Saturday, August 30, 2014

Potato zaps RLS

Potato zaps Restless Leg Syndrome

Andre Willers
30 Aug 2014
“Potatoes are masters of Maskirovka”  … General картофель .
Restless Leg Syndrome ameliorization by application of glycoalkaloids to regulate micro-ecology of skin- and other bacteria .
Discussion :
1.Fancy speak for how and why the old Potato-in-the-bed trick works .
An exotoxin is a toxin secreted by bacteria.[1] An exotoxin can cause damage to the host by destroying cells or disrupting normal cellular metabolism. They are highly potent and can cause major damage to the host. Exotoxins may be secreted, or, similar to endotoxins, may be released during lysis of the cell. Gram negative pathogens may secrete outer membrane vesicles containing lipopolysaccharide endotoxin and some virulence proteins in the bounding membrane along with some other toxins as intra-vesicular contents, thus adding a previously unforeseen dimension to the well-known eukaryote process of membrane vesicle trafficking, which is quite active at the host-pathogen interface.

3. Chemotaxis

Chemotaxis: a physiological adapation

A dramatic example of rapid adaptation is the process of "chemotaxis", i.e. the ability to movetoward nutrients (positive chemotaxis) and away from noxious compounds (negative chemotaxis).
This is a valuable trait in bacteria, since it enages them to swim to sources of nutrients and away from toxic chemicals. 
Chemotactic responses are very rapid, they do not require active gene expression. 
How is it that bacteria move directionally?  One way that some types of bacteria move involves organelles known as flagella. 
These are rotary motors powered by ion gradients across the plasma membrane.  This motor drives a helical propeller.  The basic motor can rotate in either a clockwise or a counterclockwise wise direction.

Because their three dimensional structure, when the flagellum rotates in one direction (counterclockwise), the bacteria moves forward; when rotating in the opposite direction, the bacteria tumbles.
Tumbling allows the bacteria to change direction.  When the motor switches back to the counter clockwise direction, the bacteria swims off in a new direction. 
The ability to control chemotaxis involves controlling the probability that the motor will switch its direction of rotation. 
In bacteria, motor rotation switching is controlled by a receptor system that senses changing concentrations of attractants (inhibiting switch) and repellants (induce switching).

There are two possible ways to do this. One would be to measure the difference in concentration between the two ends of the bacterium. This is difficult, since the bacteria is very small (1-2 um long) and the differences in concentration between two points separated by 1-2 um is very small (perhaps we could calculate this?)  
An alternative approach is to measure the concentration around the bacteria as a function of time.  If it is increasing, the bacteria is moving toward the source of the chemical; if it is decreasing, it is moving away from thge source. 
4. Thus, chemotaxis provides an important mechanism for establishing the high local cell densities required for quorum-dependent interactions.

5.They all sit and watch each other :
Interspecies quorum sensing is a way bacteria in a community can determine how many of their own and other species are in an area. Bacteria secrete these molecules, which increase in proportion to cell number. Once the molecule hits a certain concentration it triggers the transcription of certain genes such as virulence factors. It has been discovered that bacteria can not only interact via quorum sensing with members of their own species but that there is a kind of universal molecule that allows them to gather information about other species as well.[1] This universal molecule is called autoinducer 2 or AI-2.[1]

6. The Treatment:
Putting it all together :
6.1Chemotaxis induces the urge to move .
6.2 Clothing and bedding harbours bacteria , viruses and fungi that the body sees as threats .
6.3These threats are communicated via exotoxins and endotoxins in the bioaerosol .
6.4 Diurnal rhythm : at night , switches on because of higher risk .
6.5 Learned Helplessness :
If the system perceives threats from all directions , but no gradients in concentration over time or direction , it moves at random .
Prolonged exposure to this sort of stimulus will surface as various disorders in the brain (eg Parkinsons , akathisia , some forms of dementia , etc) .
Neuronal Networks weights get randomized by such stimuli .
7. This suggests some treatments :
7.1 Bacterial Maskirovka . (
The potato has a potent arsenal of pesticides (glycoalkaloids) (See Appendix A) that can be used to create artificial gradients in threat signals . Most are in the potato skin , so only the peel is really needed .
Soap will work as well . Anything that disturbs the threat picture .
7.2 The potato must be somewhere accessible to airborne particles (bioaerosols) .
Porous materials are no barrier . Carry the peels in your pocket in a porous plaster .

