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Engineered immune cells recognize tumor cells and call a halt to cancers we thought were incurable

Engineered immune cells recognize tumor cells and call a halt to cancers we thought were incurable | Amazing Science | Scoop.it

The latest techniques involve genetically engineering immune T-cells to target and kill cancer cells, while leaving healthy cells relatively unscathed.

 

T-cells normally travel around the body clearing sickly or infected cells. Cancer cells can sometimes escape their attention by activating receptors on their surface that tell T-cells not to attack. ALL affects another type of immune cell, the B-cells, so Sadelain takes T-cells from people with ALL and modifies them to recognise CD19, a surface protein on all B-cells – whether cancerous or healthy. After being injected back into the patient, the reprogrammed T-cells destroy all B-cells in the person's body. This means they need bone marrow transplants afterwards to rebuild their immune systems. But because ALL affects only B-cells, the therapy guarantees that all the cancerous cells are destroyed.

 

A team led by Carl June from the University of Pennsylvania in Philadelphia used the same technique to treat several children with ALL, including Emily Whitehead (pictured right). He will present the latest results in December at the American Society of Hematology meeting in New Orleans. He will also report on the progress of adults with chronic lymphocytic leukaemia, who were treated with a similar technique that targeted B-cells, including some who are still in remission three years later.

 

Other teams are developing more targeted forms of immunotherapy, engineering T-cells to recognise markers that only cancer cells possess. What gives T-cells this potential, is that they can home in on what is going on inside cells, as well as outside. This vastly expands the range of potential targets.

 

Inside all cells, proteins are routinely broken apart and the resultant debris of tiny fragments called peptides are ferried to the cell surface by molecules called human leukocyte antigens (HLAs). These peptides then get inspected by passing T-cells – a process that allows the immune system to routinely check what is going on inside cells.

 

If the peptide fragment looks normal, the T-cell gives the OK and moves on, but if it is abnormal, perhaps because of a viral invasion or cancer mutation, the T-cell will destroy the cell. But sometimes, for unknown reasons, mutated cancer peptides are seen as healthy by T-cells and are ignored. So now, researchers are reprogramming T-cells to respond specifically to peptides with hallmarks of cancer delivered to the surface from within cells.


Once such peptides are identified, there are two ways to engineer T-cells to seal cancer's fate. The first involves taking a person's T-cells and engineering them so they have new genes that make new receptors. These receptors bind exclusively to the cancer peptide, so once they are injected, the T-cells home in on and destroy all cells that contain the peptide.

 

The second way is to produce artificial T-cell receptors that are primed to recognise a cancer peptide. These receptors contain features that enable them to kill cancer cells once they have bound to them. These features include arms that summon passing native T-cells, or toxic chemicals that kill cells exposed to them.

 

The first technique has put 16 out of 20 people with myeloid myeloma into remission for two years. They had a T-cell treatment by Adaptimmune in Oxford, UK, that targets a peptide called NY-ESO created inside tumor cells.

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Who are the doctors most trusted by other doctors? Big data can tell you!

Who are the doctors most trusted by other doctors? Big data can tell you! | Amazing Science | Scoop.it

ZocDoc, Healthgrades, Vitals, Yelp and other sites can tell you what patients think of their doctors. But finding out in any aggregate way what doctors think of their peers has been much harder, if not near impossible, for patients — up until now.

 

By accessing information in government databases through FOIA (Freedom of Information Act) requests, healthcare innovators are now able to share connections between doctors that are based on millions of physician referrals — a valuable indicator of who doctors hold in esteem.

 

HealthTap, a Palo Alto-based startup that connects patients with an online network of 17,000 doctors, also this week launched a new feature based partly on Trotter’s data. Called “DOConnect,” it combines Trotter’s Medicare data with physician data from its own site and other sources to give patients a new window into their doctors’ networks.

 

“This isn’t just friendships and business connections. This is who doctors trust,” said HealthTap co-founder and CEO Ron Gutman. “If you could know who your doctor’s doctor is, if you knew who they would choose, this lets you see that for the first time.”

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100 gene transcripts can generate thousands of conotoxins in a single species of cone snail

100 gene transcripts can generate thousands of conotoxins in a single species of cone snail | Amazing Science | Scoop.it

Cone snails produce highly complex venom comprising mostly small biologically active peptides known as conotoxins or conopeptides. Early estimates that suggested 50-200 venom peptides are produced per species have been recently increased at least 10-fold using advanced mass spectrometry.


To uncover the mechanism(s) responsible for generating this impressive diversity, we used an integrated approach combining second-generation transcriptome sequencing with high sensitivity proteomics. From the venom gland transcriptome of Conus marmoreus, a total of 105 conopeptide precursor sequences from 13 gene superfamilies were identified. Over 60% of these precursors belonged to the three gene superfamilies O1, T, and M, consistent with their high levels of expression, which suggests these conotoxins play an important role in prey capture and/or defense. Seven gene superfamilies not previously identified in C. marmoreus, including five novel superfamilies, were also discovered. To confirm the expression of toxins identified at the transcript level, the injected venom of C. marmoreus was comprehensively analyzed by mass spectrometry, revealing 2710 and 3172 peptides using MALDI and ESI-MS, respectively, and 6254 peptides using an ESI-MS TripleTOF 5600 instrument.


All conopeptides derived from transcriptomic sequences could be matched to masses obtained on the TripleTOF within 100 ppm accuracy, with 66 (63%) providing MS/MS coverage that unambiguously confirmed these matches. Comprehensive integration of transcriptomic and proteomic data revealed for the first time that the vast majority of the conopeptide diversity arises from a more limited set of genes through a process of variable peptide processing, which generates conopeptides with alternative cleavage sites, heterogeneous post-translational modifications, and highly variable N- and C-terminal truncations. Variable peptide processing is expected to contribute to the evolution of venoms, and explains how a limited set of ∼ 100 gene transcripts can generate thousands of conopeptides in a single species of cone snail.

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Medical Training Goes Holographic

Medical Training Goes Holographic | Amazing Science | Scoop.it

A system called zSpace uses a large screen, glasses and pointer — like a ballpoint pen — to interact with and manipulate 3-D images. The technology allows users to look completely around the object, examine it from all angles, and zoom in and out. It’s also finding its way into universities and will be used to train med students and future surgeons. One day, the doctor operating on a wounded soldier or diagnosing a veteran may have had his humble beginnings exploring a virtual body.

