Lactose intolerance and the proteins of the synaptic cleft

What does lactose intolerance have to do with the zillions of proteins happily infesting the synaptic cleft?  Only someone whose mind was warped into very abstract thinking by rooming with philosophy majors in college would see a connection.

The synaptic cleft is of immense theoretical interest to neuroscientists, drug chemists and pharmacologists, and of great practical interest to people affected by neurologic and psychiatric disease either in themselves or someone they know (e.g. just about everyone).

Almost exactly a year ago I wrote a post about a great paper on the proteins of the synaptic cleft by Thomas Sudhof.  You may read the post after the *****

Well Dr. Sudhof is back with another huge review of just how synapses are formed [ Neuron vol. 100 pp. 276 – 293 ’18 ], which covers very similar ground.

It is clear that he’s depressed by the state of the field.  Here are a few quotes

“I believe that we may need to pay more attention to technical details than customary because the pressures on investigators have increased the tendency to publish preliminary results, especially results obtained with new methods whose limitations are not yet clear.”

Translation: a lot of the stuff coming out is junk.

“Given the abundance of papers reporting non-validated protein interactions that cannot possibly be all correct, it seems that confidence in a possible protein-protein interaction requires either isolation of a stable complex or biophysical measurements of interactions using recombinant purified proteins.”

Translation:  Oy vey !

“Pre- or postsynaptic specializations are surprisingly easy to induce by diverse signals. This was first shown in pioneering studies demonstrating that polylysine beads induce formation of presynaptic nerve terminals in cultured neurons and in brain in vivo.” Obviously this means that you have to be very careful when you claim that a given protein or two causes a synapse to form, which researchers have not been.”

Translation not needed.

Then on to the meat of the review.  “An impressive number of candidate synaptic Cell Adhesion Molecules (CAMs) has been described (9 classes are given each with multiple members). For some of these CAMs, compelling data demonstrate their presence in synapses and suggest a functional role in synapses. Others, however, are less well documented. If one looks at the results in total, the overall impression is puzzlement: how do so many CAMs contribute to shaping a synapse?”

Then from 281 – 286 he goes into the various CAMs, showing the extent and variety of proteins found in the synaptic cleft.  Which ones are necessary and what are they doing?  Can they all be important.  There must be some redundancy as knockout of some doesn’t do much.

Here is where lactose tolerance/intolerance comes in to offer succor to the harried investigator.

Bluntly, they must be doing something, and something important,  or they wouldn’t be there.

People with lactose intolerance have nothing wrong with the gene which breaks down lactose.  Babies have no problem with breast milk.  The enzyme (lactase)  produced from the gene is quite normal in all of us.  However 10,000 years ago and earlier, cattle were not domesticated, so there was no dietary reason for a human weaned from the breast to make the enzyme.  Something turned off lactase production — from my reading, it’s not clear what.   The control region (lactase enhancer) for the lactase gene is 14,000 nucleotides upstream from the gene itself.  After domestication of cattle, so that people could digest milk their entire lives a mutation arose changing cytosine to thymine in the enhancer.  The farthest back the mutation has been found is 6.500 years. 3 other mutations are known, which keep the lactase gene expressed past weaning.  They arose independently.  All 4 spread in the population, because back then our ancestors were in a semi-starved state most of the time, and carriers had better nutrition.

How does this offer succor to Dr. Sudhof?  Simply this, here is a mechanism to turn off production of an enzyme our ancestors didn’t need past weaning.  Don’t you think this would be the case for all the proteins found in and around the synapse.  They must be doing something or they wouldn’t be there.  I realize that this is teleology writ large, but evolutionary adaptations make you think this way.

*****

The bouillabaisse of the synaptic cleft

The synaptic cleft is so small ( under 400 Angstroms — 40 nanoMeters ) that it can’t be seen with the light microscope ( the smallest wavelength of visible light 3,900 Angstroms — 390 nanoMeters).  This led to a bruising battle between Cajal and Golgi a just over a century ago over whether the brain was actually made of cells.  Even though Golgi’s work led to the delineation of single neurons he thought the brain was a continuous network.  They both won the Nobel in 1906.

Semifast forward to the mid 60s when I was in medical school.  We finally had the electron microscope, so we could see synapses. They showed up as a small CLEAR spaces (e.g. electrons passed through it easily leaving it white) between neurons.  Neurotransmitters were being discovered at the same time and the synapse was to be the analogy to vacuum tubes, which could pass electricity in just one direction (yes, the transistor although invented hadn’t been used to make anything resembling a computer — the Intel 4004 wasn’t until the 70s).  Of course now we know that information flows back and forth across the synapse, with endocannabinoids (e. g. natural marihuana) being the major retrograde neurotransmitter.

Since there didn’t seem to be anything in the synaptic cleft, neurotransmitters were thought to freely diffuse across it to being to receptors on the other (postsynaptic) side e.g. a free fly zone.

Fast forward to the present to a marvelous (and grueling to read because of the complexity of the subject not the way it’s written) review of just what is in the synaptic cleft [ Cell vol. 171 pp. 745 – 769 ’17 ] http://www.cell.com/cell/fulltext/S0092-8674(17)31246-1 (It is likely behind a paywall).  There are over 120 references, and rather than being just a catalogue, the single author Thomas Sudhof extensively discusseswhich experimental work is to be believed (not that Sudhof  is saying the work is fraudulent, but that it can’t be used to extrapolate to the living human brain).  The review is a staggering piece of work for one individual.

The stuff in the synaptic cleft is so diverse, and so intimately involved with itself and the membranes on either side what what is needed for comprehension is not a chemist but a sociologist.  Probably most of the molecules to be discussed are present in such small numbers that the law of mass action doesn’t apply, nor do binding constants which rely on large numbers of ligands and receptors. Not only that, but the binding constants haven’t been been determined for many of the players.

Now for some anatomic detail and numbers.  It is remarkably hard to find just how far laterally the synaptic cleft extends.  Molecular Biology of the Cell ed. 5 p. 1149 has a fairly typical picture with a size marker and it looks to be about 2 microns (20,000 Angstroms, 2,000 nanoMeters) — that’s 314,159,265 square Angstroms (3.14 square microns).  So let’s assume each protein takes up a square 50 Angstroms on a side (2,500 square Angstroms).  That’s room for 125,600 proteins on each side assuming extremely dense packing.  However the density of acetyl choline receptors at the neuromuscular junction is 8,700/square micron, a packing also thought to be extremely dense which would give only 26,100 such proteins in a similarly distributed CNS synapse. So the numbers are at least in the right ball park (meaning they’re within an order of magnitude e.g. within a power of 10) of being correct.

