Diabetes

[roamer]

Jhananda, I have not reviewed datasets which place the prevalance around 1940, but it certainly doesn't contradict anything i've said.  1940 would have been the dramatic rise in chemical assisted monocultures which would have placed an increased amount of processed corn or grain derivatives in our diet.  It was also a period of radical mechanization of all labor which altered activity patterns.  And of course as you point out also a new era of exposure to toxic previously unencountered chemicals, medicines ect to the human genome.

I do not dismiss some influence of the later, but being a bit of a lifelong athlete and gym buff I have known many people who have maintained health into old age or reversed health and type ii diabetes at old age.  They did it through diet and activity manipulation and many of them still had exposure to numerous toxins additional from lab jobs ect and all of them lived in the same toxic environment with immunizations, antibiotics,  ect that everyone else is encountering.

[Jhanananda]

Jhananda, I have not reviewed datasets which place the prevalance around 1940, but it certainly doesn't contradict anything i've said.  1940 would have been the dramatic rise in chemical assisted monocultures which would have placed an increased amount of processed corn or grain derivatives in our diet.  It was also a period of radical mechanization of all labor which altered activity patterns.  And of course as you point out also a new era of exposure to toxic previously unencountered chemicals, medicines ect to the human genome.

While I agree with much of what you have to say here; nonetheless, Indians resident on Hopi and Tohono O'odham Indian reservations, who have eaten the same food, with they have grown for thousands of yeas, are developing type II diabetes, and its associated complex.  It suggests to me some other influence, which I believe might very well be as simple as: drinking ground water; and/or immunizations; and/or extensive use of antibiotics, all three of which were made available to those tribes in the 30s.

I do not dismiss some influence of the later, but being a bit of a lifelong athlete and gym buff I have known many people who have maintained health into old age or reversed health and type ii diabetes at old age.  They did it through diet and activity manipulation and many of them still had exposure to numerous toxins additional from lab jobs ect and all of them lived in the same toxic environment with immunizations, antibiotics,  ect that everyone else is encountering.

In my case we will not know until there is more investigation.  I have an appointment with a rheumatologist next month.  It might just be the key to my health problems starting around 15.  It just so happens that I manifest the symptomatology of both Reiter's Syndrome and Ankylosing Spondylitis‎. In both cases there are both genetic and the product of gastrointestinal infection.

[roamer]

If indeed the Hopi diet remained the same (which i suspect it did not, likely additional foods were added) I would be very curious to know how overall activity patterns changed. 

Causation becomes tricky in such complex systems as the human body.  You are probably on to something about your gut changing around 15, this has increasingly been shown to cause cascading changes in the body.  However as with dukka the only relevant question is how best to treat.  My argument is good food or minimization of environmental toxins simply is not enough.  It takes lifestyle reengineering and using a hunter gather energy diet expenditure model as a guideline seems like the most robust model to mimic.

[Jhanananda]

If indeed the Hopi diet remained the same (which i suspect it did not, likely additional foods were added) I would be very curious to know how overall activity patterns changed. 

Yes, the majority of Native Americans have embraced western diets; however, there are traditional Hopi and Tohono O'odham who live in remote areas, and who live pretty much as they have for 1000s of years, and yet they too are developing diabetes and its associated conditions.

Causation becomes tricky in such complex systems as the human body.  You are probably on to something about your gut changing around 15, this has increasingly been shown to cause cascading changes in the body.  However as with dukka the only relevant question is how best to treat.  My argument is good food or minimization of environmental toxins simply is not enough.  It takes lifestyle reengineering and using a hunter gather energy diet expenditure model as a guideline seems like the most robust model to mimic.

Well, I agree that good food and minimization of environmental toxins simply is not enough.  It takes lifestyle reengineering and using a hunter gather energy diet; all of which I have done; nonetheless, my health is still in decline.  Old-age is certainly and aspect of this decline, but the decline seems much too steep, so I am investigating the genetic component.

