Showing posts with label Cell Biology. Show all posts
Showing posts with label Cell Biology. Show all posts

Daily Science Journal (Feb. 2, 2008) — While biomedical, electronics, and other branches of research are marching steadily into the realm of the smaller-than-small nanometer scale, building needed materials at this scale has been problematic.

Recently, however, a team from The Scripps Research Institute unveiled a novel approach to the problem that yields a material with novel properties, which some might find reminiscent of Flubber. The material is produced using naturally occurring proteins as templates for uniform, self-assembled, nano-scale construction.

The material, an organic polymer described in today's early, online edition of the Proceedings of the National Academy of Sciences (PNAS), could one day find application in everything from screening for disease to microelectronics.


Using Nature's Tricks

Nature is replete with examples of molecules such as DNA that self-assemble with uniform patterns on the nanoscale, but until now researchers have had limited success duplicating such processes. The new study, however, provides one synthetic method that has effectively mimicked the templating strategy used in nature for nanoscale construction in the lab.

To create the new material, the Scripps Research team, led by Scripps Research President Richard Lerner and Assistant Professor Tobin Dickerson, began with a natural nanoscale product, a bacterial virus or phage. A nanometer is one billionth of a meter, or the width of a few average atoms.

Specifically, the product the team worked with was a phage known as M13. If scaled up, the phage is proportionally equivalent to a 4-foot-long pencil, with the tip and eraser roughly representing the active parts of the phage that infect bacteria. Other proteins that are biologically inert and analogous to the wood body of the pencil provide the filamentous phage's structure.

Having worked with phage extensively in other applications, the team decided to explore the possibility of using those structural proteins as a potential template for nanoscale construction. To do so, the team chemically modified molecular protrusions on the proteins so they would attract and bind with the components needed to form strands of polyacrylamide, a common polymer used to make laboratory gels.

The resulting polymer-phage combination, which twists into helices like DNA and RNA molecules, takes on the shape of a comb with the polymers as the teeth. These teeth in turn interlock to form a strangely resilient, rubbery solid.

Shocking Flexibility

Once the new material, known as a protein-polymer bioconjugate, was created, the group was shocked to find that it was almost impossible to break a sample apart. It could be sliced, but no matter how hard researchers compressed or squeezed it, it always bounced back to its original state, because the stable phage proteins act like rebar in concrete to provide strength.

Further analyses uncovered additional important characteristics. The combs do not grow completely uniformly—some combs grow more teeth than others, for instance, before interlocking with a nearby comb. But the combs can only be a prescribed distance apart for the chemical interlocking to occur, which leads to uniform size for the pores between the comb teeth. The pores proved to be about 4 nanometers wide and greater than 1 micrometer (one millionth of a meter) in length.

This uniformity is in stark contrast with experiments which showed that simply mixing the phages with polymers without the templating procedure produces a chaotic hodge podge at the molecular level.

Interestingly, after creating the material, the group discovered that a British scientist had done theoretical calculations about how flexible rods—like the phages—would pack together using the least amount of energy.

"It was extremely gratifying to see that the mathematics had exactly predicted what we were observing," says Dickerson

Putting It to Use

Although the work was intended mainly as a proof of concept for using the phages as templates, the researchers have a number of potential applications in mind. The phage can be produced easily and cheaply in very large quantities, and the polymer components are readily available and simple to combine to create the final material. These characteristics would make commercial application possible.

"In essence, bacteriophage-derived materials are a renewable resource," says Dickerson.

The defined channels established by the pores in the material might be adapted as pathways for electrons for use in microelectronics. Or, the pores might be used as a filter for certain molecules, for example to test blood samples for proteins whose presence is tied to particular diseases. More complex potential uses might include altering the biologically active portions of the phage to attract specific molecules, forcing them into the polymers' pores, or to block others.

"These tools can be visualized like Tinkertoys ® or Legos ®," says Dickerson of the possibilities. "You can think about this really in engineering terms using macroscopic analogs such as baskets, or lids, or holes."

To add to the list of potential applications, the team has already begun exploring additional materials that might be created using the basic phage construction scheme.

In addition to Dickerson and Lerner, authors on the paper, entitled "Biologically templated organic polymers with nanoscale order," were Bert Willis, Lisa Eubanks, Malcolm Wood, and Kim Janda, all of Scripps Research. See http://www.pnas.org/cgi/content/abstract/0711308105v1.

The work was supported by the Skaggs Institute for Chemical Biology, Worm Institute for Research and Medicine, and a National Institutes of Health Kirschstein National Research Service Award.

Adapted from materials provided by Scripps Research Institute.



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Daily Science Journal (Dec. 13, 2007) — Researchers at Rensselaer Polytechnic Institute have developed a new way to seek out specific proteins, including dangerous proteins such as anthrax toxin, and render them harmless using nothing but light. The technique lends itself to the creation of new antibacterial and antimicrobial films to help curb the spread of germs, and also holds promise for new methods of seeking out and killing tumors in the human body.

