infection. The test highlighted three antiviral genes that are induced by interferon, an important immune system protein. When the expression level of these genes increased in cortical neurons, the cells’ susceptibility to viral infection decreased.

The researchers also identified mechanisms that make some of these changes in genetic programming happen: regulatory factors known as microRNA, and differences in the way DNA is modified in the cell nucleus, both of which can affect gene expression levels.

Some of the genetic changes are only helpful against specific viral families, while others are effective against a broader spectrum of viruses and bacteria. The scientists can’t say yet if the differences in infection susceptibility are driven by the need to prevent infection or if they are a byproduct of changes that help neurons in particular brain regions perform essential functions.

To learn more about how these innate immune genes help cells resist infection, Diamond and his colleagues are disabling them in the brains of mice.

New Study Validates Longevity Pathway

A new study demonstrates what researchers consider conclusive evidence that the red wine compound resveratrol directly activates a protein that promotes health and longevity in animal models.

What’s more, the researchers have uncovered the molecular mechanism for this interaction, and show that a class of more potent drugs currently in clinical trials act in a similar fashion. Pharmaceutical compounds similar to resveratrol may potentially treat and prevent diseases related to aging in people, the authors contend.

These findings are published in the March 8 issue of Science.

For the last decade, the science of aging has increasingly focused on sirtuins, a group of genes that are believed to protect many organisms, including mammals, against diseases of aging. Mounting evidence has demonstrated that resveratrol, a compound found in the skin of grapes as well as in peanuts and berries, increases the activity of a specific sirtuin, SIRT1, that protects the body from diseases by revving up the mitochondria, a kind of cellular battery that slowly runs down as we age. By recharging the batteries, SIRT1 can have profound effects on health.

Mice on resveratrol have twice the endurance and are relatively immune from effects of obesity and aging. In experiments with yeast, nematodes, bees, flies and mice, lifespan has been extended.

“In the history of pharmaceuticals, there has never been a drug that binds to a protein to make it run faster in the way that resveratrol activates SIRT1,” said David Sinclair, Harvard Medical School professor of genetics and senior author on the paper. “Almost all drugs either slow or block them.”

In 2006, Sinclair’s group published a study showing that resveratrol could extend the lifespan of mice, and the company Sirtris Pharmaceuticals, which was started by HMS researchers, was founded to make drugs more potent than resveratrol. (Sinclair is a co-founder of Sirtris, a GlaxoSmithKline company, and remains a scientific advisor. Sirtris currently has a number of sirtuin-activating compounds in clinical trials.)

But while numerous studies, from Sinclair’s lab and elsewhere, underscored a direct causal link between resveratrol and SIRT1, some scientists claimed the studies were flawed.

The contention lay in the way SIRT1 was studied in vitro, using a specific chemical group attached to the targets of SIRT1 that fluoresces more brightly as SIRT1 activity increases. This chemical group, however, is synthetic and does not exist in cells or in nature, and without it the experiments did not work. As a response to this, a paper published in 2010 surmised that resveratrol’s activation of SIRT1 was an experimental artifact, one that existed in the lab, but not in an actual animal. SIRT1 activity in mice was, the paper claimed, at best an indirect result of resveratrol, and perhaps even a sheer coincidence.

As a result, a debate erupted over the particular pathway that resveratrol and similar compounds affected. Does resveratrol directly activate SIRT1 or is the effect indirect? “We had six years of work telling us that this was most definitely not an artifact,” said Sinclair. “Still, we needed to figure out precisely how resveratrol works. The answer was extremely elegant.”

Sinclair and Basil Hubbard, then a doctoral student in the lab, teamed up with a group of researchers from both the National Institutes of Health and Sirtris Pharmaceuticals to address this question.

First, the team addressed the problem of the fluorescent chemical group. Why was it required for resveratrol to rev up SIRT1 in the test tube? Instead of dismissing the result as an artifact, the researchers surmised that the chemical might be mimicking molecules found naturally in the cell. These turned out to be a specific class of amino acid, the building blocks of proteins. In nature, there are three amino acids that resemble the fluorescent chemical group, one of which is tryptophan, a molecule abundant in turkey and notable for inducing drowsiness. When researchers repeated the experiment, swapping the fluorescing chemical group on the substrate with a tryptophan residue, resveratrol and similar molecules were once again able to activate SIRT1.

