Tony Pantalleresco Radio Show notes – Month September 2014

Tony Pantallaresco

Welcome to Tony Pantalleresco Radio Show notes – Month September 2014

Show of September 6th 2014

What is Biofilm?

Estimating downwind concentrations of viable airborne microorganisms in dynamic atmospheric conditions

Allergic reaction to antibiotic residues in foods? You may have to watch what your fruits and veggies eat

Atmospheric Nanoparticles Impact Health

Chelant extraction of heavy metals from contaminated soils

A Citizen’s Guide to Soil Washing

How to Clean Out Polluted Soil

SOD B Extramel Improves Physical, Mental Performance

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What is Biofilm?
Biofilm (a bacterial film) is a mixture of different micro-organisms that are held together and protected by glue-like materials (carbohydrates)[F1] . The glue-like material that micro-organisms secrete allow them to attach themselves to surfaces. Listeria, Pseudomonas, Bacillus, Escherichia coli and Salmonella are some common micro-organisms found in biofilms.–Over time, bacteria on food contact surfaces (some carrying disease) can form into biofilm. In food production areas, biofilm is a sign that the area is unsanitary and needs to be properly cleaned. Unsanitary food surfaces can contaminate food products and reduce their shelf life. Biofilm in the food industry has high food residue and mineral content. It grows over time and becomes strongly attached to food contact surfaces. The greatest concern is that biofilm may contribute to the production of contaminated products from cross contamination.

Removing Biofilm
If biofilm is a concern in your facility, it’s important to clean and sanitize frequently and thoroughly. It’s also important to work with your chemical supplier to find the right products to remove and eliminate biofilms.—Removal of biofilm is achieved by a combination of four factors: 1) formulations and concentrations of cleaning and sanitizing agents 2) exposure time 3) temperature 4) mechanical activity[F2] The combination of these four things can dissolve biofilm and the organic material it sticks to. [F3] Extensive scrubbing with proper chemicals is important, because any biofilm residue can promote more biofilm growth. Talk to your chemical supplier to find the right system for your facility. The location, age, and history of biofilm formation within your facility will shape the guidelines to these four factors. Biofilm micro-organisms can have a high resistance to chemicals and you need to find the right combination to ensure effectiveness.

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Estimating downwind concentrations of viable airborne microorganisms in dynamic atmospheric conditions.
B Lighthart and
A J Mohr
1. Environmental Research Laboratory, U.S. Environmental Protection Agency, Corvallis, Oregon 97333.
ABSTRACT
A Gaussian plume model has been modified to include an airborne microbial survival term that is a best-fit function of laboratory experimental data of weather variables. The model has been included in an algorithm using microbial source strength and local hourly mean weather data to drive the model through a summer- and winter-day cycle. For illustrative purposes, a composite airborne “virus” (developed using actual characteristics from two viruses) was used to show how wind speed could have a major modulating effect on near-source viable concentrations. For example, at high wind speeds such as those occurring during the day, or with short travel times, near-source locations experience high viable concentrations because the microorganisms have not had time to become inactivated. As the travel time increases, because of slow wind speed or longer distances, die-off modulation by sunshine, relative humidity, temperature, etc., potentially becomes increasingly predominant.

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Allergic reaction to antibiotic residues in foods? You may have to watch what your fruits and veggies eat
Source-American College of Allergy, Asthma and Immunology (ACAAI)

People with food allergies always have to watch what they eat. Now, they may have to watch what their fruits and vegetables eat, as it seems it’s possible to have an allergic reaction to antibiotic residues in food.–An article published in the September issue of Annals of Allergy, Asthma and Immunology, the scientific publication of the American College of Allergy, Asthma and Immunology (ACAAI), examines the case of a 10 year-old girl who had an anaphylactic (severely allergic) reaction after eating blueberry pie. Although she had a medical history of asthma and seasonal allergies, and known anaphylaxis to penicillin and cow’s milk, she wasn’t known to be allergic to any of the ingredients in the pie.–After weeks of testing on both the young girl and a sample of the pie, the article authors decided that what had caused the reaction was a streptomycin-contaminated blueberry. Streptomycin, in addition to being a drug used to fight disease, is also used as a pesticide in fruit, to combat the growth of bacteria, fungi, and algae.–“As far as we know, this is the first report that links an allergic reaction to fruits treated with antibiotic pesticides,” said allergist Anne Des Roches, MD,FRCP, lead study author. “Certain European countries ban the use of antibiotics for growing foods, but the United States and Canada still allow them for agricultural purposes.”-[F4] The authors note that new regulations from the Food and Drug Administration may help to reduce antibiotic contaminants in food, which will help reduce antibiotic resistance and may also help reduce this type of event.–“This is a very rare allergic reaction,” said allergist James Sublett, MD, ACAAI president-elect. “Nevertheless, it’s something allergists need to be aware of and that emergency room personnel may need to know about in order to help determine where anaphylactic reactions may arise. Anyone who is at risk for a life-threatening allergic reaction should always carry epinephrine[F5] . They also need to know how to use their epinephrine [F6] in an emergency situation.”–Story Source–The above story is based on materials provided by American College of Allergy, Asthma and Immunology (ACAAI). Note: Materials may be edited for content and length

Recipe—if one has allergies sustaining a good dose of vitamin C will block or minimize the histamine release which may trigger a allergic response and sustain using nettle leaf or root as tea—utilizing as well pine bark –quercitrin and zinc ( which all would support the bronchrial area and strengthen and support the lungs as well from infection ) also things like feverfew—methionine – and Korean ginseng ( minimize use with young people since it can be overpowering) and aloe vera

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Atmospheric Nanoparticles Impact Health
Nanoparticles are atmospheric materials so small that they can’t be seen with the naked eye[F7] , but they can very visibly affect both weather patterns and human health all over the world and not in a good way, according to a study by a team of researchers at Texas A&M University.-Researchers Lin Wang, Renyi Zhang, Alexei Khalizov, Jun Zheng, Wen Xu, Yan Ma and Vinita Lal in the Departments of Atmospheric Sciences and Chemistry say that nanoparticles appear to be growing in many parts of the world[F8] , but how they do so remains a mystery.—Their work is published in the current issue of “Nature Geoscience” and was funded by the National Science Foundation and The Welch Foundation. –The team looked at how nanoparticles are formed and their relationship with certain organic vapors responsible for additional growth.–This is one of the most poorly understood of all atmospheric processes, Zhang says. But we found that certain types of organics tend to grow very rapidly. When this happens, they scatter light back into space, and that definitely has a cooling effect sort of a reverse greenhouse effect.’ It can alter Earth’s weather patterns and it also tends to have a negative effect on human health.–Persons with breathing problems, such as those who suffer from asthma, emphysema or other lung ailments, can be at risk, he notes.–Zhang says the team used new methods of measuring nanoparticles and formed new models to determine their impact on atmospheric conditions. These changes on our weather systems appear to be the most dramatic consequences of these nanoparticles, he adds.– Once these form, they can change cloud formations, which in turn can affect weather all over the world, so this can become a global problem to deal with. We’re trying to get a better understanding of these particles work and grow. They can form near areas that have petrochemical plants, such as Houston, which also has high amounts of aerosols from traffic emissions and other numerous factories.[F9] But were still trying to learn how they form and interact with the atmosphere.–Many types of trees and plants also contribute to the formation of nanoparticles[F10] , which are natural processes, Zhang says, and certain forms of organic materials can also speed up the development of the particles. But all of these ultimately affect the atmosphere, and very often, cloud formation, where the aerosols scatter light and radiation back into space and provide the seeds of cloud droplets and development.– These nanoparticles are very small about one million times smaller than a typical raindrop, Zhang says. But what they do can have a huge effect on our weather.–Contact: Renyi Zhang at (979) 845-7656� or Keith Randall, News & Information Services, at (979) 845-4644

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Chelant extraction of heavy metals from contaminated soils.
Peters RW.

Author information
Abstract
The current state of the art regarding the use of chelating agents to extract heavy metal contaminants has been addressed. Results are presented for treatability studies conducted as worst-case and representative soils from Aberdeen Proving Ground’s J-Field for extraction of copper (Cu), lead (Pb), and zinc (Zn). The particle size distribution characteristics of the soils determined from hydrometer tests are approximately 60% sand, 30% silt, and 10% clay. Sequential extractions were performed on the ‘as-received’ soils (worst case and representative) to determine the speciation of the metal forms. The technique speciates the heavy metal distribution into an easily extractable (exchangeable) form, carbonates, reducible oxides, organically-bound, and residual forms. The results indicated that most of the metals are in forms that are amenable to soil washing (i.e. exchangeable+carbonate+reducible oxides). The metals Cu, Pb, Zn, and Cr have greater than 70% of their distribution in forms amenable to soil washing techniques, while Cd, Mn, and Fe are somewhat less amenable to soil washing using chelant extraction. However, the concentrations of Cd and Mn are low in the contaminated soil. From the batch chelant extraction studies, ethylenediaminetetraacetic acid (EDTA), citric acid, and nitrilotriacetic acid (NTA) were all effective in removing copper, lead, and zinc from the J-Field soils. Due to NTA being a Class II carcinogen, it is not recommended for use in remediating contaminated soils. EDTA and citric acid appear to offer the greatest potential as chelating agents to use in soil washing the Aberdeen Proving Ground soils. The other chelating agents studied (gluconate, oxalate, Citranox, ammonium acetate, and phosphoric acid, along with pH-adjusted water) were generally ineffective in mobilizing the heavy metals from the soils. The chelant solution removes the heavy metals (Cd, Cu, Pb, Zn, Fe, Cr, As, and Hg) simultaneously. Using a multiple-stage batch extraction, the soil was successfully treated passing both the Toxicity Characteristics Leaching Procedure (TCLP) and EPA Total Extractable Metal Limit. The final residual Pb concentration was about 300 mg/kg, with a corresponding TCLP of 1.5 mg/l. Removal of the exchangeable and carbonate fractions for Cu and Zn was achieved during the first extraction stage, whereas it required two extraction stages for the same fractions for Pb. Removal of Pb, Cu, and Zn present as exchangeable, carbonates, and reducible oxides occurred between the fourth- and fifth-stage extractions. The overall removal of copper, lead, and zinc from the multiple-stage washing were 98.9%, 98.9%, and 97.2%, respectively. The concentration and operating conditions for the soil washing extractions were not necessarily optimized. If the conditions had been optimized and using a more representative Pb concentration (approximately 12000 mg/kg), it is likely that the TCLP and residual heavy metal soil concentrations could be achieved within two to three extractions. The results indicate that the J-Field contaminated soils can be successfully treated using a soil washing technique.

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A Citizen’s Guide to Soil Washing

What is soil washing?
Soil washing is a technology that uses liquids (usually water, sometimes combined with chemical additives) and a mechanical process to scrub soils. This scrubbing removes hazardous contaminants and concentrates them into a smaller volume. Hazardous contaminants tend to bind, chemically or physically, to silt and clay. Silt and clay, in turn, bind to sand and gravel particles. The soil washing process separates the contaminated fine soil (silt and clay) from the coarse soil (sand and gravel). When completed, the smaller volume of soil, which contains the majority of the fine silt and clay particles, can be further treated by other methods (such as incineration or bioremediation) or disposed of according to state and federal regulations. The clean, larger volume of soil is not toxic and can be used as backfill.

A Quick Look at Soil Washing

Separates fine-grained particles (silt and clay) from coarse-grained particles (sand and gravel).
Significantly reduces the volume of contaminated soil.
Is a relatively low-cost alternative for separating waste and minimizing volume required for subsequent treatment.
Is a transportable technology that can be brought to the site.

How does soil washing work?
A simplified drawing of the soil washing process is illustrated in Figure 1. The equipment is transportable so that the process can be conducted at the site. The first step of the process is to dig up the contaminated soil and move it to a staging area where it is prepared for treatment. The soil is then sifted to remove debris and large objects, such as rocks. The remaining material enters a soil scrubbing unit, in which the soil is mixed with a washing solution and agitated. The washing solution may be simply water or may contain additives, like detergent, which remove the contaminants from the soil. This process is very similar to washing laundry. The washwater is drained out of the soil scrubbing unit and the soil is rinsed with clean water. The larger scale soil washing equipment presently in use can process over 100 cubic yards of soil per day. The heavier sand and gravel particles in the processed soil settle out and are tested for contaminants. If clean, this material can be used on the site or taken elsewhere for backfill. If traces of contaminants are still present, the material may be run through the soil washer again or collected for alternate treatment or off-site disposal. Off-site disposal may be regulated by the Resource Conservation Recovery Act (RCRA) or the Toxic Substance Control Act (TSCA).

Figure 1
The Soil Washing Process

The contaminated silt and clay in the washwater settle out and are then separated from the washwater. The washwater, which now also contains contaminants, is treated by wastewater treatment processes so it can be recycled for further use. As mentioned earlier, the washwater may contain additives, some of which may interfere with the wastewater treatment process. If this is the case, the additives must be removed or neutralized by “pretreatment” methods before the washwater goes to wastewater treatment. Once separated from the washwater, the silt and clay are tested for contaminants. If all the contaminants were transferred to the washwater and the silt and clay are clean, they can be used at the site or taken elsewhere for use as backfill. If still contaminated, the material may be run through the soil washing process again, or collected for alternate treatment or off-site disposal in a permitted RCRA or TSCA landfill.

Not All Soil Is Created Equal

Soil is comprised of fine-grained (silt and clay) and coarse-grained (sand and gravel) particles, organic material (decayed plant and animal matter), water, and air. Contaminants tend to readily bind, chemically or physically, to silt, clay, and organic material. Silt, clay, and organic material, in turn, bind physically to sand and gravel. When the soil contains a large amount of clay and organic material, the contaminants attach more easily to the soil and, therefore, are more difficult to remove than when a small amount of clay and organic material is present.

Why consider soil washing?

Soil washing can be used as a technology by itself, but is often used in combination with other treatment technologies. Perhaps the principal use of soil washing is as a volume reduction technique in which the contaminants are concentrated in a relatively small mass of material. The larger the percentage of coarse sand and gravel in the material to be processed (which can be cleaned and perhaps returned to the site), the more cost-effective the soil washing application will be.