Transmission electron microscope images show carbon nanotubes conjugated with anthrax toxin before (a, b) and after (c, d) exposure to ultraviolet light. This light caused the adsorbed toxin to deactivate and fall off the nanotube, which is why the structures in pictures c and d are smaller in diameter than those in pictures a and b. (Credit: Rensselaer/Ravi Kane)

Scientists have long been interested in wrapping proteins around carbon nanotubes, and the process is used for various applications in imaging, biosensing, and cellular delivery. But this new study at Rensselaer is the first to remotely control the activity of these conjugated nanotubes.


A team of Rensselaer researchers led by Ravi S. Kane, professor of chemical and biological engineering, has worked for nearly a year to develop a means to remotely deactivate protein-wrapped carbon nanotubes by exposing them to invisible and near-infrared light. The group demonstrated this method by successfully deactivating anthrax toxin and other proteins.

"By attaching peptides to carbon nanotubes, we gave them the ability to selectively recognize a protein of interest -- in this case anthrax toxin -- from a mixture of different proteins," Kane said. "Then, by exposing the mixture to light, we could selectively deactivate this protein without disturbing the other proteins in the mixture."

By conjugating carbon nanotubes with different peptides, this process can be easily tailored to work on other harmful proteins, Kane said. Also, employing different wavelengths of light that can pass harmlessly through the human body, the remote control process will also be able to target and deactivate specific proteins or toxins in the human body. Shining light on the conjugated carbon nanotubes creates free radicals, called reactive oxygen species. It was the presence of radicals, Kane said, that deactivated the proteins.

Kane's new method for selective nanotube-assisted protein deactivation could be used in defense, homeland security, and laboratory settings to destroy harmful toxins and pathogens. The method could also offer a new method for the targeted destruction of tumor cells. By conjugating carbon nanotubes with peptides engineered to seek out specific cancer cells, and then releasing those nanotubes into a patient, doctors may be able to use this remote protein deactivation technology as a powerful tool to prevent the spread of cancer.

Kane's team also developed a thin, clear film made of carbon nanotubes that employs this technology. This self-cleaning film may be fashioned into a coating that -- at the flip of a light switch -- could help prevent the spread of harmful bacteria, toxins, and microbes.

"The ability of these coatings to generate reactive oxygen species upon exposure to light might allow these coatings to kill any bacteria that have attached to them," Kane said. "You could use these transparent coatings on countertops, doorknobs, in hospitals or airplanes -- essentially any surface, inside or outside, that might be exposed to harmful contaminants."

Kane said he and his team will continue to hone this new technology and further explore its potential applications.

Details of the project are outlined in the article "Nanotube-Assisted Protein Deactivation" in the December issue of Nature Nanotechnology.

Co-authors of the paper include Department of Chemical and Biological Engineering graduate students Amit Joshi and Shyam Sundhar Bale; postdoctoral researcher Supriya Punyani; Rensselaer Nanotechnology Center Laboratory Manager Hoichang Yang; and professor Theodorian Borca-Tasciuc of the Department of Mechanical, Aerospace, and Nuclear Engineering.

The group has filed a patent disclosure for their new selective nanotube-assisted protein deactivation technology. The research project was funded by the U.S. National Institutes of Health and the National Science Foundation.

Adapted from materials provided by Rensselaer Polytechnic Institute, via EurekAlert!, a service of AAAS.




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Daily Science Journal (Aug. 7, 2007) — Green tea could hold promise as a new treatment for skin disorders such as psoriasis and dandruff, Medical College of Georgia researchers say.

Dr Hsu says, "There are no cures for autoimmune diseases. But it is possible that this is a non-toxic way to regulate them. We need further study -- on humans -- to determine the full effects." (Credit: Medical College of Georgia)

Researchers studied an animal model for inflammatory skin diseases, which are often characterized by patches of dry, red, flaky skin caused by the inflammation and overproduction of skin cells. Those treated with green tea showed slower growth of skin cells and the presence of a gene that regulates the cells' life cycles.

"Psoriasis, an autoimmune disease, causes the skin to become thicker because the growth of skin cells is out of control," says Dr. Stephen Hsu, an oral biologist in the MCG School of Dentistry and lead investigator on the study published in the Aug. 18 edition of Experimental Dermatology. "In psoriasis, immune cells, which usually protect against infection, instead trigger the release of cytokines, which causes inflammation and the overproduction of skin cells."


Other autoimmune diseases with similar side effects include lupus, which can lead to skin lesions, and dandruff.

Green tea, already shown to suppress inflammation, helps by regulating the expression of Caspase-14, a protein in genes that regulates the life cycle of a skin cell.

"That marker guides cells by telling them when to differentiate, die off and form a skin barrier," Dr. Hsu says. "In people with psoriasis, that process is interrupted and the skin cells don't die before more are created and the resulting lesions form."

Animal models treated with green tea also showed reduced levels of proliferating cell nuclear antigen, a gene expressed when skin cells multiply. In psoriasis, the gene is over-expressed and speeds production of skin cells.

"Before treatment, the antigen, PCNA, was present in all layers of the skin," Dr. Hsu says. "Typically, PCNA is only found in the basal layer, the innermost layer where skin cells continually divide and new cells push the older ones to the skin surface, where they eventually slough off. After being treated with green tea, the animal models showed near-normal levels of PCNA in only the basal layers."

