Showing posts with label enzyme. Show all posts
Showing posts with label enzyme. Show all posts

Tuesday, October 2, 2007

Bacteria Breakthrough For Microdot Printed Circuits

You can see above a not-to-scale graphic showing "how catalyst (blue hollow-ended beads) dangles from patterned stamp, while dye particles (gold balls) are bonded to DNA chains to make DNA coating visible. After stamp (blue) presses into DNA coating (yellow) at center the catalyst detaches dye and DNA chain's tip (bottom right). That disruption creates patterning in DNA coating (top right)." Credits: graphic by Alexander Shestopalov, caption by Duke University - Here is a link to a larger version of this graphic.

Bacteria Breakthrough For Microdot Printed Circuits

An E. Coli infection to the human body is not a good thing. The infection may create symptoms that include severe abdominal cramping, bloody diarrhea, and sometimes nausea with vomiting.

Bacteria, however, has enzyme properties that allow one to improve the preciseness of the surface pattern of a printed circuit one-hundred fold of traditional inking methods.

This discovery will have a tremendous effect on the depth of computer processes that can be placed on a single chip, in that, this inkless technique could be used to build complex nanoscale devices with unprecedented precision to create microdevices such as labs-on-a-chip.

This excerpted from Wikipedia –

1) After the discovery of microtechnology (~1958) for realizing integrated semiconductor structures for microelectronic chips, these lithography-based technologies were soon applied in pressure sensor manufacturing (1966) as well.

Due to further development of these usually CMOS-compatibility limited processes, a tool box became available to create micrometre or sub-micrometre sized mechanical structures in silicon wafers as well: the Micro Electro Mechanical Systems (MEMS) era (also indicated with Micro System Technology - MST) had started.

2) Lab-on-a-chip (LOC) is a term for devices that integrate (multiple) laboratory functions on a single chip of only millimeters to a few square centimeters in size and that are capable of handling extremely small fluid volumes down to less than pico liters. Lab-on-a-chip devices are a subset of MEMS devices and often indicated by "Micro Total Analysis Systems" (µTAS) as well.

Microfluidics is a broader term that describes also mechanical flow control devices like pumps and valves or sensors like flowmeters and viscometers. However, strictly regarded "Lab-on-a-Chip" indicates generally the scaling of single or multiple lab processes down to chip-format, whereas "µTAS" is dedicated to the integration of the total sequence of lab processes to perform chemical analysis.

Reference Here>>

Over time, this can have an effect on the general miniaturization of electronic devices. Hand held devices just may become finger held devices.

This excerpted EurekAlert! from Duke University –

Using catalysts to stamp nanopatterns without ink
Contact: Monte Basgall, Duke University - Public release date: 26-Sep-2007

Using enzymes from E. coli bacteria, Duke University chemists and engineers have introduced a hundred-fold improvement in the precision of features imprinted to create microdevices such as labs-on-a-chip.

Their inkless microcontact printing technique can imprint details measuring close to 1 nanometer, or billionths of a meter, the Duke team reported in the Sept. 24, 2007 issue of the Journal of Organic Chemistry.

"This has a lot of potential, because we don't have the resolution issue," said Robert Clark, a professor of mechanical engineering and materials science and dean at Duke’s Pratt School of Engineering. “The really important part is that with a biological catalyst there’s no ink involved,” added Duke chemistry professor Eric Toone.

Clark, Toone and three graduate students authored the report on their study, which was funded by the National Science Foundation (NSF).

In traditional microcontact printing -- also called soft lithography or microstamping -- an elastic stamp’s end is cast from a mold created via photolithograpy – a technique used to generate microscopic patterns with light. Those patterns are then transferred to a surface by employing various biomolecules as inks, rather like a rubber stamp.
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A shortcoming of traditional microcontact printing is that pattern transfer relies on the diffusion of ink from the stamp to the surface. This same diffusion spreads out beyond the limits of the pattern as the stamp touches the surface, degrading resolution and blurring the feature edges, Clark and Toone said.

Because of this mini-blurring, the practical limit to defect-free patterning is “in excess of 100 nanometers,” said the report, whose first author, Phillip Snyder, is a former Toone graduate student now working as a postdoctoral researcher in Whitesides’ group.

A 100 nanometer limit of resolution is about 1,000 times tinier than a human hair’s width. While that seems very precise, the Duke team now reports it can boost accuracy limits to less than 2 nanometers by entirely eliminating inking.

Clark and graduate student Matthew Johannes crafted a microstamp out of a gel-like material called polyacrylamide, which compresses more uniformly than the silicone material known as PDMS which is normally used in microstamping.

In lieu of ink, Snyder, Toone and graduate student Briana Vogen suspended a biological catalyst on the stamp with a molecular “tether” of amino acids. For this proof-of-principle demonstration, Toone’s team chose as a catalyst the biological enzyme exonuclease I, derived from the bacterium E. coli.

In one set of experiments, the polyacrylamide stamp pattern bearing the tethered enzymes was then pressed on a surface of gold that had been covered with a uniform coating of single-stranded DNA molecules. The DNA molecules had also been linked to fluorescent dye molecules to make the coating visible under a microscope.

Wherever the enzyme met the DNA, the end of the DNA chain and its attached dye were broken off and removed. That created a dye-less pattern of dots on the DNA coating, each dot measuring about 10 millionths of a meter diameter each.