7.3 A fan will have an effect .

7.4 The potato peels will obviously become exhausted after a while and need replacement . The time will depend on the poison concentrations in the potato .
7.5 Electronic tumbling of the bacteria (see how chemotaxis works above) using swirling magnetic fields . Something like APStherapy machines (
Potato works for certain types of cancer cells (warts) . It can work better if combined with swirling magnetic fields .
7.6 Numerous approaches suggest themselves .
Patches , baths , showers , ointments , radiation , etc, etc .
8.Caution :
The idea is to disrupt or steer threat patterns by manipulating poisons , not to kill off the patient .
Potato : Glycoalkaloids are strong cell disruptors and neurotoxins . More is not better here .
9.Other disorders may be helped by this approach .
Akathisia , urinal tract infections , SIBO , leaky guts , swollen legs caused by cellulitis , cellulitis . it’s little sister cellulite (the potato goes OUTSIDE , not inside! ) , etc.
10. Aphrodisiac .
Hunt the wily wandering potato under the covers .
A spud for all occasions .
Regards ,

Appendix A
How Deadly Are Nightshades?
Nightshades have a reputation as bad actors in a variety of chronic conditions, such as arthritis, fibromyalgia, and IBS. But what do we really know about how these foods affect our health?
Meet the Nightshade (Solanaceae) Family: 
·         Tomatoes
·         Eggplant
·         Potatoes
·         Goji Berries
·         Tobacco
·         Peppers (bell peppers, chili peppers, paprika, tamales, tomatillos, pimentos, cayenne, etc) 
At first glance, the nightshades may look like a random collection of foods that couldn’t possibly be related.  However, every nightshade plant produces fruits that all sport that same adorable little green elfish hat.  Of the foods above, only tomatoes, eggplants, goji berries and peppers are “fruits” (the potato is a tuber and tobacco is a leaf).  The fruits of potato and tobacco plants wear the same telltale hat, but we don’t eat the fruits of those plants. 
Nightshades of all types were considered inedible prior to the 1800’s, because some varieties, such as “deadly nightshade” (atropa belladonna) were known to be so toxic.  However, today most Americans eat “edible” nightshades every day in the form of French fries, mashed potatoes, salsa, spaghetti sauce, ketchup, and many other popular foods.
What are glycoalkaloids?
Glycoalkaloids are natural pesticides produced by nightshade plants.  [They are also present in small amounts in a few non-nightshades:  cherries, apples, and sugar beets.]  Glycoalkaloids are bitter compounds which are found throughout the plant, but especially in leaves, flowers, and unripe fruits.  They defend the plants against bacteria, fungi, viruses, and insects.  How do these chemicals kill pests?
Glycoalkaloids bind strongly to the cholesterol in the cell membranes of predators, disrupting the structure of their membranes, and causing their cells to leak or burst open upon contact—acting like invisible hand grenades.
Glycoalkaloids have another powerful trick up their sleeves—they also act as neurotoxins, by blocking the enzyme cholinesterase.  This enzyme is responsible for breaking down acetylcholine, a vital neurotransmitter that carries signals between nerve cells and muscle cells.  When the enzyme is blocked, acetylcholine can accumulate and electrically overstimulate the predator’s muscle cells.  This can lead to paralysis, convulsions, respiratory arrest, and death.  Military “nerve gases” work exactly the same way.
Ok, so glycoalkaloids are clearly nightmarish compounds for tiny creatures daring to storm the nightshade’s citadel, but how much do we know about their effects on human health?
Proposed glycoalkaloid health benefits
Health benefits?  From a pesticide?  Hmmm…
Glycoalkaloids are structurally similar to glucocorticoids, such as our body’s stress hormone, cortisol.  Cortisol has many roles in the body, one of which is to reduce inflammation.  Therefore, perhaps it is not so surprising that glycoalkaloids have been shown to have anti-inflammatory properties in laboratory studies of animals.
It should also not be surprising that glycoalkaloids have been shown in laboratory studies to possess antibiotic and antiviral properties, since this is what nature designed them for.
In laboratory (in vitro) studies, glycoalkaloids can trigger cancer cells to self-destruct.  This process is called “apoptosis.”  Unfortunately, they can also cause healthy non-cancerous cells to do the same thing.  Cancer studies in live animals and humans (in vivo) have not yet been conducted.