 

“It’s not like 3-D when you go to the movie; that’s actually one dimension,” said David Lenihan, dean of preclinical medicine and associate professor of neuroanatomy at Touro College in New York. The system will be part of a virtual laboratory set to open in fall 2014 at Touro. “You have to take the field of view that it gives you. With zSpace, you can look around.”

 

Lenihan said that from an academic perspective, virtual humans modeled with 3-D holography techniques will provide some benefits over cadaver dissections. One of the problems with real materials is how things shift after death — the vascular system is not quite where it would normally be; the muscle tone isn’t quite as firm. “Your skill set is actually much better in the virtual world, because the relationships are better to real life,” he said.

 

Increases in computing power mean that processing complex, data-rich MRI scans or radiographs is now easier and faster. Computers can run the information-intensive models and present the image in robust, movable 3-D that you can seemingly pull out of the screen and spin around. While the virtual bodies dissected at Touro will be rendered objects designed with the University of Iowa, Lenihan said future applications on zSpace and similar technologies could pull data from real patients.

 

Another potential benefit of the holographic technology is the Magic School Bus-like ability to zoom down to the smallest scales and explore the body in ways you cannot experience in real life.

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Region of brain identified that is responsible for nicotine withdrawal symptoms

Region of brain identified that is responsible for nicotine withdrawal symptoms | Amazing Science | Scoop.it

Headaches, anxiety, irritability—these and other symptoms of nicotine withdrawal can significantly deter smokers from being able to kick the habit. Now, in what may be a significant step toward alleviating those symptoms, UMass Medical School neuroscientist Andrew R. Tapper, PhD, and colleagues have identified the region of the brain in which they originate.

“We were surprised to find that one population of neurons within a single brain region could actually control physical nicotine withdrawal behaviors,” said Dr. Tapper, associate professor of psychiatry and interim director of the Brudnick Neuropsychiatric Research Institute at UMMS.

 

The Tapper lab discovered that physical nicotine withdrawal symptoms are triggered by activation of GABAergic neurons (neurons that secrete GABA, the brain’s predominant inhibitory neurotransmitter), in the interpeduncular nucleus, an area deep in the midbrain that has recently been shown to be involved in nicotine intake. Their study was published in the Nov. 14 issue of the journal Current Biology.

 

“Most of the work in the field has been focused on the immediate effects of nicotine, the addictive component in tobacco smoke, on reward circuits in the brain,” Tapper explained. “But much less is known regarding what happens when you take nicotine away from someone who has been smoking for a long time that causes all these terrible withdrawal symptoms. Our main goal was to understand what brain regions are activated—or deactivated—to cause nicotine withdrawal symptoms.

 

They did this through a series of experiments performed in mouse models with sophisticated neurochemistry and brain imaging methods, including recently developed optogenetics techniques in which specific neurons can be activated by light.

 

Most surprising was their discovery that nicotine withdrawal symptoms can be activated or deactivated independent of nicotine addiction. “When we activated the GABAergic neurons in the interpeduncular nucleus, mice suffered withdrawal symptoms even if they had no previous nicotine exposure,” Tapper noted.

 

These findings are promising because existing treatments intended to help people quit smoking are not always effective. “There are very few treatments to help people quit smoking,” Tapper said. “If you can dampen the activity of this brain region chemically during nicotine withdrawal then you would hopefully be able to help someone quit smoking because you could reduce some of the withdrawal symptoms that they are experiencing.”

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Zayd El-Ali's curator insight, December 12, 2013 10:15 PM

One population of neurons in a section of the brain controls the symptoms of withdrawal. If this, based on the article, is true. Then a drug could be used to contain this area of the brain with these specific neurons in order to prevent the effects the withdrawal.  I included this article because the Importance of this new research can lead other scientist to build on the foundation provided, and even get one step closer for a cure to the effects of withdrawal. 

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Decade-long experience of retroviral-modified chimeric antigen receptor T cells for treating cancer

Decade-long experience of retroviral-modified chimeric antigen receptor T cells for treating cancer | Amazing Science | Scoop.it

One child surviving ‘incurable’ cancer is an amazing event, but there is a lot more work to be done to find out how best to use this new technology. At the moment it’s still highly experimental and expensive. It’s only being trialled in a very small number of patients, primarily to make sure it is safe, and so far we’ve seen that it doesn’t work for everyone.

 

In the case of the child whose cancer came back after treatment, the researchers found that her cancer cells had somehow stopped carrying the T cells’ target molecule. So it’s likely that other targets will need to be identified, to make the treatment more effective for more patients in the future.

 

On a positive note, there’s no reason why this type of treatment should be restricted to cancers affecting the immune system (namely leukaemia and lymphoma), although they’re much more accessible to the killer T cells. Researchers elsewhere are investigating how to target a range of different types of cancer with this approach.

 

There are several similar therapies being tested in the lab and in clinical trials around the world, including in the UK. And Cancer Research UK scientists are finding out whether harmless genetically-engineered viruses could be used as therapeutic vaccines, training the immune system to seek and destroy cancer cells.

 

It’s still early days for these exciting new approaches and there are many hurdles to jump, but we’re looking forward to the day when they can be used to treat patients on a wider scale.

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Pacemaker Okayed in Europe Is One-Tenth the Size of Those Used Now

Pacemaker Okayed in Europe Is One-Tenth the Size of Those Used Now | Amazing Science | Scoop.it

Throughout the history of medicine, many devastating illnesses were first treated with dangerous, sometimes even barbaric, methods that initially seemed almost as bad as the sickness. Over time, those treatments get refined and it becomes hard to believe that people once died of epilepsy, for example, or from what seem now to be minor heart conditions.

 

Many types of arrythmia, for example, that used to kill patients no longer do. More than 4 million people around the world wear pacemakers. But it’s still a serious matter to have life-saving pacemakers installed and a limitation to live with one.

 

But in the coming year, it will likely become significantly easier to receive and live with a pacemaker. Developed by Silicon Valley startup Nanostim, a device about the size of a AAA battery, or one-tenth the size of a conventional pacemaker, was recently approved for use in Europe. It is installed through a catheter in the femoral vein in a minimally invasive procedure. Then, for about 10 years it sits inside the ventricle of the heart and delivers its regulatory electrical pulses wirelessly.