What’s the point?

When you see how many different proteins and different varieties of the same protein reside in the cleft, the numbers for  each individual element is likely to be small, meaning that you can’t use statistical mechanics but must use sociology instead.

The review focuses on the neurExins (I capitalize the E  to help me remember that they are prEsynaptic).  Why?  Because they are the best studied of all the players.  What a piece of work they are.  Humans have 3 genes for them. One of the 3 contains 1,477 amino acids, spread over 1,112,187 basepairs (1.1 megaBases) along with 74 exons.  This means that just over 1/10 of a percent of the gene is actually coding for for the amino acids making it up.  I think it takes energy for RNA polymerase II to stitch the ribonucleotides into the 1.1 megabase pre-mRNA, but I couldn’t (quickly) find out how much per ribonucleotide.  It seems quite wasteful of energy, unless there is some other function to the process which we haven’t figured out yet.

Most of the molecule resides in the synaptic cleft.  There are 6 LNS domains with 3 interspersed EGFlike repeats, a cysteine loop domain, a transmembrane region and a cytoplasmic sequence of 55 amino acids. There are 6 sites for alternative splicing, and because there are two promoters for each of the 3 genes, there is a shorter form (beta neurexin) with less extracellular stuff than the long form (alpha-neurexin).  When all is said and done there are over 1,000 possible variants of the 3 genes.

Unlike olfactory neurons which only express one or two of the nearly 1,000 olfactory receptors, neurons express mutiple isoforms of each, increasing the complexity.

The LNS regions of the neurexins are like immunoglobulins and fill at 60 x 60 x 60 Angstrom box.  Since the synaptic cleft is at most 400 Angstroms long, the alpha -neurexins (if extended) reach all the way across.

Here the neurexins bind to the neuroligins which are always postsynaptic — sorry no mnemonic.  They are simpler in structure, but they are the product of 4 genes, and only about 40 isoforms (due to alternative splicing) are possible. Neuroligns 1, 3 and 4 are found at excitatory synapses, neuroligin 2 is found at inhibitory synapses.  The intracleft part of the neuroligins resembles an important enzyme (acetylcholinesterase) but which is catalytically inactive.  This is where the neurexins.

This is complex enough, but Sudhof notes that the neurexins are hubs interacting with multiple classes of post-synaptic molecules, in addition to the neuroligins — dystroglycan, GABA[A] receptors, calsystenins, latrophilins (of which there are 4).   There are at least 50 post-synaptic cell adhesion molecules — “Few are well understood, although many are described.”

The neurexins have 3 major sites where other things bind, and all sites may be occupied at once.  Just to give you a taste of he complexity involved (before I go on to  larger issues).

The second LNS domain (LNS2)is found only in the alpha-neurexins, and binds to neuroexophilin (of which there are 4) and dystroglycan .

The 6th LNS domain (LNS6) binds to neuroligins, LRRTMs, GABA[A] receptors, cerebellins and latrophilins (of which there are 4)_

The juxtamembrane sequence of the neurexins binds to CA10, CA11 and C1ql.

The cerebellins (of which there are 4) bind to all the neurexins (of a particular splice variety) and interestingly to some postsynaptic glutamic acid receptors.  So there is a direct chain across the synapse from neurexin to cerebellin to ion channel (GLuD1, GLuD2).

There is far more to the review. But here is something I didn’t see there.  People have talked about proton wires — sites on proteins that allow protons to jump from one site to another, and move much faster than they would if they had to bump into everything in solution.  Remember that molecules are moving quite rapidly — water is moving at 590 meters a second at room temperature. Since the synaptic cleft is 40 nanoMeters (40 x 10^-9 meters, it should take only 40 * 10^-9 meters/ 590 meters/second   60 trillionths of a second (60 picoSeconds) to cross, assuming the synapse is a free fly zone — but it isn’t as the review exhaustively shows.

It it possible that the various neurotransmitters at the synapse (glutamic acid, gamma amino butyric acid, etc) bind to the various proteins crossing the cleft to get their target in the postsynaptic membrane (e.g. neurotransmitter wires).  I didn’t see any mention of neurotransmitter binding to  the various proteins in the review.  This may actually be an original idea.

I’d like to put more numbers on many of these things, but they are devilishly hard to find.  Both the neuroligins and neurexins are said to have stalks pushing them out from the membrane, but I can’t find how many amino acids they contain.  It can’t find how much energy it takes to copy the 1.1 megabase neurexin gene in to mRNA (or even how much energy it takes to add one ribonucleotide to an existing mRNA chain).

Another point– proteins have a finite lifetime.  How are they replenished?  We know that there is some synaptic protein synthesis — does the cell body send packages of mRNAs to the synapse to be translated there.  There are at least 50 different proteins mentioned in the review, and don’t forget the thousands of possible isoforms, each of which requires a separate mRNA.

Old Chinese saying — the mountains are high and the emperor is far away. Protein synthesis at the synaptic cleft is probably local.  How what gets made and when is an entirely different problem.

A large part of the review concerns mutations in all these proteins associated with neurologic disease (particularly autism).  This whole area has a long and checkered history.  A high degree of cynicism is needed before believing that any of these mutations are causative.  As a neurologist dealing with epilepsy I saw the whole idea of ion channel mutations causing epilepsy crash and burn — here’s a link — https://luysii.wordpress.com/2011/07/17/we’ve-found-the-mutation-causing-your-disease-not-so-fast-says-this-paper/

Once again, hats off to Dr. Sudhof for what must have been a tremendous amount of work

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Cellular senescence (again, again)

As well as being involved in normal cellular function, wound healing, embryology, and warding off cancer, cellular senescence may be involved in one form of neurodegeneration according to [ Nature vol. 562 pp. 503 – 504, 578 – 582 ’18 ]

Alzheimer’s disease is characterized by two findings visible with only a light microscope — the senile plaque which occurs outside neurons, and the neurofibrillary tangle (which occurs inside them).  The latter is due to accumulation of excessively phosphorylated tau protein.  A few mutations in the tau protein are known to cause neurodegeneration.  One such is the substitution of serine (S) for proline (P) at position #301 in tau (e. g. the P301S mutation).

