Smart Economy

Longevity and Healthy Aging

Aging Gracefully without Chronic Human Diseases

Time Magazine and National Geographic both recently ran cover stories with a baby on the cover. The tagline reads: This baby could live to be 142 years old. So the question that immediately comes to mind  is: Is this possible  and can everyone age gracefully without chronic human diseases? And can we do it with today’s technology ?

To explore this question we first need to compare the mortality in 1900 versus today. In 1900, half of the deaths were caused by pneumonia, influenza, tuberculosis, gastrointestinal infections.  These diseases were cured largely with antibiotics and antivirals after world war two. Cancer, diabetes, coronary  heart disease, and Alzheimer’s were present but  not near the rates we see today.  Now, the top causes of death today, according to US statistics are heart disease, lung cancer, lung disease (asthma) , stroke, Alzheimer’s,  diabetes  and hypertension. An interesting question to ask would be: Do all these new, twenty first century diseases all have something in common, just like the top causes of death  in 1900 where from bacterial or viral sources?  A review of the medical literature shows two primary causes. In turns out that most chronic human diseases are 1) triggered by excess free radicals or oxidative stress and 2) are the result of a lack of essential minerals and trace minerals and cofactors in our diets.

Oxidative Stress

Free radicals or oxidative stress is the result of our cells metabolizing or breathing oxygen. Free radical are the “waste” products of life, and they are very destructive to cell membranes, proteins and DNA.  Our body has a natural protective system-antioxidant enzymes that neutralize excess free radicals. If this delicate balance is  disrupted,  this then leads to an inflammation response in the body and eventually, chronic diseases could ensue. As we age, our natural antioxidant protective systems decline and we are told to supplement with natural botanical antioxidants, such as blueberries , green tea or cinnamon or antioxidant supplements such as Vitamin E or coenzyme Q10. But then we run into the “antioxidant paradox”. All these food antioxidants  work great as antioxidants in a test tube in the lab, but in human clinical trials, the beneficial effects are either inconclusive or negative. This is because saliva and gut bacteria metabolise many these botanical antioxidants, before they have a chance to act beneficially.

Ukrainian scientists discovered the world’s highest antioxidant called Carbon 60 hydrated fullerenes, which is stable and inert and not metabolised by bacteria. Carbon 60, a natural product,  was discovered in 1985 and a Nobel prize in Chemistry was awarded for this discovery in 1996.  Scientists and doctors were calling it the panacea or silver bullet in medicine, but because it’s not naturally water soluble, just like diamonds, charcoal or activated charcoal, this frustrated scientists. Ukrainian scientist in Kharkiv the first to discovered how to dissolve Carbon 60 in water in 1994. After 20 years of preclinical, safety and clinical studies,  Carbon 60 hydrated fullerenes  were approved as a  “dietary supplement” by the Ukrainian Ministry of Health and has been on the Ukrainian market since 2010. There is now a US patent pending. Scientists are now claiming that most chronic human diseases are triggered by excess free radicals. Just do a search on PUBMED with the key words “oxidative stress” and your own disease and you will find a link. (http://www.ncbi.nlm.nih.gov/pubmed ). Reduce excess free radicals and reduce your disease symptoms. A study at the University of Paris in 2010 showed that rats feed a diet supplemented  with Carbon 60 in olive oil, increased the lifespan of rats by 90% from an average of 25-30 months to over 55 months.

Essential minerals and trace minerals.

The National Post ran a story on May 1- “Obese Canadians should be granted legal protection from discrimination, professor says.”  The obesity debate is totally missing the point. Most Doctors are ignoring their own medical research. The cause of the 400% increase in obesity in last 3 decades in Canada and sharp rise in most other chronic human diseases since the 1930’ is due to a chronic lack of essential minerals and trace minerals in our diet (plants, fruits and vegetables) that are needed as co-factors in the body for our biochemical pathways to work efficiently.

The soil minerals concentrations have been dropping  worldwide for the last 100 years, so less and less minerals are  absorbed in fruits, vegetables and other plants. This is due primarily to the fact that  we no longer cook and heat our homes with wood and throw away the ashes (95% minerals) back into the garden to replenish the soil with the 60 essential and trace minerals that our bodies need. Fertilizer only has 3 minerals. Even the nutritional supplements commonly found in most health food stores don't carry the full complement of 90  essential nutrients, which should include 60 essential minerals, 15 essential vitamins, 12 essential amino acids and 3 fatty acids and  the right doses of each and the correct easily absorbable mineral salts.