[roamer]

Jhanananda,

All theories aside it really sucks to hear you are experiencing like far too many a rapid decline in health.  I am saddened to hear of your difficulties and really do hope you are able to find some relief, comfort and relative stability in the inevitable aging process.  Life has always been transient but my sense is that we really are in strange times concerning general health and I am of course very sympathetic to your situation and hope for the best.   On a personal note my father is going through the same health decline with type ii diabetes at 64 years of age.  His parents and grandparents never had such a thing and its rather remarkable to me to see my father quite frail and unhealthy when I remember my grandfather at an even older age quite healthy and hard working.  In fact my grandfather lived to be 84 and milk cows and ran a dairy farm until 80 years of age, he died of a stroke he incurred while splitting wood. Modernity it seems is not a friend of health and the notion of industrial progress is for most an myth to line the pockets of extraordinarily wealthy and powerful. 

[Jhanananda]

Jhanananda,

All theories aside it really sucks to hear you are experiencing like far too many a rapid decline in health.  I am saddened to hear of your difficulties and really do hope you are able to find some relief, comfort and relative stability in the inevitable aging process.  Life has always been transient but my sense is that we really are in strange times concerning general health and I am of course very sympathetic to your situation and hope for the best.   

Thank-you for your kind thoughts; and I agree, there are one or more contributing factors to accelerated decline of people in the US American culture.

On a personal note my father is going through the same health decline with type ii diabetes at 64 years of age.  His parents and grandparents never had such a thing and its rather remarkable to me to see my father quite frail and unhealthy when I remember my grandfather at an even older age quite healthy and hard working.  In fact my grandfather lived to be 84 and milk cows and ran a dairy farm until 80 years of age, he died of a stroke he incurred while splitting wood. Modernity it seems is not a friend of health and the notion of industrial progress is for most an myth to line the pockets of extraordinarily wealthy and powerful.

I have had somewhat the same experience; however, my grandfathers, both farmers, died in their early 60s; whereas, both of my parents died almost 90, and neither farmers, but raised on farms.  However, I am inclined to agree with you, that there is something in the rapid industrialization of the USA from the depression to the present that is at the root of the decline of US American culture. 

So far my decline seems to be a genetic condition that I may have shared with at least one of my grandparents, which is exacerbated by the presence of naturally occurring radioactive elements in the water that I was drinking and cooking with.  So, not consuming ground water from here on out, might just improve my overall health, and that of others.

[Jhanananda]

It is hard to believe that there are 20 Fruits that are good For Diabetics to eat, because fruit tends to be high in carbohydrates, so I will be checking them out.  Olives are one of the only exceptions to this rule that I can think of at this time, because they have zero carbs, while black olives containing high levels of anthocynin.

According to various guidelines laid down by nutritionist and medical institutions, at least 4-5 servings of fruits needs to be consumed daily by every individual. If you are diabetic, we are sure that this question has popped up in your head, “is it safe to have fruits?” To help all diabetics with this question, we have Mrs. Kamna Desai – Nutritionist, with us. She says, "Yes, diabetics can have fruits, provided the sugar level of the patient is in control, but these fruits must be consumed in a limited quantity. Diabetics need an equivalent serving of fruits on a day to day basis. One needs to be careful about not going overboard with fruits like bananas, litchis, chickoo, and custard apples." To help diabetic patients, she lists down some fruits which will not affect blood sugar.