Transgenic expression of the mutant tau in mice mimics the human illness.  Long before neurofibrillary tangles appear in neurons, glial cells (which don’t express much tau and never have neurofibrillary tangles) develop cellular senescence.  Neurons don’t show this.

p16^INK4a is a transcription factor which turns on cellular senescence, leading to expression of a bunch of proteins known as the Senescence Associated Secretory Phenotype (SASP).  It was elevated in glia.  The authors were able to prevent the neurodegeneration using another genetic tool, which produced cell death in cells expression p16^INK4a.  There was fewer neurofibrillary tangles in the animals.

The nature of the neural signal to glia causing senescence isn’t known at this point.  How glia signal back also isn’t known.

So are drugs killing senescence cells (senolytics) a possible treatment of neurodegeneration?  Stay tuned.

As readers of this blog well know, I’ve been flogging an idea of mine — that excessive cellular senescence with release of SASP products is behind the faatigue of chronic fatigue syndrome.   I’d love it if someone would measure p16^INK4a in these people — it’s so easy to do, and if the idea is correct would lead to a rational treatment for some with the disorder.

Neurodegeneration is a far larger fish to fry than CFS, and I hope people with it don’t get lost in the shuffle.

Here’s the idea again

Not a great way to end 2017

2017 ended with a rejection of the following letter to PNAS.

As a clinical neurologist with a long standing interest in muscular dystrophy(1), I was referred many patients who turned out to have chronic fatigue syndrome (CFS) . Medicine, then and now, has no effective treatment for CFS.

A paper (2) cited In an excellent review of cellular senescence (3) was able to correlate an intracellular marker of senescence (p16^INK4a) with the degree of fatigue experienced by patients undergoing chemotherapy for breast cancer. Chemotherapy induces cellular senescence, and the fatigue was thought to come from the various cytokines secreted by senescent cells (Senescence Associated Secretory Phenotype—SASP) It seems logical to me to test CFS patients for p16^INK4a (4).
I suggested this to the senior author; however, he was nominated as head of the National Cancer Institute just 9 days later. There the matter rested until the paper of Montoya et al. (5) appeared in July. I looked up the 74 individual elements of the SASP and found that 9 were among the 17 cytokines whose levels correlated with the degree of fatigue in CFS. However, this is not statistically significant as Montoya looked at 51 cytokines altogether.

In October, an article(6) on the possibility of killing senescent cells to prevent aging contained a statement that Judith Campisi’s group (which has done much of the work on SASP) had identified “hundreds of proteins involved in SASPs”. (These results have not yet been published.) It is certainly possible that many more of Montoya’s 17 cytokines are among them.

If this is the case, a rational therapy for CFS is immediately apparent; namely, the senolytics, a class of drugs which kills senescent cells. A few senolytics are currently available clinically and many more are under development as a way to attack the aging process (6).

If Montoya still has cells from the patients in the study, measuring p16^INK4a could prove or disprove the idea. However, any oncology service could do the test. If the idea proves correct, then there would be a way to treat the debilitating fatigue of both chemotherapy and CFS—not to mention the many more medical conditions in which severe fatigue is found.
Chemotherapy is a systemic process, producing senescent cells everywhere, which is why DeMaria (2) was able to use circulating blood cells to measure p16^INK4a. It is possible that the senescent cells producing SASP in CFS are confined to one tissue; in which case testing blood for p16^INK4a would fail. (That would be similar to pheochromocytoma cells, in which a few localized cells produce major systemic effects.)

Although senolytics might provide symptomatic treatment (something worthwhile having since medicine presently has nothing for the CFS patient), we’d still be in the dark about what initially caused the cells to become senescent. But this would be research well worth pursuing.

Anyone intrigued by the idea should feel free to go ahead and test it. I am a retired neurologist with no academic affiliation, lacking the means to test it.
References

1 Robinson, L (1979) Split genes and musclar dystrophy. Muscle Nerve 2: 458 – 464

2. He S, Sharpless N (2017) Senescence in Health and Disease. Cell 170: 1000 – 1011

3. Demaria M, et al. (2014) Cellular senescence promotes adverse effects of chemotherapy and cancer relapse. Cancer Discov. 7: 165 – 176

4. https://luysii.wordpress.com/2017/09/04/is-the-era-of-precision-medicine-for-chronic-fatigue-syndrome-at-hand/

5. Montoya JG, et al., (2017) Cytokine signature associated with disease severity in chronic fatigue syndrome patients, Proc Natl Acad Sci USA 114: E7150-E7158

6. Scudellari M, (2017) To stay young, kill zombie cells Nature 551: 448 – 450

Is a rational treatment for chronic fatigue syndrome at hand?

If an idea of mine is correct, it is possible that some patients with chronic fatigue syndrome (CFS) can be treated with specific medications based on the results of a few blood tests. This is precision medicine at its finest.  The data to test this idea has already been acquired, and nothing further needs to be done except to analyze it.

Athough the initial impetus for the idea happened only 3 months ago, there have been enough twists and turns that the best way explanation is by a timeline.

First some background:

As a neurologist I saw a lot of people who were chronically tired and fatigued, because neurologists deal with muscle weakness and diseases like myasthenia gravis which are associated with fatigue.  Once I ruled out neuromuscular disease as a cause, I had nothing to offer then (nor did medicine).  Some of these patients were undoubtedly neurotic, but there was little question in my mind that many others had something wrong that medicine just hadn’t figured out yet — not that it hasn’t been trying.

Infections of almost any sort are associated with fatigue, most probably caused by components of the inflammatory response.  Anyone who’s gone through mononucleosis knows this.    The long search for an infectious cause of chronic fatigue syndrome (CFS) has had its ups and downs — particularly downs — see https://luysii.wordpress.com/2011/03/25/evil-scientists-create-virus-causing-chronic-fatigue-syndrome-in-lab/

At worst many people with these symptoms are written off as crazy; at best, diagnosed as depressed  and given antidepressants.  The fact that many of those given antidepressants feel better is far from conclusive, since most patients with chronic illnesses are somewhat depressed.