“For the past 40 years the therapeutic promise of leadless [or wireless] pacing has been discussed, but until now, no one has been able to overcome the technical challenges,” Dr. Johannes Sperzel of the Kerckhoff Clinic in Bad Nauheim, Germany, said in a news release.

 

The first pacemaker was the size of an ice hockey puck and had to be installed in the abdomen. Currently, most are about the size of a watch and are installed in a “surgical pocket” under the skin near the collarbone. The padded wires, or leads, that feed down to the heart to stimulate it cause many patients discomfort.

 

After the approval came through, Minnesota-based St. Jude Medical acquired Nanostim for $123.5 million. The company, which made the first pacemaker in 1958, had funded Nanostim’s work, and the Nanostim pacemaker uses a St. Jude Medical electrode.

 


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Carbon Nanotube Sensors For Long-Term Subdermal Glucose Biosensing

Carbon Nanotube Sensors For Long-Term Subdermal Glucose Biosensing | Amazing Science | Scoop.it

Carbon nanotubes are nano-scale, cylindrical structures composed of carbon chains, with unique properties that make them particularly attractive for use in the construction of various nanotechnologies. Nanoscale sensors and devices have shown to be increasingly relevant in the medical world, as they seem to address many of the prominent issues that modern medicine is not yet able to confront, or address cost-effectively.


Researchers at MIT are taking advantage of the unique properties of carbon nanotubes, namely natural fluorescence in a spectral region that has little interference from biological media, to construct biosensors that monitor the presence of nitric oxide, an important signaling molecule whose levels fluctuate in cancer cells. The sensor consists of a nanotube segment, wrapped in a particular DNA sequence, which binds to the target molecules, altering the fluorescent luminosity. The technology is currently being adapted for use in diabetic patients, by altering the sensor to bind to glucose. Theoretically, the technology could be adapted to any number of biomolecules by altering the DNA sequence used in the sensor.

 

What makes this technology more appealing, however, is the prospect of long term monitoring when localized subdermally. The nanosensor can be injected directly into the bloodstream for short-term use, or placed under the skin of a patient for uninterrupted monitoring for over a year, according to the research. This is accomplished by embedding the sensor in a biocompatible gel that can protect the technology for up to 400 days; the researchers believe it could potentially last longer.

 

Furthermore, when injected into the bloodstream, the nanosensor was observed to pass through the heart and lungs without causing damage or clumping, an important factor for venous sensing technology.

Hopefully, the development of this technology will be successful, as it could be extremely helpful in monitoring the progression of diabetes, cancer, or virtually any other illness that can be gauged through the presence of biomarkers.

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New Hope for a Drug that Might Work for a Variety of Cancers

New Hope for a Drug that Might Work for a Variety of Cancers | Amazing Science | Scoop.it

The most frequently mutated gene across all types of cancers is a gene called p53. Unfortunately it has been difficult to directly target this gene with drugs. Now a multi-institutional research team, led by Dr. Lewis Cantley and investigators at Weill Cornell Medical College, has identified a family of enzymes they say is crucial for the growth of cancers that have genetic aberrations in p53. Targeting these enzymes with novel agents might prevent the growth of p53 mutant cancers, thereby benefiting a broad spectrum of cancer patients, including those with breast, ovarian, lung, colorectal and brain tumors.


In the Nov. 7, 2013 issue of Cell, investigators pinpoint two cellular enzymes — Type 2 phosphatidylinositol-5-phosphate 4-kinases α and β (Type 2 PIP kinases) — as essential for cancer growth when cells have lost p53, the powerful tumor-suppressor gene long dubbed the "guardian of the genome." More than half of all cancers lose this gene, allowing these cancers to grow at will.

 

The researchers discovered that the Type 2 PIP kinases are not critical for the growth of normal cells but become essential for cell growth when p53 is lost due to mutations or deletions. The scientists showed, in animal and lab studies of human cancer cells, that targeting these molecules effectively shuts down the growth of p53 mutant cancers.

 

Although the studies were conducted in human breast cancer cells, the researchers believe Type 2 PIP kinase inhibitors could block the growth of cancers with a mutated or missing p53 gene.

 

"The fact that one can delete the Type 2 PIP kinases in normal human cells or in mice with essentially no effect on cell survival suggests that inhibitors of these enzymes should have little toxicity," says Dr. Cantley, the study's senior author and director of the Cancer Center at Weill Cornell Medical College and NewYork-Presbyterian Hospital.

 

Dr. Cantley is already leading an effort to develop drugs to shut down these kinases. "Well-designed Type 2 PIP kinase inhibitors may turn the tide on p53 mutant cancer," he says.

 

Dr. Cantley is known for his discovery of the PI 3-kinase oncogene, and pioneering work in teasing apart how the gene contributes to cancer. PI 3-kinases (PI3K) have been linked to a wide variety of cellular functions, including cell growth and proliferation, and most cancers activate PI3K by one or more mechanisms. Dr. Cantley's discovery led to promising avenues for the development of personalized cancer therapies.

 

Activity of PI3K is in some cases linked to Type 2 PIP kinases, so in this study, Dr. Cantley sought to understand the function of these enzymes. Because the researchers knew that a subset of breast cancers over-express these molecules, investigators looked at their role in HER2-positive breast cancers, which typically are more aggressive tumors.

 

The researchers, including those from Harvard Medical School, Beth Israel Deaconess Medical Center and other institutions, discovered that the enzymes are silent in cells that have healthy p53. One critical role of p53 is to "rescue" cells that are producing excess reactive oxygen species (ROS), which are byproducts of cells that are growing too rapidly. The oxidative stress produced by ROS can damage cell structures, so p53 attempts to reduce ROS in affected cells. "If, however, ROS levels exceed the capacity of p53s to rescue it, then p53 takes on a second function, which is to kill the cell," Dr. Cantley says.

 

"That is why cancers often disable p53. If p53 is mutated or gone, then the cell keeps on growing at a very high rate," he says. "And then ROS begins to damage genes, making the cancer even more aggressive."


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Smog blamed as 8-year-old girl becomes youngest lung cancer patient

Smog blamed as 8-year-old girl becomes youngest lung cancer patient | Amazing Science | Scoop.it

An eight-year-old Chinese girl has become the mainland's youngest lung cancer patient, with her illness blamed directly on environment factors.

 

The girl from Jiangsu lived by a busy road where she inhaled all kinds of dust and particles, China News Service cited Dr Feng Dongjie of Jiangsu Cancer Hospital as saying. These included superfine PM2.5 particles, less than 2.5 microns wide, that are considered the most dangerous component of smog, Feng said.