Ideally, the soil washing process would lead to a volume reduction of about 90% (which means only 10% of the original volume would require further treatment). Wastes with a high percentage of fine silt and clay will require a larger quantity of material to go on to subsequent, more expensive treatment. These soils may not be good candidates for soil washing.

Soil washing is used to treat a wide range of contaminants, such as metals, gasoline, fuel oils, and pesticides. There are several advantages to using this technology. Soil washing:

· Provides a closed system that remains unaffected by external conditions. This system permits control of the conditions (such as the pH level and temperature) under which the soil particles are treated.

· Allows hazardous wastes to be excavated and treated on-site.

· Has the potential to remove a wide variety of chemical contaminants from soils.

· Is cost-effective because it can be employed as a pre-processing step, significantly reducing the quantity of material that would require further treatment by another technology. It also creates a more uniform material for subsequent treatment technologies.

What Is An Innovative Treatment Technology?

Treatment technologies are processes applied to hazardous waste or contaminated materials to permanently alter their condition through chemical, biological, or physical means. Treatment technologies are able to alter, by destroying or changing, contaminated materials so that they are less hazardous or are no longer hazardous. This may be done by reducing the amount of contaminated material, by recovering or removing a component that gives the material its hazardous properties or by immobilizing the waste. Innovative treatment technologies are those that have been tested, selected, or used for treatment of hazardous waste or contaminated materials but still lack well-documented cost and performance data under a variety of operating conditions.

Will soil washing work at every site?
Soil washing works best when the soil does not contain a large amount of silt or clay. In some cases, soil washing is best applied in combination with other treatment technologies, rather than as a technology by itself.

Removal of contaminants can often be improved during the soil washing process by adding chemical additives to the washwater. However, the presence of these additives may cause some difficulty in the treatment of the used wastewater and the disposal of residuals from the washing process. Costs of handling and managing the additives have to be weighed against the amount of improvements in the performance of the soil washing process.

Where has soil washing been used?
At the King of Prussia site in New Jersey, soil washing was used to remove metal contamination such as chromium, copper, mercury, and lead from 19,000 tons of soil and sludge at a former industrial waste reprocessing facility. The soil washing process was able to clean the materials to meet clean-up goals for eleven metals. For example, chromium levels went from 8,000 milligrams chromium per kilogram of soil (mg/kg) to 480 mg/kg. Table 1 on page 4 lists some of the Superfund sites where soil washing has been selected.

Table 1
Examples of Superfund Sites Where Soil Washing Has Been Selected *

Name of Site
Status**
Medium
Contaminants

Myers Property, NJ
In design
Soil, sediment
Metals

Vineland Chemical, NJ
In design
Soil
Metals

GE Wiring Devices, PR
In design
Soil, sludge
Metals

Cabot Carbon/Koppers, FL
In design
Soil
Semi-volatile organic compounds (SVOCs)
polyaromatic hydrocarbons (PAHs)
metals

Whitehouse Waste Oil Pits
Predesign
Soil, sludge
Volatile organic compounds (VOCs)
PCBs
PAHs
metals

Cape Fear Wood Preserving
Design complete
Soil

Moss American, WI
Predesign
Soil
PAHs, metals

Arkwood, AR
In design
Soil, sludge

For a listing of Superfund sites at which innovative treatment technologies have been used or selected for use, contact NCEPI at the address in the box below for a copy of the document entitled Innovative Treatment Technologies: Annual Status Report (7th Ed.), EPA 542-R-95-008. Additional information about the sites listed in the Annual Status Report is available in database format. The database can be downloaded free of charge from EPA’s Cleanup Information bulletin board (CLU-IN). Call CLU-IN at 301-589-8366 (modem). CLU-IN’s help line is 301-589-8368. The database also is available for purchase on diskettes. Contact NCEPI for details

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How to Clean Out Polluted Soil
One of the biggest challenges a gardener can face is growing plants in polluted soil. Many things can pollute soil, including chemicals that remain from past uses of the ground, flooding, and sewer back-up. If you think your soil is polluted, it is important to conduct a soil test. You can buy a do-it-yourself soil test, but if you are worried about serious contamination, it might be best to have a professional evaluation of the soil.

For heavily polluted or contaminated soil, especially with deadly chemicals, it would be best to dig up the area and dispose of the existing soil, and then replace it with new. This can be very costly, but depending on the level of pollution, may be the only choice.

If you discover that the soil is only moderately polluted, there are ways you can remedy the situation on your own.

Step 1 – Do a Soil Test
Finding out what your soil lacks or has an abundance of is the first thing that must happen so you can take appropriate steps. By knowing this, you will know what to add to make the soil healthier.

Step 2 – Add Organic Materials
One way to improve your soils health is to add organic matter, such as compost, manure, leaves, or grass clippings. Use a tiller of spade to work the matter into the top 6 to 4 inches of soil. By adding the organic material, you will encourage micro organisms to move into your soil, and you will attract worms which act as natural aerators in soil.

Step 3 – Add Nutrients
Depending on the results of your soil test, add lime to lower pH levels or sulfur to raise the pH level. Add fertilizer to increase nitrogen, potassium and phosphorus.

Step 4 – Improve Drainage
If your soil has poor drainage, it will be harder for it to shed it’s toxins. The compost will help with drainage, but you can also add gypsum and coarse sand to soil to make it less dense. Pea gravel may also be used. Work the materials into the soil well.

Step 5 – Bring in the Micro Organisms
You can add your own micro organisms to the soil to help speed the process. Many of the thrive in contaminated soils, and will suck up and metabolize the pollutants for you.

Step 6 – Grow Plants that Clean the Soil
There are several plants that can help clean up the soil. Sunflowers will draw toxins out of the soil with their roots, and have been used for years as natural detoxifiers. Another plant that will help the process by the same method is ferns. Ferns and sunflowers both work as natural leeches, and will suck up many of the chemical pollutants left behind in the soil.

Step 7 – Continue to Work the Soil
Keep the soil tilled, watered, and fertilized. Between the improved drainage, the micro organisms, the pollutant eating plants, and your care of the soil, it will soon be healthy and ready to garden.

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SOD B Extramel Improves Physical, Mental Performance

AVIGNON, France—A new study confirmed the quick and lasting efficiency of Bionov’s SOD B Extramel in improving mental and physical performance. (Nutrients. 2014 July;6(6) 2348-2359. doi: 10.3390/nu6062348)

SOD B Extramel contains the high activity bioactive SuperOxide Dismutase (SOD) that fights one of the major consequences of stress and mental disorders: oxidative stress. Chronic stress,decreased cognitive performances and physical fatigue are associated with an over-production of Reactive Oxygen Species. –The results showed a significant reduction of stress and physical fatigue, as well as an improvement in cognitive performances and over all life quality. The double-blind, placebo-controlled trial was performed on 61 people. Participants reported a reduction of stress (-8.8 percent) and physical fatigue (-9.4 percent), as well as improved cognitive performances (13.9 percent) after three months of oral supplementation at 10 mg/day.—More than 450 million people worldwide have a mental disorder, according to the World Health Organization. SOD B Extramel represents another solution to the growing segment of stress and cognition. In 2008, it was awarded the European Anti-Stress Promising Ingredient of the Year Award

Recipe—THE simplest way to increase SOD is by combining Zinc ( citrate) 15mgs to copper 1mg—this would be the easiest way to increase SOD levels

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[F1]Or in todays times a polymer material as well as nanoparticle—an example would be vaccines or other methods of delivery or containment

[F2]In the case of internal exposure the same principles apply such as formulations ( TSP –Borax Peroxide- MMS-Turpentine-Vitamin C-Iodine-Herbals and Botanicals that can have an impact on the removal-exposure time –would be indicating the amount of time to utilize the components—Temperature can be utilize by either swear lodges—heat from saunas—hot tubs—baths—mechanical activity can be utilized as devices that can either send frequencies or signals or currents through the body

[F3]Which would be to our organs-tissues—skeletal structure

[F4]Why is this not alarming or surprising—do you ever get the feeling your being involved in a experiment—and you happen to be the experiment!!!

[F5]Medical uses—
Epinephrine vial 1mg (Adrenalin)

Adrenaline is used to treat a number of conditions including: cardiac arrest, anaphylaxis, and superficial bleeding.[20] It has been used historically for bronchospasm and hypoglycemia, but newer treatments for these that are selective for beta2 adrenoceptors, such as salbutamol, a synthetic epinephrine derivative, are currently preferred. Currently the maximum recommended daily dosage for patients in a dental setting requiring local anesthesia with a peripheral vasoconstrictor is 10 mg/lb of total body weight [2

[F6]One may want to maintain a study intack of tyrosine or phenyalanine to assist in the sustaining of proper adrenal function as well as B% and vitamin A

[F7]Part of the reason you cannot see them is that there size is a factor and what they are being mixed with in regard to aluminum—titanium dioxide—which actually camoflauges the particles

[F8]I find this –playing ignorant to the cause is ridiculously deceiving—almost to a level of playing innocent—when in fact they know there are self replicating and smart dust nanoparticles airborne not to mention the manufatucring processes that use the nano is leaking out of there containment

[F9]As you can see they are omitting out of the explanation aerosoling the skies with chemtrails which are nanosmart dust programmable material that self replicates and the results of this exposure can be seen on the trees and plants and insects

[F10]This is called pollen

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Show of the Month September 13 2014

Allergic reaction to antibiotic residues in foods? You may have to watch what your fruits and veggies eat
How skin falls apart- Pathology of autoimmune skin disease revealed at the nanoscale
Vibration may help heal chronic wounds
Long-term use of pills for anxiety and sleep problems may be linked to Alzheimer’s
Copper as a Anti Viral

Liposome research meets nanotechnology to improve cancer treatment

Stealth Liposomes

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Allergic reaction to antibiotic residues in foods? You may have to watch what your fruits and veggies eat
Source-American College of Allergy, Asthma and Immunology (ACAAI)

People with food allergies always have to watch what they eat. Now, they may have to watch what their fruits and vegetables eat, as it seems it’s possible to have an allergic reaction to antibiotic residues in food.–An article published in the September issue of Annals of Allergy, Asthma and Immunology, the scientific publication of the American College of Allergy, Asthma and Immunology (ACAAI), examines the case of a 10 year-old girl who had an anaphylactic (severely allergic) reaction after eating blueberry pie. Although she had a medical history of asthma and seasonal allergies, and known anaphylaxis to penicillin and cow’s milk, she wasn’t known to be allergic to any of the ingredients in the pie.–After weeks of testing on both the young girl and a sample of the pie, the article authors decided that what had caused the reaction was a streptomycin-contaminated blueberry. Streptomycin, in addition to being a drug used to fight disease, is also used as a pesticide in fruit, to combat the growth of bacteria, fungi, and algae.–“As far as we know, this is the first report that links an allergic reaction to fruits treated with antibiotic pesticides,” said allergist Anne Des Roches, MD,FRCP, lead study author. “Certain European countries ban the use of antibiotics for growing foods, but the United States and Canada still allow them for agricultural purposes.”-[F1] The authors note that new regulations from the Food and Drug Administration may help to reduce antibiotic contaminants in food, which will help reduce antibiotic resistance and may also help reduce this type of event.–“This is a very rare allergic reaction,” said allergist James Sublett, MD, ACAAI president-elect. “Nevertheless, it’s something allergists need to be aware of and that emergency room personnel may need to know about in order to help determine where anaphylactic reactions may arise. Anyone who is at risk for a life-threatening allergic reaction should always carry epinephrine[F2] . They also need to know how to use their epinephrine [F3] in an emergency situation.”–Story Source–The above story is based on materials provided by American College of Allergy, Asthma and Immunology (ACAAI). Note: Materials may be edited for content and length

Recipe—if one has allergies sustaining a good dose of vitamin C will block or minimize the histamine release which may trigger a allergic response and sustain using nettle leaf or root as tea—utilizing as well pine bark –quercitrin and zinc ( which all would support the bronchrial area and strengthen and support the lungs as well from infection ) also things like feverfew—methionine – and Korean ginseng ( minimize use with young people since it can be overpowering) and aloe vera

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How skin falls apart- Pathology of autoimmune skin disease revealed at the nanoscale
Date:

September 10, 2014

Source:

University at Buffalo

UB researchers have pinpointed important changes in cellular behavior that occur in Pemphigus Vulgaris, the rare, blistering skin disease —-University at Buffalo researchers and colleagues studying a rare, blistering disease have discovered new details of how autoantibodies destroy healthy cells in skin. This information provides new insights into autoimmune mechanisms in general and could help develop and screen treatments for patients suffering from all autoimmune diseases, estimated to affect 5-10 percent of the U.S. population.–The research, published in PLoS One on Sept. 8, has the potential to help clinicians identify who may be at risk for developing Pemphigus vulgaris (PV), an autoimmune skin disorder, by distinguishing pathogenic (disease-causing) autoimmune antibodies from other nonpathogenic autoimmune antibodies.–PV results in the often painful blistering of the skin and mucous membranes. Generally treated with corticosteroids and other immunosuppressive agents, the condition is life-threatening if untreated.–“Our work represents a unique intersection between the fields of biology and engineering that allowed for entirely new investigational strategies applied to the study of clinical disease,” says Animesh A. Sinha, MD, PhD, Rita M. and Ralph T. Behling Professor and chair of the Department of Dermatology in the UB School of Medicine and Biomedical Sciences and senior author on the study.-The immediate application of the research, Sinha explains, is in helping scientists pinpoint important changes in cell behavior.–“These changes could be the differentiation of stem cells or the development of metastases in cancer or, as we are studying it, the point at which a cell is going to implode because it’s under autoimmune attack,” he says.–Sinha’s research team, in collaboration with scientists at Michigan State University, describe the use of atomic force microscopy (AFM), a technique originally developed to study nonbiological materials, to look at cell junctions and how they rupture, a process called acantholysis.–“It has been very difficult to study cell junctions, which maintain the skin’s barrier function by keeping cells attached to each other,” says Sinha. “These junctions, micron-sized spots on cell membranes, are very complex molecular structures. Their small size has made them resistant to detailed investigation.”–Sinha’s interest lies in determining what destroys those junctions in Pemphigus Vulgaris.–“We haven’t understood why some antibodies generated by the condition cause blisters and why other antibodies it generates do not,” says Sinha.–By studying the connections between skin cells using AFM and other techniques that probe cells at the nanoscale, Sinha and his colleagues report that pathogenic antibodies change structural and functional properties of skin cells in distinct ways.–“Our data suggest a new model for the action of autoantibodies in which there are two steps or ‘hits’ in the development of lesions,” says Sinha. “The first hit results in the initial separation of cells but only the pathogenic antibodies drive further intracellular changes that lead to the breaking of the cell junction and blistering.”–The researchers examined the cells using AFM, which requires minimal sample preparation and provides three-dimensional images of cell surfaces.–The AFM tip acts like a little probe, explains Sinha. When tapped against a cell, it sends back information regarding the cell’s mechanical properties, such as thickness, elasticity, viscosity and electrical potential.–“We combined existing and novel nanorobotic techniques with AFM, including a kind of nanodissection, where we physically detached cells from each other at certain points so that we could test what that did to their mechanical and biological functions,” Sinha adds.–Those data were then combined with information about functional changes in cell behavior to develop a nanomechanical profile, or phenotype, for specific cellular states.–He also envisions that this kind of nanomechanical phenotyping should allow for the development of predictive models for cellular behavior for any kind of cell.–“Ultimately, in the case of autoimmunity, we should be able to use these techniques as a high-throughput assay to screen hundreds or thousands of compounds that might block the effects of autoantibodies and identify novel agents with therapeutic potential in given individuals,” says Sinha. “Such strategies aim to advance us toward a new era of personalized medicine.”–Story Source–The above story is based on materials provided by University at Buffalo. The original article was written by Ellen Goldbaum. Note: Materials may be edited for content and length.–Journal Reference-Kristina Seiffert-Sinha, Ruiguo Yang, Carmen K. Fung, King W. Lai, Kevin C. Patterson, Aimee S. Payne, Ning Xi, Animesh A. Sinha. Nanorobotic Investigation Identifies Novel Visual, Structural and Functional Correlates of Autoimmune Pathology in a Blistering Skin Disease Model. PLoS ONE, 2014; 9 (9): e106895 DOI: 10.1371/journal.pone.0106895

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Vibration may help heal chronic wounds
Date:

March 31, 2014

Source:

University of Illinois at Chicago

Eileen Weinheimer-Haus, first author, and Timothy Koh, principal investigator.

Wounds may heal more quickly if exposed to low-intensity vibration, report researchers at the University of Illinois at Chicago.–The finding, in mice, may hold promise for the 18 million Americans who have type 2 diabetes, and especially the quarter of them who will eventually suffer from foot ulcers. Their wounds tend to heal slowly and can become chronic or worsen rapidly.–Timothy Koh, UIC professor of kinesiology and nutrition in the UIC College of Applied Health Sciences, was intrigued by studies at Stony Brook University in New York that used very low-intensity signals to accelerate bone regeneration.–“This technique is already in clinical trials to see if vibration can improve bone health and prevent osteoporosis,” Koh said.–Koh and his coworkers at UIC collaborated with Stefan Judex of Stony Brook to investigate whether the same technique might improve wound healing in diabetes. The new study, using an experimental mouse model of diabetes, is published online in the journal PLOS One.–The low-amplitude vibrations are barely perceptible to touch.–“It’s more like a buzz than an earthquake,” said Eileen Weinheimer-Haus, UIC postdoctoral fellow in kinesiology and nutrition, the first author of the study.–The researchers found that wounds exposed to vibration five times a week for 30 minutes healed more quickly than wounds in mice of a control group.–Wounds exposed to vibration formed more granulation tissue, a type of tissue important early in the wound-healing process. Vibration helped tissue to form new blood vessels — a process called angiogenesis — and also led to increased expression of pro-healing growth factors and signaling molecules called chemokines, Weinheimer-Haus said.–“We know that chronic wounds in people with diabetes fail to form granulation tissue and have poor angiogenesis, and we believe these factors contribute to their wounds’ failure to heal,” said Koh. He and his colleagues want to determine whether the changes they see in cell populations and gene expression at wound sites underlie the observed improvement in healing.—“The exciting thing about this intervention is how easily it could be translated to people,” Koh said. “It’s a procedure that’s non-invasive, doesn’t require any drugs, and is already being tested in human trials to see if it’s protective of bone loss.” A clinical study, in collaboration with Dr. William Ennis, director of the Wound Healing Clinic at UIC, is planned, Koh said.–Story Source–The above story is based on materials provided by University of Illinois at Chicago. The original article was written by Jeanne Galatzer-Levy. Note: Materials may be edited for content and length.–Journal Reference-Eileen M. Weinheimer-Haus, Stefan Judex, William J. Ennis, Timothy J. Koh. Low-Intensity Vibration Improves Angiogenesis and Wound Healing in Diabetic Mice. PLoS ONE, 2014; 9 (3): e91355 DOI: 10.1371/journal.pone.0091355

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Long-term use of pills for anxiety and sleep problems may be linked to Alzheimer’s
Date:

September 9, 2014

Source:

BMJ-British Medical Journal

Taking benzodiazepines — widely prescribed drugs to treat anxiety and insomnia — is associated with an increased risk of developing Alzheimer’s disease, particularly for long-term users, suggests a new study.—Taking benzodiazepines — widely prescribed drugs to treat anxiety and insomnia — is associated with an increased risk of developing Alzheimer’s disease, particularly for long-term users, suggests a new study.—The researchers warn that unwarranted long-term use should be considered a public health concern. Dementia currently affects about 36 million people worldwide and this number is expected to double every 20 years, reaching 115 million by 2015. Although a increased risk of dementia has been identified in benzodiazepine users, the nature of this association, whether causal or not, remains unclear.–So a team of researchers based in France and Canada set out to investigate the relationship between the risk of Alzheimer’s disease and benzodiazepine exposure over a several years, as well as a potential dose-response relationship.–Using data from the Quebec health insurance program database (RAMQ), they tracked the development of Alzheimer’s disease in a sample of elderly residents living in Quebec, Canada who had been prescribed benzodiazepines.–Over a period of at least six years, they identified 1,796 cases of Alzheimer’s disease. They then compared each case with 7,184 healthy people matched for age, sex, and duration of follow-up.–Results show that past use of benzodiazepines for three months or more was associated with an increased risk (up to 51%) of Alzheimer’s disease. The strength of association increased with longer exposure and with use of long-acting benzodiazepines rather than short-acting ones.–Further adjustment for symptoms that might indicate the start of dementia, such as anxiety, depression or sleep disorders, did not meaningfully alter the results.–In this large case-control study, benzodiazepine use was associated with an increased risk of Alzheimer’s disease, say the authors. They emphasise that the nature of the link is still not definitive, but say the stronger association seen with long-term exposures “reinforces the suspicion of a possible direct association, even if benzodiazepine use might also be an early marker of a condition associated with an increased risk of dementia[F8] .”–Benzodiazepines are “indisputably valuable tools for managing anxiety disorders and transient insomnia” they write, but warn that treatments “should be of short duration and not exceed three months.”–They conclude that their findings are of “major importance for public health, especially considering the prevalence and chronicity of benzodiazepine use in elderly populations and the high and increasing incidence of dementia in developed countries.”–In view of the evidence, they conclude that “it is now crucial to encourage physicians to carefully balance the benefits and risks when initiating or renewing a treatment with benzodiazepines and related products in elderly patients.”–In an accompanying editorial, Professor Kristine Yaffe of the University of California at San Francisco and Professor Malaz Boustani of the Indiana University Center for Aging Research, point out that in 2012 the American Geriatrics Society updated its list of inappropriate drugs for older adults to include benzodiazepines, precisely because of their unwanted cognitive side effects.–Yet almost 50% of older adults continue to use these drugs, they say. And without any formal monitoring system, the potential long term consequences on brain health are likely to be missed, adding to the growing prevalence of cognitive impairment among older people, they suggest.–Given the expanding numbers of older people likely to be treated with several drugs at a time, and/or who are at risk of Alzheimer’s disease, this gap needs to be plugged, they say.–Story Source–The above story is based on materials provided by BMJ-British Medical Journal. Note: Materials may be edited for content and length.-Journal References–S. Billioti de Gage, Y. Moride, T. Ducruet, T. Kurth, H. Verdoux, M. Tournier, A. Pariente, B. Begaud. Benzodiazepine use and risk of Alzheimer’s disease: case-control study. BMJ, 2014; 349 (sep09 2): g5205 DOI: 10.1136/bmj.g5205 -K. Yaffe, M. Boustani. Benzodiazepines and risk of Alzheimer’s disease. BMJ, 2014; 349 (sep09 6): g5312 DOI: 10.1136/bmj.g5312

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Copper as a Anti Viral

In 1964, Yamamoto and colleagues [53] reported on the inactivation of bacteriophages by copper. Jordan and Nassar [54] in 1971 showed that copper (0.2 mg/l) present as copper carbonate or colloidal copper inactivated infectious bronchitis virus. Totsuka and Ohtaki [55] in 1974 showed that the effect of copper sulfate on poliovirus RNA is proportional to its concentration, and that most amino acids except cysteine had a protective effect as did Fe2+ and Al3+. Similarly, Coleman and colleagues [56] in 1974 reported that herpes simplex virus (HSV) type I was quite sensitive to silver. Sagripanti and colleagues [57,58] found in 1992 that cupric and ferric ions were by themselves able to inactivate five enveloped or nonenveloped, single- or double-stranded DNA or RNA viruses (phi X174, T7, phi 6, Junin, and HSV). The metals were even more effective than glutaraldehyde in inactivating the viruses. The metal virucidal effect was enhanced by the addition of peroxide, particularly for Cu2+. In every case, the viruses were more resistant to iron-peroxidase than copper-peroxidase on a metal concentration basis. The inactivation of HSV by copper was enhanced by the following reducing agents at the indicated relative level: ascorbic acid >> hydrogen peroxide > cysteine. Treatment of HSVinfected cells with combinations of Cu2+ and ascorbate completely inhibited virus plaque formation to below 0.006% of the infectious virus input. The logarithm of the surviving fraction of HSV mediated by 1 mg of Cu2+ per liter and 100 mg of reducing agent per liter followed a linear relationship with reaction time. The kinetic rate constant for each reducing agent was – 0.87 min-1 (r = 0.93) for ascorbate[F9] , -0.10 min-1 (r = 0.97) for hydrogen peroxide, and -0.04 min-1 (r = 0.97) for cysteine. The protective effects of metal chelators and catalase, the lack of effect of superoxide dismutase, and the partial protection conferred by free-radical scavengers suggest that the mechanism of copper-mediated HSV inactivation is similar to that reported for copper mediated DNA damage [59].Sagripanti and Lightfoote [60] reported that Human Immunodeficiency Virus Type 1 (HIV-1) was inactivated by both cupric or ferric ions when the virus was free in solution and also 3 hr after cell infection. Fifty percent inactivation of cell-free HIV-1 was achieved with Cu2+ at a concentration between 0.16 and 1.6 mM, or by 1.8 to 18 mM Fe3+. Thus, the dose to inactivate 50% of infectious HIV-1 (IC50) by Cu2+ or Fe3+ is higher than that reported for glutaraldehyde (0.1 mM), for sodium hypochlorite (1.3 mM), and for sodium hydroxide (11.5 mM). It is however significantly lower than that required for HIV-1 inactivation by ethanol (360 mM). Treatment of infected cells for 30 min at 20°C with 6 mM Cu2+ or Fe3+ completely inhibited the formation of syncytia and the synthesis of virus-specific p24 antigen in HIV-infected cells, while still preserving cell viability. We have recently reported the use of copper in free flow filters that deactivate HIV-1 and West Nile Virus. The copper filters reduced the infectious titers of both viruses by 5 to 6 log [19]. Wong et al [61] reported a 106-fold reduction in bacteriophage R17 infectivity due to RNA degradation in the presence of both ascorbate and Cu2+. A study published in 2001 reported on the inactivation of poliovirus and bacteriophage MS-2 in copper pipes containing tap water as a result of a synergistic effect between copper and free chlorine [62]. It was found that the log reduction/hour of the bacteriophage MS-2 in the presence of 400 μg/l of leached copper was 0.385, in 20 mg/l free chlorine 7.605 and with both copper and chlorine, 10.906. This suggests that an oxidizing agent such as chlorine or hydrogen peroxide is necessary to break open the virus protein coat and allow the copper to bind to and denature the nucleic acid. The International Copper Association, Ltd. investigated the effects of copper on the survival of waterborne viruses [63]. They found that poliovirus was completely inactivated by copper sulfate (20 mg/l) in the presence of hydrogen peroxide (10 μM), confirming observations of a synergistic effect of copper ions in the presence of oxidizing agents. The effect was reduced by the presence of a protective agent, L-histidine. Other proposed protective agents, disodium hydrogen orthophosphate, bovine serum albumin and catalase, were found to be relatively ineffective. Similarly, they found that copper reduced coxsackie virus types B2 & B4, echovirus 4 and simian rotavirus SA11 infectivity by over 98%. They concluded that there does not appear to be any significant difference between the capacity of copper to inhibit the different types of virus tested. The polio, coxsackie and echo viruses may be expected to behave similarly as they are have a similar size and are common members of the enterovirus group. However, the rotaviruses are considerably larger (75 nm diameter as opposed to 28 nm) and belong to the Reovirus group, a completely different family of viruses. They thus suggest that whatever mechanism is removing or inactivating the viruses, it is not dependent on subtle properties associated with the surface of the viruses. In another study, the efficacy of copper and silver ions, in combination with low levels of free chlorine, was evaluated for the disinfection of hepatitis A virus, human rotavirus, human adenovirus, and poliovirus in water [64]. There was little inactivation of Hepatitis A virus and human rotavirus under all conditions. Poliovirus showed more than a 4 log titer reduction in the presence of copper and silver combined with 0.5 mg of free chlorine per liter or in the presence of 1 mg of free chlorine per liter alone. Human adenovirus remained active longer than poliovirus with the same treatments, although it remained active significantly less than hepatitis A virus and human rotavirus. The addition of 700 μg of copper and 70 μg of silver per liter did not enhance the inactivation rates after exposure to 0.5 or 0.2 mg of free chlorine per liter, although on some occasions it produced a level of inactivation similar to that induced by a higher dose of free chlorine alone. This data indicates that copper and silver ions alone in water systems may not provide a reliable alternative to high levels of free chlorine for the disinfection of viral pathogens.