The 1 June 2017 Cell had a long and interesting review of cellular senescence by Norman Sharpless [ vol. 169 pp. 1000 – 1011 ].  Here is some background about the entity.  If you are familiar with senescent cell biology skip to the paragraph marked **** below

Cells die in a variety of ways.  Some are killed (by infections, heat, toxins).  This is called necrosis. Others voluntarily commit suicide (this is called apoptosis).   Sometimes a cell under stress undergoes cellular senescence, a state in which it doesn’t die, but doesn’t reproduce either.  Such cells have a variety of biochemical characteristics — they are resistant to apoptosis, they express molecules which prevent them from proliferating and — most importantly — they secrete a variety of proinflammatory molecules collectively called the Senescence Associated Secretory Phenotype — SASP).

At first the very existence of the senescent state was questioned, but exist it does.  What is it good for?  Theories abound, one being that mutation is one cause of stress, and stopping mutated cells from proliferating prevents cancer. However, senescent cells are found during fetal life; and they are almost certainly important in wound healing.  They are known to accumulate the older you get and some think they cause aging.

Many stresses induce cellular senescence of which mutation is but one.  The one of interest to us is chemotherapy for cancer, something obviously good as a cancer cell turned senescent has stopped proliferating.   If you know anyone who has undergone chemotherapy, you know that fatigue is almost invariable.

****

One biochemical characteristic of the senescent cell is increased levels of a protein called p16^INK4a, which helps stop cellular proliferation.  While p16^INK4a can easily be measured in tissue biopsies, tissue biopsies are inherently invasive. Fortunately, p16^INK4a can also be measured in circulating blood cells.

What caught my eye in the Cell paper was a reference to a paper about cancer [ Cancer Discov. vol. 7 pp. 165 – 176 ’17 ] by M. Demaria, in which the levels of p16^INK4a correlated with the degree of fatigue after chemotherapy.  The more p16^INK4a in the blood cells the greater the fatigue.

I may have been the only reader of both papers with clinical experience wth chronic fatigue syndrome.  It is extremely difficult to objectively measure a subjective complaint such as fatigue.

As an example of the difficulty in correlating subjective complaints with objective findings, consider the nearly uniform complaint of difficulty thinking in depression, with how such patients actually perform on cognitive tests — e. g. there is  little if any correlation between complaints and actual performance — here’s a current reference — Scientific Reports 7, Article number: 3901(2017) —  doi:10.1038/s41598-017-04353.

If the results of the Cancer paper could be replicated, p16^INK4 would be the first objective measure of a patient’s individual sense of fatigue.

So I wrote both authors, suggesting that the p16^INK4a test be run on a collection of chronic fatigue syndrome (CFS) patients. Both authors replied quickly, but thought the problem would be acquiring patients.  Demaria said that Sharpless had a lab all set up to do the test.

Then fate (in the form of Donald Trump) supervened.  A mere 9 days after the Cell issue appeared, Sharpless was nominated to be the head of the National Cancer Institute by President Trump.  This meant Dr. Sharpless had far bigger fish to fry, and he would have to sever all connection with his lab because of conflict of interest considerations.

I also contacted a patient organization for chronic fatigue syndrome without much success.  Their science advisor never responded.

There matters stood until 22 August when a paper and an editorial about it came out [ Proc. Natl. Acad. Sci. vol. 114 pp. 8914 – 8916, E7150 – E7158 ’17 ].  The paper represented a tremendous amount of data (and work).  The blood levels of 51 cytokines (measures of inflammation) and adipokines (hormones released by fat) were measured in both 192 patients with CFS (which can only be defined by symptoms) and 293 healthy controls matched for age and gender.

In this paper, levels of 17 of the 51 cytokines correlated with severity of CFS. This is a striking similarity with the way the p16^INK4 levels correlated with the degree of fatigue after chemotherapy).  So I looked up the individual elements of the SASP (which can be found in Annu Rev Pathol. 21010; 5: 99–118.)  There are 74 of them. I wondered how many of the 51 cytokines measured in the PNAS paper were in the SASP.  This is trickier than it sounds as many cytokines have far more than one name.  The bottom line is that 20 SASPs are in the 51 cytokines measured in the paper.

If the fatigue of CFS is due to senescent cells and the SASPs  they release, then they should be over-represented in the 17 of the 51 cytokines correlating with symptom severity.  Well they are; 9 out of the 17 are SASP.  However although suggestive, this increase is not statistically significant (according to my consultants on Math Stack Exchange).

After wrote I him about the new work, Dr. Sharpless noted that CFS is almost certainly a heterogeneous condition. As a clinician with decades of experience, I’ve certainly did see some of the more larcenous members of our society who used any subjective diagnosis to be compensated, as well as a variety of individuals who just wanted to withdraw from society, for whatever reason. They are undoubtedly contaminating the sample in the paper. Dr. Sharpless thought the idea, while interesting, would be very difficult to test.

But it wouldn’t at all.  Not with the immense amount of data in the PNAS paper.

Here’s how. Take each of the 9 SASPs and see how their levels correlate with the other 16 (in each of the 192 CSF patients). If they correlate better with SASPs than with nonSASPs, than this would be evidence for senescent cells being the cause some cases of CFS. In particular, patients with a high level of any of the 9 SASPs should be studied for such correlations.  Doing so should weed out some of the heterogeneity of the 192 patients in the sample.

This is why the idea is testable and, even better, falsifiable, making it a scientific hypothesis (a la Karl Popper).  The data to refute it is in the possession of the authors of the paper.

Suppose the idea turns out to be correct and that some patients with CFS are in fact that way because, for whatever reason, they have a lot of senescent cells releasing SASPs.

This would mean that it would be time to start trials of senolyic drugs which destroy senescent cells on the group with elevated SASPs. Fortunately, a few senolytics are currently inc linical use.  This would be precision medicine at its finest.

Being able to alleviate the symptoms of CFS would be worthwhile in itself, but SASP levels could also be run on all sorts of conditions associated with fatigue, most notably infection. This might lead to symptomatic treatment at least.  Having gone through mono in med school, I would have loved to have been able to take something to keep me from falling asleep all the time.

The New York Times loves terrorism if it’s the right sort

The following is a verbatim quote of an article which appeared on the front page of the New York Times 11 September 2001 (yes that 11 September).

