Low Bone Density in Autism and Brain Calcification (Bone-Vascular Axis + Altered Calcium Homeostasis), – a role for Vitamin K2, or something more potent?

Today’s post with a long tittle is a spin-off from looking at the health benefits of the Mediterranean Diet.  This often quoted diet really does make you live longer and healthier; scientists are again trying to understand exactly why.  This sent me looking at various things, one of which was vitamin k, which is abundant in the Mediterranean diet.  It turns out that another healthy diet, one found in Japan, may have incorporated an even better source of this vitamin, since it is high in vitamin K2 rather than K1.

I thought this post would just end up being about general health, rather than autism specifically, but as I did more digging it seems highly credible that some people’s autism could be improved simply by adjusting their calcium homeostasis.

This will not come as a surprise to one of our readers who discovered that giving oral calcium supplements to her son with Asperger’s triggered a major regression towards Classic autism. Fortunately it was reversed by stopping the supplementation.

You likely have an older relative with osteoporosis, which is caused by decreased bone density.  Osteoporosis is defined as a bone density of 2.5 standard deviations below that of a young adult.

Osteoporosis is a condition caused by loss of calcium homeostasis, meaning that bones are losing too much calcium to the blood.  Not surprisingly this calcium has to go somewhere and researchers have come up with the idea of the bone-vascular axis, to explain that this calcium ends up causing vascular calcification, particularly in the heart.

Because so many Americans have heart disease, the condition is very well funded and studied.  You can measure the level of calcium deposits (calcification) in the heart and you can measure bone density.

Many people with osteoporosis (loss of calcium in the bones) suffer from vascular calcification.

People who have a diet high in vitamin K and particularly vitamin K2 have much lower incidence of diseases of the bone-vascular axis and therefore live longer.

In Japan high dose vitamin K2 is a registered drug to treat osteoporosis.  In the West K2 exists as a drug, but not for osteoporosis or calcification.

In the rest of the world it is available as a supplement in very low doses.

In the Western world of evidence-based medicine it appears Japanese evidence does not count.  This is not the first time I have encountered this.

In the west people with osteoporosis might be prescribed calcium supplements that have added vitamin D to promote absorption.

Fortunately there also some interesting drugs that have been developed to affect calcium homeostasis.  Some are now cheap generics.

Bone-vascular axis in Autism

This is an autism blog, so we already know that in autism there is excess calcium found in those samples held in brain banks.

There was also a very recent study:-

Dystrophy with Calcification within Brains of Autistic and Control Subjects

Background: Intracranial calcifications are observed in many diseases including those with viral and bacterial infections, vascular pathology, toxic injury, brain tumors, teratomas, lissencephaly, in children with Fahr’s disease, and very often in parasitic infections (Rabbitt et al 1969).

Objectives: Our neuropathological studies of autistic subjects brains have revealed the presence of dystrophic changes with calcification. The aim of this study was to determine the prevalence of this type of encephalopathy in autistic and control cohorts.

Methods: The brain hemispheres of 13 autistic and 14 control subjects 4 to 64 years of age were fixed in 10% formalin, dehydrated and embedded in celloidin and cut into 200 μm- or 50 μm-thick coronal serial sections

Results: Dystrophy with calcification was found in all of the 13 autistic and 14 control brains examined. Dystrophic changes disrupt the continuity of the cortical ribbon and white matter in the frontal, temporal and occipital lobes but only on the lateral side of the brain. The pathology spreads from the leptomeningeal vessels to the cortex and white matter and was detectable by postmortem MRI and histopathological examination. Microscopic examination revealed linear dystrophic lesions free of neurons but with signs of neuronal degeneration at the border between the dystrophic and normal cortex. There was no sign of activation of astrocytes or macrophages within the dystrophic and adjacent brain tissue. The dominant component of the dystrophic lesions was calcium deposits.

Conclusions: Similar morphology of lesions in control and autistic subjects 4 to 64 years of age suggests that dystrophic calcifications undergo relatively limited modifications with age. However, the presence of degenerated neurons and vessels with degenerated smooth muscle cells in the border zone between the lesion and cortex suggests the process of brain tissue damage continues to progress decades after the original causative events. Multifocal dystrophy with calcification in all the examined brains of autistic and control subjects reflects a common pathological mechanism with yet undetermined subclinical or clinical manifestations.

What about reduced bone density in autism?  Well I thought nobody would have looked, but they have.

Studies Link Autism to Low Bone Density and Increased Fractures

The increased risk was greatest among girls and women affected by autism spectrum disorder:

* Girls with autism had eight times the hip-fracture rate of other girls.

* Women with the disorder had ten times the rate of spinal fracture of other women.

* Boys with autism had double the hip-fracture rate of other boys.

* Men and women with autism (ages 23 to 50) had nearly 12 times the hip fracture rate of other adults.

* Women with autism also had double the rate of arm, wrist and hand fractures.

Bone Density in Peripubertal Boys with Autism Spectrum Disorders

Brief Report: Bone Fractures in Children and Adults with Autism Spectrum Disorders

So it looks like more severe autism (autistic girls have 8 times higher fracture rate) in particular is linked with reduced bone density. Girls with autism tend to have more severe autism, at least until recently. This is what you would have expected, the more severe the autism the more disturbed the calcium homeostasis and likely bone-vascular axis.

Is there excess calcium in the hearts of people with autism? I guess nobody thought to look.  People will severe autism tend not to live into old age and so data will be limited.