How do we know that we need 90 essential nutrients? Just talk to any veterinarian. Vets have cured over 600 chronic human diseases in farm animals and in zoo animals by supplementing their food with nutritional pellets. When was the last time you saw a cow with arthritis and a pig with Alzheimer’s?  Vets have to cure an animal after the first time otherwise beef would cost over $500 a pound or eggs $50 a dozen.  Why? Because animals don’t have health insurance. Doctors are quite content to treat your disease symptoms for the rest of your life, billing health insurance and not curing your disease after a few visits. Human  clinical studies in the past 40 years have shown that most chronic human diseases are also caused by essential mineral deficiencies and can be controlled with the proper essential and trace minerals. Diabetes has been controlled with the right amount of chromium and vanadium and other essential cofactors. Arthritis is a lack of proper calcium absorption and cofactors such as Vitamin D and magnesium. Greying hair is a copper deficiency in the diet.

So can we live to be 100 or over without chronic human diseases? Yes by reducing excess oxidative stress or free radicals in our body and ensuring that we get the right daily balance of 90 essential nutrients including 60 minerals and trace minerals.

If you would like more information on the above or a copy of my presentation on Aging Gracefully without Chronic Human Diseases that  I gave at Ukrainian Canadian Social Services last week, send me an email  [email protected] or call (416) 819-9667 or download it from this link  http://bit.ly/1KzZvm8  For Ukrainian listeners,  catch  my Ukrainian radio interview on Radio KONTAKT archives from Saturday April 25, 2015 http://www.kontaktglobal.com/radio-saturday.html

 

Walter Derzko is the president of the startup C60 Water North America

UPDATE on the Carbon[60] Hydrated Fullerenes-The Highest Antioxidant in the World

Scientists  have made a  significant leap in cancer research.

You never change things by fighting the existing reality. To change something, build a new model that makes the existing model obsolete.

R. Buckminster Fuller

90% of all cancer deaths come from cancer spreading in the body from one organ to another, called metastasis. Stop metastasis, stop many unnecessary premature cancer deaths.

Belgium researches in 2014 determined that cancer metastasis  can only occur if there are excess free radicals in the body. Specifically they conclude: “under certain conditions, the mitochondria produce more free radicals known as superoxide ions (O2.-). It is this overproduction of superoxide that leads to the formation of metastasis and, consequently, the growth of a tumor. “ See “A world first: Researchers identify a treatment that prevents tumor metastasis (Université catholique de Louvain)”  http://www.eurekalert.org/pub_releases/2014-07/ucdl-awf072414.php

So, logically, if you can reduce excess oxidative stress and free radical levels in the body back to  normal levels, theoretically, you  should be able to prevent metastasis and reduce the risk of cancer  growth and eliminate most cancer deaths. 

This not only applies to cancer, but many other chronic human diseases, which appear to be triggered by excess free radicals too.

Just do a search on PUBMED using the term “oxidative stress” the name of any chronic human disease and you will see a connection/link  to free radicals and most diseases. See http://www.ncbi.nlm.nih.gov/pubmed

In fact, researcher in Ukraine have already reduced oxidative stress  in cancer  cases and many other chronic human diseases with a safe, natural, organic molecule, called Carbon [60] or specifically the water-soluble version, called Carbon 60 Hydrated Fullerenes, C60HYFNs. It was approved by the Ukrainian Ministry of Health in 2010, as a dietary supplement and antioxidant, after 20 years of preclinical and clinical studies.  It’s the world’s highest antioxidant and free radical scavenger,  neutralizing excess free radicals. It was discovered in 1985 and a Nobel prize in Chemistry was awarded for the discover in 1996. Two years before that,  Ukrainian researched discovered how to make Carbon[60] water-soluble, since most Carbon allotropes do not dissolve  in water.

 (see  the medical and biological properties in Carbon 60 Hydrated Fullerenes   http://www.ipacom.com/index.php/en/fullerenes-and-water-left/74 )

Most recently (Dec. 2014), Ukrainian researchers showed that C60HYFNs combined with traditional chemotherapy (Doxorubicin) , eliminates tumors in animal models without the toxic effects seen with traditional chemo. They showed that this combination: “resulted in tumor growth inhibition, prolongation of life, metastasis inhibition, and increased number of apoptotic tumor cells and was more effective than the corresponding course of Dox treatment alone.”

See C60 fullerene as synergistic agent in tumor-inhibitory Doxorubicin treatment. http://www.ncbi.nlm.nih.gov/pubmed/25504158

Carbon 60, a radiation protector,  also reduces the negative effects of traditional radiation therapy. See  Peculiarities of the antioxidant and radioprotective effects of hydrated C60 fullerene nanostuctures in vitro and in vivo.  http://www.ncbi.nlm.nih.gov/pubmed/19539750

The molecule C60 HYFNs is now US patent pending (March 2014). http://www.freepatentsonline.com/y2014/0079746.html

We are in the process of submitting the clinical studies to the FDA to get a GRAS designation to sell C60HYFNs as an antioxidant  dietary supplement in the USA, Canada and Mexico.