“We discovered a signature for activation that is in fact found in the cell and doesn’t require these other synthetic groups,” said Hubbard, first author of the study. “This was a critical result, which allowed us to bridge the gap between our biochemical and physiological findings.

“Next, we needed to identify precisely how resveratrol presses on SIRT1’s accelerator,” said Sinclair. The team tested approximately 2,000 mutants of the SIRT1 gene, eventually identifying one mutant that completely blocked resveratrol’s effect. The particular mutation resulted in the substitution of a single amino acid residue, out of the 747 that make up SIRT1. The researchers also tested hundreds of other molecules from the Sirtris library, many of which are far more powerful than resveratrol, against this mutant SIRT1. All failed to activate it.

The authors propose a model for how resveratrol works: When the molecule binds, a hinge flips, and SIRT1 becomes hyperactive.

Although these experiments occurred in a test tube, once the researchers identified the precise location of the accelerator pedal on SIRT1—and how to break it—they could test their ideas in a cell. They replaced the normal SIRT1 gene in muscle and skin cells with the accelerator-dead mutant. Now they could test precisely whether resveratrol and the drugs in development work by tweaking SIRT1 (in which case they would not work) or one of the thousands of other proteins in a cell (in which they would work). While resveratrol and the drugs tested revved up mitochondria in normal cells (an effect caused activating by SIRT1), the mutant cells were completely immune.

“This was the killer experiment,” said Sinclair. “There is no rational alternative explanation other than resveratrol directly activates SIRT1 in cells. Now that we know the exact location on SIRT1 where and how resveratrol works, we can engineer even better molecules that more precisely and effectively trigger the effects of resveratrol.”

The researchers plan on continuing academic-industry collaborations with the goal of bringing to fruition drugs that treat diseases associated with aging.

Obesity makes fat cells act like they’re infected

The inflammation of fat tissue is part of a spiraling series of events that leads to the development of type 2 diabetes in some obese people. But researchers have not understood what triggers the inflammation, or why.

In Cell Metabolism this month (cover), scientists from The Methodist Hospital report fat cells themselves are at least partly to blame — high calorie diets cause the cells to make major histocompatibility complex II, a group of proteins usually expressed to help the immune system fight off viruses and bacteria. In overweight mice and humans the fat cells, or adipocytes, are issuing false distress signals — they are not under attack by pathogens. But this still sends local immune cells into a tizzy, and that causes inflammation.

"We did not know fat cells could instigate the inflammatory response," said principal investigator and Methodist Diabetes & Metabolism Institute Director Willa Hsueh, M.D. "That’s because for a very long time we thought these cells did little else besides store and release energy. But what we have learned is that adipocytes don’t just rely on local resident immune cells for protection — they play a very active role in their own defense. And that’s not always a good thing."

In pinpointing major histocompatibility complex II (MHCII) as a cause of inflammation, the researchers may have also identified a new drug target for the treatment of obesity. Blocking the MHCII response of adipocytes wouldn’t cure obesity, Hsueh said, “but it could make it possible for doctors to alleviate some of obesity’s worst consequences while the condition itself is treated.”

Could the inflammation caused by a high fat diet serve any purpose, or is it a senseless response to an unnaturally caloric diet?

"The expression of MHCII in adipocytes does not seem to be helpful to the body," said co-lead author Christopher Lyon, Ph.D. "It is not at all clear what the advantage would be, given all the negative long-term consequences of fat tissue inflammation in people who are obese, including insulin resistance and, eventually, full diabetes. This just appears to be a runaway immune response to a modern high calorie diet."

Hsueh added, “The bottom line is, you’re feeding and feeding these fat cells and they’re turning around and biting you back. They’re doing the thing they’re supposed to do — storing energy — but reacting negatively to too much of it.”

The scientists studied fat cells from obese, female humans (via biopsy) and overfed male mice. The researchers said that while they expect similar MHCII expression to occur in overweight male humans and female mice, further studies are needed to establish this.

The immunology of adipocyte inflammation is complex. It begins with the import of excess nutrients from the bloodstream, which are converted and stored as fat and stimulate the production of the hormone leptin. Excess leptin, spurred by a high calorie diet, excites CD4 T cells to produce a second signaling molecule, interferon gamma, which causes adipocytes to produce MHCII. This dialogue between adipocytes and T cells appears to initiate the inflammatory response to high fat diet — Hsueh and her group found that overfed mice lacking MHCII experienced less inflammation.