”I don’t regret setting bombs,” Bill Ayers said. ”I feel we didn’t do enough.” Mr. Ayers, who spent the 1970’s as a fugitive in the Weather Underground, was sitting in the kitchen of his big turn-of-the-19th-century stone house in the Hyde Park district of Chicago. The long curly locks in his Wanted poster are shorn, though he wears earrings. He still has tattooed on his neck the rainbow-and-lightning Weathermen logo that appeared on letters taking responsibility for bombings. And he still has the ebullient, ingratiating manner, the apparently intense interest in other people, that made him a charismatic figure in the radical student movement.

Now he has written a book, ”Fugitive Days” (Beacon Press, September). Mr. Ayers, who is 56, calls it a memoir, somewhat coyly perhaps, since he also says some of it is fiction. He writes that he participated in the bombings of New York City Police Headquarters in 1970, of the Capitol building in 1971, the Pentagon in 1972. But Mr. Ayers also seems to want to have it both ways, taking responsibility for daring acts in his youth, then deflecting it.

”Is this, then, the truth?,” he writes. ”Not exactly. Although it feels entirely honest to me.”

But why would someone want to read a memoir parts of which are admittedly not true? Mr. Ayers was asked.

”Obviously, the point is it’s a reflection on memory,” he answered. ”It’s true as I remember it.”

Mr. Ayers is probably safe from prosecution anyway. A spokeswoman for the Justice Department said there was a five-year statute of limitations on Federal crimes except in cases of murder or when a person has been indicted.

Mr. Ayers, who in 1970 was said to have summed up the Weatherman philosophy as: ”Kill all the rich people. Break up their cars and apartments. Bring the revolution home, kill your parents, that’s where it’s really at,” is today distinguished professor of education at the University of Illinois at Chicago. And he says he doesn’t actually remember suggesting that rich people be killed or that people kill their parents, but ”it’s been quoted so many times I’m beginning to think I did,” he said. ”It was a joke about the distribution of wealth.”

He went underground in 1970, after his girlfriend, Diana Oughton, and two other people were killed when bombs they were making exploded in a Greenwich Village town house. With him in the Weather Underground was Bernardine Dohrn, who was put on the F.B.I.’s 10 Most Wanted List. J. Edgar Hoover called her ”the most dangerous woman in America” and ”la Pasionara of the Lunatic Left.” Mr. Ayers and Ms. Dohrn later married.

In his book Mr. Ayers describes the Weathermen descending into a ”whirlpool of violence.”

”Everything was absolutely ideal on the day I bombed the Pentagon,” he writes. But then comes a disclaimer: ”Even though I didn’t actually bomb the Pentagon — we bombed it, in the sense that Weathermen organized it and claimed it.” He goes on to provide details about the manufacture of the bomb and how a woman he calls Anna placed the bomb in a restroom. No one was killed or injured, though damage was extensive.

Between 1970 and 1974 the Weathermen took responsibility for 12 bombings, Mr. Ayers writes, and also helped spring Timothy Leary (sentenced on marijuana charges) from jail.

Today, Mr. Ayers and Ms. Dohrn, 59, who is director of the Legal Clinic’s Children and Family Justice Center of Northwestern University, seem like typical baby boomers, caring for aging parents, suffering the empty-nest syndrome. Their son, Malik, 21, is at the University of California, San Diego; Zayd, 24, teaches at Boston University. They have also brought up Chesa Boudin, 21, the son of David Gilbert and Kathy Boudin, who are serving prison terms for a 1981 robbery of a Brinks truck in Rockland County, N.Y., that left four people dead. Last month, Ms. Boudin’s application for parole was rejected.

So, would Mr. Ayers do it all again, he is asked? ”I don’t want to discount the possibility,” he said.

”I don’t think you can understand a single thing we did without understanding the violence of the Vietnam War,” he said, and the fact that ”the enduring scar of racism was fully in flower.” Mr. Ayers pointed to Bob Kerrey, former Democratic Senator from Nebraska, who has admitted leading a raid in 1969 in which Vietnamese women and children were killed. ”He committed an act of terrorism,” Mr. Ayers said. ”I didn’t kill innocent people.”

Mr. Ayers has always been known as a ”rich kid radical.” His father, Thomas, now 86, was chairman and chief executive officer of Commonwealth Edison of Chicago, chairman of Northwestern University and of the Chicago Symphony. When someone mentions his father’s prominence, Mr. Ayers is quick to say that his father did not become wealthy until the son was a teenager. He says that he got some of his interest in social activism from his father. He notes that his father promoted racial equality in Chicago and was acceptable as a mediator to Mayor Richard Daley and the Rev. Dr. Martin Luther King Jr. in 1966 when King marched in Cicero, Ill., to protest housing segregation.

All in all, Mr. Ayers had ”a golden childhood,” he said, though he did have a love affair with explosives. On July 4, he writes, ”my brothers and I loved everything about the wild displays of noise and color, the flares, the surprising candle bombs, but we trembled mostly for the Big Ones, the loud concussions.”

The love affair seems to have continued into adulthood. Even today, he finds ”a certain eloquence to bombs, a poetry and a pattern from a safe distance,” he writes.

He attended Lake Forest Academy in Lake Forest, Ill., then the University of Michigan but dropped out to join Students for a Democratic Society.

In 1967 he met Ms. Dohrn in Ann Arbor, Mich. She had a law degree from the University of Chicago and was a magnetic speaker who often wore thigh-high boots and miniskirts. In 1969, after the Manson family murders in Beverly Hills, Ms. Dohrn told an S.D.S. audience: ”Dig it! Manson killed those pigs, then they ate dinner in the same room with them, then they shoved a fork into a victim’s stomach.”

In Chicago recently, Ms. Dohrn said of her remarks: ”It was a joke. We were mocking violence in America. Even in my most inflamed moment I never supported a racist mass murderer.”

Ms. Dohrn, Mr. Ayers and others eventually broke with S.D.S. to form the more radical Weathermen, and in 1969 Ms. Dohrn was arrested and charged with resisting arrest and assaulting a police officer during the Days of Rage protests against the trial of the Chicago Eight — antiwar militants accused of conspiracy to incite riots at the 1968 Democratic National Convention.

In 1970 came the town house explosion in Greenwich Village. Ms. Dohrn failed to appear in court in the Days of Rage case, and she and Mr. Ayers went underground, though there were no charges against Mr. Ayers. Later that spring the couple were indicted along with others in Federal Court for crossing state lines to incite a riot during the Days of Rage, and following that for ”conspiracy to bomb police stations and government buildings.” Those charges were dropped in 1974 because of prosecutorial misconduct, including illegal surveillance.