Since you can study and measure calcification non-invasively, some researcher with time on his/her hands might want to correlate reduced bone density with calcification in the brain/heart.

Given the critical role calcium signaling plays in signaling within the brain, it is clear that excess physical calcium has the potential to disturb all the finely balance flows of Ca²⁺ ions that control many aspects of brain function.

In particular the excess Ca²⁺ affects mitochondria, which is known to be disturbed in many people with autism.  The mechanism here is the mitochondrial aspartate/ glutamate carrier (AGC).

Altered calcium homeostasis in autism-spectrum disorders: Evidence from biochemical and genetic studies of the mitochondrial aspartate/glutamate carrier AGC1

Autism is a severe developmental disorder, whose pathogenetic underpinnings are still largely unknown. Temporocortical gray matter from six matched patient–control pairs was used to perform post-mortem biochemical and genetic studies of the mitochondrial aspartate/ glutamate carrier (AGC), which participates in the aspartate/malate reduced nicotinamide adenine dinucleotide shuttle and is physiologically activated by calcium (Ca ²⁺). AGC transport rates were significantly higher in tissue homogenates from all six patients, including those with no history of seizures and with normal electroencephalograms prior to death. This increase was consistently blunted by the Ca ²⁺ chelator ethylene glycol tetraacetic acid; neocortical Ca ²⁺ levels were significantly higher in all six patients; no difference in AGC transport rates was found in isolated mitochondria from patients and controls following removal of the Ca ²⁺ -containing postmitochondrial supernatant. Expression of AGC1, the predominant AGC isoform in brain, and cytochrome c oxidase activity were both increased in autistic patients, indicating an activation of mitochondrial metabolism. Furthermore, oxidized mitochondrial proteins were markedly increased in four of the six patients. Variants of the AGC1-encoding SLC25A12 gene were neither correlated with AGC activation nor associated with autism-spectrum disorders in 309 simplex and 17 multiplex families, whereas some unaffected siblings may carry a protective gene variant. Therefore, excessive Ca ²⁺ levels are responsible for boosting AGC activity, mitochondrial metabolism and, to a more variable degree, oxidative stress in autistic brains. AGC and altered Ca ²⁺ homeostasis play a key interactive role in the cascade of signaling events leading to autism: their modulation could provide new preventive and therapeutic strategies.

Other diseases of brain calcification

There are conditions known to be caused by brain calcification.

Vascular Calcification

Vascular Calcification

Clinically, vascular calcification is now accepted as a valuable predictor of coronary heart disease.  Achieving control over this process requires understanding mechanisms in the context of a tightly controlled regulatory network, with multiple, nested feedback loops and cross talk between organ systems, in the realm of control theory. Thus, treatments for osteoporosis such as calcitriol, estradiol, bisphosphonates, calcium supplements, and intermittent PTH are likely to affect vascular calcification, and, conversely, many treatments for cardiovascular disease such as statins, antioxidants, hormone replacement therapy, angiotensin-converting enzyme inhibitors, fish oils, and calcium channel blockers may affect bone health. As we develop and use treatments for cardiovascular and skeletal diseases, we must give serious consideration to the implications for the organ at the other end of the bone-vascular axis.

Fahr disease

Idiopathic Basal Ganglia Calcification, also known as Fahr disease, is a rare, genetically dominant, inherited neurological disorder characterized by abnormal deposits of calcium in areas of the brain that control movement. Through the use of CT scans, calcifications are seen primarily in the basal ganglia and in other areas such as the cerebral cortex

Brain calcifications induce neurological dysfunction that can be reversed by a bone drug

Perivascular calcifications within the brain form in response to a variety of insults. While considered by many to be benign, these calcium phosphate deposits or "brain stones" can become large and are associated with neurological symptoms that range from seizures to parkinsonian symptoms. Here we hypothesize that the high concentrations of calcium in these deposits produce reversible, toxic effects on neurons that can be overcome with "bone" drugs that chelate solid phase calcium phosphates. We present preliminary findings that suggest a direct association between progressive neurological symptoms and brain calcification and the symptomatic improvement of seizures, headaches, and parkinsonian symptoms in patients treated with the bisphosphonate drug disodium etidronate, normally used to treat bone diseases. Future, longitudinal epidemiological studies and randomized trials will be needed to determine the true relationship between brain stones and neurological disorders as well as the utility of bisphosphonates in their prevention and treatment.

Possible therapies for brain calcification

Etidronic Acid

Etidronic acid (Didronel ®) is a bisphosphonate used to strengthen bone, treat osteoporosis, and treat Paget's disease of bone.

Bisphosphonates primarily reduce osteoclastic activity, which prevents bone resorption, and thus moves the bone resorption/formation equilibrium toward the formation side and hence makes bone stronger on the long run. Etidronate, unlike other bisphosphonates, also prevents bone calcification. For this reason, other bisphosphonates, like alendronate, are preferred when fighting osteoporosis. To prevent bone resorption without affecting too much bone calcification, etidronate must be administered only for a short time once in a while, for example for two weeks every 3 months. When given on a continuous basis, say every day, etidronate will altogether prevent bone calcification. This effect may be useful and etidronate is in fact used this way to fight heterotopic ossification. But in the long run, if used on a continuous basis, it will  cause osteomalacia.