1] Kiwi. Many researchers have shown a positive correlation between kiwi consumption and lowering of blood sugar level
2] Black jamun. Undoubtedly, this is one of the best fruits for diabetics. It is known to improve blood sugar control. Seeds of these fruit can be powdered and consumed by patients to control diabetes.
3] White jamun. Diabetic patients can consume jamun fruit daily, to control their sugar levels.  White jamuns are also high in fiber.
4] Starfruit. Similar to jamuns, starfruits are good for diabics too as they help improve blood sugar control. But caution needs to be exercised in case the person has diabetes neuropathy.
5] Guava. Guava controls diabetes and it is good for constipation. Guavas are high in vitamin A and vitamin C and contain high amounts of dietary fiber. This fruit has a reasonably low GI.
6] Cherries. Their GI value is 20 (or even less in some varieties) which makes it a good healthy snack for diabetes patients at any time of the day.
7] Peaches. These tasty fruits are a great healthy treat, with a low GI and good for diabetics.
8] Berries. Numerous variety of berries are available throughout the world and almost all seem to be a rich source of antioxidants. Diabetics can include a serving of different berries to keep their sugars in check. To name a few: Strawberries, blueberries and blackcurrants, raspberries, cranberries, chokeberries, blackberries, and acai berries are good for diabetes patients.
9] Apples. Apples contain antioxidants, which help to reduce cholesterol levels, cleanse the digestive system, and boost the immune system. Apples also contain nutrients that help in the digestion of fats.
10] Pineapples. Good for diabetics, pineapples also benefit the body as they are rich in anti-viral, anti-inflammatory and anti-bacterial properties.
11] Pears. These delicious fruits are a good snacking option for diabetics as they are rich vitamins and fiber.
12] Papaya. They are good for diabetics because they are rich in vitamin and other minerals.
13] Figs. Their richness in fibre helps with insulin function in diabetes patients.
14] Oranges. These citrus fruits can be consumed on a daily basis by diabetics, as they are rich in vitamin C.
15] Watermelon. Although watermelons have a high GI value, their glycemic load is low, making them good fruits for diabetes patients. However, consume in moderation.
16] Grapefruit. It is a good option for diabetics as it slows down the blood sugar peak.
17] Pomegranate. These tiny red rubies help diabetic people improve their blood sugar statistics.
18] Cantaloupe. This fruit is high in the Glycemic Index, but has a good amount of fibre. Therefore, within moderation, it can easily be include
19] Jackfruit. Jackfruit contains vitamin A, vitamin C, thiamin, riboflavin, niacin, calcium, potassium, iron, manganese and magnesium among many other nutrients. Good for diabetes as they improve insulin resistance.
20] Amla. A fruit loaded with vitamin C and fiber, Amla is a healthy addition to the diabetic diet.

[bodhimind]

I've recently heard about alpha lipoic acid for diabetes, which apparently helps lower sugar levels. I donated some help over for GWV, but I hope some can be used to acquire some healthy supplements/food for you too.

[Jhanananda]

Thanks, bodhimind.

[Jhanananda]

Could a Diabetes Drug Help Beat Alzheimer's Disease?

Metformin may slow or reverse dementia and cognitive impairment, even in nondiabetics

Most of the 20 million people diagnosed with type 2 diabetes in the U.S. take metformin to help control their blood glucose. The drug is ultrasafe: millions of diabetics have taken it for decades with few side effects beyond gastrointestinal discomfort. And it is ultracheap: a month's supply costs $4 at Walmart. And now new studies hint that metformin might help protect the brain from developing diseases of aging, even in nondiabetics.

Diabetes is a risk factor for neurodegenerative diseases, but using metformin is associated with a dramatic reduction in their incidence. In the most comprehensive study yet of metformin's cognitive effects, Qian Shi and her colleagues at Tulane University followed 6,000 diabetic veterans and showed that the longer a patient used metformin, the lower the individual's chances of developing Alzheimer's disease, Parkinson's disease, and other types of dementia and cognitive impairment.

[Jhanananda]

I have been reviewing my log of health data.  A year ago in November I was admitted into ER with a blood sugar of 350 and high blood pressure, and pulse rates, and a very low blood oxygen level.  Since then I acquired a pulse oxymeter. 

I have been trying to make sense of my blood sugar that seems to be steady on a good value for weeks at a time, then it wanders up, as high as 350.  Now that I can examine a year of data regarding blood sugar and blood oxygen I saw that when I am having seasonal allergies, which is something that I have had since childhood, my blood oxygen goes down, but what was surprising was my blood sugar goes up. 

So, now I may have another causal relationship to my diabetes to deal with, which is good.  I just need to move to a place that does not have plant polens that I am allergic to.

[DDawson]

I've been watching videos by a Dr. Glidden and its his believe that diabetes is caused by a chromium and vanadium deficiency among other possible deficiencies.  You may be already aware of this, but check him out.  His tutorials are well done and eye opening.

[Jhanananda]

Thank-you, DDawson, for posting your comment and suggestion on this thread.

Yes, I have read numerous research reports since the 50s that chromium (cr-3) helped diabetics control their blood sugar.  I started cr-3 supplementation on December 3rd of last year at 1000micro grams per day.  The next day my blo0od sugar reading was almost normal and the lowest in months. 