 

The country's breakneck urbanisation and industrialisation has created some of the world's worst urban pollution, which is blamed for soaring rates of cancer and respiratory diseases.

 

In Beijing, which has suffered frequent, severe smog in recent years, deaths from lung cancer rose by 56 per cent from 2001 to 2010. A fifth of all cancer patients suffer lung cancer, figures from the Beijing Health Bureau show. It became the leading cause of cancer deaths among men in the capital and the second-biggest among women, after breast cancer, in 2010.

 

The World Health Organisation's "2010 Global Burden of Disease" study found that air pollution accounted for 1.2 million premature deaths worldwide in 2010, including 140,000 deaths from lung cancer.

 

Last month the WHO's International Agency for Research on Cancer said that air pollution from traffic and industrial fumes caused lung cancer and was also linked to bladder cancer.

 

Air pollution, mostly caused by transport, power generation, industrial or agricultural emissions and residential heating and cooking, was found to pose similar health risks to breathing in second-hand tobacco smoke.

 

The WHO said that in 2010, 223,000 people died from lung cancer worldwide resulting from air pollution.

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Christian Troiani's curator insight, November 10, 2013 1:40 AM

This article is based on lung cancer being mainly caused by air pollution from traffic and industry. It conveys this message effectively by stating at the beginning of the article that an eight year old girl is the youngest patient with lung cancer. The article suggests the girl contracted the disease because she lived on a busy road where she inhaled various dangerous pollutants. It goes on to emphasise its point by stating various alarming statistics based on traffic and industrial air pollutants being responsible for lung cancer. The World Health Organisation found that air pollutants accounted for 1.2 million premature deaths in 2010 worldwide which included 140,000 lung cancer deaths. A noted oncologist states that lung cancer is the  most common cause of cancer in Asia followed by stomach and liver cancer. This article is blaming air pollution for various cancers, especially lung cancer.

Nicholas Grozdanov's curator insight, November 13, 2013 8:02 PM

An eight year old girl becomes china's youngest lung cancer patient .Lung cancer is the most common cancer in Asia, Dr Hao Xishan, a noted oncologist. this is because of the pollution which is produced by thetransport, power generation, industrial or agricultural emissions and residential heating and cooking.


This is a deadly result as this could happen to anyone who lives in a heavy polluted region.

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DNA testing reveals that herbal supplements are often not what they seem

DNA testing reveals that herbal supplements are often not what they seem | Amazing Science | Scoop.it
A study using DNA testing offers perhaps the most credible evidence to date of adulteration, contamination and mislabeling in the herbal supplement industry.

 

Herbal products available to consumers in the marketplace may be contaminated or substituted with alternative plant species and fillers that are not listed on the labels. According to the World Health Organization, the adulteration of herbal products is a threat to consumer safety. Our research aimed to investigate herbal product integrity and authenticity with the goal of protecting consumers from health risks associated with product substitution and contamination.

 

A team of researchers used DNA barcoding to conduct a blind test of the authenticity for (i) 44 herbal products representing 12 companies and 30 different species of herbs, and (ii) 50 leaf samples collected from 42 herbal species. Their laboratory also assembled the first standard reference material (SRM) herbal barcode library from 100 herbal species of known provenance that were used to identify the unknown herbal products and leaf samples. They recovered DNA barcodes from most herbal products (91%) and all leaf samples (100%), with 95% species resolution using a tiered approach (rbcL + ITS2). Most (59%) of the products tested contained DNA barcodes from plant species not listed on the labels. Although they were able to authenticate almost half (48%) of the products, one-third of these also contained contaminants and or fillers not listed on the label. Product substitution occurred in 30/44 of the products tested and only 2/12 companies had products without any substitution, contamination or fillers. Some of the contaminants they found pose serious health risks to consumers.

 

Conclusion: Most of the herbal products tested were of poor quality, including considerable product substitution, contamination and use of fillers. These activities dilute the effectiveness of otherwise useful remedies, lowering the perceived value of all related products because of a lack of consumer confidence in them. The research team suggests that the herbal industry should embrace DNA barcoding for authenticating herbal products through testing of raw materials used in manufacturing products. The use of an SRM DNA herbal barcode library for testing bulk materials could provide a method for 'best practices' in the manufacturing of herbal products. This would provide consumers with safe, high quality herbal products.

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Carbon Nanotube-Based Sensors Can be Implanted Under the Skin for a Year

Carbon Nanotube-Based Sensors Can be Implanted Under the Skin for a Year | Amazing Science | Scoop.it

Hollow, one-nanometer-thick carbon nanotubes have drawn great interest as sensors. Led by postdoc Nicole Iverson, Strano's lab has built a sensor that can monitor NO in living animals for more than a year.


Strano's lab has recently developed carbon nanotube sensors for a variety of molecules, including hydrogen peroxide and toxic agents such as the nerve gas sarin. Such sensors take advantage of carbon nanotubes' natural fluorescence, by coupling them to a molecule that binds to a specific target. When the target is bound, the tubes' fluorescence brightens or dims.Strano's lab has previously shown that carbon nanotubes can detect NO if the tubes are wrapped in DNA with a particular sequence. In the new paper, the researchers modified the nanotubes to create two different types of sensors: one that can be injected into the bloodstream for short-term monitoring, and another that is embedded in a gel so it can be implanted long-term under the skin.


To make the particles injectable, Iverson attached PEG, a biocompatible polymer that inhibits particle-clumping in the bloodstream. She found that when injected into mice, the particles can flow through the lungs and heart without causing any damage. Most of the particles accumulate in the liver, where they can be used to monitor NO associated with inflammation.

 

"So far we have only looked at the liver, but we do see that it stays in the bloodstream and goes to kidneys. Potentially we could study all different areas of the body with this injectable nanoparticle," Iverson says. The longer-term sensor consists of nanotubes embedded in a gel made from alginate, a polymer found in algae. Once this gel is implanted under the skin of the mice, it stays in place and remains functional for 400 days; the researchers believe it could last even longer. This kind of sensor could be used to monitor cancer or other inflammatory diseases, or to detect immune reactions in patients with artificial hips or other implanted devices, according to the researchers.


Once the sensors are in the body, the researchers shine a near-infrared laser on them, producing a near-infrared fluorescent signal that can be read using an instrument that can tell the difference between nanotubes and other background fluorescence.