Walter Derzko

President

C60 Water North America

416-819-9667

Carbon 60 or Fullerenes -the template for life on Earth?

Fullerenes -the template for life on Earth?

 

Isr Med Assoc J. 2012 Oct;14(10):602-6.

Fullerene and the origin of life.

Goodman G¹, Gershwin ME, Bercovich D.

Author information

• ¹Human Molecular Genetics and Pharmacogenetics,      Migal Biotechnology Institute, Kiryat Shmona, Israel.

Abstract

The role of carbon in the development of life and as the structural backbone of all organisms is universally accepted and an essential part of evolution. However, the molecular basis is largely unknown and the interactions of carbon with nitrogen and oxygen in space are enigmatic. In 1985, the previously unknown form of carbon, coined fullerene, was discovered. We hypothesize that by virtue of the unique properties of fullerene, this hollow, ultra-robust, large, purely carbon molecule was the earliest progenitor of life. It acted as a stable universal biologic template on which small molecules spontaneously assembled and then formed, by further assembly, a surface mantle (here termed rosasome) of larger molecules. We submit that this process, by its inherent flexibility, initiated evolution, allowing the emergence of parallel diverse rosasome lines responding selectively to varying spatial environments. For example, rosasomal lines mantled with nucleotide and peptide layers are conceived as primordial forerunners of the ubiquitous ribosome. Moreover, the parallel independent and interdependent evolution of rosasome lines would be more rapid than sequential development, refute precedence of either DNA or RNA, and explain the evolution of integration of two subunits with different structures and functions in ribosomes and of the triplet nature of the codon. Based on recent astronomical data, this hypothesis supports the concept that life is not a singularity. This concept also suggests a potential vehicle for therapeutics, biotechnology and genetic engineering.

Stanford scientists develop 'playbook' for reverse engineering tissue

 

 

STANFORD, Calif. — Consider the marvel of the embryo. It begins as a glob of identical cells that change shape and function as they multiply to become the cells of our lungs, muscles, nerves and all the other specialized tissues of the body.

Now, in a feat of reverse tissue engineering, Stanford University researchers have begun to unravel the complex genetic coding that allows embryonic cells to proliferate and transform into all of the specialized cells that perform myriad biological tasks.

A team of interdisciplinary researchers took lung cells from the embryos of mice, choosing samples at different points in the development cycle. Using the new technique of single-cell genomic analysis, they recorded what genes were active in each cell at each point. Though they studied lung cells, their technique is applicable to any type of cell.

"This lays out a playbook for how to do reverse tissue engineering," said Stephen Quake, PhD, the Lee Otterson Professor in the School of Engineering and a Howard Hughes Medical Institute investigator.

The researchers' findings are described in a paper published online April 13 in Nature. Quake, who also is a professor of bioengineering and of applied physics, is the senior author. The lead authors are postdoctoral scholars Barbara Treutlein, PhD, and Doug Brownfield, PhD.

The researchers used the reverse-engineering technique to study the cells in the alveoli, the small, balloon-like structures at the tips of the airways in the lungs. The alveoli serve as docking stations where blood vessels receive oxygen and deliver carbon dioxide.

Treutlein and Brownfield isolated 198 lung cells from mouse embryos at three stages of gestation: 14.5 days, 16.5 days and 18.5 days (mice are usually born at 20 days). They also took some lung cells from adult mice.

They used standard enzymatic techniques to dissolve the proteins that hold the lung cells together in tissue form, then sorted out the specific alveolar cell types that were the focus of their study.

Their next steps involved newer techniques at the heart of their reverse-engineering process.

Recall how eyedroppers work. Squeeze the rubber bulb to evacuate the air; plunge it into a solution to fill it with fluid; squeeze the bulb again to force the fluid out. In recent years, biotechnologists have used those basic principles to develop microfluidic devices of such precision that they can suck a single cell out of solution and isolate it in a chamber to study its genetic material.

Quake's lab has pioneered the use of microfluidic devices to study single cells. In this study, they used microfluidic devices to capture their 198 sample lung cells. Then they used single-cell genomic sequencing to measure which genes were active in each cell at each time.

How did they decode genomic activity in a single cell? DNA in the nucleus of every cell contains the full genome for that organism. That's why it's possible to build an organism from a single cell. But only some of those genes are active in any given cell at any given time. That's why lung cells are different than hair cells; each cell has a different set of active genes directing its functions.

Genes direct cellular activity by making or "expressing" messenger RNA. Each mRNA instructs the cell to make a particular protein. Cells are essentially a group of interacting proteins. Therefore knowing which mRNAs are active offers a lens into the function of that cell at the point when it was captured in the microfluidic device.