“…the undifferentiating destruction of both cancer and noncancerous cell lines…leads to questions of therapeutic uses of glycoalkaloids due to safety considerations. However, it is difficult to translate the results of an in vivo trial in vitro. Therefore, both animal and human experiments are essential to confirm or disprove the in vivo data observed in these studies.”  [Milner 2011].
The other side of the sword: 
Research has shown that glycoalkaloids can burst open the membranes of red blood cells and mitochondria (our cells’ energy generators).
Some scientists have wondered whether glycoalkaloids could be one potential cause for “leaky gut” syndromes due to their ability to poke holes in cells:
“…glycoalkaloids, normally available while eating potatoes, embed themselves and disrupt epithelial barrier integrity in a dose-dependent fashion in both cell culture models and in sheets of mammalian intestine…animals with the genetic predisposition to develop IBD, demonstrated a greater degree of small intestinal epithelial barrier disruption and inflammation when their epithelium was exposed to the potato glycoalkaloids chaconine and solanine.”
Glycoalkaloids are also known to cause birth defects in laboratory animals.
Fruits vs vegetables:  here we go again!
Those of you who are familiar with my philosophy about plant foods know that I believe vegetables are far less trustworthy when it comes to health effects than edible fruits, and nightshades make this point nicely.  The only nightshade vegetable humans eat is the potato; the rest of the nightshades (other than tobacco, which is smoked, not eaten) are fruits, because they contain seeds—eggplant, tomatoes, goji and peppers.  As you will see below, even though fruits contain glycoalkaloids, they are far less likely to harm us. [To watch my Ancestral Health Symposium video about vegetables vs. fruits, click HERE.]
Potato glycoalkaloids 
Nightshade potatoes include all potatoes except for sweet potatoes and yams.
Potatoes make two glycoalkaloids:  alpha-chaconine and alpha-solanine.  These are the most toxic glycoalkaloids found in the edible nightshade family.  Alpha-chaconine is actually more potent than alpha-solanine, but solanine has been studied much more thoroughly, and is therefore more familiar.
Most of us do not associate potatoes with illness, probably because the amount of glycoalkaloid most of us eat every day is not very high.  There are numerous cases of livestock deaths from eating raw potatoes, potato berries, and potato leaves, but people do not eat these things.  However, there are well-documented reports of people getting glycoalkaloid poisoning from potatoes, typically from eating improperly stored, green, or sprouting potatoes.  At low doses, humans can experience gastrointestinal symptoms, such as vomiting and diarrhea.  At higher doses, much more serious symptoms can occur, including fever, low blood pressure, confusion, and other neurological problems.  At very high doses, glycoalkaloids are fatal.
Another reason why many people may not be bothered by potatoes is that glycoalkaloids are very poorly absorbed by the gastrointestinal tract, so, if you have a healthy digestive tract, most of the glycoalkaloid won’t make it into your bloodstream.  However, if you eat potatoes every day, levels can build up over time and accumulate in the body’s tissues and organs, because it takes many days for them to be cleared.  Also, since glycoalkaloids have the ability to burst cells open, they can theoretically cause damage to the cells that line your digestive system as they are passing through (this has been proven in animal studies but there are no human studies, to my knowledge).
Due to known toxicity, the FDA limits the glycoalkaloid content in potatoes to a maximum of 200 mg/kg potatoes (91 mg per pound).  Human studies show that doses as low as 1 mg glycoalkaloid per kg body weight can be toxic, and that doses as low as 3 mg/kg can be fatal.  This means that, if you weigh 150 lbs, then doses as low as 68 mg could be toxic, and doses as low as 202 mg could be fatal.
Potato processing 101 
The vast majority of glycoalkaloid is in the potato skin, so peeling will remove virtually all of it.  Glycoalkaloid levels can be dangerously high in unripe and sprouting potatoes; any greenish areas or “eyes” should be removed or avoided.
Glycoalkaloids survive most types of cooking and processing.  In fact, deep frying will increase levels if the oil isn’t changed frequently, so fried products such as potato skins and french fries can contain relatively high amounts:
“Mechanical damage to potato tissue increases the concentration of glycoalkaloids available for consumption. In addition, frying potatoes at high temperatures does not inactivate but instead serves to preserve and concentrate glycoalkaloids within the potato, leaving them available for ingestion and delivery to the intestine…” [Patel 2002]