 

Iverson is now working on adapting the technology to detect glucose, by wrapping different kinds of molecules around the nanotubes. Most diabetic patients must prick their fingers several times a day to take blood glucose readings. While there are electrochemical glucose sensors available that can be attached to the skin, those sensors last only a week at most, and there is a risk of infection because the electrode pierces the skin.

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UC Develops Intelligent Nano Carrier to Target Drug Delivery to Cancer Cells

UC Develops Intelligent Nano Carrier to Target Drug Delivery to Cancer Cells | Amazing Science | Scoop.it

University of Cincinnati researchers have developed the first-of-its-kind nanostructure which is unusual because it can carry a variety of cancer-fighting materials on its double-sided (Janus) surface and within its porous interior.

 

Because of its unique structure, the nano carrier can do all of the following:

 

• Transport cancer-specific detection nanoparticles and biomarkers to a site within the body, e.g., the breast or the prostate. This promises earlier diagnosis than is possible with today’s tools. 

 

• Attach fluorescent marker materials to illuminate specific cancer cells, so that they are easier to locate and find for treatment, whether drug delivery or surgery. 

 

• Deliver anti-cancer drugs for pinpoint targeted treatment of cancer cells, which should result in few drug side effects. Currently, a cancer treatment like chemotherapy affects not only cancer cells but healthy cells as well, leading to serious and often debilitating side effects.

 

This recently developed Janus nanostructure is unusual in that, normally, these super-small structures (that are much smaller than a single cell) have limited surface. This makes is difficult to carry multiple components, e.g., both cancer detection and drug-delivery materials. The Janus nanocomponent, on the other hand, has functionally and chemically distinct surfaces to allow it to carry multiple components in a single assembly and function in an intelligent manner.
 
“In this effort, we’re using existing basic nano systems, such as carbon nanotubes, graphene, iron oxides, silica, quantum dots and polymeric nano materials in order to create an all-in-one, multidimensional and stable nano carrier that will provide imaging, cell targeting, drug storage and intelligent, controlled drug release,” said UC’s Shi, adding that the nano carrier’s promise is currently greatest for cancers that are close to the body’s surface, such as breast and prostate cancer.

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Adrian Vallejo Blanco's curator insight, November 2, 2013 1:16 PM

Una nanoestructura como nave nodriza para vehiculizar drogas y marcadores contra células tumorales. Esta estructura posee una superficie de doble cara y un espacio interior hueco donde alberga la droga.

 

En su superficie, puede transportar anclados varios complejos como marcadores fluorescentes junto con receptores específicos para señales características de un tipo concreto de tumor, de modo que cuando la molécula se ancla a la célula cancerígena sea capaz de liberar la droga y no solo eso, sino emitir fluorescencia marcando y delimitando la masa tumoral facilitando tanto el diagnóstico como hasta una posible cirugía.

 

Una gran ventaja aparte de que la masa tumoral pueda ser delimitada y señalizada con fluorescencia, es que la droga contra las células cancerosas sea liberada justo en esas células cancerosas afectando así minimamente a las células sanas. Actualmente con la radio y quimioterapia se ataca a las células tumorales pero las sanas son gravemente debilitadas.

Alexis Meneses Arévalo's curator insight, November 3, 2013 6:26 AM

DALCAME

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Diamond ‘flaws’ may pave the way for nanoscale MRI and thermometry of single cells

Diamond ‘flaws’ may pave the way for nanoscale MRI and thermometry of single cells | Amazing Science | Scoop.it

Breakthrough offers high-sensitivity nanoscale sensors, and could lead to magnetic imaging of neuron activity and thermometry on a single living cell. - See more at:

 

By exploiting flaws in miniscule diamond fragments, researchers say they have achieved enough coherence of the magnetic moment inherent in these defects to harness their potential for precise quantum sensors in a material that is 'biocompatible'.

 

Nanoscopic thermal and magnetic field detectors - which can be inserted into living cells - could enhance our understanding of everything from chemical reactions within single cells to signalling in neural networks and the origin of magnetism in novel materials.

 

Atomic impurities in natural diamond structure give rise to the colour seen in rare and coveted pink, blue and yellow diamond. But these impurities are also a major research focus in emerging areas of quantum physics.

 

One such defect, the Nitrogen-vacancy Centre (NVC), consists of a gap in the crystal lattice next to a nitrogen atom. This system tightly traps electrons whose spin states can be manipulated with extreme precision.

Electron coherence - the extent to which the spins of these particles can sustain their quantum mechanical properties - has been achieved to high levels in the NVCs of large 'bulk' diamonds, with coherence times of an entire second in certain conditions - the longest yet seen in any solid material.

 

However in nanodiamonds - nanometer sized crystals that can be produced by milling conventional diamond - any acceptable degree of coherence has, until now, proved elusive.

 

Nanodiamonds offer the potential for both extraordinarily precise resolution, as they can be positioned at the nano-scale, and biocompatibility - as they have can be inserted into living cells. But without high levels of coherence in their NVCs to carry information, these unique nanodiamond benefits cannot be utilised.

 

By observing the spin dynamics in nanodiamond NVCs, researchers at Cambridge's Cavendish Laboratory, have now identified that it is the concentration of nitrogen impurities that impacts coherence rather than interactions with spins on the crystal surface.

 

By controlling the dynamics of these nitrogen impurities separately, they have increased NVC coherence times to a record 0.07 milliseconds longer than any previous report, an order of significant magnitude - putting nanodiamonds back in play as an extremely promising material for quantum sensing.

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DLR Institute of Robotics and Mechatronics: Telemanipulation in minimally invasive surgery

DLR Institute of Robotics and Mechatronics: Telemanipulation in minimally invasive surgery | Amazing Science | Scoop.it

Conventional minimally invasive surgery (MIS) is per­formed through small incisions in the patient’s skin, pre­serving healthy tissue. The surgeon works with long slender instruments, and is separated from the operation area. This arrangement challenges the surgeon’s skills due to lost hand-eye-coordination and missing direct manual con­tact to the operation area. Therefore, many sophisticated procedures still cannot be performed minimally invasive. To overcome the drawbacks of conventional MIS, telepresence and telemanipulation techniques play an im­portant role: In case of minimally invasive robotic surgery (MIRS) the instruments are not directly manipulated anymore.