Using this process the Stanford researchers revealed for the first time precisely which genes regulate the development of these particular lung cells at each step along the way to mature alveoli.

One important finding involved the development of two key cell types at the tip of the alveoli, where the lung meets blood to perform the gas exchange that keeps us alive.

Alveolar type-1 cells are the flattest cells in the body. Blood cells dock alongside them to deliver oxygen or pick up carbon dioxide. The thinness of the cell is vital to facilitating this gas transfer.

Alveolar type-2 cells are compact and cuboidal. They secrete proteins to keep the alveoli from collapsing like empty balloons, so as to maintain the inner space through which oxygen and carbon dioxide can move.

Using single-cell genomics allowed the researchers to reverse engineer the development process to show how a single progenitor cell type gives rise to both of these different, mature alveolar cells.

The researchers also captured cells in transition from the progenitor to mature cell state, gaining crucial insights into the mechanism of alveolar cell differentiation.

Although this study focused on lung cells, the technique — capturing individual cells at different stages of embryonic development and assessing gene activity through mRNA sequencing — can be used to reverse-engineer other tissues.

In addition to studying embryonic development, the technique could be used in clinical settings. For instance, researchers could study differences between individual cells in a tumor, improving our understanding of the stages of cancers and leading to better, more targeted therapies.

"This technology represents a quantal leap forward in our ability to apprehend the full diversity of cell types in a given population, including rare ones that could have special functions," said Tushar Desai, MD, MPH, assistant professor of pulmonary and critical care medicine at Stanford and co-author of the paper. "Because a comprehensive molecular characterization of each type is achieved, including the signals they send and receive, a snapshot of the communication between individual cells will also emerge and may suggest attractive therapeutic targets in disease."

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The other Stanford co-authors of the paper are Mark Krasnow, MD, PhD, professor of biochemistry and an HHMI investigator; postdoctoral scholar Angela Wu, PhD; Norma Neff, PhD, genomics core director in the Department of Bioengineering; research assistant Gary Mantalas; and F. Hernan Espinoza, PhD, an HHMI research specialist.

The research was supported by the National Heart Lung and Blood Institute, the National Institutes of Health, the Parker B. Francis Foundation and the Howard Hughes Medical Institute.

The work was also supported by the Department of Bioengineering, which is jointly operated by the School of Engineering and the School of Medicine.

The Stanford University School of Medicine consistently ranks among the nation's top medical schools, integrating research, medical education, patient care and community service. For more news about the school, please visit http://mednews.stanford.edu. The medical school is part of Stanford Medicine, which includes Stanford Hospital & Clinics and Lucile Packard Children's Hospital Stanford. For information about all three, please visit http://stanfordmedicine.org/about/news.html.

The Stanford School of Engineering has been at forefront of innovation for nearly a century, creating pivotal technologies and businesses that have transformed the worlds of technology, medicine, energy and communications and laid the foundation for Silicon Valley. The school advances modern science and engineering through teaching and research. The school is home to nine departments, 245 faculty and more than 4,000 students, tackling the world's most pressing problems in areas like human health and environmental sustainability. For more information, visit http://engineering.stanford.edu.

Print media contact: Tom Abate at (650) 736-2245 ([email protected]) Broadcast media contact: M.A. Malone at (650) 723-6912 ([email protected])

Superbright and fast X-rays image single layer of proteins

In biology, a protein's shape is key to understanding how it causes disease or toxicity. Researchers who use X-rays to takes snapshots of proteins need a billion copies of the same protein stacked and packed into a neat crystal. Now, scientists using exceptionally bright and fast X-rays can take a picture that rivals conventional methods with a sheet of proteins just one protein molecule thick.

Department of Energy, National Institutes of Health, National Science Foundation

Newest method for determining a protein's shape based on XFEL technology significantly broadens number and type of proteins that researchers can study

• 

  Caught by XFEL: X-ray free-electron lasers can create images (left) that accurately reflect the known structure of proteins determined by conventional methods (right), in this case, three bacteriorhodopsin proteins.

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RICHLAND, Wash. –          In biology, a protein's shape is key to understanding how it causes disease or toxicity. Researchers who use X-rays to take snapshots of proteins need a billion copies of the same protein stacked and packed into a neat crystal. Now, scientists using exceptionally bright and fast X-rays can take a picture that rivals conventional methods with a sheet of proteins just one protein molecule thick.

Using a type of laser known as XFEL, the technique opens the door to learning the structural details of almost 25 percent of known proteins, many of which have been overlooked due to their inability to stack properly. The team of researchers led by the Department of Energy's Pacific Northwest and Lawrence Livermore National Laboratories report their results with this unique form of X-ray diffraction in the March issue of the International Union of Crystallography Journal.