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Liposome research meets nanotechnology to improve cancer treatment
Date:

September 11, 2014

Source:

Boise State University

In the race to find more effective ways to treat cancer, Boise State University biophysicist Daniel Fologea is working outside the rules of general mathematics that say one plus one equals two. In his world, one plus one adds up to a whole lot more.—While radiotherapy can precisely target just the tumor site, systemic chemotherapy spreads a wide net, sending drugs speeding throughout the entire body in an attempt to kill cancer cells while also killing many healthy cells. Neither of these methods is highly effective when applied alone, therefore separated sessions of chemo and radiotherapy are required when fighting against solid tumors.–Reports have shown that ideally, both methods would be employed at the same time. But doing so produces levels of toxicity that often are deadly[F1] . To reduce the remote toxicity inherent to chemotherapy, the drugs can be administered into solid tumors by using liposomes, which are nanoscale vesicles made from fats and loaded with anti-cancer drugs. Liposomes self-accumulate within the tumor but the loaded drugs will be released very slowly from their encasing.—-A new patent awarded to Fologea, a professor in the Department of Physics, and co-researchers from the University of Arkansas in August 2014 holds promise of a way to combine the oomph of chemotherapy with the precision of radiotherapy, without harm to healthy cells.—In the new approach, said Fologea, “The liposomes are designed to release their precious cargo upon exposure to x-ray[F2] . Not only does this target where the medication goes, it also allows for a huge concentration of the drug to be released [F3] at once at the tumor site, thus increasing its efficacy. In addition, this combined modality of treatment employing concomitant radio and chemotherapy is supra-additive, which means it is several times more efficient than each therapy applied alone.”–Here’s how it works: liposomes have small scintillating[F4] nanoparticles embedded within them. When hit with the x-ray, they emit ultraviolet (UV) light. UV light triggers the release of Ca2+ entrapped into a photolabile cage inside the liposomes. The free Ca2+ activates an enzyme called phospholipase A2 that starts chewing the fats in the wall of the liposomes and triggers the fast release of the drug.–Now that they have a patent on the technique, researchers still expect several years of testing before the method is approved and available for cancer patients.–In the meantime, Fologea completed the initial phase of another method to provide similar results by using only materials previously approved by the FDA for treatment of cancer and other diseases. This approach will pave the way for earlier translational studies.

Working with Boise State biology professor Cheryl Jorcyk, he is looking for ways to put antibodies on the surface of the liposome, allowing them to recognize and attack cancer cells that are circulating in the body. A distinct approach to develop liposomes useful for treatment of diabetes is under development with Boise State biology professor Denise Wingett.—Story Source–The above story is based on materials provided by Boise State University. The original article was written by Kathleen Tuck. Note: Materials may be edited for content and length.

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Stealth Liposomes

Further advances in liposome research have been able to allow liposomes to avoid detection by the body’s immune system, specifically, the cells of reticuloendothelial system (RES). These liposomes are known as “stealth liposomes”, and are constructed with PEG[F5] (Polyethylene Glycol) [F6] studding the outside of the membrane. The PEG coating, which is inert in the body, allows for longer circulatory life for the drug delivery mechanism. However, research currently seeks to investigate at what amount of PEG coating the PEG actually hinders binding of the liposome to the delivery site. In addition to a PEG coating, most stealth liposomes also have some sort of biological species attached as a ligand to the liposome in order to enable binding via a specific expression on the targeted drug delivery site. These targeting ligands could be monoclonal antibodies (making an immunoliposome), vitamins, or specific antigens. Targeted liposomes can target nearly any cell type in the body and deliver drugs that would naturally be systemically delivered. Naturally toxic drugs can be much less toxic if delivered only to diseased tissues. Polymersomes, morphologically related to liposomes, can also be used this way.

In case of tumor development, certain anticancer drugs such as doxorubicin (Doxil) and daunorubicin are provided through liposomes. Liposomal cisplatin has received orphan drug designation for pancreatic cancer from EMEA

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[F1]Cancer therapy from the medical is dangerious

[F2]This is still not a good idea anything nano never reacts the same way chemically and to addd Xray to this is again counter productive —

[F3]Anything nano would not need to be with anything high volume

[F4]scintillating [F4] :brilliantly lively, stimulating, or witty

[F5]PEG compounds themselves show some evidence of genotoxicity vi,vii and if used on broken skin can cause irritation and systemic toxicity.— Also, PEG functions as a “penetration enhancer,” increasing the permeability of the skin to allow greater absorption of the product — including harmful ingredients.

[F6]Depending on manufacturing processes, PEGs may be contaminated with measurable amounts of ethylene oxide and 1,4-dioxane. i The International Agency for Research on Cancer classifies ethylene oxide as a known human carcinogen and 1,4-dioxane as a possible human carcinogen. Ethylene oxide can also harm the nervous system ii and the California Environmental Protection Agency has classified it as a developmental toxicant based on evidence that it may interfere with human development. iii 1,4-dioxane is also persistent. In other words, it doesn’t easily degrade and can remain in the environment long after it is rinsed down the shower drain

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A liposome encapsulates a region of aqueous solution inside a hydrophobic membrane; dissolved hydrophilic solutes cannot readily pass through the lipids. Hydrophobic chemicals can be dissolved into the membrane, and in this way liposome can carry both hydrophobic molecules and hydrophilic molecules

Liposomes can also be designed to deliver drugs in other ways. Liposomes that contain low (or high) pH can be constructed such that dissolved aqueous drugs will be charged in solution (i.e., the pH is outside the drug’s pI range). As the pH naturally neutralizes within the liposome (protons can pass

Recipe—take any mineral or antioxidant and fuse this is any sunflower lecithin or even fats —and when you fused them in a blender the intake of what your using will not only improve but released longer in the system

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[F1]Cancer therapy from the medical is dangerious

[F2]This is still not a good idea anything nano never reacts the same way chemically and to addd Xray to this is again counter productive —

[F3]Anything nano would not need to be with anything high volume

[F4]scintillating [F4] :brilliantly lively, stimulating, or witty

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[F1]Why is this not alarming or surprising—do you ever get the feeling your being involved in a experiment—and you happen to be the experiment!!!

[F2]Medical uses—
Epinephrine vial 1mg (Adrenalin)

Adrenaline is used to treat a number of conditions including: cardiac arrest, anaphylaxis, and superficial bleeding.[20] It has been used historically for bronchospasm and hypoglycemia, but newer treatments for these that are selective for beta2 adrenoceptors, such as salbutamol, a synthetic epinephrine derivative, are currently preferred. Currently the maximum recommended daily dosage for patients in a dental setting requiring local anesthesia with a peripheral vasoconstrictor is 10 mg/lb of total body weight [2

[F3]One may want to maintain a study intack of tyrosine or phenyalanine to assist in the sustaining of proper adrenal function as well as B% and vitamin A

[F4]Part of the reason you cannot see them is that there size is a factor and what they are being mixed with in regard to aluminum—titanium dioxide—which actually camoflauges the particles

[F5]I find this –playing ignorant to the cause is ridiculously deceiving—almost to a level of playing innocent—when in fact they know there are self replicating and smart dust nanoparticles airborne not to mention the manufatucring processes that use the nano is leaking out of there containment

[F6]As you can see they are omitting out of the explanation aerosoling the skies with chemtrails which are nanosmart dust programmable material that self replicates and the results of this exposure can be seen on the trees and plants and insects

[F7]This is called pollen

[F8]Don’t you like the dance—they are saying a 51% increase on one hand and on the other they are saying this may be and there are strong indicators butt…and there is always a butttt

[F9]Vitamin C mix

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Show of the Month September 20 2014

Vitamin E intake Critical to being healthy—- Vitamin E
Four new quassinoids from the roots of Eurycoma longifolia Jack

Anti-tumor activity of Eurycoma longifolia root extracts against K-562 cell line

Aspartame Toxicity Information

Ultrafine Particles Cross Cellular Membranes by Nonphagocytic Mechanisms in Lungs and in Cultured Cells
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Vitamin E intake Critical to being healthy
Date:

September 15, 2014

Source:

Oregon State University

Sunflower seeds and oil are a particularly good dietary source of vitamin E.–Amid conflicting reports about the need for vitamin E and how much is enough, a new analysis published today suggests that adequate levels of this essential micronutrient are especially critical for the very young, the elderly, and women who are or may become pregnant.-A lifelong proper intake of vitamin E is also important, researchers said, but often complicated by the fact that this nutrient is one of the most difficult to obtain through diet alone. It has been estimated that only a tiny fraction of Americans consume enough dietary vitamin E to meet the estimated average requirement.-Meanwhile, some critics have raised unnecessary alarms about excessive vitamin E intake while in fact the diet of most people is insufficient[F1] , said Maret Traber, a professor in the College of Public Health and Human Sciences at Oregon State University, principal investigator with the Linus Pauling Institute and national expert on vitamin E.-“Many people believe that vitamin E deficiency never happens,” Traber said. “That isn’t true. It happens with an alarming frequency both in the United States and around the world. But some of the results of inadequate intake are less obvious, such as its impact on things like nervous system and brain development, or general resistance to infection.”-Some of the best dietary sources of vitamin E — nuts, seeds, spinach, wheat germ and sunflower oil — don’t generally make the highlight list of an average American diet. One study found that people who are highly motivated to eat a proper diet consume almost enough vitamin E, but broader surveys show that 90 percent of men and 96 percent of women don’t consume the amount currently recommended, 15 milligrams per day for adults.–In a review of multiple studies, published in Advances in Nutrition, Traber outlined some of the recent findings about vitamin E. Among the most important are the significance of vitamin E during fetal development and in the first years of life; the correlation between adequate intake and dementia later in life; and the difficulty of evaluating vitamin E adequacy through measurement of blood levels alone.

Findings include:

Inadequate vitamin E is associated with increased infection, anemia, stunting of growth and poor outcomes during pregnancy for both the infant and mother.
Overt deficiency, especially in children, can cause neurological disorders, muscle deterioration, and even cardiomyopathy.
Studies with experimental animals indicate that vitamin E is critically important to the early development of the nervous system in embryos, in part because it protects the function of omega-3 fatty acids, especially DHA, which is important for brain health. The most sensitive organs include the head, eye and brain.
One study showed that higher vitamin E concentrations at birth were associated with improved cognitive function in two-year-old children.
Findings about diseases that are increasing in the developed world, such as non-alcoholic fatty liver disease and diabetes, suggest that obesity does not necessarily reflect adequate micronutrient intake.
Measures of circulating vitamin E levels in the blood often rise with age as lipid levels also increase, but do not prove an adequate delivery of vitamin E to tissues and organs.
Vitamin E supplements do not seem to prevent Alzheimer’s disease occurrence, but have shown benefit in slowing its progression.
A report in elderly humans showed that a lifelong dietary pattern that resulted in higher levels of vitamins B,C, D and E were associated with a larger brain size and higher cognitive function.
Vitamin E protects critical fatty acids such as DHA throughout life, and one study showed that people in the top quartile of DHA concentrations had a 47 percent reduction in the risk of developing all-cause dementia.
“It’s important all of your life, but the most compelling evidence about vitamin E is about a 1000-day window that begins at conception,” Traber said. “Vitamin E is critical to neurologic and brain development that can only happen during that period. It’s not something you can make up for later.”–Traber said she recommends a supplement for all people with at least the estimated average requirement of vitamin E, but that it’s particularly important for all children through about age two; for women who are pregnant, nursing or may become pregnant; and for the elderly.-Story Source–The above story is based on materials provided by Oregon State University. Note: Materials may be edited for content and length.—Journal Reference–Maret Traber. Vitamin E Inadequacy in Humans: Causes and Consequences. Advances in Nutrition, September 2014

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Vitamin E1
Maret G. Traber*
1. School of Biological & Population Health Sciences and the Linus Pauling Institute, Oregon State University, Corvallis, OR
Danny Manor
+ Author Affiliations

1. Departments of Nutrition and of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH
Vitamin E refers to the plant-derived, lipid-soluble antioxidants: tocopherols and tocotrienols. They terminate the chain reaction of lipid peroxidation. Vitamin E biological activity is different from its antioxidant activity, and there is a preference for α-tocopherol. This preference is achieved through the selective degradation and excretion of other vitamin E forms and the selective retention of α-tocopherol, mediated by the hepatic α-tocopherol transfer protein (α-TTP). Hepatic α-TTP facilitates the selective incorporation of α-tocopherol into circulating lipoproteins that distribute the vitamin to nonhepatic tissues. α-TTP is therefore considered to be the major regulator of vitamin E status in humans.

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Deficiencies:
Vitamin E deficiency occurs in humans as a result of genetic abnormalities in α-TTP or in lipoprotein synthesis or occurs secondary to fat malabsorption. Genetic defects in α-TTP are associated with a characteristic syndrome, ataxia with vitamin E deficiency (AVED). More than 20 mutations in α-TTP have been identified in human AVED patients.

Diet recommendations:
The vitamin E dietary reference intake (DRI) for adult men and women (and individuals 14–18 y) was set in 2000 at a daily estimated average requirement (EAR) of 12 mg α-tocopherol and an recommended daily allowance (RDA) of 15 mg.[F2] There are no increases for pregnancy, but for lactation the RDA is 19 mg/d. The adequate intake (AI) for infants (0–6 mo) was estimated to be 4 mg and for 7 through 12 mo to be 5 mg. The RDA for children 1 to 3 y is 6 mg; for those 4–8 y, it is 7 mg and 11 mg for those 9 to 13 y. –Plant-synthesized α-tocopherol is RRR-α-tocopherol; chiral carbons are in the R-conformation at positions 2, 4′, and 8′. Chemical synthesis results in an equal mixture of 8 different stereoisomers (RRR, RSR, RRS, RSS, SRR, SSR, SRS, SSS) that is called all-rac-α-tocopherol. The 2 position of α-tocopherol, the junction of the ring and tail, is critical for in vivo α-tocopherol vitamin activity. Only 2R forms meet human requirements.