Alendronic acid

Alendronic acid  — sold as Fosamax by Merck — is a bisphosphonate drug used for osteoporosis, osteogenesis imperfecta, and several other bone diseases. It is marketed alone as well as in combination with vitamin D (2,800 IU and 5,600 IU, under the name Fosamax+D). Merck's U.S. patent on alendronate expired in 2008 and the drug is now available as a generic. This is the most widely prescribed bisphosphonate medicine in the United States .

Vitamin K2

Vitamin K is a group of structurally similar, fat-soluble vitamins the human body requires for complete synthesis of certain proteins that are prerequisites for blood coagulation that the body needs for controlling binding of calcium in bones and other tissues. The vitamin K-related modification of the proteins allows them to bind calcium ions, which they cannot do otherwise. Without vitamin K, blood coagulation is seriously impaired, and uncontrolled bleeding occurs. Low levels of vitamin K also weaken bones and promote calcification of arteries and other soft tissues.

Vitamin K2 is an approved drug therapy in Japan for dysfunctional calcium homeostasis where calcium is lost from your bones (osteoporosis)  and added to the lining of your arteries.

The mechanism involves something called osteocalcin, but is not fully understood.

Osteocalcin originates from osteoblastic synthesis and is deposited into bone or released into circulation, where it correlates with measures of bone formation. The presence of 3 vitamin K-dependent γ carboxyglutamic acid residues is critical for osteocalcin’s structure, which appears to regulate the maturation of bone mineral. In humans, the percentage of the circulating osteocalcin that is not γ-carboxylated (percent ucOC) is used as a biomarker of vitamin K status.

Osteocalcin also plays a yet to be understood role in the glucose metabolism and insulin sensitivity.  Indeed a clinical trial in humans has confirmed this effect exists.

Vitamin K2 Supplementation Improves Insulin Sensitivity via Osteocalcin Metabolism: A Placebo-Controlled Trial

To summarize, we have demonstrated for the first time that vitamin K2 supplementation for 4 weeks increased insulin sensitivity in healthy young men, which seems to be related to increased cOC rather than modulation of inflammation. Small sample size limits firm interpretation on β-cell function. Our results are consistent with previous studies that demonstrated improved glucose intolerance or relieved insulin resistance by treatment with vitamin K1  or vitamin K2 , respectively.

 

 

So while the mechanism remains unclear, vitamin K2 does much more than is commonly thought.

It is thought that the amount of vitamin K2 in diet is too low to keep calcium where it should be and the suggested daily amount in diet is too low.

Vitamin K in the treatment and prevention of osteoporosis and arterial calcification

PURPOSE:

The role of vitamin K in the prevention and treatment of osteoporosis and arterial calcification is examined.

SUMMARY:

Vitamin K is essential for the activation of vitamin K-dependent proteins, which are involved not only in blood coagulation but in bone metabolism and the inhibition of arterial calcification. In humans, vitamin K is primarily a cofactor in the enzymatic reaction that converts glutamate residues into gamma-carboxyglutamate residues in vitamin K-dependent proteins. Numerous studies have demonstrated the importance of vitamin K in bone health. The results of recent studies have suggested that concurrent use of menaquinone and vitamin D may substantially reduce bone loss. Menaquinone was also found to have a synergistic effect when administered with hormone therapy. Several epidemiologic and intervention studies have found that vitamin K deficiency causes reductions in bone mineral density and increases the risk of fractures. Arterial calcification is an active, cell-controlled process that shares many similarities with bone metabolism. Concurrent arterial calcification and osteoporosis have been called the "calcification paradox" and occur frequently in postmenopausal women. The results of two dose-response studies have indicated that the amount of vitamin K needed for optimal gamma-carboxylation of osteocalcin is significantly higher than what is provided through diet alone and that current dosage recommendations should be increased to optimize bone mineralization. Few adverse effects have been reported from oral vitamin K.

CONCLUSION:

Phytonadione and menaquinone may be effective for the prevention and treatment of osteoporosis and arterial calcification.

Vitamin K2 reduces coronary heart disease:-

Dietary Intake of Menaquinone (Vitamin K2) Is Associated with a Reduced Risk of Coronary Heart Disease: The Rotterdam Study

In conclusion, our findings suggest a protective effect of menaquinone intake against CHD, which could be mediated by inhibition of arterial calcification. Adequate intake of foods rich in menaquinones, such as curds and (low-fat) cheese, may contribute to CHD prevention.

Reduce AGC Activity

Another option would be to reduce activity of AGC (mitochondrial aspartate/glutamate carrier) in the brain.  This is the realm of  mouse experiments.

In most neurodegenerative diseases there is too little AGC activity.   AGC is necessary for neuronal functions and is involved in myelinogenesis, so we again have to think about multiple sclerosis (MS).  MS is characterized by the loss of the ability to regenerate the myelin layer, so called remyelination. Autism is characterized by unusual myelination. 

Sulfatide is a major component in the nervous system and is found in high levels in the myelin sheath in both the peripheral nervous system and the central nervous system. Myelin is typically composed of about 70 -75% lipids, and sulfatide comprises 4-7% of this 70-75%.^[2] When lacking sulfatide, myelin sheath is still produced around the axons; however, when lacking sulfatide the lateral loops and part of the nodes of Ranvier are disorganized, so the myelin sheath does not function properly.^[5] Thus, lacking sulfatide can lead to muscle weakness, tremors, and ataxia

Dysregulation of myelin sulfatides is a risk factor for cognitive decline with age. Vitamin K is present in high concentrations in the brain and has been suggested to  regulate the sulfatide metabolism.  That would suggest that low levels of vitamin K (from diet and produced by bacteria in the intestines) might reduce sulfatide levels and hence impair myelination.