During his fugitive years, Mr. Ayers said, he lived in 15 states, taking names of dead babies in cemeteries who were born in the same year as he. He describes the typical safe house: there were usually books by Malcolm X and Ho Chi Minh, and Che Guevara’s picture in the bedroom; fermented Vietnamese fish sauce in the refrigerator, and live sourdough starter donated by a Native American that was reputed to have passed from hand to hand over a century.

He also writes about the Weathermen’s sexual experimentation as they tried to ”smash monogamy.” The Weathermen were ”an army of lovers,” he says, and describes having had different sexual partners, including his best male friend.

”Fugitive Days” does have moments of self-mockery, for instance when Mr. Ayers describes watching ”Underground,” Emile De Antonio’s 1976 documentary about the Weathermen. He was ”embarrassed by the arrogance, the solipsism, the absolute certainty that we and we alone knew the way,” he writes. ”The rigidity and the narcissism.”

In the mid-1970’s the Weathermen began quarreling. One faction, including Ms. Boudin, wanted to join the Black Liberation Army. Others, including Ms. Dohrn and Mr. Ayers, favored surrendering. Ms. Boudin and Ms. Dohrn had had an intense friendship but broke apart. Mr. Ayers and Ms. Dohrn were purged from the group.

Ms. Dohrn and Mr. Ayers had a son, Zayd, in 1977. After the birth of Malik, in 1980, they decided to surface. Ms. Dohrn pleaded guilty to the original Days of Rage charge, received three years probation and was fined $1,500. The Federal charges against Mr. Ayers and Ms. Dohrn had already been dropped.

Mr. Ayers and Ms. Dohrn tried to persuade Ms. Boudin to surrender because she was pregnant. But she refused, and went on to participate in the Brink’s robbery. When she was arrested, Ms. Dohrn and Mr. Ayers volunteered to care for Chesa, then 14 months old, and became his legal guardians.

A few months later Ms. Dohrn was called to testify about the robbery. Ms. Dohrn had not seen Ms. Boudin for a year, she said, and knew nothing of it. Ms. Dohrn was asked to give a handwriting sample, and refused, she said, because the F.B.I. already had one in its possession. ”I felt grand juries were illegal and coercive,” she said. For refusing to testify, she was jailed for seven months, and she and Mr. Ayers married during a furlough.

Once again, Chesa was without a mother. ”It was one of the hardest things I did,” said Ms. Dohrn of going to jail.

In the interview, Mr. Ayers called Chesa ”a very damaged kid.” ”He had real serious emotional problems,” he said. But after extensive therapy, ”became a brilliant and wonderful human being.” .

After the couple surfaced, Ms. Dohrn tried to practice law, taking the bar exam in New York. But she was turned down by the Bar Association’s character committee because of her political activities.

Ms. Dohrn said she was aware of the contradictions between her radical past and the comforts of her present existence. ”This is where we raised our kids and are taking care of our aging parents,” she said. ”We could live much more simply, and well we might.”

And as for settling into marriage after efforts to smash monogamy, Ms. Dohrn said, ”You’re always trying to balance your understanding of who you are and what you need, and your longing and imaginings of freedom.”

”Happily for me, Billy keeps me laughing, he keeps me growing,” she said.

Mr. Ayers said he had some of the same conflicts about marriage. ”We have to learn how to be committed,” he said, ”and hold out the possibility of endless reinventions.”

As Mr. Ayers mellows into middle age, he finds himself thinking about truth and reconciliation, he said. He would like to see a Truth and Reconciliation Commission about Vietnam, he said, like South Africa’s. He can imagine Mr. Kerrey and Ms. Boudin taking part.

And if there were another Vietnam, he is asked, would he participate again in the Weathermen bombings?

By way of an answer, Mr. Ayers quoted from ”The Cure at Troy,” Seamus Heaney’s retelling of Sophocles’ ”Philoctetes:” ” ‘Human beings suffer,/ They torture one another./ They get hurt and get hard.’ ”

He continued to recite:

History says, Don’t hope

On this side of the grave.

But then, once in a lifetime

The longed-for tidal wave

Of justice can rise up

And hope and history rhyme.

Thinking back on his life , Mr. Ayers said, ”I was a child of privilege and I woke up to a world on fire. And hope and history rhymed.”

The rehabilitation and eventual beatification of Cesar Sayoc

New York Times  27 Oct 2038 Columbia University announced today that Cesar Sayoc having served his debt to society would assume the newly created Kathy Boudin chair of Social Justice and Explosives.  The chair was named for Boudin who 20 years ago actively served as an adjunct professor at the Columbia University School of Social Work after her release from prison. Some faculty objected on the grounds that, unlike Boudin, his explosives never actually killed anyone — https://en.wikipedia.org/wiki/Kathy_Boudin.  Members of the Bill Ayers society (https://en.wikipedia.org/wiki/Bill_Ayers) were also upset, even though Sayoc, like Bill back then is a good friend of the current president.

Thomas Gold lives !

Thomas Gold was a scientific jack of all trades being involved in physics, cosmology and geochemistry, the latter of interest to us here.  He thought petroleum and other hydrocarbons were actually produced by micro-organisms below the surface of the earth, providing us with a replenishable supply (how ecological !)  Here’s part of a Wiki article about him —  Hydrocarbons are not biology reworked by geology (as the traditional view would hold), but rather geology reworked by biology.– https://en.wikipedia.org/wiki/Thomas_Gold.

Why bring him up now?  Because [ Proc. Natl. Acad. Sci. vol. 115 pp. 10702 – 10707 ’18 ] (http://www.pnas.org/content/115/42/10702) showed that 600 meters below the surface (where light and molecular oxygen never go) Cyanobacteria were found.  They use the electrons stripped from hydrogen (which is said to be produced in the subsurface by several (unspecified) abiotic mechanisms) as an energy source.  The electrons have to go somewhere, and they postulate that the electron acceptors are iron or manganese oxides. Wherever microorganisms have been found in deep continental settings hydrogen concentration decreases.

Basically life acts as the middleman, taking an energy cut from the flow of electrons from reductant to oxidant.