Food sources:
Major dietary vitamin E sources, as commonly eaten portions [USDA National Nutrient Database for Standard Reference, Release 24 (January 2012; http://www.nal.usda.gov/fnic/foodcomp/search/)] are fortified ready-to-eat cereals; nuts, especially almonds; seeds, such as sunflower seeds; greens, such as spinach; and vegetable oils, especially sunflower and safflower.

Clinical uses:
Humans with defects in the TTPA gene (encoding α-TTP) have extraordinarily low (1/100 of normal) plasma vitamin E concentrations, but if they are given vitamin E supplements, plasma concentrations normalize within hours; if supplementation is halted, plasma vitamin E concentrations decrease within days to deficient levels. A daily α-tocopherol dose (800–1200 IU) is usually sufficient to prevent further deterioration of neurologic function, and in some cases, improvements have been noted. Postmortem analysis of a brain from a vitamin E–supplemented AVED patient demonstrated vitamin E accumulation and prevention of Purkinje cell loss. -Vitamin E deficiency due to impaired lipoprotein synthesis or fat malabsorption syndromes (e.g., abetalipoprotenemia, cystic fibrosis, short bowl disorder, choleastasis, and inherited defects in bile acid synthesis) is also treated with daily vitamin E supplements (100 mg/kg body weight). –The benefit of vitamin E supplements in individuals who are not vitamin E deficient is controversial. In the elderly, impaired immune function was improved with vitamin E supplementation. Patients with macular degeneration benefited from a supplement cocktail that included vitamin E. Vitamin E supplements have decreased heart attack risk in those with the haptoglobin 2–2 genotype, which results in a dysfunctional protein and causes increased oxidation by free heme. A follow-up study of the Alpha-Tocopherol Beta-Carotene Cancer Prevention trial showed that men at baseline who consumed 12 mg α-tocopherol daily (equivalent to the vitamin E EAR) had considerably lower risk of total and cause-specific mortality 13 y later.

Toxicity:
The upper limit of tolerable intakes (UL) is 1000 mg/day, equivalent to 1100 iu synthetic or 1500 iu natural vitamin E. The UL was based on the adverse effect of an increased bleeding tendency observed in rat studies. This tendency to bleed was found to have beneficial effects in preventing venous thrombosis in a trial of vitamin E supplements in women. –Subsequent to the publication of the DRI, there have been several meta-analyses evaluating the outcomes of human vitamin E supplementation trials. Thus far, there have been no uniform mechanisms identified for the claim in meta-analyses of increased mortality associated with vitamin E supplements.

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Recent research
Vitamin E in the central nervous system:
Vitamin E deficiency manifests primarily as cerebellar ataxia in humans, underscoring the unique sensitivity of the central nervous system to oxidative stress. Interestingly, α-TTP is expressed in the cerebellum. α-Tocopherol supplementation in α-TTP knockout mice normalizes its status in all tissues except the brain. Thus, a unique relationship exists between localized vitamin E levels, expression of α-TTP, oxidative stress, and optimal cerebellar function. The detailed description of this relationship and the molecular mechanisms that underlie it are critical research questions.

Homeostasis:
An important fundamental question in vitamin E biology is “are there compensatory mechanisms in response to oxidative stress to increase α-tocopherol concentrations or distribution?.” Provocatively, brain α-TTP expression levels respond to oxidative stress, [F3] but liver α-TTP does not.

Fertility:
Vitamin E was discovered as an essential dietary factor for reproductive health in female rodents. Surprisingly, very little is known regarding human vitamin E status and reproductive health. In mice, α-tocopherol sufficiency is essential for placentation. α-TTP is expressed in the uterine wall of pregnant female mice and in the human placenta. Research in this field is of great importance because 96% of U.S. women do not meet vitamin E EAR.

Interactions with other nutrients:
The untoward effects of vitamin E supplements on blood clotting may result from vitamin E and K interactions because these supplements increase undercarboxylation of prothrombin, suggesting lower vitamin K activity. More than 90% of dietary, plant-derived vitamin K is phylloquinone (vitamin K1) with a 20-carbon phytyl side chain that is identical to that of tocopherol. Phylloquinone is converted to menadione and then to MK-4 (extrahepatic tissue vitamin K). Based on their similar structures, vitamins E and K likely share the same pathways for metabolism and excretion. Possible mechanisms for the vitamin E and K interaction have been proposed, but none have been proven.

The interactions of vitamins E and C are likely dependent on their roles as antioxidants; vitamin C can regenerate tocopherol from the tocopheroxyl radical. When α-tocopherol kinetics were evaluated in cigarette smokers, smokers had higher F2-isoprostane concentrations and faster plasma α-tocopherol disappearance rates than nonsmokers. When smokers received vitamin C supplementation (500 mg twice daily) for 2 weeks, α-tocopherol disappearance rates were normalized. Thus, in smokers with greater oxidative stress, additional vitamin C is needed to restore the α-tocopheroxyl radical to its reduced form. Importantly, vitamin C supplementation did not change F2-isoprostane concentrations, showing that α-tocopherol does not prevent radical formation or the initial oxidation of fatty acids, but halts the chain reaction of lipid peroxidation.

For further information
Food and Nutrition Board, and Institute of Medicine. Dietary reference intakes for vitamin C, vitamin E, selenium, and carotenoids. Washington, DC: National Academy Press; 2000.

Institute of Medicine. Dietary reference intakes: the essential guide to nutrient requirements. Washington, DC: National Academy Press; 2006.

Manor D, Morley S. The alpha-tocopherol transfer protein. Vitam Horm. 2007;76:45–65.

Traber MG, Atkinson J. Vitamin E, antioxidant and nothing more. Free Radic Biol Med. 2007;43:4–15.

Traber MG, Stevens JF. Beneficial effects from a mechanistic perspective. Free Radic Biol Med. 2011;51:1000–13.

Traber MG. Vitamin E. In: Erdman J, Macdonald I, Zeisel S, editors. Present knowledge in nutrition. 9th ed. Washington, DC: ILSI Press.

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Four new quassinoids from the roots of Eurycoma longifolia Jack.

Fitoterapia. 2014 Jan;92:105-10

Authors: Meng D, Li X, Han L, Zhang L, An W, Li X

Abstract
Seven compounds were isolated from the roots of Eurycoma longifolia, and characterized by comprehensive analysis of 1D and 2D NMR experiments along with single crystal X-ray diffraction. Among them, four new quassinoids were identified and three of them were diastereomers[F4] for each other. Compounds 1-7 were evaluated for cytotoxicities against HT-29, MCF-7, LOVO, BGC-823, MGC-803, HepG2, HeLa, and A549 cancer cell lines. Compounds 2 and 5 exhibited the lowest IC50 values [F5] of 24.9 μM, 11.8 μM, and 44.1 μM, 14.1 μM towards MCF-7, MGC-803 cancer cell lines, respectively, while compound 6 exhibited moderate cytotoxicity towards all the selected cancer cell lines.–PMID: 24513570 [PubMed – indexed for MEDLINE]

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Anti-tumor activity of Eurycoma longifolia root extracts against K-562 cell line: in vitro and in vivo study.

PLoS One. 2014;9(1):e83818

Authors: Al-Salahi OS, Ji D, Majid AM, Kit-Lam C, Abdullah WZ, Zaki A, Jamal Din SK, Yusoff NM, Majid AS

Eurycoma longifolia Jack has been widely used in traditional medicine for its antimalarial, aphrodisiac, anti-diabetic, antimicrobial and anti-pyretic activities. Its anticancer activity has also been recently reported on different solid tumors, however no anti-leukemic activity of this plant has been reported. Thus the present study assesses the in vitro and in vivo anti-proliferative and apoptotic potentials of E. longifolia on K-562 leukemic cell line. The K-562 cells (purchased from ATCC) were isolated from patients with chronic myelocytic leukemia (CML) were treated with the various fractions (TAF273, F3 and F4) of E. longifolia root methanolic extract at various concentrations and time intervals and the anti-proliferative activity assessed by MTS assay. Flow cytometry was used to assess the apoptosis and cell cycle arrest. Nude mice injected subcutaneously with 10(7) K-562 cells were used to study the anti-leukemic activity of TAF273 in vivo. TAF273, F3 and F4 showed various degrees of growth inhibition with IC50 [F6] values of 19, 55 and 62 µg/ml, respectively. TAF273 induced apoptosis in a dose and time dependent manner. TAF273 arrested cell cycle at G1 and S phases. Intraperitoneal administration of TAF273 (50 mg/kg) resulted in a significant growth inhibition of subcutaneous tumor in TAF273-treated mice compared with the control mice (P = 0.024). TAF273 shows potent anti-proliferative activity in vitro and in vivo models of CML and therefore, justifies further efforts to define more clearly the potential benefits of using TAF273 as a novel therapeutic strategy for CML management.–PMID: 24409284 [PubMed – indexed for MEDLINE]

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Aspartame Toxicity Information

12 East Side Dr., Suite 2-18
Concord, NH 03301
603-225-2110

Date: January 12, 2002

Please find below Evidence File #4: Reported Aspartame Toxicity Effects

Reported Aspartame Toxicity Effects
———————————–

Q. What are the reported reactions to aspartame ingestion?

How often are such effects seen?

Answer
——

==> What are the reported reactions to aspartame ingestion?

We will limit our discussion in this FAQ to reported toxicity
reactions to aspartame ingestion. Controlled studies showing
problems with aspartame ingestion will be discussed in another
FAQ. Toxicity reactions to aspartame can be divided into three
types:

1. Acute toxicity reactions occuring within 48 hours of ingestion of
an aspartame-containing product.

2. Chronic toxicity effects occuring anywhere from several days of
use to appearing a number of years (i.e., 1-20+ years) after the
beginning of aspartame use.

3. Potential toxicity effects that would be nearly impossible for
the user to recognize the link to aspartame.

In an epidemiological survey which appeared in the Journal of
Applied Nutrition (Roberts 1988), 551 persons who have
reported toxicity effects from aspartame ingestion were
surveyed. The adverse effects found cover a subset of reported
acute and chronic toxicity effects from aspartame.
What follows is a listing of the adverse health effects
which were found.

——————-
# of
people (%)
Eye
– Decreased vision and/or other eye problems 140 (25%)
(blurring, “bright flashes,” tunnel vision)
– Pain (or or both eyes) 51 (9%)
– Decreased tears, trouble with contact lens, 46 (8%)
or both
– Blindness (one or both eyes) 14 (3%)

Ear
– Tinnitus (“ringing,” “buzzing”) 73 (13%)
– Severe intolerance for noise 47 (9%)
– Marked impairment of hearing 25 (5%)

Neurologic
– Headaches 249 (45%)
– Dizziness, unsteadiness, or both 217 (39%)
– Confusion, memory loss, or both 157 (29%)
– Severe drowsiness and sleepiness 93 (17%)
– Paresthesias (“pins and needles,” “tingling”) 82 (15%)
or numbness of the limbs
– Convulsions (grand mal epileptic attacks) 80 (15%)
– Petit mal attacks and “absences” 18 (3%)
– Severe slurring of speech 64 (12%)
– Severe tremors 51 (9%)
– Severe “hyperactivity” and “restless legs” 43 (8%)
– Atypical facial pain 38 (7%)

Psychologic-Psychiatric
– Severe depression 139 (25%)
– “Extreme irritability” 125 (23%)
– “Severe anixiety attacks” 105 (19%)
– “Marked personality changes” 88 (16%)
– Recent “severe insomnia” 76 (14%)
– “Severe aggravation of phobias” 41 (7%)

Chest
– Palpitations, tachycardia (rapid heart action), 88 (16%)
of both
– “Shortness of breath” 54 (10%)
– Atypical chest pain 44 (8%)
– Recent hypertension (high blood pressure) 34 (6%)

Gastrointestinal
– Nausea 79 (14%)
– Diarrhea 70 (13%)
Associated gross blood in the stools (12)
– Abdominal pain 70 (13%)
– Pain on swallowing 28 (5%)

Skin and Allergies
– Severe itching without a rash 44 (8%)
– Severe lip and mouth reactions 29 (5%)
– Urticaria (hives) 25 (5%)
– Other eruptions 48 (9%)
– Aggravation of respiratory allergies 10 (2%)

Endocrine and Metabolic
– Problems with diabetes: loss of control; 60 (11%)
precipitation of clinical diabetes;
aggravation or simulation of diabetic
complications
– Menstrual changes 45 (6%)
Severe reduction or cessation of periods (22)
– Paradoxic weight gain 34 (5%)
– Marked weight loss 26 (6%)
– Marked thinning or loss of the hair 32 (6%)
– Aggravated hypoglycemia (low blood sugar 25 (5%)
attacks)

Other
– Frequency of voiding (day and night), burning 69 (13%)
on urination (dysuria), or both
– Excessive thirst 65 (12%)
– Severe joint pains 58 (11%)
– “Bloat” 57 (10%)
– Fluid retention and leg swelling 20 (4%)
– Increased susceptibility to infection 7 (1%)

——————-

There are other clinical reports in the scientific literature of
aspartame-caused toxicity reactions including Blumenthal (1997),
Drake (1986), Johns (1986), Lipton (1989), McCauliffe (1991),
Novick (1985), Watts (1991), Walton (1986, 1988), and Wurtman
(1985).