·         Boiling—reduces glycoalkaloids by a few percentage points
·         Microwaving—reduces glycoalkaloids by 15%
·         Deep frying at 150C (300F)—no effect (McDonald’s uses 340F degree oil)
·         Deep frying at 210C (410F)—reduces glycoalkaloid content by 40%
Glycoalkaloid levels of a few prepared potato products are available [Milner 2006]:
·         Potato chips, 1 oz bag:   0.36 to 0.88 mg chaconine and 0.29 to 1.4 mg solanine. Total glycoalkaloid concentration ranges from 2.7 to 12.4 mg per bag.
·         Fried potato skins, 4 oz:   4.4 to 13.6 mg chaconine and 2.0 to 9.5 mg solanine.  Total glycoalkaloid concentration ranges from 6.4 to 23.1 mg per 4 oz serving.
Tomato glycoalkaloids
Tomato nightshades include all types of tomatoes:  cherry tomatoes, green tomatoes, yellow tomatoes and ripe red tomatoes.
Tomatoes produce two glycoalkaloids:  alpha-tomatine and dehydrotomatine.  The majority is in the form of alpha-tomatine, so we’ll focus on that one here.
As tomatoes ripen, alpha-tomatine levels drop dramatically, from about 500 mg/kg in green tomatoes to about 5 mg/kg in ripe red tomatoes, or 2.3 mg/lb.  [For those of you keeping score at home—that’s Fruits: 1, Veggies: 0.] Artificially ripened fruits may contain higher amounts than sun-ripened fruits.
Tomato glycoalkaloids are about 20 times less toxic than potato glycoalkaloids. (Fruits: 2, Veggies: 0).  There are no dosage studies of tomatine in humans, but studies in mice tell us that 500 mg tomatine per 1 kg body weight (or 227 mg per pound) is the median lethal dose (“LD50”).  This doesn’t tell us how much it would take to kill a 150 lb person; all we know is that it would take 34 grams of tomatine to kill a 150 pound mouse.  Since ripe tomatoes contain 5 mg/kg or 2.3 mg/lb of tomatine, it would take nearly 15,000 pounds of tomatoes to kill this Mighty Mouse (probably many fewer pounds if you were to simply hurl them in his general direction from across the room).  Since green tomatoes contain 100 times more tomatine, it would only take 150 pounds of green tomatoes to kill the overgrown rodent.  We do not understand the effect of low doses of tomatine on any type of animal, including humans, over time.
Centuries ago, the common eggplant was referred to as “mad apple” due to belief that eating it regularly would cause mental illness.  Eggplants produce two glycoalkaloids:  alpha-solamargine and alpha-solasonine.  Solamargine is more potent than solasonine.
Whereas potato glycoalkaloids are located mainly in the skin, in eggplants, glycoalkaloids are found primarily within the seeds and flesh; the peel contains negligible amounts.
The common eggplant (solanum melongena) contains 10-20 mg of glycoalkaloid per kg (or 4.5 to 9 mg per pound of eggplant).  Eggplant glycoalkaloids are considered relatively nontoxic compared to potato glycoalkaloids (Fruits: 3, Veggies: 0).
The median lethal dose (LD50) in rodents is 1.75 mg/kg.  This means that it would take at least 13 pounds of eggplant to kill a 150 lb monster mouse.  [Note to self—when facing a giant rodent in a dark alley, go for the eggplants, not the tomatoes].
What about peppers and goji berries? 
Your guess is as good as mine…I could not locate any scientific information about glycoalkaloids in these foods.  Peppers, because they are fruits and because they are in a different subfamily than the rest of the nightshade foods, may contain much less glycoalkaloid?  Or none at all?  Peppers are famous for containing hot and spicy “capsaicinoids”, not glycoalkaloids (I’ll write about peppers and capsaicinoids in a future article).
Nightshades and Nicotine 
Nightshade foods also contain small amounts of nicotine, especially when unripe.  Nicotine is much higher in tobacco leaves, of course.  Scientists think that nicotine is a natural plant pesticide, although it is unclear exactly how it works to protect plants from invaders. The amount of nicotine in ripe nightshade foods ranges from 2 to 7 micrograms per kg of food. Nicotine is heat-stable, therefore, it is found in prepared foods such as ketchup and French fries.  The health effects of these small doses is not known, but some scientists wonder whether the nicotine content of these foods is why some people describe feeling addicted to them.
Do you have nightshade sensitivity? 