Instead, they are held by specialized robot arms and remotely commanded by the surgeon who comfortably sits at an input console. The surgeon virtually regains direct access to the operating field by having 3D endoscopic sight, force feedback, and restored hand-eye-coordination.

 

The DLR telesurgery scenario MIROSURGE includes an input (or master) console as well as a teleoperator consisting of 3 surgical robots (MIRO). Usually two MIROs carry surgical instruments (MICA) equipped with miniatur­ized force/torque sensors to capture reaction forces with manipulated tissue. One more MIRO can (automatically) guide a stereo video laparoscope. Both the stereo video stream and the measured forces are displayed to the surgeon at the master console. So users are not limited to see but can also feel what they are doing. An Omega.7 input device is used as force display.

 

Our ultimate ambition is robot supported surgery on the beating heart. The application of the heart-lung machine would become obsolete for a whole variety of procedures that way. Collaterally, the very traumatizing effects of the heart-lung machine on the patient could be avoided (e.g. blood contact with extrinsic surfaces, inevitable blood clotting attenuation, typical generalized inflammation reaction). Therefore, performance characteristics of the MIROs are designed to follow a stabilized beating heart motion. Additionally, the endoscopic video stream can be stabilized by optical tracking in real time so that a virtually stationary video picture can be consistently presented to the surgeon.

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People with highly superior memory powers of recall are also vulnerable to false memories

People with highly superior memory powers of recall are also vulnerable to false memories | Amazing Science | Scoop.it
People who can accurately remember details of their daily lives going back decades are as susceptible as everyone else to forming fake memories, psychologists and neurobiologists have found.

 

Persons with highly superior autobiographical memory (HSAM, also known as hyperthymesia) -- which was first identified in 2006 by scientists at UC Irvine's Center for the Neurobiology of Learning & Memory -- have the astounding ability to remember even trivial details from their distant past. This includes recalling daily activities of their life since mid-childhood with almost 100 percent accuracy.

 

The lead researcher on the study, Patihis believes it's the first effort to test malleable reconstructive memory in HSAM individuals. Working with neurobiology and behavior graduate student Aurora LePort, Patihis asked 20 people with superior memory and 38 people with average memory to do word association exercises, recall details of photographs depicting a crime, and discuss their recollections of video footage of the United Flight 93 crash on 9/11. (Such footage does not exist.) These tasks incorporated misinformation in an attempt to manipulate what the subjects thought they had remembered.

 

"While they really do have super-autobiographical memory, it can be as malleable as anybody else's, depending on whether misinformation was introduced and how it was processed," Patihis said. "It's a fascinating paradox. In the absence of misinformation, they have what appears to be almost perfect, detailed autobiographical memory, but they are vulnerable to distortions, as anyone else is."

 

He noted that there are still many mysteries about people with highly superior autobiographical memory that need further investigation. LePort, for instance, is studying forgetting curves (which involve how many autobiographical details people can remember from one day ago, one week ago, one month ago, etc., and how the number of details decreases over time) in both HSAM and control participants and will employ functional MRI to better understand the phenomenon.

 

"What I love about the study is how it communicates something that memory distortion researchers have suspected for some time: that perhaps no one is immune to memory distortion," Patihis said. "It will probably make some nonexperts realize, finally, that if even memory prodigies are susceptible, then they probably are too. This teachable moment is almost as important as the scientific merit of the study. It could help educate people -- including those who deal with memory evidence, such as clinical psychologists and legal professionals -- about false memories."

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Edible sensors that fit inside pills and tell your doctor when pills are taken predicted to be released in 2014

Edible sensors that fit inside pills and tell your doctor when pills are taken predicted to be released in 2014 | Amazing Science | Scoop.it

"If you look at every healthcare system in the world, it's finished," says Don Cowling, VP at Proteus Digital Health. "Instead of spending $10 billion (£6.4billion) trying to find a new molecule, why not spend half a billion getting today's products working properly?" That's what he is doing at California and London-based Proteus Digital Health, which harvests biological data using ingestible sensors and skin patches, to improve diagnosis and treatments already available. It's making edible sensors that fit inside pills and tell your doctor when pills are taken. They're expected to come to market in late 2014.

 

When a patient takes pills erratically and their condition worsens, a doctor may simply up the dose. Proteus is building silicon, copper and magnesium chips of about 1mm squared that can be inserted into tablets -- these report via Bluetooth when a pill's been taken.

 

In May, the firm announced a $62.5 million (£38.9 million) funding round, including investment from Oracle. But smart pills are just the start, says Cowling. Proteus's patch sensor can gather dozens of other data points, including heart rate, to present a sophisticated picture of patient health -- like a medical-grade FuelBand. "We can now get a formal classification of what disability looks like -- we can measure it." 

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Modern Medicine,  is breaking your privacy?

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Interactive Live Holography - From Science Fiction to Science Fact

Introducing live medical holography - the world's first 3D holographic display and interface system, initially for medical imaging applications. To learn more visit: http://www.realviewimaging.com/


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By reactivating a dormant gene, Lin28a, researchers could regrow hair, cartilage, bone and soft tissues

By reactivating a dormant gene, Lin28a, researchers could regrow hair, cartilage, bone and soft tissues | Amazing Science | Scoop.it
Young animals are known to repair their tissues effortlessly, but can this capacity be recaptured in adults? A new study from researchers at the Stem Cell Program at Boston Children's Hospital suggests that it can.

 

By reactivating a dormant gene called Lin28a, which is active in embryonic stem cells, researchers were able to regrow hair and repair cartilage, bone, skin and other soft tissues in a mouse model.

The study also found that Lin28a promotes tissue repair in part by enhancing metabolism in mitochondria—the energy-producing engines in cells—suggesting that a mundane cellular "housekeeping" function could open new avenues for developing regenerative treatments. Findings were published online by the journal Cell on November 7, 2013.

 

Lin28, first discovered in worms, functions in all complex organisms. It is abundant in embryonic stem cells, expressed strongly during early embryo formation and has been used to reprogram skin cells into stem cells. It acts by binding to RNA and regulating how genes are translated into proteins.


To better understand how Lin28a promotes tissue repair, the researchers systematically looked at what specific RNAs it binds to. They initially had their sights on a tiny RNA called Let-7, which is known to promote cell maturation and aging.

 

"We were confident that Let-7 would be the mechanism," says Shyh-Chang. "But there was something else involved." Specifically, the researchers found that Lin28a also enhances the production of metabolic enzymes in mitochondria, the structures that produce energy for the cell. By revving up a cell's bioenergetics, they found, Lin28a helps generate the energy needed to stimulate and grow new tissues.