Many pilots appear to be particularly susceptible to the effects of
aspartame ingestion. They have reported numerous serious toxicity
effects including grand mal seizures in the cockpit (Stoddard 1995).
Nearly 1,000 cases of pilot reactions have been reported to the
Aspartame Consumer Safety Network Pilot Hotline (Stoddard 1995).
This susceptibility may be related to ingesting methanol at altitude
as suggested in a letter from Dr. Phil Moskal, Professor of
Microbiology, Biochemistry, and Pathology, Chairman of the Department
of Pathology, Director of Public Health Laboratories (Moskal 1990),
or it may simply be that some pilots tend to ingest large quantities
of aspartame during a flight. Whatever the case, numerous warnings
about aspartame dangers have appeared in piloting journals including
The Aviation Consumer (1988), Aviation Medical Bulliten (1988),
Pacific Flyer (1988), CAA General Aviation (1989), Aviation Safety
Digest (1989), General Aviation News (1989), Plane & Pilot (1990),
Canadian General Aviation News (1990), National Business Aircraft
Association Digest (NBAA Digest 1993), International Council of
Air Shows (ICAS 1995), and the Pacific Flyer (1995). Both the U.S.
Air Force’s magazine “Flying Safety” and the U.S. Navy’s magazine,
“Navy Physiology” published articles warning about the many dangers
of aspartame including the cumlative deliterious effects of methanol
and the greater likelihood of birth defects. The articles note that
the ingestion of aspartame may make pilots more susceptible to
seizures and vertigo (US Air Force 1992).

Countless other toxicity effects have been reported to the FDA (DHHS
1995), other independent organizations (Mission Possible 1996,
Stoddard 1995), and independent scientists (e.g., 80 cases of
seizures were reported to Dr. Richard Wurtman, Food (1986)).
Samples of some aspartame toxicity reactions reported on the
Internet can be found on the Aspartame (NutraSweet) Toxicity Info
Center web page:

http://www.tiac.net/users/mgold/aspartame/

Frequently, aspartame toxicity is misdiagnosed as a specific disease.
This has yet to be reported in the scientific literature, yet it has
been reported countless times to independent organizations and
scientists (Mission Possible 1994, Stoddard 1995). In other cases,
it has been reported that chronic aspartame ingestion has triggered
or worsened certain chronic illnesses. Nearly 100% of the time, the
patient and physician assume that these worsening conditions are
simply a normal progression of the illness. Sometimes that may be
the case, but many times it is chronic aspartame poisoning.

According to researchers and physicians studying the adverse
effects of aspartame, the following list contains a selection
of chronic illnesses which may be caused or worsened by the chronic,
long-term ingestion of aspartame. (Mission Possible 1994, Stoddard
1995)*:

Brain tumors Multiple sclerosis
Epilepsy Chronic faigue syndrome
Parkinson’s Disease Alzheimer’s
Mental retardation Lymphoma
Birth defects Fibromyalgia
Diabetes Arthritis (including Rheumatoid)
Chemical Sensitivities Attention Deficit Disorder

*Note: In some cases such as MS, the severe symptoms mimic the illness or exacerbate the
illness, but do not cause the disease.

Also, please note that this is an incomplete list. Clearly,
ingestion of a very slow poison (as discussed in other FAQs) is not
beneficial to anyone who has a chronic illness.

Finally, potential toxicity effects from aspartame including brain
cancer (as seen in pre-approval research) and effects on fetal brain
and nervous system development will be discussed in other FAQs.

==> How often are such effects seen?

Until recently approximately 90% of aspartame sales were in the
United States (Monsanto 1994). Other countries are now being inundated
with aspartame, but it will be some time until they begin to feel the
full effects of aspartame toxicity on the general population. Since the
U.S. has some history of significant use, we will limit the discussion
to the frequency of effects in the U.S.

There have been well over 7,000 aspartame toxicity reactions officially received by the U.S.
Food and Drug Administration between 1982 (after aspartame was first approved) until 1995
(DHHS 1993, DHHS 1995). From this figure, we can estimate the number of actual toxicity
reactions observed.

***************************************************************************

References Cited

Aviation Consumer 1988. “SafeGuard,” June 15, 1988.

Aviation Medical Bulletin 1988. “Pilots and Aspartame,”
October 1988.

Aviation Safety Digest 1989. “Aspartame — not for the
dieting pilot?” Aviation Safety Digest, ASD 142, Spring
1989 (Australia – 062/5841111).

Blumenthal, H.J., D.A. Vance, 1997, “Chewing Gum Headaches,”
Headache, Volume 37, Number 10, pages 665-666.

Butchko, Harriett H., Frank N. Kotsonis 1994. “Postmarketing
Surveillance in the Food Industry: The Aspartame Case
Study,” in Nutritional Toxicology, edited by Frank N.
Kotsonis, Maureen Macky and Jerry Hjelle, Raven Press,
Ltd., New York, c1994.

CAA General Aviation (1989). Safety Information Leaflet,
April 1989, Great Britain.

Canadian General Aviation News 1990. “Fit to fly” Canadian
General Aviation News, March 1990, page 28.

DHHS 1993. “Adverse Reactions Associated With Aspartame
Consumption,” Department of Health & Human Services
Memorandum, April 1, 1993, Reprinted in preface of
“Bittersweet Aspartame: A Diet Delusion” by Barbara
Alexander Mullarkey, NutriVoice, P.O. Box 946, Oak
Park, Illinois 60303, (708) 848-0116.

DHHS 1995. Department of Health and Human Services. “Report
on All Adverse Reactions in the Adverse Reaction
Monitoring System.” (April 20, 1995).

Drake, M.E., 1986. “Panic Attacks and Excessive Aspartame Ingestion”
(Letter), Lancet, September 13, 1986, page 631.

Food 1986. Food Chemical News, July 28, 1986, page 44.

Food 1995. “Aspartame Adverse Reaction Reports Down in 1994
From 1985 Peak: FDA,” Food Chemical News, June 12, 1995,
page 27.

GAO 1986. “Six Former HHS Employees’ Involvement in
Aspartame’s Approval,” United States General Accounting
Office, GAO/HRD-86-109BR, July 1986.

General Aviation News 1989. “NutraSweet…too good to be
true?” by Megan Hicks, General Aviation News, July 31,
1989.

Gordon, Gregory, 1987. “NutraSweet: Questions Swirl,” UPI
Investigative Report, 10/12/87. Reprinted in US Senate
U.S. Senate Committee on Labor and Human Resources,
November 3, 1987 regarding “NutraSweet Health and Safety
Concerns.” Document # Y 4.L 11/4:S.HR6.100, page 499.

ICAS 1995. “Aspartame Side Effects: Fact or Fiction?”
International Council of Air Shows, February 1995.

Johns, Donald R., 1986. “Migraine Provoked By Aspartame,” (Letter),
New England Journal of Medicine, Volume 314, August 14, 1986,
page 456.

Kessler, David A. 1993, “Introducing MEDWatch: A New
Approach to Reporting Medication and Device Adverse
Effects and Product Problems” Journal of the American
Medical Association 269:2765-68.

Lipton, Richard B., et al., 1989. “Aspartame as a Dietary Trigger of
Headache,” Headache, Volume 29, pages 90-92.

McCauliffe, D.P., K. Poitras, 1991. “Aspartame-Induced Lobular
Panniculitis,” Journal of the American Academy of Dermitology, Volume
24, page 298-300.

Mission Possible 1994. Compiled by researchers, physicians,
and artificial sweetner experts for Mission Possible, a
group dedicated to warning consumers about aspartame.
Available from Mission Possible, 9270 River Club Pkwy,
Duluth, Georgia 30155, 770-242-2599, [email protected]

Mission Possible 1996. Conversations between Betty Martini of
Mission Possible and Mark D. Gold.

Monsanto 1994. “Monsanto Annual Report,” 1994.

Moskal, Phil, 1990. Letter from Dr. Phil Moskal to George
Leighton, June 19, 1990, Reprinted in “The Deadly Deception”
Compiled by the Aspartame Consumer Safety Network for volumes
of available published information, ACSN, P.O. Box 780634,
Dallas, Texas 75378, (800) 969-6050.

NBAA Digest 1993. “Operationally Speaking” by G. Dennis
Wright, Vice President of Operations. NBAA Digest, Volume
6, Number 6, June 1993. Available from National Business
Aircraft Association, Inc., 1200 Eighteenth St., NW,
Suite 200, Washington, DC 20036-2506, (202) 783-9000.

Novick, Nelson Lee, 1995. “Aspartame-Induced Granulomatous
Panniculitis,” Annals of Internal Medicine, Volume 102, Number 2,
pages 206-207.

Pacific Flyer 1988. “This Could Save Your Life” Pacific
Flyer Aviation News, November 1988, 3355 Mission Ave.,
Oceanside, CA 92054.

Pacific Flyer 1995. “ICAS Issues Warning To Its Members About Diet
Drinks,” March 1995.

Pauli, George, 1995. FDA Center for Food Safety and Applied
Nutrition (CFSAN). Radio broadcast: “Aspartame,” The
Derek McGinty Show, WAMU Radio (88.5 FM), Brandywine
Building, The American University, Washington, DC 20016-
8082, (202) 885-1200, August 29, 1995.

Plane & Pilot 1990. “Getting High” Plane & Pilot, January
1990, page 36-37.

Roberts, H.J., 1988. “Reactions Attributed to Aspartame-
Containing Products: 551 Cases,” Journal of Applied
Nutrition, Volume 40, page 85-94.

Stoddard, Mary Nash, 1995. Conversations between Mary Nash
Stoddard of the Aspartame Consumer Safety Network and
Mark D. Gold.

Turner, James, Leonard, Rodney, 1984. Letter from Rodney E. Leonard
and James S. Turner of Community Nutrition Institute to Dr. Fank E.
Young, FDA Commissioner, September 13, 1984. Reprinted in
“Aspartame Safety Act,” Congressional Record, Volume 131, No. 106,
August 1, 1985, page S10841.

Turner, James, 1987. Testimony of James Turner, Esq.,
Community Nutrition Institute before the U.S. Senate
Committee on Labor and Human Resources, November 3, 1987
regarding “NutraSweet Health and Safety Concerns.”
Document # Y 4.L 11/4:S.HR6.100, page 316.

US Air Force 1992. “Aspartame Alert.” Flying Safety 48(5):
20-21 (May 1992).

USDA 1988. “1988 United States Department of Agriculture
Situation and Outlook Report; Sugar and Sweeteners.”
Washington, DC: U.S. Government Printing Office, pp. 51.

WSJ 1996. “Aspartame Critic Seeks More Research On Possibility of
Links to Brain Tumors,” The Wall Street Journal, November 8, 1996.

Walton, Ralph G., 1986. “Seizure and Mania After High Intake
of Aspartame,” Psychosomatics, Volume 27, page 218-220.

Walton, Ralph G., 1988. “The Possible Role of Aspartame in
Seizure Induction,” Presented at “Dietary Phenylalanine
and Brain Function.” Proceedings of the First
International Meeting on Dietary Phenylalanine and Brain
Function, Washington, D.C., May 8-10, 1987. Center for
Brain Sciences and Metabolism Charitable Trust, P.O. Box
64, Kendall Square, Cambridge, MA 02142. Reprinted in
“Dietary Phenyalalnine and Brain Function,” c1988,
Birkhauser, Boston, MA USA, page 159-162.

Watts, Richard S., 1991. “Aspartame, Headaches and Beta Blockers”
(Letter to the Editor), Headache, March, 1991, Page 181-182.

Wilson, Steve, 1985. “Sweet Suspicions,” Television
broadcast and interviews regarding aspartame. Transcript
in Congressional Record, Volume 131, No. 106, August 1,
1985, page S10826-S10827.

Wurtman, Richard J., 1985. “Aspartame: Possible Effect on
Seizure Susceptibility” (Letter), The Lancet, Volume 2, page 1060.

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Ultrafine Particles Cross Cellular Membranes by Nonphagocytic Mechanisms in Lungs and in Cultured Cells
Marianne Geiser,1 Barbara Rothen-Rutishauser,1 Nadine Kapp,1 Samuel Schürch,1,2 Wolfgang Kreyling,3 Holger Schulz,3 Manuela Semmler,3 Vinzenz Im Hof,4 Joachim Heyder,3 and Peter Gehr1

Author information ► Article notes ► Copyright and License information ►

See “Particles in Practice: How Ultrafines Disseminate in the Body” on page A758a.

See letter “Translocation of Ultrafine Particles” in volume 114 on page A211b.

This article has been cited by other articles in PMC.