So this would appear to suggest an overlap in the effect of vitamin K and AGC activity. 

We also discover that AGC is regulated by CREB in response to pathological inflammation.  Inflammation is a recurring theme in autism.

It turns out that CREB regulates numerous genes/proteins that are dysfunctional in autism, including:-

·        Somatostatin, also known as growth hormone–inhibiting hormone (GHIH)
·        Brain-derived neurotrophic factor BDNF

·        VGF nerve growth factor.  VGF expression is induced by NGF, CREB and BDNF and regulated by neurotrophin-3.

·        genes involved in the mammalian circadian clock(PER1, PER2).

CREB (cAMP response element-binding protein) is a cellular transcription factor. It binds to certain DNA sequences calledcAMP response elements (CRE), thereby increasing or decreasing the transcription of the downstream genes. CREB was first described in 1987 as a cAMP-responsive transcription factor regulating the somatostatin gene.

Genes whose transcription is regulated by CREB include: c-fos, BDNF, tyrosine hydroxylase, numerous neuropeptides (such  assomatostatin,  enkephalin, VGF, corticotropin-releasing hormone),^[2] and genes involved in the mammalian circadian clock(PER1, PER2).

CREB is closely related in structure and function to CREM (cAMP response element modulator) and ATF-1 (activating transcription factor-1) proteins. CREB proteins are expressed in many animals, including humans.

CREB has a well-documented role in neuronal plasticity and long-term memory formation in the brain and has been shown to be integral in the formation of spatial memory.^[5] CREB downregulation is implicated in the pathology of Alzheimer's disease and increasing the expression of CREB is being considered as a possible therapeutic target for Alzheimer’s disease.^[6] CREB also has a role in photoentrainment in mammals.

Somatostatin, also known as growth hormone–inhibiting hormone (GHIH) or by several other names, is a peptide hormone that regulates the endocrine system and affects neurotransmission and cell proliferation via interaction with G protein-coupled somatostatin receptors and inhibition of the release of numerous secondary hormones. Somatostatin inhibits insulin and glucagon secretion.

For our reader in Gdansk and parents of kids who do not sleep :-

Involvement in Circadian Rhythms

Entrainment of the mammalian circadian clock is established via light induction of PER. Light excites melanopsin-containing photosensitive retinal ganglion cellswhich signal to the suprachiasmatic nucleus (SCN) via the Retinohypothalamic tract (RHT). Excitation of the RHT signals the release of glutamate which is received by NMDA receptors on SCN, resulting in a calcium influx into the SCN. Calcium induces the activity of Ca²⁺/calmodulin-dependent protein kinases, resulting in the activation of PKA, PKC, and CK2.  These kinases then phosphorylate CREB in a circadian manner that further regulates downstream gene expression. The phosphorylated CREB recognizes the cAMP Response Element and serves as a transcription factor for Per1 and Per2, two genes that regulate the mammalian circadian clock. This induction of PER protein can entrain the circadian clock to light/dark cycles inhibits its own transcription via a transcription-translation feedback loop which can advance or delay the circadian clock. However, the responsiveness of PER1 and PER2 protein induction is only significant during the subjective night.

Altered calcium homeostasis in autism-spectrum disorders: evidence from biochemical and genetic studies of the mitochondrial aspartate/glutamate carrier AGC1.

Autism is a severe developmental disorder, whose pathogenetic underpinnings are still largely unknown. Temporocortical gray matter from six matched patient-control pairs was used to perform post-mortem biochemical and genetic studies of the mitochondrial aspartate/glutamate carrier (AGC), which participates in the aspartate/malate reduced nicotinamide adenine dinucleotide shuttle and is physiologically activated by calcium (Ca(2+)). AGC transport rates were significantly higher in tissue homogenates from all six patients, including those with no history of seizures and with normal electroencephalograms prior to death. This increase was consistently blunted by the Ca(2+) chelator ethylene glycol tetraacetic acid; neocortical Ca(2+) levels were significantly higher in all six patients; no difference in AGC transport rates was found in isolated mitochondria from patients and controls following removal of the Ca(2+)-containing postmitochondrial supernatant. Expression of AGC1, the predominant AGC isoform in brain, and cytochrome c oxidase activity were both increased in autistic patients, indicating an activation of mitochondrial metabolism. Furthermore, oxidized mitochondrial proteins were markedly increased in four of the six patients. Variants of the AGC1-encoding SLC25A12 gene were neither correlated with AGC activation nor associated with autism-spectrum disorders in 309 simplex and 17 multiplex families, whereas some unaffected siblings may carry a protective gene variant. Therefore, excessive Ca(2+) levels are responsible for boosting AGC activity, mitochondrial metabolism and, to a more variable degree, oxidative stress in autistic brains. AGC and altered Ca(2+) homeostasis play a key interactive role in the cascade of signaling events leading to autism: their modulation could provide new preventive and therapeutic strategies.

