Masahiro Bando

Obesity-induced DNA released from adipocytes stimulates chronic adipose tissue inflammation and insulin resistance.

DNA released from obesity-induced degenerated adipocytes stimulates inflammation in adipose tissue and insulin resistance. Obesity stimulates chronic inflammation in adipose tissue, which is associated with insulin resistance, although the underlying mechanism remains largely unknown. Here we showed that obesity-related adipocyte degeneration causes release of cell-free DNA (cfDNA), which promotes macrophage accumulation in adipose tissue via Toll-like receptor 9 (TLR9), originally known as a sensor of exogenous DNA fragments. Fat-fed obese wild-type mice showed increased release of cfDNA, as determined by the concentrations of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) in plasma. cfDNA released from degenerated adipocytes promoted monocyte chemoattractant protein-1 (MCP-1) expression in wild-type macrophages, but not in TLR9-deficient (Tlr9−/−) macrophages. Fat-fed Tlr9−/− mice demonstrated reduced macrophage accumulation and inflammation in adipose tissue and better insulin sensitivity compared with wild-type mice, whereas bone marrow reconstitution with wild-type bone marrow restored the attenuation of insulin resistance observed in fat-fed Tlr9−/− mice. Administration of a TLR9 inhibitory oligonucleotide to fat-fed wild-type mice reduced the accumulation of macrophages in adipose tissue and improved insulin resistance. Furthermore, in humans, plasma ssDNA level was significantly higher in patients with computed tomography–determined visceral obesity and was associated with homeostasis model assessment of insulin resistance (HOMA-IR), which is the index of insulin resistance. Our study may provide a novel mechanism for the development of sterile inflammation in adipose tissue and a potential therapeutic target for insulin resistance.

Long‐chain monounsaturated fatty acid‐rich fish oil attenuates the development of atherosclerosis in mouse models

SCOPE Fish oil-derived long-chain monounsaturated fatty acids (LCMUFA) containing chain lengths longer than 18 were previously shown to improve cardiovascular disease risk factors in mice. However, it is not known if LCMUFA also exerts anti-atherogenic effects. The main objective of the present study was to investigate the effect of LCMUFA on the development of atherosclerosis in mouse models. METHODS AND RESULTS LDLR-KO mice were fed Western diet supplemented with 2% (w/w) of either LCMUFA concentrate, olive oil, or not (control) for 12 wk. LCMUFA, but not olive oil, significantly suppressed the development of atherosclerotic lesions and several plasma inflammatory cytokine levels, although there were no major differences in plasma lipids between the three groups. At higher doses 5% (w/w) LCMUFA supplementation was observed to reduce pro-atherogenic plasma lipoproteins and to also reduce atherosclerosis in ApoE-KO mice fed a Western diet. RNA sequencing and subsequent qPCR analyses revealed that LCMUFA upregulated PPAR signaling pathways in liver. In cell culture studies, apoB-depleted plasma from LDLR-K mice fed LCMUFA showed greater cholesterol efflux from macrophage-like THP-1 cells and ABCA1-overexpressing BHK cells. CONCLUSION Our research showed for the first time that LCMUFA consumption protects against diet-induced atherosclerosis, possibly by upregulating the PPAR signaling pathway.

Investigation of Uric Acid and Cystatin C on Low-Carbohydrate Diet (LCD)

Background: As to nutritional therapy, continuous discussions were observed concerning calorie restriction (CR) and low-carbohydrate diet (LCD). Authors and colleagues have applied LCD for lots of diabetic patients and reported the detail relationship with ketone bodies and Morbus (M) value. Methods: Ninety-three patients with type 2 diabetes mellitus (T2DM) were considered as subjects in the study, among which 51 were male and 52 were female, 58.3±13.2 years old on average, 60 years old in median. Methods were as follows: 1) patients were admitted and provided formular diet, which included CR diet (60% carbohydrates, 1400 kcal/day) on day 1-2, and LCD (12% carbohydrate, 1400 kcal/day) on day 3-14; 2) several biomarkers on fasting were measured on day 2, 4 and 14; 3) daily profile of blood glucose were done on day 2 and day 4. Results: According to the M-value, subjects were classified into 4 groups, which were less than 25, 26-100, 101-250, more than 251, and number was 24, 24, 24, 21, respectively. The average HbA1c in 4 groups were 6.6%, 7.4%, 8.5% and 9.5% respectively. The median M-values decreased from day 2 to 4, which were 10.4 to 9.1, 53.5 to 7.7, 150 to 19.1 and 438 to 87, respectively. The average uric acid in each group revealed significant increase from day 2 to day 14. There were significant correlation between uric acid increment and creatinine increment, and among creatinine, creatinine clearance (CCr) and Cystatin C. Conclusion: LCD showed efficacy for glucose variability with significant decrease in glucose and M-value. Renal study showed increase of serum uric acid. In addition to correlations of Cystatin C and biomarkers, current results would be from some dehydrated state and/or relative decrease of total calorie intake. These findings would become the fundamental data of efficacy of LCD and its physiological influences for renal function.