As with any food sensitivity, the only way to find out is to remove nightshades from your diet for a couple of weeks or so to see if you feel better.  There are ZERO scientific articles about nightshade sensitivity, chronic pain, or arthritis in the literature, however, the internet is full of anecdotal reports of people who have found that nightshades aggravate arthritis, fibromyalgia, or other chronic pain syndromes.  I personally am very sensitive to nightshades; they cause me a variety of symptoms, most notably heartburn, difficulty concentrating, pounding heart, muscle/nerve/joint pain, and profound insomnia.  Everyone is different, so as always, you’ll need to discover for yourself whether these foods may pose problems for your individual chemistry.  However, given what we know about nightshade chemicals, common sense tells us that these foods are well worth exploring as potential culprits in pain syndromes, gastrointestinal syndromes, and neurologic/psychiatric symptoms.
Friedman M. Tomato glycoalkaloids: role in the plant and in the diet. J Agric Food Chem2002; 50:5751-5780.  UDSA, Albany California.
Hansen AA.  Two fatal cases of potato poisoning.  Science 1925; 61(1578): 340-341.
Korpan YI et al.  Potato glycoalkaloids: true safety or false sense of security?  Trends in Biotechnology 2004; 22(3): 147-151. 
McMillan M and Thompson JC.  An outbreak of suspected solanine poisoning in schoolboys: examinations of criteria of solanine poisoning.  Q J Med 1979; 48(190): 227-243. 
Mensinga TT et al.  Potato glycoalkaloids and adverse effects in humans: an ascending dose study. Regulatory Toxicology and Pharmacology 2005;41: 66-72.  University of Utrecht, The Netherlands.
Milner SE et al.  Bioactivities of glycoalkaloids and their aglycones from Solanum species. J Agric Food Chem 2011; 59: 3454–3484. University College, Cork Ireland.
Patel B et al.  Potato glycoalkaloids adversely affect intestinal permeability and aggravate inflammatory bowel disease.  Inflammatory Bowel Diseases 2002; 8 (5): 340-346.
Sanchez-Mata MC et al.  r-Solasonine and r-Solamargine Contents of Gboma (Solanum macrocarpon L.) and Scarlet (Solanum aethiopicum L.) Eggplants J Agric Food Chem 2010; 58: 5502–5508.
Siegmund B et al.  Determination of the nicotine content of various edible nightshades (Solanaceae) and their products and estimation of the associated dietary nicotine intake.  J Agric Food Chem 1999;47: 3113−3120.
Appendix B
The study, which was published in the journal Movement Disorders, found that sufferers of Parkinson’s disease had a higher prevalence of a condition called small intestinal bacterial overgrowth, or SIBO.
In SIBO, normally harmless bacteria proliferate in large numbers in the small intestine.
Symptoms include excess gas, abdominal bloating, diarrhoea and abdominal pain. Nobody is sure how many people have it,  as it often goes undiagnosed or  is confused with irritable bowel syndrome, but estimates are of  up to 300,000 British sufferers.
And the researchers suggest that the bacteria may produce chemicals that affect the nerves in the gut, which pass on the damage to the brain – and result in Parkinson’s and MS.
Dr Emmanuel said: ‘We now think that neurological diseases such as MS and Parkinson’s are linked to the gut being more  leaky, permitting pathogens into the bloodstream and causing  an antibody response. Either  the pathogens, directly, or the immune response, indirectly, may damage nerve tissue.’
The damaged nerves then transmit these detrimental signals to the brain.
Rumbly tummy: The 'butterflies in the stomach' feeling could be linked to Parkinson's disease and multiple sclerosis
In Parkinson’s, a small part of the brain becomes progressively damaged over many years, something that affects mainly older people. MS is the most common neurological disease in young adults and occurs when the immune system attacks the nervous system.
The two diseases affect about 100,000 and 127,00 people in the UK respectively. Both have wide-ranging symptoms, affecting movement as well as causing tiredness, pain and depression. There are drugs that can ease symptoms, but no cure or even treatments that significantly slow down progression of the diseases.
Scientists hope that this discovery will pave the way for new treatments for both these disabling conditions.
They are now mapping the ‘bacterial genome’, which will identify the bacteria in an individual’s gut – something they hope will ultimately allow doctors to prescribe tailored treatments for leakiness of the gut, improving neurological symptoms in turn.
Until then, the doctors’ body United European Gastroenterology urges people to maintain a healthy diet, including foods that boost good bacteria and encourage efficient digestion. This may have an especially positive effect on mood disorders such as anxiety and depression.
They recommend eating plenty of fibre and probiotics such as live yogurt, as well as limiting sugar, processed foods, animal fat and the use of antibiotics, antacids and anti-inflammatories, as these cause imbalances in the gut.