"We already know that accumulated defects in mitochondrial metabolism can lead to aging in many cells and tissues," says Shyh-Chang. "We are showing the converse—that enhancement of mitochondrial metabolism can boost tissue repair and regeneration, recapturing the remarkable repair capacity of juvenile animals."

 

Further experiments showed that bypassing Lin28a and directly activating mitochondrial metabolism with a small-molecule compound also had the effect of enhancing wound healing. This suggests the possibility of inducing regeneration and promoting tissue repair with drugs.

 

"Since Lin28 itself is difficult to introduce into cells, the fact that we were able to activate mitochondrial metabolism pharmacologically gives us hope," Shyh-Chang says.

 

Lin28A didn't universally induce regeneration in all tissues. Heart tissue showed little effect, and while the researchers were able to enhance the regrowth of finger tips in newborn mice, they could not in adults.

 

"Lin28a could be a key factor in constituting a healing cocktail," says Shyh-Chang, "but there are other embryonic factors that remain to be found."

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How nanotechnology can advance regenerative medicine

How nanotechnology can advance regenerative medicine | Amazing Science | Scoop.it

Nanotechnology may provide new strategies for regenerative medicine, including better tools to improve or restore damaged tissues, according to a review paper that summarizes the current state of knowledge on nanotechnology with application to stem cell biology.

 

Researchers have found that the adhesion, growth, and differentiation of stem cells are likely controlled by their surrounding microenvironment, which contains both chemical and physical cues. These cues include the “nanotopography” of the complex extracellular matrix or architecture that forms a network for human tissues.

 

In their review paper published in the journal Science and Technology of Advanced Materials (open access), Yang-Kao Wang and colleagues describe studies showing how this nanotopography (which includes nanosized pores, grooves, ridges, etc.) plays important roles in the behavior and fate of stem cells.

 

The authors also discuss the application of nanoparticles to stem cell isolation, tracking and imaging; how to translate nanotechnology from two to three dimensions; and the potential limitations of using nanomaterials in stem cell biology.

 

The paper concludes that “understanding [the] interactions of nanomaterials with stem cells may provide knowledge applicable to [the development of improved] cell-scaffold combinations in tissue engineering and regenerative medicine.”

 


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Microenvironmental Regulation of Tumor Progression and Metastasis

Microenvironmental Regulation of Tumor Progression and Metastasis | Amazing Science | Scoop.it

"Cancers develop in complex tissue environments, which they depend on for sustained growth, invasion and metastasis. Unlike tumor cells, stromal cell types within the tumor microenvironment (TME) are genetically stable and thus represent an attractive therapeutic target with reduced risk of resistance and tumor recurrence. However, specifically disrupting the pro-tumorigenic TME is a challenging undertaking, as the TME has diverse capacities to induce both beneficial and adverse consequences for tumorigenesis. Furthermore, many studies have shown that the microenvironment is capable of normalizing tumor cells, suggesting that re-education of stromal cells, rather than targeted ablation per se, may be an effective strategy for treating cancer. Here we discuss the paradoxical roles of the TME during specific stages of cancer progression and metastasis, as well as recent therapeutic attempts to re-educate stromal cells within the TME to have anti-tumorigenic effects."

 

Bidirectional communication between cells and their microenvironment is critical for both normal tissue homeostasis and tumor growth. In particular, interactions between tumor cells and the associated stroma represent a powerful relationship that influences disease initiation and progression and patient prognosis. The link between chronic inflammation and tumorigenesis was first proposed by Rudolf Virchow in 1863 after the observation that infiltrating leukocytes are a hallmark of tumors. Since then, a plethora of studies have contributed to the characterization of the TME, further complicating the already challenging task of understanding and treating cancer. Whereas cancer was previously viewed as a heterogeneous disease involving aberrant mutations in tumor cells, it is now evident that tumors are also diverse by nature of their microenvironmental composition and their stromal cell proportions or activation states. In response to evolving environmental conditions and oncogenic signals from growing tumors, the TME continually changes over the course of cancer progression, underscoring the need to consider the influences of the TME on metastasis as a dynamic process and understand how tumor cells drive the construction of their own niche.


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Cancer Commons's curator insight, November 8, 2013 1:50 PM

Quail DF, Joyce JA. Nature Medicine. Nov 7, 2013.

Cancer Commons's curator insight, November 8, 2013 1:51 PM

Quail DF, Joyce JA. Nature Medicine. Nov 7, 2013.

Cancer Commons's curator insight, November 8, 2013 1:51 PM

Quail DF, Joyce JA. Nature Medicine. Nov 7, 2013.

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Glioblastoma multiforme: Incurable brain cancer gene BCL2L12 is silenced

Glioblastoma multiforme: Incurable brain cancer gene BCL2L12 is silenced | Amazing Science | Scoop.it

Scientists at Northwestern University say they were able to demonstrate the successful delivery of a drug that turns off a critical gene in glioblastoma multiforme (GBM), increasing survival rates significantly in animals with the deadly disease. This form of brain cancer, which ended Sen. Edward Kennedy’s life, kills approximately 13,000 Americans a year.

 

According to the investigators, the novel therapeutic, which is based on nanotechnology, is small and nimble enough to cross the blood-brain barrier and get to where it is needed—the brain tumor.

 

Designed to target a specific cancer-causing gene in cells, the drug flips the switch of the oncogene to “off,” silencing the gene, they added. This knocks out the proteins that keep cancer cells immortal.

 

In a study of mice (“Spherical Nucleic Acid Nanoparticle Conjugates as an RNAi-Based Therapy for Glioblastoma”), the nontoxic drug was delivered by intravenous injection. In animals with GBM, the survival rate increased nearly 20%, and tumor size was reduced three to four fold, as compared to the control group. The results were published October 30 in Science Translational Medicine.

 

“We preclinically evaluate an RNA interference (RNAi)–based nanomedicine platform, based on spherical nucleic acid (SNA) nanoparticle conjugates, to neutralize oncogene expression in GBM,” wrote the scientists. “In vivo, the SNAs penetrated the blood-brain barrier and blood-tumor barrier to disseminate throughout xenogeneic glioma explants. SNAs targeting the oncoprotein Bcl2Like12 (Bcl2L12)—an effector caspase and p53 inhibitor overexpressed in GBM relative to normal brain and low-grade astrocytomas—were effective in knocking down endogenous Bcl2L12 mRNA and protein levels, and sensitized glioma cells toward therapy-induced apoptosis by enhancing effector caspase and p53 activity.”