Go to:

Abstract
High concentrations of airborne particles have been associated with increased pulmonary and cardiovascular mortality, with indications of a specific toxicologic role for ultrafine particles (UFPs; particles < 0.1 μm). Within hours after the respiratory system is exposed to UFPs, the UFPs may appear in many compartments of the body, including the liver, heart, and nervous system. To date, the mechanisms by which UFPs penetrate boundary membranes and the distribution of UFPs within tissue compartments of their primary and secondary target organs are largely unknown. We combined different experimental approaches to study the distribution of UFPs in lungs and their uptake by cells. In the in vivo experiments, rats inhaled an ultrafine titanium dioxide aerosol of 22 nm count median diameter. The intrapulmonary distribution of particles was analyzed 1 hr or 24 hr after the end of exposure, using energy-filtering transmission electron microscopy for elemental microanalysis of individual particles. In an in vitro study, we exposed pulmonary macrophages and red blood cells to fluorescent polystyrene microspheres (1, 0.2, and 0.078 μm) and assessed particle uptake by confocal laser scanning microscopy. Inhaled ultrafine titanium dioxide particles were found on the luminal side of airways and alveoli, in all major lung tissue compartments and cells, and within capillaries. Particle uptake in vitro into cells did not occur by any of the expected endocytic processes, but rather by diffusion or adhesive interactions.[F7] Particles within cells are not membrane bound and hence have direct access to intracellular proteins, organelles, and DNA, which may greatly enhance their toxic potential. Keywords: aerosol, erythrocytes, lungs, macrophages, microscopy, nanoparticles, rats, surfactant High concentrations of airborne particles have been associated with increased pulmonary and cardiovascular mortality, with indications of a specific toxicologic role for ultrafine particles (UFPs; particles with diameters < 0.1 μm) (Peters et al. 1997). UFPs may induce inflammatory and prothrombotic responses, promoting atherosclerosis, thrombogenesis, and the occurrence of other cardiovascular events (Schulz et al. 2005). Human data suggest that inhaled UFPs influence lung physiology (Pietropaoli et al. 2004). UPFs may also affect the autonomic nervous system or act directly on cells in various organs and induce mutations (Harder et al. 2005; Samet et al. 2004). After exposure of the respiratory system to UFPs, the UFPs may appear within hours in many compartments of the body, including the liver, heart, and nervous system (Brown et al. 2002; Kreyling et al. 2002; Oberdörster et al. 2004). UFPs are formed by gas-to-particle conversion or by incomplete fuel combustion. Despite considerable efforts to reduce air pollution, the environmental burden by UFPs may have increased rather than decreased over time (Kreyling et al. 2003). Moreover, the fast-growing nanotechnology industry generates new UFPs daily, which may become aerosolized at some stage and may present additional health risks. UFPs possess increased toxicity compared with larger particles composed of the same materials [F8] (Ferin et al. 1992). Their environmental burden is characterized by high number concentrations but low mass concentrations. Thus, a relatively large surface area per unit mass facilitates adsorption of various organic compounds from the ambient air and enhances interaction with biological molecules within the organism. Deposition of UFPs in the respiratory system is caused by diffusional displacement. Depending on particle size, deposition occurs efficiently in the nose, the conducting airways, and the alveoli. Although particles with diameters > 1 μm usually remain on the epithelial surface upon their deposition (Gehr et al. 1990; Geiser et al. 2003; Schürch et al. 1990) and are subjected to clearance by cough, mucociliary transport, and/or phagocytosis by macrophages, UFPs seem to penetrate the boundary membranes of the lungs rapidly—a unique feature for insoluble particles (Brown et al. 2002; Kreyling et al. 2002; Oberdörster et al. 2002). In addition, transport across the olfactory epithelium and accumulation in the brain were reported for various UFP types (Oberdörster 2004). In vitro experiments revealed penetration of UFPs into mitochondria of macrophages and epithelial cells that was associated with oxidative stress and mitochondrial damage (Li et al. 2003).

Because everyone on earth inevitably inhales thousands to millions of UFPs with each breath, it is important to assess health risks by UFP air pollution. The costs of actions to be taken to reduce ambient aerosol particles are high and will affect the economy greatly, presenting an urgent need to clarify the fate of inhaled UFPs. To date, the mechanisms by which UFPs penetrate boundary membranes and the distribution of UFPs within tissue compartments of their primary and secondary target organs are largely unknown.—This study is the first to investigate the distribution of inhaled UFPs within lungs at the individual particle level and combines different experimental approaches[F9] —an in vivo inhalation study in rats and an in vitro cell exposure study on pulmonary macrophages and red blood cells (RBCs).—-In the in vivo experiments, rats inhaled an ultrafine titanium dioxide aerosol of 22 nm count median diameter (CMD) during 1 hr, resulting in a deposition of 4–5 μg TiO2 per animal. The intrapulmonary distribution of deposited particles was analyzed immediately or 24 hr after the end of exposure, using energy-filtering transmission electron microscopy (EFTEM) to allow elemental microanalysis of individual particles (Kapp et al. 2004).–In the in vitro study, we exposed cultured porcine pulmonary macrophages and human RBCs to fluorescent polystyrene microspheres with diameters of 1, 0.2, and 0.078 μm and assessed particle uptake by confocal laser scanning microscopy (CLSM).

Materials and Methods
Animals.
The animal experiments were conducted under federal guidelines for the use and care of laboratory animals (German Animal Protection Law) and were approved by the District of Upper Bavaria (Approval No. 211-2531-108/99) and by the GSF Institutional Animal Care and Use Committee, as well as in accordance with the Swiss Federal Act on Animal Protection and the Swiss Animal Protection Ordinance. Ten young, adult, male WKY/NCrl BR rats [body weight (bw) 250 ± 10 g (mean ± SD; Charles River, Sulzfeld, Germany] were housed under standard conditions, with access to food and water ad libitum, in a room controlled for humidity (55% relative humidity) and temperature (22°C) and with lighting on a 12-hr day/night cycle. Animals were anesthetized by intramuscular injection of a mixture of medetomidine (Domitor; 150 μg/100 g bw; Pfizer GmbH, Karlsruhe, Germany), midazolam (Dormicum; 0.2 mg/100 g bw; Hoffmann-La Roche AG, Grenzach-Wyhlen, Germany), and Fentanyl (0.5 μg/100 g bw; Janssen-Cilag GmbH, Neuss, Germany) for inhalation and lung fixation and anticoagulated by intra-peritoneal injection of 2,000 IU heparin (Heparin-Natrium-ratiopharm, ratiopharm GmbH, Ulm/Donautal, Germany) for lung fixation (Kapp et al. 2004). Anesthesia was antagonized by subcutaneous injection of atipamezole (Antisedan; Pfizer GmbH), flumazenil (Anexate, Hoffmann-La Roche AG), and naloxone (Narcanti, Janssen Animal Health, Neuss, Germany).

Aerosols and inhalation.
Titanium is suitable for EFTEM because it does not interfere with the heavy metals used for tissue preparation (Kapp et al. 2004). TiO2 is inert and nonpathogenic (Templeton 1994), and ultrafine aerosols thereof remain as insoluble particles (Donaldson et al. 2002). Generally, commercially available TiO2 particles (Degussa, Düsseldorf, Germany) show a positive Zeta potential of 20–30 mV as measured by Malvern Zetasizer 3000 HS (Malvern Instruments Ltd., Malvern, Worcestershire, UK) The generation and inhalation of the TiO2 aerosol used in this study has been described in detail by Kapp et al. (2004). Briefly, ultrafine TiO2 aerosols were generated with a Palas spark generator (Palas GmbH, Karlsruhe, Germany) in a pure argon plus 0.1% oxygen stream. The aerosol was quasi-neutralized by a radioactive 85Kr source, diluted, and conditioned for inhalation. Particle size distribution and number concentration were monitored continuously by a differential electrical mobility particle sizer and a condensation particle counter. An aerosol of 22 nm CMD (geometric SD of 1.7) was produced. The mean number concentration was 7.3 × 106 particles/cm3 (SD 0.5 × 106 particles/cm3), corresponding to a mass concentration of 0.11 mg/m3. The 22 nm particles measured as aerosol particles are already agglomerates of smaller primary structures formed immediately after spark ignition and condensation. The estimated size of the primary structures was 4 nm [F10] as derived from the measured specific surface area of 330 m2/g of the UFPs produced in this study and the TiO2 bulk density of 4.2 g/cm3.

Each anesthetized rat was placed in an airtight plethysmograph box. The animals inhaled the aerosol in pairs (one each for the 1-hr and 24-hr examinations) for 1 hr via an endotracheal tube by negative-pressure ventilation (−1.5 Pa) at a breathing frequency of 45/min, resulting in a minute volume of about 200 cm3/min (Kreyling et al. 2002).

From breathing and aerosol parameters, the deposited amount of TiO2 was calculated to be 4–5 μg in each rat. After aerosol exposure, anesthesia of one of the paired rats was antagonized as described above, and the animal was returned to its cage for 24 hr.

Lung fixation and tissue preparation.
Lungs were fixed either 1 hr or 24 hr after the aerosol inhalation by sequential intravascular perfusion with buffered 2.5% glutaraldehyde (Agar Scientific Ltd., Plano GmbH, Wetzlar, Germany), 1.0% osmium tetroxide (Simec, Zofingen, Switzerland), and 0.5% uranyl acetate (Fluka Chemie GmbH, Sigma-Aldrich, Buchs, Switzerland). Lungs were then subjected to systematic tissue sampling, dehydration in a graded series of ethanol, and embedding in Epon (Fluka) (Im Hof et al. 1989; Kapp et al. 2004). Ultrathin (≤50 nm) sections were cut from five to eight tissue blocks per animal, mounted onto uncoated 600-mesh copper grids, and stained with uranyl acetate and lead citrate (Ultrostain, Leica, Glattbrugg, Switzerland).

Particle localization and elemental micro-analysis in situ.
On ultrathin sections, 12 fields, corresponding to an area of 1,820 μm2 each, were systematically sub-sampled and investigated for the presence and localization of TiO2 particles in a LEO 912 transmission electron microscope (LEO, Oberkochen, Germany) equipped with an in-column energy filter allowing energy dispersion for element specific contrast. TiO2 particles were identified by parallel electron energy-loss spectroscopy (parallel-EELS), electron spectroscopic imaging, and image-EELS (Kapp et al. 2004). For elemental micro-analysis, we used the L 2 , 3 edge of Ti at 456 eV energy loss. We obtained bright-field and structure-sensitive micrographs (recorded at 250 eV), as well as element-specific contrast for TiO2, by digital acquisition.

Cell culture experiments.
Porcine lung macrophages (provided by K. McCullough and H. Gerber, Institute for Virus Diseases and Immune Prophylaxis, Mittelhäusern, Switzerland) were cultured at 106 cells/mL in two-chamber slides (VWR International AG, Dietikon, Switzerland) for 24 hr at 37°C and 5% CO2 in RPMI 1640 medium (containing 25 mM Hepes; LabForce AG, Nunningen, Switzerland) with 10% fetal bovine serum (LabForce), 1% l-glutamine (LabForce), and 1% penicillin/streptomycin (Gibco BRL, Invitrogen AG, Basel, Switzerland). Human RBCs, always freshly isolated from the same donor, were cultured at 8 × 106 cells/mL and for 6–24 hr, as described above.

For CLSM, fluorescent polystyrene microspheres with diameters of 1, 0.2, and 0.078 μm (Fluoresbrite plain yellow green; Polysciences, Chemie Brunschwig AG, Basel, Switzerland) were added to the cells at 1010 particles/mL in supplement-free RPMI 1640 medium for 4 hr. To inhibit phagocytic uptake, cells were pretreated with cytochalasin D (cytD, 10 μg/mL; Fluka) for 30 min and during incubation with particles (Thiele et al. 2001). Macrophages were then washed in phosphate-buffered saline (PBS), fixed in 3% paraformaldehyde/PBS, treated with 0.1 M glycine/PBS, permeabilized with 0.2% Triton X-100/PBS, and stained for F-actin with rhodamine-phalloidin (1:100; Molecular Probes, VWR International AG, Lucerne, Switzerland) for 60 min at 37°C. RBCs were fixed in 2.5% glutaraldehyde/PBS, resulting in a red autofluorescence. Preparations were mounted in PBS:glycerol (2:1) containing 170 mg/mL Mowiol 4-88 (Calbiochem, VWR International AG).

For TEM analysis, RBCs were incubated with 0.025 μm gold particles (ANAWA Trading SA, Wangen, Switzerland) at 6.6 × 1010 particles/mL. RBCs were fixed with buffered 2.5% glutaraldehyde, 1% osmium tetroxide, and 0.5% uranyl acetate, and prepared for TEM as described above for lung tissue.

Microscopic analysis of cultured cells.
We used a MicroRadiance system from Bio-Rad (Hemel Hempstead, UK) combined with an inverted Nikon microscope (Eclipse TE3000; lasers: GHe/Ne 543 nm and Ar 488 nm) (Nikon, Küsnacht, Switzerland). Optical sections with a voxel dimension of 50 × 50 × 200 nm were taken with a 100× /1.4 plan-apochromate objective. We performed image processing and visualization using IMARIS software (Bitplane AG, Zurich, Switzerland). We applied a deconvolution algorithm using Huygens2 software (Scientific Volume Imaging B.V., Hilversum, Netherlands) to increase axial and lateral resolutions and to decrease noise (Rothen-Rutishauser et al. 1998). Experiments were performed in triplicate or quadruplicate, and 30–50 cells were scanned by CLSM for each data point. We examined ultrathin sections of RBCs incubated with gold particles in a Philips 300 TEM at 60 kV (Philips, Zurich, Switzerland).

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Results
Distribution of inhaled UFPs in lungs.
We found TiO2 particles on the luminal side of airways and alveoli as well as within each tissue compartment of the lung (Figures 1 and ​and2).2). Particles were localized within epithelial and endothelial cells, within fibroblasts and between collagen fibrils in the connective tissue, within blood capillaries, and even within RBCs (Figure 1). Intracellular particles were localized most often in the cytoplasm and rarely within the nucleus. Particles within cells were not membrane bound. On average, 79.3 ± 7.6% (mean ± SD) of the particles were found on the luminal side of airways and alveoli, 4.6 ± 2.5% were within epithelial or endothelial cells, 4.8 ± 4.5% within the connective tissue, and 11.3 ± 3.9% within the capillaries. The relative distributions of particles among different lung compartments at 1 hr and at 24 hr after inhalation were not different from each other and correlated with the volume fractions of the respective compartments (Figure 2).

Figure 1

EFTEM micrographs of particles (arrows) in the lung parenchyma. Abbreviations: AL, alveolar lumen; C, collagen fibril; CL, capillary lumen; EC, erythrocyte; EN, capillary endothelial cell; EOS, eosinophil granulocyte; EP, epithelium; F, fibroblast; FC, …

Figure 2

Relative distribution of particles localized in the different lung compartments at 1 hr and 24 hr after inhalation. Volume densities (Vv) for lung tissue compartments from Burri et al. (1973), Pinkerton et al. (1992), and Tschanz et al. (1995, 2003).