Appendix C
Motion to Form a Quorum
Sungsu Park,1 Peter M. Wolanin,2 Emil A. Yuzbashyan,1
Pascal Silberzan,3 Jeffry B. Stock,2* Robert H. Austin1
Bacterial gene expression is frequently regulated
by small molecules secreted into the
surrounding medium. These autoinducers
build up with increasing cell number until a
critical population density or “quorum” is
achieved, at which point the cells produce a
response such as virulence or biofilm formation
that requires the coordinated activity of
large numbers of individuals. It has generally
been assumed that quorum formation derives
primarily from conditions favorable for
growth to high cell density (1).
Numerous studies have shown that motility
promotes biofilm formation, but this
dependence has been attributed to random
transport of cells from
the bulk medium to
a surface (2). Such
experiments have
been conducted with
smooth surfaces that
have no preferred surface
sites or flow
chambers where gradients
of attractant
chemicals will be dispersed.
Here we show
that, given appropriate
surface topologies,
bacteria can use chemotaxis
to associate
and form a quorum.
Thus, chemotaxis provides
an important
mechanism for establishing
the high local cell densities required
for quorum-dependent interactions.
A culture of Escherichia coli grown to moderate
density (approximately 2 _ 108 cells/ml) in
either rich media or in minimal media was used
to uniformly fill a microfluidic chamber (7 mm
by 3 mm by 30 _m) with a small central enclosure
(250 _m by 250 _m) constructed from
silicone elastomer. Over the course of 1 to 3
hours (depending on the media), the cells migrate
from the chamber into the central enclosure
through a narrow (40 _m) channel (Fig. 1A).
This behavior is not observed with a mutant
strain that is motile but deficient in chemotaxis.
Cells accumulate in the enclosure because
they are attracted to each other due to their
secretion of amino acids, such as glycine, that
are chemoattractants. We detected this secretion
by analysis of the free amino acid content of the
growth media over time. The serine receptor,
Tsr, is the most abundant chemotaxis receptor in
E. coli, whereas the aspartate receptor, Tar, is
also present at relatively high levels (3). The
redox-sensing aerotaxis receptor, Aer, is present
at low levels but still effectively modulates chemotaxis.
Tsr binds L-serine with the highest
affinity, but it also binds L-alanine, L-cysteine,
and glycine (4). Whereas strains with tsr deleted
were unable to accumulate in the enclosure, tar
or aer deletion had little or no effect. Moreover,
addition of saturating levels of L-serine (0.5
mM), which effectively competes with glycine,
completely blocked accumulation of wild-type
cells. Saturating concentrations of L-aspartate
had no effect. The particular amino acid that is
most important in mediating self attraction
seems to depend on the conditions of growth
before nutrient depletion. It has previously been
shown that E. coli grown in succinate secrete
aspartate, which acts through Tar to cause cells
to associate into dense colonies in soft agar (5).
Chemotaxis has generally been considered as a
mechanism for cell dispersal. In nutrient-depleted
environments, however, the cells themselves
become sources of attractant molecules. Movement
toward the amino acids secreted by the
cells is enhanced by the ability of the chemotaxis
system to adjust its sensitivity so that it can
respond to very low concentrations of attractant
chemicals (6). Accumulation of a high local
density of cells may offer advantages such as
enhanced genetic exchange or communal degradation
of antibiotics, as well as the enabling of
quorum-dependent behaviors.
The sites at which cells accumulate depend
on the geometry of their surroundings. Results
with cells in percolated lattices formed from silicone
elastomer indicate that E. coli tend to accumulate
in any areas of these mazes where the
geometry provides a sufficiently enclosed space,