My blood sugar continued to go up and down, but mostly close to normal for the next month, then it went up at the same time I was experiencing juniper fever.  So, my conclusion is cr-3 is something that I need, but allergies seems to have a causal relationship to my diabetes.

Another point needing to be made here is, I had avoided taking benadryl for my seasonal allergies until after my visit to the emergency room a year ago in November.  I have found that taking benadryl for my allergies has lowered my blood sugar during allergy season; however, it still rises to about 200 instead of 350.

[Jhanananda]

Faux fasting diet regenerates pancreas to reverse diabetes

Using stem cells to create insulin-producing beta cells that could be transplanted into diabetics is being investigated as a possible cure for type 1 diabetes and treatment for type 2, but new research suggests that a special diet could reprogram cells in the pancreas to do the same thing. Researchers at the University of Southern California (USC) claim that a diet that mimics the effects of fasting spurs the growth of new insulin-producing beta cells in the pancreases of mice, essentially reversing the disease.

Both type 1 and type 2 diabetes center around insulin. Put very simply, in type 1 diabetes, the body – specifically, the pancreas – can't produce enough insulin, while in type 2 diabetes, the body doesn't use insulin properly and eventually is unable to produce enough insulin to compensate. In both type 1 and late-stage type 2 diabetes, insulin-producing beta cells in the pancreas are lost, meaning many diabetics need to take insulin to replace what's not being made by the pancreas.

Looking to discover the effects of a fasting-mimicking diet (FMD) on diabetes sufferers, USC researchers used mice with type 2 diabetes and another group in which type 1 diabetes had been simulated by giving them high doses of a drug to kill their insulin-producing beta cells. They found that mice in both groups – even those in the later stages of the disease – regained healthy insulin production, had a reduction in insulin resistance, and had more stable blood glucose levels.

The researchers say the brief, periodic diet, which was designed to mimic the effects of a water-only fast, activated genes that are normally only switched on in the developing pancreases of fetal mice. These genes prompted the production of neurogenin-3 (Ngn3), a protein that led to the generation of healthy new beta cells in the adult mice.

Providing an indication that similar results could be expected in humans, the team, led by Valter Longo, the director of the Longevity Institute at USC's Leonard Davis School of Gerontology, found that fasting also increased the expression of Ngn3 and sped up insulin production in pancreatic cell cultures from human donors with type 1 diabetes.

Such a diet could also have wider health benefits, with a previous study from the team demonstrating that human participants who followed the special FMD for five days each month in a three-month span cut their risks for not only diabetes, but also cancer, heart disease and other age-related diseases. Additionally, in another study that preceded that, the team says diet also showed potential for reducing visceral fat, increasing the effectiveness of chemotherapy for cancer treatments, and alleviating the symptoms of multiple sclerosis.

The FMD, which is being marketed commercially under the name "Prolon," involves cutting one's calorie intake by around two-thirds for five days, during which the person's body doesn't recognize that they are eating, before returning to normal intake for the remainder of the month. The researchers are pursuing a larger FDA trial on the potential of the FMD to treat diabetes in humans, but it should be made clear that no one should embark on such a diet without first consulting their doctor.

The team's most recent study was published in the journal Cell.

[Jhanananda]

Fasting-Mimicking Diet Promotes Ngn3-Driven β-Cell Regeneration to Reverse Diabetes

Highlights

    •    Fasting mimicking diet induces prenatal-development gene expression in adult pancreas
    •    FMD promotes Ngn3 expression to generate insulin-producing β cells
    •    Cycles of FMD reverse β-cell failure and rescue mice from T1D and T2D
    •    Inhibition of PKA or mTOR promotes Ngn3-driven β-cell regeneration in human T1D islets

Summary

Stem-cell-based therapies can potentially reverse organ dysfunction and diseases, but the removal of impaired tissue and activation of a program leading to organ regeneration pose major challenges. In mice, a 4-day fasting mimicking diet (FMD) induces a stepwise expression of Sox17 and Pdx-1, followed by Ngn3-driven generation of insulin-producing β cells, resembling that observed during pancreatic development. FMD cycles restore insulin secretion and glucose homeostasis in both type 2 and type 1 diabetes mouse models. In human type 1 diabetes pancreatic islets, fasting conditions reduce PKA and mTOR activity and induce Sox2 and Ngn3 expression and insulin production. The effects of the FMD are reversed by IGF-1 treatment and recapitulated by PKA and mTOR inhibition. These results indicate that a FMD promotes the reprogramming of pancreatic cells to restore insulin generation in islets from T1D patients and reverse both T1D and T2D phenotypes in mouse models.