Interferon gamma from T cells exacerbates the inflamed adipocytes’ behavior and causes another type of immune cell, M2 macrophages, to be converted to their pro-inflammatory (M1) version.

"It was known that macrophages and T cells are major players," said lead author Tuo Deng, Ph.D. "But no one knew what the start signals were to ignite inflammation.

RNA was extracted from adipocytes purified from fat tissue biopsies and subjected to microarray analysis, which allowed the researchers to see what genes were increased in overweight subjects. The researchers found high expression of most MHCII complex and MHCII antigen processing genes. Similar gene expression patterns were observed in mice within two weeks of starting a high-fat diet, and this mirrored pro-inflammatory changes in fat tissue CD4 T cells. Hsueh says her group plans to investigate whether the inflammatory response in overfed mice can be blocked when MHCII expression is specifically reduced in adipocytes.

Hsueh says that if she and her group can identify the antigen(s) that MHCII is presenting to T cells in fat tissue, medical researchers would have a new approach to target adipose inflammation in obese patients. The hypothesis is that if a treatment can interfere with the production or MHCII presentation of these antigens, this would reduce the activation of fat tissue immune cells and thus reduce inflammation. Determining the MHCII antigen(s) involved in the inflammatory response of fat tissue to weight gain is one of her group’s next goals, she says.

Human brain treats prosthetic devices as part of the body

People with spinal cord injuries show strong association of wheelchairs as part of their body, not extension of immobile limbs injuries.

The human brain can learn to treat relevant prosthetics as a substitute for a non-working body part, according to research published March 6 in the open access journal PLOS ONE by Mariella Pazzaglia and colleagues from Sapienza University and IRCCS Fondazione Santa Lucia of Rome in Italy, supported by the International Foundation for Research in Paraplegie.

The researchers found that wheelchair-bound study participants with spinal cord injuries perceived their body’s edges as being plastic and flexible to include the wheelchair, independent of time since their injury or experience with using a wheelchair. Patients with lower spinal cord injuries who retained upper body movement showed a stronger association of the wheelchair with their body than those who had spinal cord impairments in the entire body.

According to the authors, this suggests that rather than being thought of only as an extension of the immobile limbs, the wheelchairs had become tangible, functional substitutes for the affected body part. As Pazzaglia explains, “The corporeal awareness of the tool emerges not merely as an extension of the body but as a substitute for, and part of, the functional self.”

Previous studies have shown that people with prosthetic devices that extend or restore movement may make such tools part of their physical identity, but whether this integration was due to prolonged use or a result of altered sensory input was unclear. Based on the results of this study, the authors suggest that it may be the latter, as the brain appears to continuously update bodily signals to incorporate these tools into a sense of the body. The study concludes that this ability may have applications in rehabilitation of physically impaired people.

(Image: University of Miami)

Researchers look to breath to identify stress

According to a new pilot study, published in IOP Publishing’s Journal of Breath Research, there are six markers in the breath that could be candidates for use as indicators of stress.

The researchers hope that findings such as these could lead to a quick, simple and non-invasive test for measuring stress; however, the study, which involved just 22 subjects, would need to be scaled-up to include more people, over a wider range of ages and in more “normal” settings, before any concrete conclusions can be made, they state.

Lead-author of the study, Professor Paul Thomas, said: “If we can measure stress objectively in a non-invasive way, then it may benefit patients and vulnerable people in long-term care who find it difficult to disclose stress responses to their carers, such as those suffering from Alzheimer’s.”

The study, undertaken by researchers at Loughborough University and Imperial College London, involved 22 young adults (10 male and 12 female) who each took part in two sessions: in the first, they were asked to sit comfortably and listen to non-stressful music; in the second, they were asked to perform a common mental arithmetic test that has been designed to induce stress.

A breath test was taken before and after each session, whilst heart-rates and blood pressures were recorded throughout. The breath samples were examined using a technique known as gas chromatography-mass spectrometry, and then statistically analysed and compared to a library of compounds.

Two compounds in the breath – 2-methyl, pentadecane and indole – increased following the stress exercise which, if confirmed, the researchers believe could form the basis of a rapid test.

A further four compounds were shown to decrease with stress, which could be due to changes in breathing patterns.

“What is clear from this study is that we were not able to discount stress. It seems sensible and prudent to test this work with more people over a range of ages in more normal settings.