"In this paper, we're proving it's possible to use an XFEL to study individual monolayers of protein," said PNNL microscopist James Evans. "Just being able to see any diffraction is brand new."

Evans co-led the team of two dozen scientists with LLNL physicist Matthias Frank. The bright, fast X-rays were produced at the Linac Coherent Light Source at SLAC National Accelerator Laboratory in Menlo Park, Calif., the newest of DOE's major X-ray light source facilities at the national laboratories. LCLS, currently the world's most powerful X-ray laser, is an X-ray free-electron laser. It produces beams millions of times brighter than earlier X-ray light sources.

Coming in at around 8 angstrom resolution (which can make out items a thousand times smaller than the width of a hair), the proteins appear slightly blurry but match the expected view based on previous research. Evans said this level of clarity would allow researchers, in some cases, to see how proteins change their shape as they interact with other proteins or molecules in their environment.

To get a clearer view of protein monolayers using XFEL, the team will need to improve the resolution to 1 to 3 angstroms, as well as take images of the proteins at different angles, efforts that are currently underway.

Not Your Family's Crystal

Researchers have been using X-ray crystallography for more than 60 years to determine the shape and form of proteins that form the widgets and gears of a living organism's cells. The conventional method requires, however, that proteins stack into a large crystal, similar to how oranges stack in a crate. The structure of more than 80,000 proteins have been determined this way, leading to breakthroughs in understanding of diseases, pathogens, and how organisms grow and develop.

But many proteins found in nature do not stack easily. Some jut from the fatty membranes that cover cells, detecting and interacting with other cells and objects, such as viruses or bacteria, in the surrounding area. These proteins are not used to having others of their kind stack on top. These so-called membrane proteins make up about 25 percent of all proteins but only 2 percent of proteins that researchers have determined structures for.

Wafer Thin

Researchers in the last decade have been pursuing the idea that one sheet of proteins could be visualized if the X-rays were bright enough but flashed on and off quickly enough to limit the damage wrought by the powerful X-rays. Two years ago, scientists demonstrated they could use XFEL technology on crystals of proteins about 15 to 20 sheets thick.

Evans, Frank and their team wanted to push this further. The team worked on a way to create one-sheet-thick crystals of two different proteins — a protein called streptavidin and a membrane protein called bacteriodopsin. The structures of both proteins are well-known to scientists, which gave the team something to compare their results to.

The team shined the super-bright X-rays for a brief moment — about 30 femtoseconds, a few million billionths of a second — on the protein crystals. They created so much data in the process that it took them more than a year to analyze all of it.

The resulting images look like the known structures, validating this method. Next, the researchers will try to capture proteins changing shape as they engage in a chemical reaction. For this, even shorter flashes of X-rays might be needed to see the action clearly.

If successful, shorter flashes of XFEL might mean longer lines at SLAC.

Evans is based at EMSL, the Environmental Molecular Sciences Laboratory at PNNL. This work was supported by the Department of Energy Office of Science, National Institutes of Health, National Science Foundation, Lawrence Livermore National Laboratory, and Pacific Northwest National Laboratory.

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Reference: Matthias Frank, David B. Carlson, Mark S. Hunter, Garth J. Williams, Marc Messerschmidt, Nadia A. Zatsepin, Anton Barty, W. Henry Benner, Kaiqin Chu, Alexander T. Graf, Stefan P. Hau-Riege, Richard A. Kirian, Celestino Padeste, Tommaso Pardini, Bill Pedrini, Brent Segelke, M. Marvin Seibert, John C. H. Spence, Ching-Ju Tsai, Stephen M. Lane, Xiao-Dan Li, Gebhard Schertler, Sebastien Boutet, Matthew Coleman and James E. Evans. Femtosecond X-ray Diffraction from Two-Dimensional Protein Crystals, International Union of Crystallography Journal, Feb. 10, 2014, DOI: 10.1107/S2052252514001444.

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SLAC's LCLS is the world's most powerful X-ray free-electron laser. A DOE national user facility, its highly focused beam shines a billion times brighter than previous X-ray sources to shed light on fundamental processes of chemistry, materials and energy science, technology and life itself.

Proteases fight bacteria

New mechanisms of action open alternative approaches to antibiotics development:

Two new weapons in the battle against bacteria

 

The target: bacterial protease ClpP - Image: M. Gersch / TUM

13.02.2014,  Research news

Proteases are vital proteins that serve for order within cells. They break apart other proteins, ensuring that these are properly synthesized and decomposed. Proteases are also responsible for the pathogenic effects of many kinds of bacteria. Now chemists at the Technische Universitaet Muenchen (TUM) have discovered two hitherto unknown mechanisms of action that can be used to permanently disarm an important bacterial protease.