 

“This is a beautiful marriage of a new technology with the genes of a terrible disease,” said Chad A. Mirkin, Ph.D., a nanomedicine expert and a senior co-author of the study.

 

“This proof-of-concept further establishes a broad platform for treating a wide range of diseases, from lung and colon cancers to rheumatoid arthritis and psoriasis.”

 

The power of gene regulation technology is that a disease with a genetic basis can be attacked and treated if scientists have the right tools, pointed out Dr. Mirkin. Thanks to the Human Genome Project and genomics research over the last two decades, there is an enormous number of genetic targets; having the right therapeutic agents and delivery materials has been the challenge, he explained.

 

“The RNA interfering-based SNAs are a completely novel approach in thinking about cancer therapy,” said Alexander H. Stegh, Ph.D., a co-author on the study. “One of the problems is that we have large lists of genes that are somehow disregulated in glioblastoma, but we have absolutely no way of targeting all of them using standard pharmacological approaches.

 

That's where we think nanomaterials can play a fundamental role in allowing us to implement the concept of personalized medicine in cancer therapy.”

 

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Gut Bacteria have been implicated in Allergies and Obesity, now also in Rheumatoid Arthritis

Gut Bacteria have been implicated in Allergies and Obesity, now also in Rheumatoid Arthritis | Amazing Science | Scoop.it

“It’s been suspected for years and years, both in humans and in the animal model, that the development of autoimmune diseases like arthritis is dependent on the gut microbiota,” says immunologist Diane Mathis of Harvard Medical School in Boston. Now, she says, those suspicions are beginning to be confirmed in humans. “It’s a very striking finding.”

 

Rheumatoid arthritis is a mysterious disease. It can strike at any age, typically beginning in young and middle-aged adults and causing painfully stiff, swollen joints in the hands and feet. It can also destroy bone and cartilage and damage organs like the lungs and kidneys. Scientists aren’t sure what causes rheumatoid arthritis, but they do know that it’s an autoimmune disorder, meaning that the body’s immune system is attacking its own tissues. And that’s where gut bacteria come in.

 

Gut bacteria have an intricate relationship with our immune system. We need to be able to tolerate helpful microbes while still recognizing and fighting invaders. Immunologist Dan Littman of New York University knew that gut microbes are important to the development of a particular type of immune cell his team studies, known as a Th17 cell. Mice that are reared in sterile conditions produce very few of these cells, and his group had previously found that mice bought from one supplier had far more Th17 cells than those that came from a different supplier. The difference turned out to be due to the rodents’ gut microbes.

 

When Littman presented that result at a conference several years ago, Mathis, who was in the audience, told him that she had seen a change in her lab animals when they were moved to a lab in a different town. Instead of spontaneously developing a mouse version of arthritis, they remained healthy. Littman and Mathis collaborated to find out why and tracked down the difference to a particular type of bacterium that, when present in the intestines, trains the immune system to produce Th17 cells, which in turn release molecules that cause inflammation and bone damage in arthritis.

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Dual-action drug shows promise against diabetes and also promotes weight loss in rodents

Dual-action drug shows promise against diabetes and also promotes weight loss in rodents | Amazing Science | Scoop.it

An experimental diabetes treatment that packs the action of two natural hormones into a single injectable agent has been shown to successfully lower blood sugar in humans, monkeys and rodents. Marking a new approach in the treatment of the disease, the currently unnamed molecule also seems likely to cause fewer gastrointestinal side effects in humans than did other diabetes medicines.

 

“We aimed for achieving the best glycaemic control with as little effect on the gut as possible,” says Richard DiMarchi, a biomolecular scientist at Indiana University in Bloomington, and a member of the international team that publishes the results today in Science Translational Medicine.


The molecule, which targets receptors for the two hormones, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), was developed in DiMarchi’s lab. Swiss pharmaceutical company Roche, based in Basel, supported the research and has licensed the agent.


As part of the study, 44 patients with type 2 diabetes received once-weekly injection of the dual-action molecule at various doses for six weeks while nine others received placebo injections. Blood tests showed a dose-dependent response; at the highest doses, a standard marker of blood glucose levels dropped an average of 1.1 percentage points from the baseline (which ranged from 7.4% to 7.9%; normal levels are below 5.7% in non-diabetic patients). In the placebo group, the marker dropped by just 0.16 points.

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United States to approve cocktail of new oral antiviral drugs to fight hepatitis C

United States to approve cocktail of new oral antiviral drugs to fight hepatitis C | Amazing Science | Scoop.it

Improved treatments offer hope for eradication of viral liver infection. For decades, people with hepatitis C virus (HCV) have had to endure gruelling treatment regimens that include injections of the drug interferon, which can cause severe nausea and depression. But with the imminent approval of several highly effective oral antiviral drugs, and more on the way, researchers say that eradicating the infection worldwide is now a realistic goal.


Unlike previous HCV treatments, which sought to enhance the immune system with interferon and other drugs, the latest group of oral medications interferes with the virus’s ability to replicate and make proteins. A US Food and Drug Administration (FDA) board recommended two such drugs — simeprevir, made by Johnson & Johnson in New Brunswick, New Jersey, and sofosbuvir from Gilead Sciences in Foster City, California — for approval last week. When each is taken in combination with a drug called ribavirin, the treatment eliminates hepatitis C in around 80% of people.


“This is the first time in the history of humankind that we have a cure for a viral disease,” says pharmacologist Raymond Schinazi of Emory University in Atlanta, Georgia.

 

Findings from trials of different drug combinations are set to be released this week. A phase II study called COSMOS tested a combination of sofosbuvir and simeprevir in 197 people with HCV who had either not responded to interferon or who had advanced liver fibrosis caused by the virus. After 12 weeks of treatment, the drugs completely cleared the virus in more than 90% of participants.

 

Another study, led by physician Kazuaki Chayama at Hiroshima University in Japan, treated 220 people with a combination of daclatasvir and asunaprevir, two new drugs from Bristol-Myers Squibb in New York. The cocktail cured 85% of participants. Eric Hughes, lead global medical researcher at the company, says that it plans to submit the drugs for FDA approval in 2014.


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