The mitochondrial aspartate/glutamate carrier isoform 1 gene expression is regulated by CREB in neuronal cells

The aspartate/glutamate carrier isoform 1 is an essential mitochondrial transporter that exchanges intramitochondrial aspartate and cytosolic glutamate across the inner mitochondrial membrane. It is expressed in brain, heart and muscle and is involved in important biological processes, including myelination. However, the signals that regulate the expression of this transporter are still largely unknown. In this study we first identify a CREB binding site within the aspartate/glutamate carrier gene promoter that acts as a strong enhancer element in neuronal SH-SY5Y cells. This element is regulated by active, phosphorylated CREB protein and by signal pathways that modify the activity of CREB itself and, most noticeably, by intracellular Ca²⁺ levels. Specifically, aspartate/glutamate carrier gene expression is induced via CREB by forskolin while it is inhibited by the PKA inhibitor, H89. Furthermore, the CREB-induced activation of gene expression is increased by thapsigargin, which enhances cytosolic Ca²⁺, while it is inhibited by BAPTA-AM that reduces cytosolic Ca²⁺ or by STO-609, which inhibits CaMK-IV phosphorylation. We further show that CREB-dependent regulation of aspartate/glutamate carrier gene expression occurs in neuronal cells in response to pathological (inflammation) and physiological (differentiation) conditions. Since this carrier is necessary for neuronal functions and is involved in myelinogenesis, our results highlight that targeting of CREB activity and Ca²⁺ might be therapeutically exploited to increase aspartate/glutamate carrier gene expression in neurodegenerative diseases.

Vitamin K2 and Myelin

Age- and brain-region-specific effects of dietary vitamin K on myelin sulfatides

Dysregulation of myelin sulfatides is a risk factor for cognitive decline with age. Vitamin K is present in high concentrations in the brain and has been implicated in the regulation of sulfatide metabolism. Our objective was to investigate the age-related interrelation between dietary vitamin K and sulfatides in myelin fractions isolated from the brain regions of Fischer 344 male rats fed one of two dietary forms of vitamin K: phylloquinone or its hydrogenated form, dihydrophylloquinone for 28 days. Both dietary forms of vitamin K were converted to menaquinone-4 in the brain. The efficiency of dietary dihydrophylloquinone conversion to menaquinone-4 compared to dietary phylloquinone was lower in the striatum and cortex, and was similar to those in the hippocampus. There were significant positive correlations between sulfatides and menaquinone-4 in the hippocampus (phylloquinone-supplemented diet -12mo and 24mo; dihydrophylloquinone -supplemented diet - 12mo) and cortex (phylloquinone-supplemented diet -12mo and 24 mo). No significant correlations were observed in the striatum. Furthermore, sulfatides in the hippocampus were significantly positively correlated with MK-4 in serum. This is the first attempt to establish and characterize a novel animal model that exploits the inability of dietary dihydrophylloquinone to convert to brain menaquinone-4 to study the dietary effects of vitamin K on brain sulfatide in brain regions controlling motor and cognitive functions. Our findings suggest that this animal model may be useful for investigation of the effect of the dietary vitamin K on sulfatide metabolism, myelin structure, and behavior functions.

Low sulfatide content in brain myelin has been recently linked with the disruption of myelin integrity [14,21], whereas the disruption of myelin integrity was implicated as an essential contributor to cognitive deficit [6, 7, 43, 44]. Although our findings of dietary-associated decreases in myelin sulfatides suggest a potential disruption in myelin integrity in evaluated brain regions, it is currently unknown whether such disruption would be sufficient to modify motor and cognitive functions controlled by these brain regions.

In summary, this is the first study to demonstrate the effect of dietary vitamin K on sulfatides and MK-4 in the purified brain myelin. It remains to be determined whether long-term and/or higher dietary dK consumption would be sufficient to affect brain-region-specific changes in the: (a) number and/or metabolic activity of oligodendrocytes; (b) rate of myelin formation and loss, (c) activity of genes responsible for the synthesis of myelin constituents. Furthermore, the behavioral consequences of altered sulfatide concentrations through manipulation of dietary vitamin K remain to be assessed.

Fat-soluble vitamins as disease modulators in multiple sclerosis

Vitamin K Biological properties relevant for an effect in MS – Vitamin K is a group of fat-soluble vitamins, needed for posttranslational modification of proteins involved in blood coagulation and bone metabolism. It includes two natural groups of vitamer chemicals: K1 (phylloquinone) and K2 (menaquinone). In addition to its effects of coagulation and bone metabolism, it has been demonstrated that oligodendrocyte precursors and immature neurons are protected from oxidative injury by vitamin K2 (61). Vitamin K has no known function in the immune system in humans. Trials in animal models – One study has been performed in the EAE-model (62). The authors reported that the severity of EAE was significantly ameliorated by the prophylactic administration of vitamin K2, although it was not effective when given after the onset. The authors reported that the vitamer seemed to work by inhibition of inflammatory cellular infiltration. Human trials – No human trials have been performed on the effect of vitamin K on MS disease activity or prevention.

Vitamin K as an antioxidant

Novel Role of Vitamin K in Preventing Oxidative Injury to Developing Oligodendrocytes and Neurons

Oxidative stress is believed to be the cause of cell death in multiple disorders of the brain, including perinatal hypoxia/ischemia. Glutamate, cystine deprivation, homocysteic acid, and the glutathione synthesis inhibitor buthionine sulfoximine all cause oxidative injury to immature neurons and oligodendrocytes by depleting intracellular glutathione. Although vitamin K is not a classical antioxidant, we report here the novel finding that vitamin K₁ and K₂ (menaquinone-4) potently inhibit glutathione depletion-mediated oxidative cell death in primary cultures of oligodendrocyte precursors and immature fetal cortical neurons with EC₅₀ values of 30 nM and 2 nM, respectively. The mechanism by which vitamin K blocks oxidative injury is independent of its only known biological function as a cofactor for γ-glutamylcarboxylase, an enzyme responsible for posttranslational modification of specific proteins. Neither oligodendrocytes nor neurons possess significant vitamin K-dependent carboxylase or epoxidase activity. Furthermore, the vitamin K antagonists warfarin and dicoumarol and the direct carboxylase inhibitor 2-chloro-vitamin K₁ have no effect on the protective function of vitamin K against oxidative injury. Vitamin K does not prevent the depletion of intracellular glutathione caused by cystine deprivation but completely blocks free radical accumulation and cell death. The protective and potent efficacy of this naturally occurring vitamin, with no established clinical side effects, suggests a potential therapeutic application in preventing oxidative damage to undifferentiated oligodendrocytes in perinatal hypoxic/ischemic brain injury.