Proteins are made up of a chain of amino acids and are vital for all cell processes. Proteases are among the most important types of protein. Like “molecular scissors”, they cut other proteins at given positions and thereby execute important cell functions. By cutting the amino acid chains to the right length or breaking proteins apart they, for example, activate or deactivate proteins, decompose defective ones or switch signal sequences that serve to transport proteins to their proper position within a cell.

But proteases are important not only for human cells – bacteria also rely on them. There are hardly any effective antibiotics left in the fight against pathogens like multi-resistant strains of Staphylococcus aureus bacteria or Mycobacterium tuberculosis which causes tuberculosis. 

Researchers around the world are thus working ardently to find new ways of disarming the proteases in these strains to combat them. At the heart of this effort lies the so-called ClpP protease. It comprises 14 subunits and has a central regulatory function. The usual approach to deactivating this protease is to block all active centers of the ClpP. These are effectively the “cutting edges of the scissors”, i.e. the portions of the protein that are responsible for breaking apart other proteins. 

“However, the inhibitors used in the past have one decisive disadvantage,” explains Stephan Sieber who heads the Chair for Organic Chemistry II at the Technische Universitaet Muenchen (TUM). “They don’t permanently disarm the proteins, but only work for a few hours. On top of that, to be effective they must attack all active centers of the protein.” 

New strategies against bacteria

In collaboration with Professor Michael Groll, who heads the Chair for Biochemistry, Malte Gersch and Roman Kolb, doctoral candidates at Professor Sieber’s chair, have succeeded in uncovering two completely new mechanisms that can be used to permanently deactivate these important bacteriological proteases – in one case even without having to attack all active centers of the protein. 

The first mechanism disrupts the arrangement of amino acids required for the cohesion of the protease subunits. As a result the protease breaks into two parts. The second acts directly on the core of the active center. It converts the amino acid that does the actual splitting into another kind of amino acid – the “scissors” lose their edge and the protein is rendered inoperable. Both approaches inhibit the protease in completely novel ways and are thus very promising for the development of new forms of medication. 

The scientists also found a whole series of inhibitors that initiate the two mechanisms. “Knowing the ways in which substances deactivate the proteases is a huge advance,” says Gersch. “We can now optimize the substances and possibly also apply the principle to other proteases.” 

In further research, Gersch and Sieber plan to test their substances on living bacterial strains to determine if these are truly inhibited in growth and pathogenic effect. “Although the bacteria are not completely disarmed, they produce significantly fewer toxins that are conducive to inflammation,” says Gersch. “The basic idea is that we give the immune system more time to handle the pathogens on its own while the formation of new resistances is suppressed.” 

The research was funded by the Deutsche Forschungsgemeinschaft (DFG; SFB 749 and SFB 1035, Cluster of Excellence Center for Integrated Protein Research Munich (CIPSM)), by the European Research Council (ERC starting grant), the Fund of the German Chemical Industry and the German National Academic Foundation. The crystal structures were determined in collaboration with the synchrotron radiation source of the Paul Scherrer Institute in Villigen (Switzerland). 

Publication:

Malte Gersch, Roman Kolb, Ferdinand Alte, Michael Groll, Stephan Sieber. Disruption of Oligomerization and Dehydroalanine Formation as Mechanisms for ClpP Protease Inhibition. Journal of the American Chemical Society (JACS), 2014, 136 (4), pp 1360-1366 –DOI: 10.1021/ja4082793

Contact:

Prof. Stephan A. Sieber Chair for Organic Chemistry II Technische Universitaet Muenchen Lichtenbergstr. 4, 85747 Garching, Germany Tel.: +49 89 289 13302 / 13301 (Sekr.) E-Mail – Web

Food processors beware: Salmonella biofilms incredibly resistant to powerful disinfectants

 

		IMAGE:   This image shows photos of biofilms on surfaces at low magnification (300)   and high magnification (3000) using electron microscope. Click here for more information.

Once Salmonella bacteria get into a food processing facility and have an opportunity to form a biofilm on surfaces, it is likely to be extraordinarily difficult, if not impossible, to kill it, according to research published ahead of print in the journal Applied and Environmental Microbiology.

Researchers from National University of Ireland, Galway conducted a study in which they attempted to kill Salmonella biofilms on a variety of hard surfaces, using three types of disinfectant.