View File

Introduction

The ability of animals to survive food deprivation is an adaptive response accompanied by the atrophy of many tissues and organs to minimize energy expenditure. This atrophy and its reversal following the return to a normal diet involve stem-cell-based regeneration in the hematopoietic and nervous systems (Brandhorst et al., 2015, Cheng et al., 2014). However, whether prolonged fasting and refeeding can also cause pancreatic regeneration and/or cellular reprogramming leading to functional lineage development is unknown. β cells residing in pancreatic islets are among the most sensitive to nutrient availability. Whereas type 1 and type 2 diabetes (T1D and T2D) are characterized by β-cell dedifferentiation and trans-differentiation (Cnop et al., 2005, Dor and Glaser, 2013, Talchai et al., 2012, Wang et al., 2014), β-cell reprogramming, proliferation and/or stepwise re-differentiation from pluripotent cells are proposed as therapeutic interventions (Baeyens et al., 2014, Chera et al., 2014, Maehr et al., 2009, Pagliuca et al., 2014, Sneddon et al., 2012, Zhou et al., 2008, Ben-Othman et al., 2017, Li et al., 2017), suggesting that lineage conversion is critical in both diabetes pathogenesis and therapy (Weir et al., 2013).

Although dietary intervention with the potential to ameliorate insulin resistance and type II diabetes has been studied extensively for decades, whether this has the potential to promote a lineage-reprogramming reminiscent of that achieved by iPSCs-based engineering remains unknown. We previously showed that cycles of prolonged fasting (2–3 days) can protect mice and humans from toxicity associated with chemotherapy and can promote hematopoietic stem-cell-dependent regeneration (Cheng et al., 2014, Laviano and Rossi Fanelli, 2012, Raffaghello et al., 2008). In consideration of the challenges and side effects associated with prolonged fasting in humans, we developed a low-calorie, low-protein and low-carbohydrate but high-fat 4-day fasting mimicking diet (FMD) that causes changes in the levels of specific growth factors, glucose, and ketone bodies similar to those caused by water-only fasting (Brandhorst et al., 2015) (see also Figure S1 for metabolic cage studies). Here, we examine whether cycles of the FMD are able to promote the generation of insulin-producing β cells and investigate the mechanisms responsible for these effects.

Results
Cycles of a FMD Rescue Mice from Late-Stage T2D

As a consequence of insulin resistance, the decrease in the number of functional insulin-producing β cells contributes to the pathophysiology of T2D by eventually leading to insulin deficiency (Cnop et al., 2005, Dor and Glaser, 2013). Previously, we showed that a 4-day fasting mimicking diet (FMD) could induce metabolic changes similar to those caused by prolonged fasting and could reduce insulin and glucose levels while increasing ketone bodies and igfbp1 (Brandhorst et al., 2015; Figure S1). Although the role of periodic fasting and fasting mimicking diets on insulin secretion is unknown, the effects of intermittent fasting and chronic calorie restrictions (CR) on insulin sensitivity have been previously reported (Barnosky et al., 2014). Here, our focus is on the putative effects of the FMD in promoting β-cell regeneration, although we have also investigated the effects of the FMD on insulin resistance.