In summary, we demonstrate for the first time that oxidative cell death induced by GSH depletion in primary OL precursors and in primary cortical neurons can be prevented by nanomolar concentrations of vitamin K₁ and MK-4. The cytoprotective effect of K vitamins in this model is independent of their known biological role in carboxylation. They do not prevent the loss of intracellular GSH caused by cystine depletion but markedly inhibit ROS accumulation and, thus, cell death. These results suggest a new approach to developing potential preventative and therapeutic strategies for neurological diseases in which GSH depletion-induced oxidative stress plays a role.

L-Carnitine and Calcium Chelation

I think we have established the link between excess calcium and some types of mitochondrial dysfunction.

Regular readers will know that one important element in autism mitochondrial therapies, like Dr Kelley’s and others, is the supplement L-carnitine, which in responders seems to show effect very quickly.

Is it a coincidence that one of the properties of this supplement is as a chelator of calcium?

L-carnitine is a calcium chelator: a reason for its useful and toxic effects in biological systems

Chelation normally refers to removing harmful metals from the body.  In the case of calcium we just want to put it back in the bones, not remove it from the body.

The study earlier in this post appear to show that the brain calcium deposits do not grow over time, for some reason calcium got deposited very early in life and just stays there.  The deposits do not grow but do continue to do damage.  So considering them like brain stones might be helpful.   Therapies do exist for such brain stones, as we saw using drugs developed for osteoporosis.

Conclusion

Excess calcium maybe one of those few simple concepts in autism that you do not need a PhD to fully understand.  It may also be at the root cause of further complex dysfunctions where that PhD really would be useful.

I think some of those CREB-associated dysfunctions and indeed some mitochondrial problems might just disappear if any existing excess calcium was removed.

If you can go to the doctor to measure calcification in your heart, why not do it for your brain?  Coronary Calcium Scans are common and take about 10 minutes.

 If there is no brain calcification, great. 

If it brain calcification exists, then treat it, just like the doctor would treat Grandma’s osteoporosis.

Measure bone density; all women over 65 are recommended to have a DXA scan.  So the technology is already here.

Vitamin K2 is seen as very safe, but you might need to eat a lot of Natto if you have calcification, probably better used for prevention.

Why do the Harvard researchers who have noted low bone density in autism not make a few further connections and understand the implications and treatment options?

There were also interesting issues that arose regarding multiple sclerosis (MS), but that is not really an issue for this blog.

Vitamin K2 looks like yet another good thing for people with type 1 or type 2 diabetes.

I have to add vitamin K2 to my growing list of possible dementia therapies, before I forget. It affects myelin sulfatides, which are one cause cognitive decline in the elderly.

Final Words

This did become rather a lengthy post.

Vitamin K2 is likely highly beneficial for many people, but just how much you need to decalcify a brain is unknown.  I suspect far more than in your average supplement.
 

Perhaps the dosage in the Japanese K2 drug would have an impact.  The Western RDA is 0.075 mg a day;  in Japan they used 45mg in trials, a dose 600 times larger.

Cheap generic bisphosphonate drugs might be better and then K2 for maintenance therapy?

Some serious scientific investigation looks warranted, given the therapies are sitting on the shelf.   Don’t hold your breath.

Intranasal Insulin for Some Autism vs IGF-1 and NNZ-2566

 

Very often the simplest solutions are the best and very often, when fault finding a problem, people overlook the obvious.  

I seem to be forever having to mend things and I find this all the time.

Back in 2013, when I knew much less about autism, I wrote about the experimental use of insulin like growth factor 1 (IGF-1) in autism.  

It’s a Small World – IGF-1 and NNZ-2566 in Autism

It turned out that in autism the many different growth factors can be disturbed (too much, or too little) and this variation does indeed define some specific types of autism.  For example in Rett Syndrome there are very low levels of Nerve Growth Factor (NGF); low levels of NGF in some older people is the cause of their dementia.  In more common types of autism NGF is actually elevated.

IGF-1 is very well studied.

 

IGF-1 is a primary mediator of the effects of growth hormone (GH). 

Growth hormone is made in the anterior pituitary gland, is released into the blood stream, and then stimulates the liver to produce IGF-1. IGF-1 then stimulates systemic body growth, and has growth-promoting effects on almost every cell in the body, especially skeletal muscle, cartilage, bone, liver,kidney, nerves, skin, hematopoietic cell, and lungs. This would explain why adults abusing GH may end up needing hip and knee replacements.

Before getting into the science, IGF-1 has long been available as a drug to treat children with growth delays.  In the US this drug is being used on children with a type of autism called Phelan-McDermid Syndrome.