This research is important because some treatments for psoriasis and dandruff can have dangerous side effects, he says.

"The traditional treatment of ultraviolet light and medication, while it can control the lesions and be used long term, may cause squamous cell carcinoma – the second most common form of skin cancer," Dr. Hsu says. "Some of the most effective anti-dandruff shampoos also have carcinogens in them. While the U.S. Food and Drug Administration allows that in small amounts, the bottom line is that we don't know the long-term effects of using those products continuously."

Green tea, which is plant-derived, may be an alternative, he says. But scientists must work to overcome some barriers with the treatment.

The chemicals in green tea are so active that they are oxidized too quickly when mixed with other ingredients. They also dissolve in water, which cannot penetrate the skin's barrier.

Researchers are looking for a balanced formula that can dissolve in fats, which can permeate the skin, Dr. Hsu says.

Adapted from materials provided by Medical College of Georgia.

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Tea Extracts Help Treat Damaged Skin In Cancer Patients

Tea extracts work as an effective treatment for patients who suffer from damaged skin following radiation treatment for cancer. Researchers show that this might partly be due to the anti-inflammatory properties of tea.

In a study published in the open access journal BMC Medicine, researchers show that tea acts at the cellular level, by inhibiting inflammatory pathways, to reduce inflammation. They also show that tea extracts reduce the duration of radiation-induced skin damage by up to 10 days in patients who received radiation treatment.

Frank Pajonk, from the University of California in Los Angeles, USA, and colleagues from the University of Freiburg, Germany, studied the effects of green tea and black tea extracts on patients who had been treated with radiotherapy, which can damage the skin. The authors then analysed the effects of the same tea extracts on human and mouse white blood cells in culture.

Pajonk et al. find that tea extracts reduce the duration of skin toxicity following radiotherapy by 5 to 10 days. Green tea extracts are more effective than black tea extracts in some patients. Pajonk et al. also show that tea extracts reduce the release of pro-inflammatory cytokines, such as IL-1beta, IL-6, IL-8, TNFalpha and PGE2, in human white blood cells in culture, with green tea having higher anti-inflammatory properties than black tea. Both black tea and green tea inhibit one major inflammatory pathway in mouse white blood cells.

Pajonk et al. add that tea's high content of polyphenols is likely to be responsible for its high anti-inflammatory activity, but that other pathways are probably involved in its clinical effectiveness.

Adapted from materials provided by BioMed Central, via EurekAlert!, a service of AAAS.




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Daily Science Journal (Jun. 15, 2007) — They are the largest group of white blood cells: neutrophil granulocytes kill microorganisms. Neutrophils catch microbes with extracellular structures nicknamed Neutrophil Extracellular Traps (NETs) that are composed of nucleic acid and aggressive enzymes.

Neutrophil granulocytes have trapped Shigella bacteria in NETs. (Image: Dr. Volker Brinkmann, Max Planck Institute for Infection Biology)

A group of scientists lead by Arturo Zychlinsky at the Max-Planck-Institute for Infectious Biology in Berlin, Germany discovered, how the neutrophils form this snaring network (Journal of Cell Biology, online, January 8, 2007). Once triggered, the cells undergo a novel program leading to their death. While they perish, the cells release the content of their nuclei. The nucleic acid, mingled with bactericidal enzymes, forms a lethal network outside the cell. Invading bacteria and pathogenic fungi get caught and killed in the NETs.


Every minute, several million neutrophils leave the bone marrow and are ready to defend the body of invading germs. They are the immune system’s first line of defence against harmful bacteria and migrate into the tissue at the site of infection to combat pathogens. For more than hundred years it was known that neutrophil granulocytes kill bacteria very efficiently by devouring them. After eating the germs neutrophils kill tehm with antimicrobial proteins.

The group of scientists lead by Arturo Zychlinsky at the Max-Planck-Institute for Infectious Biology discovered a second killing mechanism: neutrophil granulocytes can form web-like structures outside the cells composed of nucleic acid and enzymes which catch bacteria and kill them. The scientists were able to generate impressive micrographs of these nets. But it remained a mystery how the granulocytes could mobilise the contents of their nuclei and catapult it out of the cells.Only after lengthy live cell imaging and biochemical studies it became clear how neutrophils make NETs. The cells get activated by bacteria and modify the structure of their nuclei and granules, small enzyme deposits in the cytoplasm.

"The nuclear membrane disintegrates, the granules dissolve, and thus the NET components can mingle inside the cells", explains Volker Brinkmann, head of the microscopy group. At the end of this process, the cell contracts until the cell membrane bursts open and quickly releases the highly active melange. Once outside the cell, it unfolds and forms the NETs which then can trap bacteria.

Surprisingly, this process is as effective as devouring bacteria: "NETs formed by dying granulocytes kill as many bacteria as are eaten up by living blood cells", says Arturo Zychlinsky. Thus, neutrophils fulfil their role in the defence battle even after their deaths.

Adapted from materials provided by Max Planck Society.



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