Given that β cells replicate at an extremely low rate in the adult pancreas (Meier et al., 2008, Teta et al., 2005) and that β-cell neogenesis occurs rarely (Xiao et al., 2013), depletion of β cells and the consequent loss of insulin secretion during late-stage diabetes have often been considered conditions whose reversals require islet and stem cell transplantation (Fiorina et al., 2008, Kroon et al., 2008, Milanesi et al., 2012, Pipeleers et al., 2002, Villani et al., 2014). To determine whether the FMD could affect the β-cell deficiency associated with T2D, we studied its effect on mice with a point mutation in the leptin receptor gene (Leprdb/db), which causes insulin resistance in the early stages and failure of β-cell function in the late stages. As reported by others, db/db mice developed hyperglycemia at 10 weeks of age, which we refer to as baseline (BL) (Figure 1A). The insulin levels first increased to compensate for insulin resistance but drastically declined after 2 weeks of severe hyperglycemia (Figures 1B and 1C) (Arakawa et al., 2001). As a result, db/db mice began to die at around 4 months of age. We attempted to reverse these late-stage T2D phenotypes by treating 12-week-old mice (14 days after the hyperglycemia stabilized, baseline) with weekly cycles of the 4-day FMD (Figure 1A). FMD cycles caused a major reduction and return to nearly normal levels of blood glucose in db/db mice by day 60 (Figure 1B). The FMD cycles also reversed the decline in insulin secretion at day 30 and improved plasma insulin levels at day 90 (Figure 1C). A homeostasis model assessment (HOMA) was performed to estimate steady-state β-cell function (%B) and insulin sensitivity (%S), as previously described (Hsu et al., 2013, Matthews et al., 1985). The results indicate that the reversal of hyperglycemia was mainly caused by an induction of steady-state β-cell function (%B) (Figure 1D). Nevertheless, mice receiving the FMD showed improved glucose tolerance and insulin tolerance compared to the ad libitum (AL) fed controls (Figure 1E). Notably, although db/db mice on the FMD diet gained less weight compared to those on the regular diet, they maintained a weight that was ∼22% higher than that of their healthy wild-type (WT) littermates during the entire experiment (Figure S2C). Altogether, these results indicate that FMD cycles rescued mice from late-stage T2D by restoring insulin secretion and reducing insulin resistance, leading to a major improvement in survival (Figure 1F, ∗p < 0.05, log-rank test for trend).

Cycles of FMD Reverse β-Cell Failure in T2D

Dedifferentiation of β cells, which results in increased non-hormone-producing cells within pancreatic islets, is a feature of diabetic β-cell failure (Dor and Glaser, 2013, Kim-Muller et al., 2014, Talchai et al., 2012). Similar to what was previously reported by others, we found an increase in cells producing neither insulin nor glucagon (i.e., non-α/β) and a decrease in β-cell number in pancreatic islets of late-stage T2D mice, but not in age-matched WT controls (Figures 1G and S2, db/db BL compared to WT/AL). We also found that β-cell proliferation was low in the late stage of the disease (Figure 1H, AL day 60 compared to BL). Whereas db/db mice fed ad libitum (db/db:AL) showed a 60%–80% reduction in β-cell count at day 60, db/db mice receiving FMD cycles (db/db:FMD) displayed a major improvement in the number and proliferation of insulin-generating β cells (comparing db/db BL, Figures 1G-1I). Pancreatic islets collected from db/db mice treated with FMD cycles (day 60) displayed increased glucose-stimulated insulin secretion (GSIS), compared to that of islets from db/db:AL mice (Figure 1J). We also determined that a longer exposure time (time point 120) was necessary to distinguish between the functionality of islets from db/db:AL and db/db:FMD group mice (Figure 1J). Overall, these results suggest that, in addition to improving insulin sensitivity, FMD induced β-cell regeneration to reverse β-cell loss, which may alleviate late-stage T2D symptoms and mortality.

FMD Cycles Restore Insulin-Dependent Glucose Homeostasis in STZ-Induced T1D

To examine further the role of FMD cycles in stimulating β-cell regeneration, we applied FMD cycles on a T1D model in which high-dose streptozotocin (STZ) treatment causes the depletion of insulin-secreting β cells (Wu and Huan, 2008, Yin et al., 2006). Starting 5 days after STZ treatment, which we refer to as baseline (STZ BL), hyperglycemia (>300 mg/dl) was observed in both mice fed AL and those subjected to multiple cycles of the 4-day FMD every 7 days (4 days of FMD followed by 3 days of re-feeding, every 7 days per cycle) (Figures 2A and 2B ). Levels of blood glucose continued to increase in STZ-treated mice receiving the AL diet and reached levels above 450 mg/dl at both days 30 and 50. On the other hand, in mice receiving FMD cycles, hyperglycemia and insulin deficiency were both significantly alleviated on day 30 (Figure 2B, sample size indicated in parentheses). Remarkably, the levels of these physiological parameters returned to a nearly normal range at days 50–60 after the FMD cycles (Figures 2B and 2C, sample size indicated in parentheses). Intraperitoneal glucose tolerance tests (IPGTTs) at day 50 confirmed that STZ-treated mice undergoing the FMD cycles have improved capacity to clear exogenous blood glucose (STZ+ FMD), compared to mice on the regular chow (STZ) (Figure 2D).