“We will need to think carefully about experimental design in order to explore this potential relationship further as there are ethical issues to consider when deliberately placing volunteers under stress. Any follow up study would need to be led by experts in stress,” Professor Thomas continued.

Breath profiling has become an attractive diagnostic method for clinicians, and recently researchers have found biomarkers associated with tuberculosis, multiple cancers, pulmonary disease and asthma. It is still unclear how to best manage external factors, such as diet, environment and exercise, which can affect a person’s breath sample.

“It is possible that stress markers in the breath could mask or confound other key compounds that are used to diagnose a certain disease or condition, so it is important that these are accounted for,” said Professor Thomas.

The researcher’s initial assumptions are that stressed people breathe faster and have increased pulse rates and an elevated blood-pressure, which is likely to change their breath profile. They emphasise, however, that it is too soon to postulate the biological origins and the roles of the compounds as part of a stress-sensitive response.

Is it a Stroke or Benign Dizziness? A Simple Bedside Test Can Tell

A bedside electronic device that measures eye movements can successfully determine whether the cause of severe, continuous, disabling dizziness is a stroke or something benign, according to results of a small study led by Johns Hopkins Medicine researchers.

"Using this device can directly predict who has had a stroke and who has not," says David Newman-Toker, M.D., Ph.D., an associate professor of neurology and otolaryngology at the Johns Hopkins University School of Medicine and leader of the study described in the journal Stroke. “We’re spending hundreds of millions of dollars a year on expensive stroke work-ups that are unnecessary, and probably missing the chance to save tens of thousands of lives because we aren’t properly diagnosing their dizziness or vertigo as stroke symptoms.”

Newman-Toker says if additional larger studies confirm these results, the device could one day be the equivalent of an electrocardiogram (EKG), a simple noninvasive test routinely used to rule out heart attack in patients with chest pain. And, he adds, universal use of the device could “virtually eliminate deaths from misdiagnosis and save a lot of time and money.”

To distinguish stroke from a more benign condition, such as vertigo linked to an inner ear disturbance, specialists typically use three eye movement tests that are essentially a stress test for the balance system. In the hands of specialists, these bedside clinical tests (without the device) have been shown in several large research studies to be extremely accurate — “nearly perfect, and even better than immediate MRI,” says Newman-Toker. One of those tests, known as the horizontal head impulse test, is the best predictor of stroke. To perform it, doctors or technicians ask patients to look at a target on the wall and keep their eyes on the target as doctors move the patients’ heads from side to side. But, says Newman-Toker, it requires expertise to determine whether a patient is making the fast corrective eye adjustments that would indicate a benign form of dizziness as opposed to a stroke.

For the new study, researchers instead performed the same test using a small, portable device — a video-oculography machine that detects minute eye movements that are difficult for most physicians to notice. The machine includes a set of goggles, akin to swimming goggles, with a USB-connected webcam and an accelerometer in the frame. The webcam is hooked up to a laptop where a continuous picture of the eye is taken. Software interprets eye position based on movements and views of the pupil, while the accelerometer measures the speed of the movement of the head.

Newman-Toker says the test could be easily employed to prevent misdiagnosis of  as many as 100,000 strokes a year, leading to earlier stroke diagnosis and more efficient triage and treatment decisions for patients with disabling dizziness. Overlooked strokes mean delayed or missed treatments that lead to roughly 20,000 to 30,000 preventable deaths or disabilities a year, he says. The technology, he adds, could someday be used in a smartphone application to enable wider access to a quick and accurate diagnosis of strokes whose main symptom is dizziness, as opposed to one-sided weakness or garbled speech.

The diagnosis of stroke in patients with severe dizziness, vomiting, difficulty walking and intolerance to head motion is difficult, Newman-Toker says. He estimates there are 4 million emergency department visits annually in the United States for dizziness or vertigo, at least half a million of which involve patients at high risk for stroke. The most common causes are benign inner ear conditions, but many emergency room doctors, Newman-Toker says, find it nearly impossible to tell the difference between the benign conditions and something more serious, such as a stroke. So they often rely on brain imaging - usually a CT scan, an expensive and inaccurate technology for this particular diagnosis.

The Hopkins-led study enrolled 12 patients at The Johns Hopkins Hospital and the University of Illinois College of Medicine at Peoria, who later underwent confirmatory MRI. Six were diagnosed with stroke and six with a benign condition using video-oculography. MRI later confirmed all 12 diagnoses.