such as dead ends and cul-de-sacs. Self-attractive
behavior in chemotaxis has been modeled using
the Keller-Segel equations (7). Our analysis of
these equations indicates that for a small volume
connected by a small opening to a large volume,
such as our enclosure within a relatively large
microfluidic chamber (Fig. 1A) or the dead ends
in a maze (Fig. 1B), random fluctuations in cell
number that cause an increase in the density of
bacteria inside the small volume can increase irreversibly
to produce a dense accumulation of cells.
Our results suggest that self attraction could
readily produce local cell densities that exceed
the threshold necessary for quorum-dependent
processes. This notion was supported by the observation
that, after coming together within the
chamber, wild-type E. coli tended to exhibit a
further association to form dense granular aggregates.
A proposed E. coli quorum sensing signal,
AI-2, is produced by the LuxS enzyme (8). A
strain with luxS deleted accumulated into the
enclosure just like wild type, but was never observed
to form dense aggregates. Vibrio harveyi,
a highly motile marine bacterium, exhibits a similar
tendency to accumulate in confined spaces.
These cells produce light in regions of high population
density (Fig. 1, B and C). This bioluminescence,
which is one of the most well-studied
of quorum-dependent responses (8), confirms
that chemotaxis-mediated associations facilitated
by closed geometries can lead to activation of
quorum sensing–dependent genes and their associated
References and Notes
1. S. Swift et al., Adv. Microb. Physiol. 45, 199 (2001).
2. J. W. McClaine, R. M. Ford, Biotechnol. Bioeng. 78,
179 (2002).
3. S. Clarke, D. E. Koshland Jr., J. Biol. Chem. 254, 9695
4. J. Adler, Annu. Rev. Biochem. 44, 341 (1975).
5. E. O. Budrene, H. C. Berg, Nature 376, 49 (1995).
6. J. E. Segall, S. M. Block, H. C. Berg, Proc. Natl. Acad.
Sci. U.S.A. 83, 8987 (1986).
7. M. P. Brenner, L. S. Levitov, E. O. Budrene, Biophys. J.
74, 1677 (1998).
8. M. G. Surette, M. B. Miller, B. L. Bassler, Proc. Natl.
Acad. Sci. U.S.A. 96, 1639 (1999).
9. Materials and Methods are available as supporting
online material on Science Online.
10. We thank E. C. Cox for insightful comments on the
manuscript; J. S. Parkinson for strains RP437, RP2361,
RP5700, and UU1117, and for helpful advice; H. C. Berg
for strain HCB317 and for comments and advice; and B.
Bassler for strains BB120 and BB170 and for the gift of
AI-2 produced in vitro. We also gratefully acknowledge
M. Taga and K. Xavier for advice and assistance; N. C.
Darnton, P. Silberzan, H. Lin, and C. Gabel for discussions;
W. Austin for technical assistance; and J. Chen for
swarm plate assays. Supported by grants from DARPA
(MDA972-00-1-0031), NIH (R01 HG001506 and F32
GM064228 to P.M.W.), and the State of New Jersey
(NJCST 99-100-082-2042-007).
Supporting Online Material
Materials and Methods
28 October 2002; accepted 16 April 2003
1Department of Physics, 2Department of Molecular
Biology, Princeton University, Princeton, NJ 08544,
USA. 3Institut Curie, 75005 Paris, France.
*To whom correspondence should be addressed. Email:
Fig. 1. E. coli and V. harveyi accumulation and quorum sensing. (A) Epifluorescence
images of green fluorescent protein (GFP)–labeled E. coli in M9 minimal
media as they accumulate into a central 250 _m by 250 _m enclosure via a
40-_m-wide channel through 100-_m-wide walls. After 3 hours the density of
cells is more than seven times greater inside than outside. The rectangles are
silicone pillars that support the roof of the chamber. (B) Dark-field image of V.
harveyi after 8 hours in the maze. The narrowest passages are 100 _m wide.
Lines corresponding to the walls of the maze are overlaid for clarity. (C)
Photon-counting image of the intrinsic luminescence, indicating active quorum
sensing in areas where the cells have accumulated at high density (9).
188 11 JULY 2003 VOL 301 SCIENCE


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