Levels of certain circulating cytokines have been used as indicators to determine islet pathological status in patients with recent-onset T1D (Baeyens et al., 2014, Grunnet et al., 2009, Lebastchi and Herold, 2012). We performed a 23-plex immunoassay to determine the effects of the FMD on inflammatory markers. We found that FMD cycles not only suppressed the circulating cytokines associated with β-cell damage (e.g., TNFα and IL-12), but also increased circulating cytokines associated with β-cell regeneration (e.g., IL-2 and IL-10) (Figure 2E, day 30) (Dirice et al., 2014, Rabinovitch, 1998, Zhernakova et al., 2006).

Taken together, these results indicate that FMD cycles reduce inflammation and promote changes in the levels of cytokines and other proteins, which may be beneficial for the restoration of insulin secretion and the reversal of hyperglycemia.

FMD Cycles Reverse STZ-Induced β-Cell Depletion

The characterization of pancreatic islet cells indicates a strong association between restored glucose homeostasis and the replenishment of pancreatic β cells in animals undergoing FMD cycles. STZ treatments resulted in an increase of non-α/β cells (Figure S3) and an ∼85% depletion of insulin-secreting β cells (Figure 2F, STZ BL). The transient increase of non-α/β cells was reversed by day 30 in both groups (Figures S2D and S2E). Mice receiving weekly cycles of the FMD showed a major increase in proliferative β cells followed by a return to a nearly normal level of insulin-generating β cells by d50 (Figures 2F–2H). In contrast, mice that received ad libitum access to regular chow remained depleted of β cells for >50 days (Figures 2F and 2H). Overall, the increase of non-α/β prior to β-cell proliferation raises the possibility that weekly cycle of the FMD might mediate the fate conversion of non-α/β cells to β cells to reverse the STZ-induced β-cell depletion, although other scenarios are possible.

FMD and Post-FMD Re-feeding Promote β-Cell Regeneration in Non-diabetic Mice

We investigated whether and how the FMD and the post-FMD re-feeding period could regulate the cell populations within the islets to promote β-cell regeneration independently of diabetes, with a focus on the non-α/β cells and proliferative β cells. To characterize cellular and hormonal changes, pancreatic samples and peripheral blood of wild-type C57Bl6 mice fed with the FMD for 4 days were collected before the diet (BL), at the end of the diet (day 4), and 1 or 3 days after mice returned to the normal diet (RF1d or RF3d). The FMD caused a trend of decrease in the number and size of pancreatic islets (Figure 3A) and reduced the proportion of β cells by 35% (Figures 3B and 3C; see also Figure S4 for absolute numbers). These effects were reversed within 3 days of re-feeding (Figures 3A and 3C). Non-α/β cells began to proliferate at the end of the FMD, and this proliferation persisted until 1 day after re-feeding (RF1d), leading to a 2.5-fold increase in non-α/β cells (proportion per islet) at RF1d (Figure S4). By RF3d, the number of non-α/β cells had dropped and that of β cells returned to basal levels (BL), although β cells remained in a much more proliferative state in the FMD group (Figure 3B and 3C). The expression of the proliferation marker PCNA was elevated in β cells, but not α cells, after re-feeding post FMD (Figures 3B and 3C and S4). Despite the number of α cells per islet remaining the same, the transitional α-to-β or β-to-α cells that co-express both α (i.e., glucagon) and β cell (i.e., Pdx-1 or insulin) markers were increased in mice that received the FMD (Figure 3D). In summary, the FMD promotes a decrease in the numbers of differentiated or committed cells, followed by the induction of transitional cells and major increases in the proliferation and number of insulin-generating β cells (Figure 3E).