Now, regular readers will recall from my last post on intranasal insulin that it was in this very syndrome that there was a successful intranasal insulin.

So most likely without delving into the science at all it looks like IGF-1 and intranasal insulin are both options to treat the same dysfunction.

Using IGF-1

Clinical Trial in 22q13 Deletion Syndrome (Phelan-McDermid Syndrome)

Using Intranasal Insulin

Intranasal insulin to improve developmental delay in children with 22q13 deletion syndrome: an exploratory clinical trial.

NNZ-2566

This is an Australian drug that is a modified version of IGF-1 (a so called analog).  They modified it so that it can be taken orally rather than by injection.  The developer has a very thorough presentation showing why they think it should be effective in autism.  

  

NNZ-2566 - Rationale for use in Autistic Spectrum Disorders

The Science

The first thing to note is that insulin and IGF-1 act as messengers.  Disruption in growth factor signaling can have serious consequences.

Insulin and IGF-1 both activate the same insulin receptor (IR).

Most people think that insulin is a just a hormone produced in their pancreas that regulates the amount of glucose (sugar) in their blood.  It does of course do that, but it actually does much more.

  

Insulin receptors are expressed all over the body including the brain.

Here is a relatively simple presentation explaining the role of insulin signaling in the brain:-

The role of insulin receptor signaling in the brain

Now for the diehard scientists among you that have been reading about all those signaling pathways that lie behind autism, cancer and many other hard to treat conditions, look at the graphic below.

We know the importance of RAS.  Impaired RAS signaling underlies the RASopathies, one feature of which is cognitive loss (MR/ID), another is autism.

We also know the importance of Akt (PKB/protein kinase B) in some types of autism.  PTEN appears again.

So irrespective of an undoubtedly important effect on glucose and insulin resistance, we should expect activation of insulin receptors in the brain, in some types of autism, to have a further positive effect.

It would seem to be a potential therapy for RASopathies.

As is often the case, there are extreme dysfunctions of RAS and I suggest there are more mild dysfunctions.

I suggest that some people with autism and some cognitive dysfunction have a partial RASopathy.

Since autism contains both extremes of many dysfunctions, there will undoubtedly be types of autism that respond negatively, or not at all, to activation of insulin receptors in the brain.

Practicalities

Nobody likes injections and that is necessary to give IGF-1.

NNZ-2566 is an experimental autism drug and on past performance that means it will take decades to reach the market, if ever.

That leaves insulin which was sitting all along in your local pharmacy.

Intranasal insulin was once investigated for use in diabetics, but it did not work.  It is not absorbed into the blood stream.

This is of course the huge advantage for people with autism, since we only want to activate the insulin receptors in the brain.  If you are not diabetic why would you want to have any effects in the rest of the body?

Indeed there are known major side effects of injecting IGF-1 or GH (growth hormone) into adults.  All kinds of things start growing and this can lead to terrible results.

The fact that all the studies show that intranasal insulin does not enter the blood stream and so lower blood glucose levels, makes it a much better drug for autism than IGF-1 or indeed NNZ-2566.

Insulin

There are various types of insulin and the main difference is that some are modified to be longer acting.

The basic insulin is soluble or clear insulin, and nowadays is synthetic rather than derived from pigs.  Examples include Humulin Regular/R/S by Lilly.

The standard concentration is 100 IU/ml.

The trials in Alzheimer’s and other conditions varied in dosage but generally used about 20 to 40 IU per day.

This is not a trivial dose.  If injected, rather than inhaled, that dose would have a significant effect on lowering blood sugar and would be dangerous.

My antihistamine nasal spray gives a metered dose of 0.14 ml.

So without any dilution, if filled with off the shelf insulin it would dispense 14 IU per spray.

So no special high tech drugs, dilutants/diluents or dispensers appear to be necessary. Some trials do use fancy inhalers, like the one in the video at the end of this post.

To be prudent it might be wise to dilute the insulin so as to gradually increase the dose.  Maybe in some people the nasal membrane is more permeable than in others.  Some of the trials did this, but most did not.

A fridge is required, because insulin needs to be kept chilled.

I do wonder why nobody seems to be researching this in autism.  Silly point, as one insulin researcher commented on the earlier post; there is no big money to be made, hence no interest.

Insulin & Alzheimer’s

The reasons that intranasal insulin improves Alzheimer’s, and likely will Down Syndrome, may differ to those help in (some) autism.

Beta amyloid is key to Alzheimer’s (and early onset Alzheimer’s in Down Syndrome) but is not a known issue in autism.  Central insulin resistance is an issue in Alzheimer’s and might well be in autism.  

Perhaps people with mitochondrial dysfunction (an energy conversion dysfunction) might particularly benefit from increased glucose uptake in the brain.  It appears that mitochondrial dysfunction plays a role in insulin resistance. 

Role of Mitochondrial Dysfunction in Insulin Resistance

The activation of the RAS pathway might be highly beneficial to some people with autism.  

Here is a good film, which refers to the studies from previous posts and shows the effect on one man with Alzheimer's. 

Insulin and Alzheimer's (approx. 22 min.)

 You also see their fancy inhaler device.

Interview Series with Leading Autism Researchers

Seth Bittker, a regular reader of this blog and our occasional guest blogger, is creating an excellent interview series with leading autism researchers.

These interviews will be of interest to all readers of this blog.  There is something for everyone, whether you are the type that reads the literature in detail, or is more interested in the lay summary.

There are three interviews, all with podcasts, and more will be added later.