Seriously, life may have actually arisen in such situations.

Has the holy grail for Parkinson’s disease been found?

Will the horribly named SynuClean-D treat Parkinsonism?  Here is the structure described  verbally.  Start with pyridine.  In the 2 position put benzene with a nitrogroup in the meta position, position 3 on pyridine NO2, position 4 CF3, position 5 CN (is this trouble?) position 6 OH.  That’s it.  Being great chemists you can immediately see what it does.

Back up a bit.  One of the pathologic findings in parkinsonism in the 450,000 dopamine neurons we have in the pars compacta at birth, is the Lewy body, which is largely made of the alpha-synuclein protein.  This is thought to kill the neurons in some way (just which form of alpha-synuclein is the culprit is still under debate — the monomer, the tetramer etc. etc).  Even the actual conformation of the monomer is still under debate (intrinsically disordered) etc. etc.

The following paper [ Proc. Natl. Acad. Sci. vo. 115 pp. 10481 – 10486 ’18 ] claims that SynuClean-D inhibits alpha-synuclein aggregation, disrupts mature amyloid fibrils made from it, prevents fibril propagation and abolishes the degeneration of dopamine neurons in an animal model of Parkinsonism.  Wow ! ! !

Time for some replication — look at the disaster from Harvard Med School about cardiac stem cells, with 30+ papers retracted. https://www.nytimes.com/2018/10/15/health/piero-anversa-fraud-retractions.html.  Ghastly.

You can’t con an honest man, but you can con yourself

How could Kavanaugh and Ford tell two diametrically opposed stories, which both sincerely believed to be true? Here are 3 examples of exactly how it could happen, the first from clinical neurologic practice, the other two from the New York Times and the Journal of the American Medical Association.

As a neurologist I was asked to do an Independent Medical Evaluation (IME) on an unfortunate man who was electrocuted at work (he worked on high voltage transmission lines). He went into cardiac arrest and sustained severe brain damage. The issue was not fault, which the power company readily admitted, but whether in what appeared to be a vegetative state, with no visible response to verbal commands, he was in fact conscious but unable to respond. In the latter case the reward to the family would have been substantially larger (for pain and suffering in addition to loss of consortium, etc. etc.). It was claimed that facilitated communication showed that he was able to write the answer to simple calculations given verbally, not visually.

Reviewing the chart before seeing the man, showed that he and his wife were truly admirable people, adopting children that no one else wanted and raising them despite limited income. He was seen at the rehab facility, with attorneys for the insurer for the power company and his family present. It was apparent that the people caring for him were quite devoted, both to him and his wife and were very sincere, especially one of his young therapists.

The neurologic exam showed that although he did react to deep pain (sternal compression), he did not follow simple commands (e.g. blink). He appeared to be in a coma. Following the neurologic examination the young therapist then demonstrated how when he held the man’s hand to which a pencil was attached, the man could actually perform calculations — add 2 and 2 produce a 4, etc. etc. Several such calculations were produced all with correct answer.

What do you think I did next?

No peeking. Think about it.

 

 

 

 

 

I took the first sheet of paper away, placed a clean sheet under the man’s hand and asked for a repeat (this time with the therapist’s eyes closed).

This produced a bunch of random lines, nothing more.  When the therapist opened his eyes and saw the results, he was visibly shaken and close to tears.

Was therapist faking the whole time? At any time? I seriously doubt it. A fraudster could easily have produced a reasonable number with his eyes shut. Try it yourself. He didn’t.

“You can’t con an honest man” — http://www.amazon.com/The-Sting-Man-Inside-Abscam/dp/0143125273

True, but you certainly can con yourself.

The second example is of a highly educated woman (a tenured professor of ethics at Rutgers Newark) using ‘facilitated communication’ who convinced herself that a severely retarded individual could communicate, and was in fact in love with her.  The jury convicted her of sexual assault and sent her to prison.

The article appeared in 25 October 2015 New York Times Magazine — here is a link

https://www.nytimes.com/2015/10/25/magazine/the-strange-case-of-anna-stubblefield.html.

The third example is the product of the youngest author ever to appear in the Journal of the American Medical Association (Emily Rosa age 11). She put a definitive end to “Therapeutic Touch”— http://jama.jamanetwork.com/article.aspx?articleid=187390

The point of all this is that highly intelligent people can con themselves.  I take no position on whether Ford or Kavanaugh (or both) conned themselves.  Syracuse, when I was practicing in the area in the 90s, was a hotbed of facilitated memory recollection, usually resulting in claims of sexual abuse, and I saw several parents whose life had been destroyed by it.  It would be of great interest to find out if Dr. Ford’s therapist used the technique.  We’ll likely never know and the Ford Kavanaugh affair will be the Patty Hearst affair of the decade.

Who doesn’t want to be smarter?

I’ve never met anyone (even future Nobel laureates) who didn’t wish they were smarter.  So cognitive training should do the trick.  Right?  Not so fast.  In a very well written (and even funny in parts) article in PNAS vol 115 pp. 9897 – 9904 ’18 titled “How to play 20 questions with nature and lose: Reflections on 100 years of brain training research” all the pitfalls of setting up a study to prove or disprove the benefits of cognitive training are laid out.  The paper is worth reading for anyone considered any sort of manipulation to change human behavior it (including medication which is why drug chemists should be interested in it).  You can read it for free at

http://www.pnas.org/content/early/2018/09/26/161702114.full

This didn’t work for someone —

Try this one — http://www.pnas.org/content/115/40/9897

An enormous number of pitfalls of the work already done on the efficacy of cognitive training are laid out, far too numerous to summarize here.

I’ve written about one such pitfall (expectancy effects) earlier — here it is

Science proves cognitive training will raise your IQ 5 – 10 points

Who among you doesn’t want to be smarter? A placebo controlled study with 25 people in each group showed that cognitive training raised IQ 5 – 10 points [ Proc. Natl. Acad. Sci. vol. 113 pp. 7470 – 7474 ’16 ].

You know that there has to be a catch and there is. The catch points to a problem with every placebo controlled trial ever done, particularly those with drugs, so drug chemists pay attention.

What was the placebo? It was the way subjects are recruited for these studies. Of 19 previous studies in the literature, 17 recruited patients using terms like ‘cognition’ or ‘brain training’, so the authors put out two ads for subjects.