"We found that it was not possible to kill the Salmonella cells using any of the three disinfectants, if the biofilm was allowed to grow for seven days before the disinfectant was applied," says Mary Corcoran, a researcher on the study. Even soaking the biofilms in disinfectant for an hour and a half failed to kill them

The impetus for the study was a European outbreak in which 160 people in 10 countries became ill with gastroenteritis (vomiting and diarrhea) from the Agona serotype of Salmonella, says Corcoran. That outbreak was traced to meat from a major food-processing facility.

"It seems that Salmonella Agona entered into the environment in the part of the facility where meat that was already cooked was being handled, and it had survived and contaminated the cooked meat," says Corcoran. "We were interested in determining if this particular Salmonella, that caused the outbreak, might have something special about it that makes it better at surviving in the environment of a food processing facility. Was it better at forming a dense biofilm or was it more resistant to disinfectants than other Salmonella?"

The research uncovered nothing special about that specific strain.

"We found that all of the types of Salmonella we looked at were able to adopt the specialized biofilm lifestyle on all of the surfaces we looked at, including glass, stainless steel, glazed tile, and plastic, and that the biofilm of Salmonella gets more dense over time, and becomes more firmly attached to the surface," she says.

Corcoran warns that food processing facilities must take strict care to keep Salmonella out of the clean areas where cooked foods get further processing and packaged

"People need to question whether disinfectants that are promoted as killing various types of bacteria are really as effective in real life situations where biofilms can form as they are claimed to be based on experiments that do not use biofilms. A lot of the time, the disinfectant may add very little, if anything, to good cleaning and appropriate food handling practices," says Corcoran. "There is a need for more research to define better methods for killing Salmonella biofilms."

In the US, an estimated million-plus cases of Salmonella occur annually, with 23,000 hospitalizations and 450 fatalities reported each year, according to the Centers for Disease Control and Prevention.

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A copy of the manuscript can be found online at http://bit.ly/asmtip0114e. The final version of the article is scheduled for the February 2014 issue of Applied and Environmental Microbiology.

Applied and Environmental Microbiology is a publication of the American Society for Microbiology (ASM). The ASM is the largest single life science society, composed of over 39,000 scientists and health professionals. Its mission is to advance the microbiological sciences as a vehicle for understanding life processes and to apply and communicate this knowledge for the improvement of health and environmental and economic well-being worldwide.

 

http://www.eurekalert.org/pub_releases/2014-01/asfm-fpb011514.php

PUBLIC RELEASE DATE:

15-Jan-2014
Contact: Jim Sliwa [email protected] 202-942-9297 American Society for Microbiology

Gene activity and transcript patterns visualized for the first time in thousands of single cells

Biologists of the University of Zurich have developed a method to visualize  the activity of genes in single cells. The method is so efficient that, for the  first time, a thousand genes can be studied in parallel in ten thousand single human cells. Applications lie in fields of basic research and medical
diagnostics. The new method shows that the activity of genes, and the spatial organization of the resulting transcript molecules, strongly vary between single cells.

Whenever cells activate a gene, they produce gene specific transcript molecules, which make the function of the gene available to the cell. The measurement of gene activity is a routine activity in medical diagnostics, especially in cancer medicine. Today's technologies determine the activity of
genes by measuring the amount of transcript molecules. However, these technologies can neither measure the amount of transcript molecules of one thousand genes in ten thousand single cells, nor the spatial organization of transcript molecules within a single cell. The fully automated procedure,
developed by biologists of the University of Zurich under the supervision of Prof. Lucas Pelkmans, allows, for the first time, a parallel measurement of the amount and spatial organization of single transcript molecules in ten thousands single cells. The results, which were recently published in the scientific journal Nature Methods, provide completely novel insights into the variability of gene activity of single cells.

Robots, a fluorescence microscope and a supercomputer

The method developed by Pelkmans' PhD students Nico Battich and Thomas Stoeger is based upon the combination of robots, an automated fluorescence microscope and a supercomputer. "When genes become active, specific transcript molecules are produced. We can stain them with the help of a robot", explains Stoeger. Subsequently, fluorescence microscope images of brightly glowing transcript molecules are generated. Those images were analyzed with the supercomputer Brutus, of the ETH Zurich. With this method, one thousand human genes can be studied in ten thousand single cells. According to Pelkmans, the advantages of this method are the high number of single cells and the
possibility to study, for the first time, the spatial organization of the transcript molecules of many genes.

New insights into the spatial organization of transcript molecules

The analysis of the new data shows that individual cells distinguish themselves in the activity of their genes. While the scientists had been suspecting a high variability in the amount of transcript molecules, they were surprised to discover a strong variability in the spatial organization of transcript molecules within single cells and between multiple single cells. The transcript molecules adapted distinctive patterns.