1)    Dr. Richard Frye on folate receptor alpha antibodies and folinic supplementation in autism. 

2)    Dr. Robert Hendrenon his recent double blind placebo controlled trial of methyl-B12 in autism.  

3)    Dr. Derrick Lonsdale on thiamine deficiency and supplementation of thiamine derivatives in autism.   

Good work Seth!  I am sure he will be interested in your suggestions for future interview candidates. 

By the way, in the third interview, Dr Lonsdale refers to his preferred thiamine supplement called TTFD, this has been used for decades in Japan.   It is also called Fursultiamine and the Japanese brand name is Alinamin, which is made by Takeda, a major Japanese company.  The Japanese website (in English) is here.

Takeda have various combinations with other B vitamins and some are sold on Amazon/eBay.  Some with very high amounts of B12.  

In interview number two above, Dr Hendrenon clearly does believe in the merits of extra B12 in autism.  His trial did inject the vitamin, rather than take it orally. 

It is best to only supplement at high doses the B vitamins that really help in your specific case; there are known  negative reactions to some B vitamins, so best to go through them one by one.

There is also a version of TTFD sold as Allithiamine by a small US company, Ecological Formulas, which Seth is investigating.

So as long as swallowing pills is possible, you have the opportunity to replicate Dr Lonsdale's trial and see if you have a responder or not. 


Side Effects of high dose B vitamins

According to the University of Maryland taking any one of the B vitamins for a long period of time can result in an imbalance of other important B vitamins, they suggest taking a B-complex vitamin which includes all B vitamins.

This might explain why some people who initially respond well to high doses of biotin, vitamin B7, later experience a negative response. 

Many people do not respond well to high doses of multiple B vitamins as prescribed by some DAN-type doctors.

Dr Frye, from interview number one, is also a big believer in B vitamin supplementation.

Clearly B vitamin supplementation needs to be much better thought out, to keep any good effects, without developing any bad effects. 

Dad! Can I cheat?

This week at school is a talent show and Monty, aged 12 with ASD, has signed up to play the piano.

Last month was a poetry evening in the senior school, where the older pupils recited poetry in a wide variety of European languages.  Monty did attend to support his big brother and did not do anything to embarrass him.

I think the talent show is more for the juniors, even though it is open to the seniors.

Nine years ago when Monty got his diagnosis, we were given some rather eclectic tips, including he might develop epilepsy and not to expect him to participate in school events.  I always thought the latter was an odd thing to say at the end of your autism assessment, however true it might be.

Nonetheless we have endeavoured to overcome the odds and make sure he does participate in school events.  Monty is the only one with autism in the junior school, while in the senior school there is one with Asperger’s, who proudly recited his poem the other evening.  The school is very good and supports everyone to participate in events.  It is a very small school, which does make things easier.

Talent Show Preparation

The only question with the piano recital is what to play and whether to use the music book or play from memory.

I am surprised that much of the music Monty plays he has actually memorised, but now he plays longer pieces and so the scope for slip ups is greater.

So I asked him to play his latest piece, Phantom of the Keys, from memory and after a couple of minutes he got stuck, picked up the music book turned around and said “Dad! Can I cheat?”

So he won’t be playing from memory.

More General Preconceptions

I think quite often people with autism are held back by preconceived limits on what they can achieve.

Some young adults with autism achieve far more than others and the limiting factor does not seem to be their inherent abilities, much more what they have done in their first twenty years of preparation for life.

Some quite able people with Asperger’s cannot use public transport and, assuming they cannot drive, how are they ever going to have a job? The problem often is anxiety, but in the previous twenty years was it not possible to address that issue?

Monty used to really hate the sound of babies crying.  The effective therapy was to be exposed to that very sound he found excruciating and not to hide him away from it. Now he is perfectly OK with that sound.

The same seems to apply to going to the cinema/theatre, if you start going at a young age you get used to the light and sound.  If you accept sensory overload as barrier, then you may just lower your sights accordingly.

The other day I was talking to a parent of a boy a few years older than Monty and I was really impressed that he goes to his special school alone, by the regular city bus, in the middle of one of Europe’s biggest cities.   He does not get lost, or wander off.  Good for him and you can see him being able to do some kind of adapted job in the future.

Other people develop intellectually, but early problems like toileting are left unaddressed, so you will have adult that cannot care for himself.

Some kind of job, or structured daytime activity, seems a prerequisite for all adults (with, or without autism) but if you cannot get to that job and care for yourself while you are there, you have ruled out that option.

The key does seem to be higher aspirations earlier on, rather than just accepting the many difficulties as immovable barriers.

Outcomes vary widely because each person is an experiment, there is very little sharing of accumulated knowledge.  Basic things like doing some extra school work at home often do not a happen.  In special schools, parents complain that their child cannot do even basic maths or write, while teachers comment “you can always tell which ones get some help at home”. 

The ones that get the help at home are the ones likely to get the extra support at school.

Common sense would suggest that parents must realize that if a child has great learning difficulties and there are six kids to one teacher, not much learning is going to take place at school and, if none takes place at home, the result is not going to be good.

You might think that some learned professional would right an “Autism for Dummies” so that many common mistakes could be avoided.  Such people do indeed exist, who can quickly spot errors that you make, not realizing the eventual long term consequence.

In the ideal world if you receive an autism diagnosis, you would be assigned a mentor who would give you occasional guidance and support through to adulthood. That way more people would achieve their potential.