The microdots are very precise because the catalyst that created them could not shift its position more than the length of its chemical tether -- less than 1 nanometer, the Duke team reported. "Whether the stamp was left on for a short period of time, or for days, the pattern did not change,” Clark said.

The inkless microstamp could also re-use the same suspended catalyst molecule repeatedly. “Enzymes can deteriorate with extended use,” Clark acknowledged. “But because of our tether attachment chemistry, we can easily wash the old enzyme off, put on a new one and keep going,” Clark said.

In follow-up research, Clark and Toone are now evaluating more durable microstamping materials attached to longer lasting catalysts that are non-enzymatic.
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“Soft lithography has really revolutionized the field of surface science over the last 30 years,” said Toone. “And I honestly believe that using catalysts instead of diffusive processes is going to become the way that soft lithography is done in the future.”
Reference Here>>
(ht: SlashDot-One Decade Old)

This discovery illustrates that it's a very small Oblate Spheroid, after all!

Thursday, August 30, 2007

Remember This! … Fibroblast-Neprilysin

UNTREATED Alzheimer's Brain Cells


TREATED Alzheimer's Brain Cells

Plaques comprised of amyloid-beta are the hallmark pathology of Alzheimer's disease. In this study, the scientists used an amyloid-degrading enzyme to clear these amyloid cobwebs from the brain - as illustrated in these untreated (top) versus treated (bottom) brain images. Image Credit: Harvard Photographic Services

Remember This! … Fibroblast-Neprilysin

In a report that first appeared August 27 on the Web site of the Public Library of Science, experiments centered on new methods to relieve the damaging effect of Alzheimer’s disease are showing great promise.

Simply stated, it is believed that when some humans grow older, the ability to control the build up of protein based plaques in the brain becomes reduced. These proteins create toxic clumps and tangled fibers that ultimately kill cells and interfere with the brains’ ability to recall memories and think (a situation similar to the build up of cholesterol in arteries that reduce the flow of blood).

An enzyme named Neprilysin has shown great promise in breaking down the toxic clumps of fibrous protein in the brains of mice. What has been found through these tests, and may be unique, is the method of the delivery of this enzyme to the effected portions of the brain under the toxic grip of Alzheimer’s protein-fiber goo.

Skin from the patient may actually provide the best method to first create the Neprilysin enzyme and further, through a process termed Fibroblast create a combo that can be implanted back into the patient for the desired result – a freer thinking brain.

This Fibroblast-Neprilsin combo type of tratment process has shown and may prove to have a positive benefit to the treatment of cancers, blood, muscle, and eye diseases, spinal cord injuries, stroke, Parkinson’s and Huntington diseases, and amyotrophic lateral sclerosis (Lou Gehrig’s disease).

This from Harvard University Gazette Online -

Brain implants relieve Alzheimer’s damage
Toxic plaques cleared away

William J. Cromie - Harvard News Office - August 28, 2007

Genetically engineered cells implanted in mice have cleared away toxic plaques associated with Alzheimer’s disease.

The animals were sickened with a human gene that caused them to develop, at an accelerated rate, the disease that robs millions of elderly people of their memories. After receiving the doctored cells, the brain-muddling plaques melted away. If this works in humans, old age could be a much happier time of life.

Alzheimer’s involves a protein called amyloid-beta.
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“Delivery of genes that led to production of an enzyme that breaks up amyloid showed robust clearance of plaques in the brains of the mice,” notes Dennis Selkoe, Vincent and Stella Coates Professor of Neurologic Diseases at Harvard Medical School. “These results support and encourage further investigation of gene therapy for treatment of this common and devastating disease in humans.”
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The first published report of the experiments, done by Selkoe and other researchers from Harvard-affiliated Brigham and Women’s and McLean hospitals, appeared Aug. 27 on the Web site of the Public Library of Science.

The gene delivery technique employed by the research team has been used in several other trials with animals that model human diseases, including cancers. The procedure involves removing cells from patients, making genetic changes, and then putting back the modified cells, which should treat a disease or disability.
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“Several of these potential treatments have advanced to human trials, with encouraging outcomes for patients,” says Matthew Hemming, lead author of the report and a graduate student in Selkoe’s lab.
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The Harvard team used skin cells from the animal’s own body to introduce a gene for an amyloid-busting enzyme known as neprilysin. The skin cells, also known as fibroblasts, “do not form tumors or move from the implantation site,” Hemming notes. “They cause no detectable adverse side effects and can easily be taken from a patient’s skin.” In addition, other genes can be added to the fibroblast-neprilysin combo, which will eliminate the implants if something starts to go wrong.
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The experiments proved that the technique works, but will it work in humans?

One major obstacle, Selkoe says, is the larger size of a human brain compared to that of a mouse. That difference will require an increase of amyloid-busting activity throughout a much larger space.

One solution might involve implanting the genes and fibroblasts where they have the best access to amyloid-beta, in the spinal fluid for example, instead of trying to inject them into a small target. The amyloid-killing combo might be put into capsules that would secrete neprilysin into the blood circulating in the brain, eliminating the need to hit an exact spot.

This or some other clever maneuver that does not require surgery might eliminate the gooey plaques, but will that improve a person’s memory? And will the change be long-lasting? “Further work is needed to determine if reducing the plaque burden has cognitive benefits over a long period,” notes Hemming, “but there’s a wealth of evidence arguing that it will.”
Reference Here>>

(re-posted at MAXINE)