Here are the two ads they used

Ad #1

Brain Training and Cognitive Enhancement
Numerous studies of ahown that working memory training can increase fluid intelligences (several references cited)
Participate in a study today !
EMail for more information GMUBrainTraining@Gmail.com

Ad #2

EMail Today and Participate in a study
Need SONA credits? (I have no idea what they are)
Sign up for a study today and earn up to 5 credits
Participate in a study today !
cforough@masonlive.gmu.edu

I might mention that the two ads were identical in total size, font sizes, coloration used etc. etc.

” Two individual difference metrics regarding beliefs about cognition and intelligence were also collected as potential moderators. The researchers who interacted with participants were blind to the goal of the experiment and to the experimental condition”  Not bad. Not bad at all.

The results: those recruited with ad #1 showed the increase in IQ, those recruited with ad #2 showed no improvement.

It was an expectancy effect. Those who thought intelligence could be raised by training, showed the greatest IQ improvement.   Every sick patient wants to get better, and any drug trial simply must mention what it is for, the risks and rewards, so this effect is impossible to avoid. It probably explains the high placebo response rate for migraine and depression (over 30% usually).

What is really impressive (to me at least) is that the improvement was not in a subjective rating scale (such as is used for depression), but in something as objective as it gets. IQ questions have a right and wrong answers. You can argue about whether they ‘really’ measure intelligence, but they measure what they measure and fluid intelligence is one of them.

Medicine is full of fads and fashions, sugar is poison, fat is bad (no it’s good) etc. etc. and this is true in spades for treatments, particularly those touted in the press. Next time you’re in a supermarket, look at the various nostrums mentioned in the magazines at the checkout stand.

When I first started out in practice, one particular headache remedy was getting great results. The rationale behind it seemed bizarre, so I asked a very smart  old GP about it — his advice — “use it while it works”. Rest in peace, Herb

Triplets and TADs

Neurologists have long been interested in triplet diseases — https://en.wikipedia.org/wiki/Trinucleotide_repeat_disorder.  The triplet is made of a string of 3 nucleotides.  Example —  cytosine adenosine guanosine or CAG — which accounts for a lot of them.  We have lots of places in our genome where such repeats normally occur, with the triplets repeated up to 42 times.  However in diseases like Huntington’s chorea the repeats get to be as many as 250 CAGs in a row.  You normally are quite fine as long as you have under 36 of them, and no one has fewer than 6 at this particular location.

Subsequently, expansions of 4, 5, and 6 nucleotide repeats have also been shown to cause disease, bring the total of repeat expansion diseases to over 40.  Why more than half of them should affect the nervous system entirely or for the most part is a mystery.  Needless to say there are plenty of theories.

This leads to three questions (1) there are repeats all over the genome, why do only 40 or so of them expand (2) since we all have repeats in front of the genes where they cause disease why don’t we all have the diseases (3) why do the number of repeats expand with each succeeding generation — the phenomenon is called anticipation.  I saw one such example where a father brought his son to my muscular dystrophy clinic.  The boy had moderately severe myotonic dystrophy.  When I shook the father’s hand, it was clear that he had mild myotonia, which had in no way impaired his life (he was a successful banker).

A recent paper in Cell may help answer the first question and has a hint about the second [ Cell vol. 175 pp. 38 – 40, 224 – 238 ’18 ].  21 of 27 disease associated short tandem repeats (daSTRs) localize to something called a topologically associated domain (TAD) or subdomain (subTAD) boundary. These are defined as contiguous intervals in the genome in which every pair has an elevated interaction frequency compared to loci out side the domain.  TADs and subTADs are measured using chromosome conformation capture assays (acronyms for them include 3C, CCC, 4C, 5C, Hi-C).

Briefly they are performed as follows.  Intact nuclei are isolate from live cel cultures.  These are subjected to paraformaldehye crosslinking to fix segment of genome in close physical proximity. The crosslinked genomic DNA is digested with a restriction endonuclease, and the products expanded by PCR using primers in all possible combinations.  Then having a complete genome sequence in hand, you see what regions of the genome got close enough together to show up in the assay.

This may help explain question one, and the paper gives some speculation about question two — we don’t all have these diseases, because unlike the unfortunates with them, we don’t have problems in our genes for DNA replication, repair and recombination.  There is some evidence for this;  studies in model organisms with these mutations do have short tandem repeat instability.

Unfortunately the paper doesn’t discuss anticipation, because no clinicians appear to be among the authors, even though they’re from Penn which 50+ years ago was very strong in clinical neurology.

None of this work discusses the fascinating questions of how the expanded repeats cause disease, or why so many of them affect the nervous system.

The Kavanaugh Ford confrontation will be to this decade what the Patty Hearst kidnapping was to a previous one  — https://en.wikipedia.org/wiki/Patty_Hearst.  Since I suffered 4 episodes of physical (not sexual) abuse as a kid, and dealt with this extensively as a neurologist, I’m trying to decide whether to write about it.  Emotions are high and there are a lot of nuts out there on the net. There is even a reasonable possibility that both Ford and Kavanaugh are right and not lying.

How art and science differ

The difference between art and science was really brought home to me on a visit to a museum on our recent trip to Venice.  I’d never heard about the Memphis Group of artists before — https://en.wikipedia.org/wiki/Memphis_Group.  They mostly made wacky looking furniture, as a revolt against the austere Bauhaus style –https://en.wikipedia.org/wiki/Bauhaus.   Back in the day, people living in the Harvard Law dorms Gropius (one of the Bauhaus founders) designed hated them as unlivable.  Similarly one can regard Pop art as a revolt against abstract expressionism, impressionism as a revolt against classicism, etc. etc.   The very notion of progress and building on the past is antithetical.

Science also produces a succession of theories, but they build on previously successful theories extending and incorporating them.  Special and general relativity subsume Newtonian mechanics, Maxwell’s laws merge electricity and magnetism.  Current molecular biology extends the classic dogma of the 60s and 70s — DNA makes RNA makes protein.  None of the older theories are rejected just improved.

So progress is inherent to the scientific enterprise, and is unheard of in the arts.  In one sense the arts are closer to the creative destruction of capitalism than its practitioners would like to admit.

I’ll be back to writing more scientific posts shortly, but here’s a question to be answered in a later post.

What extremely famous artist, spent more of his life as a mechanical and military engineer than as an artist?