Uptake of UFPs by macrophages and RBCs.
We studied the uptake of fine (1–0.2 μm) and ultrafine (< 0.1 μm) fluorescent microspheres in cultured macrophages as a model for phagocytic cells and in human RBCs as a model for nonphagocytic cells. Additionally, we treated macrophage cultures with cytD to block phagocytosis. Cells were prepared for CLSM, and a deconvolution algorithm was applied to increase lateral as well as axial resolution. We found particles of all sizes within macrophages (Figure 3A), however, with different percentages of cells involved in particle uptake. On average, 77 ± 15% (mean ± SD) of the macrophages contained UFPs, 21 ± 11% contained 0.2 μm particles, and 56 ± 30% contained 1 μm particles (Figure 3B). CytD did not inhibit the uptake of ultrafine and 0.2 μm particles but blocked the uptake of 1 μm particles by these cells (Figure 3B). We found ultrafine and 0.2 μm, but not 1 μm, particles in RBCs (Figure 4A). TEM analysis of RBCs incubated with 0.025 μm gold particles showed that intracellular particles were not membrane bound (Figure 4B). Figure 3 (A) CLSM micrographs of fluorescent polystyrene spheres (1, 0.2, and 0.078 μm) taken up by macrophages in the absence (−) or presence (+) of cytD (bar = 2 μm); F-actin is shown in red, and particles are green. The xy and xz projections … Figure 4 (A) CLSM micrographs of fluorescent polystyrene spheres taken up by RBCs. Autofluorescence of the cells is shown in red, and particles are green (bar = 2 μm). The xy and xz projections allow clear differentiation between internalized (arrows) … Go to: Discussion The ultrastructural analyses of lung tissue demonstrated that 1 hr after aerosol inhalation, 24% of ultrafine TiO2 particles, on average, were located within and beyond the epithelial barrier (i.e., in the main lung tissue compartments, in the cytoplasm, and in the nucleus of the cells). These results confirm data from human studies, where the inhalation of ultrafine carbon particles affected pulmonary diffusing capacity (Pietropaoli et al. 2004), suggesting that particles in the interstitium have physiologic effects. This study also provides evidence for some particle translocation into the micro-vasculature. In previous studies on iridium particles, we found minute fractions of particles translocated into secondary target organs (Kreyling et al. 2002; Semmler et al. 2004b). Different particle materials—iridium versus TiO2—may have resulted in different particle translocation patterns, and we do not know the exact composition and structure of the ultrafine particle surface, which is likely to influence particle translocation. The present study focuses on particle distribution within the primary target organ, the lung, and the results do not determine which fraction of particles may have escaped the lung micro-vasculature to be systemically circulated. Particles found within cells were not membrane bound, indicating a nonendocytic uptake. In addition, the overall distribution pattern of the particles in the lungs (i.e., the percentages of particles in the different lung compartments) at 1 hr and at 24 hr after particle inhalation was the same and was correlated with the volume densities of the corresponding lung compartments, implying that ultrafine TiO2 particles can move between tissue compartments without restraint. Our results are in contrast with those obtained by Stearns et al. (2001), who studied the uptake of ultrafine TiO2 particles in vitro in the A549 epithelial cell line. They found that membrane-bound vesicles contained mostly large aggregates of TiO2. Sometimes vesicles with clusters consisting of as few as two to three particle profiles were observed. In a pilot study in vitro with porcine macrophages, we obtained similar results (data not shown). Whether these clusters contained few particles only or were sectioned through the top of larger ones is not known. However, because ultrafine TiO2 aggregate very quickly in polar liquids such as cell culture medium and because particle concentration was fairly high, as seen from micrographs, it is very likely that particles aggregated within the cell culture medium and that cells engulfed these large clusters by an endocytic pathway. Particle agglomeration on the lung epithelium during the 1-hr inhalation, however, is very unlikely because of the size of the inner lung surface and the number of deposited UFPs. The fact that 80% of the retained TiO2 particles were still on the luminal side of the epithelium even 24 hr after inhalation is surprising and in contrast to previous studies on the lavageability of ultrafine iridium particles, where only 20% of the particles could be lavaged[F11] from the epithelial surfaces 24 hr after inhalation (Kreyling et al. 2002). The low lavageability of ultrafine iridium particles in contrast to high lavageability of 80% of 0.5–10-μm particles (Oberdörster et al. 2002) was interpreted as either higher adhesion of UFPs to epithelial membranes or epithelial uptake and penetration of UFPs into the interstitium. The large fraction of TiO2 particles on the luminal side of the epithelium in the present study strongly supports higher adhesion of UFPs to epithelial structures. However, it must also be considered that proteins are very likely to bind rapidly to the particles, which then may affect the further metabolic fate of the particles in terms of their adhesion, residence time on the epithelium or uptake, and even penetration through the epithelium. Along this line, the difference of the two particle materials and surfaces may have led to binding of those proteins,[F12] which then may have mediated major uptake and penetration of iridium particles into and through the epithelium. We already have first evidence that ultrafine commercial TiO2 particles bind more readily to other proteins in the lung-lining fluid than carbonaceous and amorphous silica particles (Semmler et al. 2004a). Microscopic analyses of phagocytic and nonphagocytic cells incubated with different particle types showed that macrophages take up fine and ultrafine polystyrene microspheres and that treatment with cytD inhibits the uptake of 1.0-μm particles, but not uptake of the smaller particles, by these cells. Ultrafine polystyrene and gold particles also entered RBCs and were not membrane bound. The mechanisms of intracellular uptake of macromolecules, particles, and even cells are subsumed as endocytosis[F13] . Material to be ingested is progressively enclosed by the plasma membrane, which eventually detaches to form an endocytic vesicle. Phagocytosis and pinocytosis are distinguished by the size of endocytic vesicles formed. Phagocytosis, a receptor-mediated, actin-based process, is characteristic for neutrophils, macrophages, and dendritic cells. It is the main mechanism for the clearance of insoluble 1- to 3-μm particles from the alveoli. Pinocytosis involves the ingestion of fluid and solutes via vesicles of about 100 nm in diameter. There are at least four basic mechanisms, most of which can be demonstrated in lungs and involve specific receptor–ligand interactions: a) macro-pinocytosis, b) clathrin-mediated, actin-based endocytosis, c) caveolae-mediated endo- or transcytosis, and d) clathrin- and caveolae-independent endocytosis (Conner and Schmid 2003). Kruth et al. (1999) described an additional endocytic process, patocytosis, in which hydrophobic polystyrene particles < 0.5 μm are transported through induced plasma membrane channels into an extensive labyrinth of interconnected membrane-bound compartments. None of these endocytic pathways, all of which include vesicle formation, is likely to account for the translocation of UFPs in our study, as intracellularly localized particles were not membrane bound. Moreover, because RBCs contained UFPs and cytD treatment of macrophages did not prevent UFP translocation into these cells, particle uptake by any actin-based mechanism can also be excluded. Transport via pores, as suggested for lung–blood substance exchange (Conhaim et al. 1988; Hermans and Bernard 1999), is another potential mechanism for UFP translocation. TiO2 particles may diffuse through such pores. A transport mechanism by diffusion is consistent with the observed spatial distribution of UFPs in our inhalation study. Thus far, signal-mediated transport via pores has been demonstrated only for ultrafine gold particles of up to 39 nm in diameter, through the nuclear pore complex in Xenopus oocytes, where transport velocities depended on particle size (Panté and Kann 2002). Passive uptake (not triggered by receptor–ligand interactions) may also occur by electrostatic, Van der Waals, or steric interactions, subsumed under “adhesive interactions” (Rimai et al. 2000). Rimai et al. showed that 8-μm glass particles were approximately 90% engulfed by a polystyrene substrate, compared with 22-μm particles, which were only 30% engulfed. However, the influence of particle size on their engulfment was not clarified in these publications. Several concepts for the nonspecific engulfment of particles through interfacial structures (including cell membranes) have been suggested. A thermodynamic model using the “wettability criterion” was successful in predicting passive particle uptake, although it did not take into account the elastic properties of the cell membrane (Chen et al. 1997). In another thermodynamic analysis combined with a molecular dynamics simulation, Bresme and Quirke (1999) showed that line tension influences the wetting behavior of nanoparticles at liquid–vapor and liquid–liquid interfaces. These authors found negative line tension values for particles of a few nanometers in diameter, but positive values for those an order of magnitude larger. A negative line tension favors the initial wetting of a spherical particle after its approach to an interface. Shanahan (1990) studied the engulfment of solid-surface heterogeneities equivalent to particles in the nanometer range by unbalanced capillary forces (free energy perturbations). Thermal capillary waves cause fluid droplets to coalesce with a fluid substrate by film drainage at the interface, breakage of the film, and intrusion of the particle into the bulk phase (Aarts et al. 2004). Thus, thermal capillary fluctuations may enhance particle transport through cell membranes. Experimental results demonstrate consistently greater immersion of smaller particles than larger ones into a liquid substrate covered by surfactant film in vitro as well as in situ in airways. These results support the concept that line tension plays a significant role in particle displacement (Geiser et al. 2000; Schürch et al. 1999). It remains to be determined which chemical and physical properties of membranes and particles are responsible for the translocation of UFPs in vivo. Interestingly, we did not see any difference in particle uptake in vitro with respect to differing surface charges or surface chemistry when we used three different particle types: a metal, a metal oxide, and a synthetic polymeric material (data not shown). However, these particles were added to the cells in suspension and did not approach the cells from the air or require passage through a surfactant film first, as in the in vivo inhalation experiments. In these in vivo experiments, electrostatic interactions are likely important for particle deposition and subsequent retention. In summary, UFPs of various materials can cross any cellular membrane, but neither endocytosis, which is based on vesicle formation, nor any actin-based mechanisms are likely to account for UFP translocation into the cell. Our results from the inhalation experiments with TiO2 particles point to a transport mechanism that includes adhesive interactions or, in terms of thermodynamics, interfacial and line tension effects. In addition, particle diffusion and uptake promoted by thermal capillary waves might play a role in particle transport through membranes. After the deposition of nanometer-size particles, their further fate may be largely independent from particle surface chemistry and charge. Consequently, it is a possible fate of inhaled ambient UFPs that they are transferred from the lungs to most other organs. In a first analysis of ultrathin sections from hearts of the same rats, we found ultrafine TiO2 particles in the connective tissue, that is, within fibroblasts. There may be no means on the cellular level to prevent, influence, or direct their uptake. Moreover, the toxic potential of UFPs is greatly enhanced by their free location and movement within cells, which promote interactions with intracellular proteins and organelles and even the nuclear DNA. Potential health implications of our findings are related not only to ambient UFPs but also to engineered “nanoscaled particles,” which may be released into our environment during their production, transport, and aging, or during waste disposal (The Royal Society 2004). Routes of exposure to nanomaterials include oral, cutaneous, and inhalative uptake, the latter being addressed in the present study. Although the number of particles translocated into the cells may vary substantially (Nanosafe 2004) according to their physicochemical properties, the data of the present study strongly suggest that adverse health outcomes associated with the uncontrolled presence of nanoscale particles in tissues require further attention. Footnotes We thank S. Frank, B. Haenni, B. Kupferschmid, and B. Tschirren for excellent technical assistance and L.M. Cruz-Orive for his help with the lung sampling design. This study was supported by the Swiss National Science Foundation; the Swiss Agency for the Environment, Forest, and Landscape; and the Silva Casa Foundation. References Aarts DG, Schmidt M, Lekkerkerker HN. Direct visual observation of thermal capillary waves. Science. 2004;304:847–850. [PubMed] Bresme F, Quirke N. Nanoparticulates at liquid/liquid interfaces. Phys Chem Chem Phys. 1999;1:2149–2155. Brown JS, Zeman KL, Bennett WD. 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[PubMed] TOP B ——————————————————————————– [F1]AS per usual anything that is of any benefit will be considered a evil and the concern is about some imaginary non issue—there are some exceptions and amounts to be used are always need to be explored—there are always extreme’s in this 2 sides to every element in excess or or in deficits [F2]This is the medical side but in the alternative 200IU to 400 IU of Vitamin E was base—- To convert IUs of vitamin E into milligrams of alpha-tocopherol: multiply the number of IUs by 0.67. To convert milligrams of alpha-tocopherol to IUs: multiply the number of milligrams by 1.5. So 200IU X 0.67 = 13.4 mgs 400 IU X 0.67 = 26.8 ,gs [F3]Vitamin E protects the brain of oxidation —that is crucial to fat damage in the brain—they call this liperfuscion which is an oxidative damage of fats that can collect in the brain this is one method of protection [F4]an isomer that is a stereoisomer of a compound having two or more chiral centers and that is not a mirror image of another stereoisomer of the same compound [F5]IC50 is the acronym for “half maximal inhibitory concentration”. IC50 value indicates the concentration needed to inhibit a biological or biochemical function by half (e.g. inhibition of enzymes, affinity to cell receptors). In pharmaceutical research, it is a frequently used unit to specify the in vitro potency of a drug or a NCE. Amongst others, determination of IC50 is commonly calculated via linear interpolation: The activity of an enzyme is determined after exposure to a series of inhibitor concentrations. IC50 is calculated by the following formula: IC50 = (50%-LowInh%)/(HighInh%-LowInh%)x(HighConc-LowConc) + LowConc LowInh% / HighInh%: % inhibition directly below / above 50% inhibition LowConc / HighConc: Corresponding concentrations of test compound [F6]IC50 is the acronym for “half maximal inhibitory concentration”. IC50 value indicates the concentration needed to inhibit a biological or biochemical function by half (e.g. inhibition of enzymes, affinity to cell receptors). In pharmaceutical research, it is a frequently used unit to specify the in vitro potency of a drug or a NCE. Amongst others, determination of IC50 is commonly calculated via linear interpolation: The activity of an enzyme is determined after exposure to a series of inhibitor concentrations. IC50 is calculated by the following formula: IC50 = (50%-LowInh%)/(HighInh%-LowInh%)x(HighConc-LowConc) + LowConc LowInh% / HighInh%: % inhibition directly below / above 50% inhibition LowConc / HighConc: Corresponding concentrations of test compound [F7]Interactivity—with proteins and DNA [F8]Base metals do not react the same way as nano particles —nanosilver does not react as colloidal silver—nanomagnesium does not react the same way as magnesium oxide—when in nano they collect internally and then bind to anything –protein—fat—dna—tissue—and can cause a compromise on the immune function and making you less resistant to being infected by any means [F9]These are just one particle not a combination [F10]Which can penetrate topically [F11]A washing, especially of a hollow organ such as the stomach or lower bowel, with repeated injections of water [F12]What happens to nano particles when entered into us they can spread and stick inside and as well mix with other proteins or materials within the body to produce new proteins—indicating growth [F13]In endocytosis, the cell engulfs some of its extracellular fluid (ECF) including material dissolved or suspended in it. A portion of the plasma membrane is invaginated and pinched off forming a membrane-bounded vesicle called an endosome