"We realized that genes with a similar function also have a similar variability in the transcript patterns," explains Battich. "This similarity exceeds the variability in the amount of transcript molecules, and allows us to predict the function of individual genes." The scientists suspect that
transcript patterns are a countermeasure against the variability in the amount of transcript molecules. Thus, such patterns would be responsible for the robustness of processes within a cell.

The importance of these new insights was summarized by Pelkmans: "Our method will be of importance to basic research and the understanding of cancer tumors because it allows us to map the activity of genes within single tumor cells."

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Literature:

Nico Battich, Thomas Stoeger, Lucas Pelkmans. Image-based transcriptomics in thousands of single human cells at single-molecule resolution. Nature Methods. DOI: 10.1038/nmeth.2657

 

Human skin wound dressings to treat cutaneous ulcers

 

Quebec City, October 2, 2013—Researchers from Université Laval's Faculty of
Medicine and CHU de Québec have shown that it is possible to treat venous ulcers
unresponsive to conventional treatment with wound dressings made from human skin
grown in vitro. A study published recently in the journal Advances in Skin
and Wound Care demonstrates how this approach was successfully used to treat
venous lower-extremity ulcers in patients who had been chronically suffering
from such wounds.

About 1% of the population suffers from lower-extremity ulcers. These wounds
regularly become inflamed or infected and are very slow to heal, if they do at
all. They are frequently associated with aging, diabetes, and circulatory system
disorders such as varicose veins and oedema. "Obese individuals and those who
work constantly standing up are especially vulnerable. These ulcers can persist
for years. It can be a hellish clinical situation when standard treatments don't
work," noted Dr. François A. Auger, director of both the study and LOEX, the
tissue engineering and regenerative medicine laboratory where it was
conducted.

Standard treatment for ulcers involves methodically cleaning these wounds and
applying compression bandages. Drugs became available around 20 years ago but
they are expensive and their efficacy has been somewhat limited. A graft using
the patient's own skin can be effective but is problematic because it requires a
significant amount of skin to be removed from elsewhere on the body.

This very problem inspired LOEX researchers to use their expertise with in
vitro skin culture to create biomaterial-free biological wound dressing. The
process is complex and requires several steps: removing 1 cm2 of skin from the
patient, isolating the appropriate cells, growing them in vitro, and creating a
skin substitute with both dermis and epidermis. After eight weeks of growth the
self-assembled sheets of skin substitute can be applied over the ulcers, much
like bandages, and replaced weekly as long as necessary. "This totally
biological bandage is much more than a physical barrier," stresses Dr. Auger.
"The cells secrete molecules that speed up healing by helping to set natural
healing processes in motion. It would be hard to imagine a model closer to the
human body's natural physiology."

Tests were successfully carried out on five patients. It took only an average
of seven weeks to cure 14 ulcers that had been affecting patients for at least
six months, and in some cases, several years. "This is a last recourse once all
other treatment options have been exhausted," notes François A. Auger.

Dr. Auger sees another promising application for these biological bandages:
"We have shown that this is effective for patients with leg ulcers. Now, we
intend to carry out a clinical study to demonstrate that the same treatment
works for patients with serious burns, as soon as we get the necessary
approvals."

 

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In addition to Dr. Auger, the study's co-authors are: Olivier Boa, Chanel
Beaudoin Cloutier, Hervé Genest, Raymond Labbé, Bertrand Rodrigue, Jacques
Soucy, Michel Roy, Frédéric Arsenault, Carlos E. Ospina, Nathalie Dubé,
Marie-Hélène Rochon, Danielle Larouche, Véronique J. Moulin, and Lucie
Germain.

Information:
Dr. François A. Auger
Faculty of Medicine
Université
Laval
418 527-4546
[email protected]

 

 

Scientists Force Mature Cells to Revert to Stem Cells

Scientists Force Mature Cells to Revert to Stem Cells – (Bloomberg
– September 11, 2013)
Scientists have turned back the hands of time in  cells within a living creature. Researchers in Spain used a technique created  seven years ago to force mature cells in mice to revert to an original form of
stem cell with the potential to change into any type of living tissue.
Previously, scientists were only been able to achieve this change in a petri  dish. The newest experiment may one day let doctors work entirely inside the  body to regenerate tissue and, perhaps, more complex organs. This could include  reconnecting a severed spinal cord or generating healthy heart cells. “This is
the next step along a continuum,” said Daley, a professor at Harvard Medical  School in Boston who wrote an accompanying editorial on the work, which he  wasn’t involved with. “What this is hinting at is that maybe we can, by  regressing tissues in the patient, regenerate this embryonic potential and, with
direction, regenerate a particular tissue.” The reverted mouse cells were also found to be more primitive than stem cells taken from embryos or created in the lab. This means they can be turned into a placenta and other embryonic-support membranes, a factor beyond the capacity of the other cells, the researchers wrote.