Effect of Low Carbohydrate Diet on Type 2 Diabetic Patients and Usefulness of M-Value

Background: Debate between the Calorie Restriction (CR) and Low-Carbohydrate Diet (LCD) has been continued for several years. We have started LCD therapy for diabetic patients since 1999, and experienced good response to control hyperglycemia. Recently the risk of postprandial hyperglycemia for cardiovascular events is reported, and the range of glucose level is focused. Morbus (M) value is proposed to be a good index to show the range of glucose fluctuation. Usefulness of LCD and usefulness of M-value were shown by combined analysis. Methods: Twenty-six patients with Type 2 Diabetes Mellitus (T2DM), who admitted to the Takao Hospital, were the subjects. They were 12 males and 14 females, and their age ranged from 38 to 78 years old. A few patients were new patients, but mostly referred from other hospitals to receive LCD treatment. All patients started from CR therapy with 60% carbohydrate for 2 days, and then carried out LCD dietary therapy with 12% carbohydrates for 10-12 days. On day 2 and 12, blood glucose level was measured at 7 points a day to see the fluctuation from morning to night. Blood and urinary samples were corrected on the same day to get ordinary biochemical data and metabolic marker. Results: To know the effect of LCD on different level of HbA1c, the patients were grouped to low (L; 6.1±0.5%), middle (M; 7.9±0.4%) and high (H; 9.2±0.7%) by HbA1c value. The M-values after 2 day CR were 26.1 (L), 94.1 (M) and 343 (H), respectively. The LCD therapy decreased M-value to 10.4 (L), 18.8 (M) and 84.2 (H), respectively. The significant reduction of M-values was a reflection of the improved postprandial glucose level. LCD was effective even in the bad controlled diabetic patients. In addition to the decreased triacylglycerol and 24 hr urinary C-peptide, uric acid increased in all patients. Conclusion: Ten to 12 day-dietary therapy with LCD was quite effective to improve blood glucose profile. Patients with high HbA1c could be safely treated by LCD. Blood glucose improvement was well correlated to the M-value.

Clinical Effect of Low Carbohydrate Diet (LCD): Case Report

Background: The discussion concerning calorie restriction (CR) and low carbohydrate diet (LCD) has been continued for long yeas. As to LCD, we have reported lots of experience and research for glucose variability and ketone bodies. Subjects and methods: Three cases with type 2 diabetes mellitus (T2DM) were on LCD and studied. Methods included 3 patterns of LCD meal which are super, standard and petit LCD, with including carbohydrate ratio 12%, 26% and 40%, respectively. Case 1 (61, M) showed hyperglycemia about 150 mg/dL to 300 mg/dL in daily profile with 12.5% in HbA1c. Starting super LCD therapy, the glucose profile decreased to less than 150 mg/dL and HbA1c decreased to 6.7% in 3 months. Case 2 (53, M) revealed HbA1c 8.3%, weight 110 kg and body mass index 34.5. By super LCD, his weight was decreased 17 kg in 5 months, with normalized HbA1c and elevated serum 3-hydroxybutyric acid (3-OHBA). Case 3 (72, M) had always extremely elevated fasting triglyceride for 5 years. Starting petit LCD for 2 years, triglyceride and weight were decreased moderately. Successively, changing to standard LCD for 1 year, weight was decreased 6 kg, and triglyceride was normalized. Discussion and conclusion: Each case suggests the characteristic beneficial effect of LCD. From our clinical experience and research, super LCD method, which is one of the very low-carbohydrate ketogenic diet (VLCKD), has evident efficacy of weight reduction. Super-LCD is characterized by strictly limitation for carbohydrate. Standard-LCD and petit-LCD method can be useful and applicable in response to the status of the patients. Thus, LCD treatment have beneficial effects for patients with various status.

Homeostasis Model Assessment (HOMA) and M Value in Daily Profile of Glucose

Background: Low Carbohydrate Diet (LCD) and Calorie Restriction (CR) have been on discussion for years. Authors have continued diabetic research about LCD, CR, Morbus (M) value and insulin secretion. In this study, homeostasis model assessment (HOMA) was investigated. Subjects and Methods: Subjects enrolled were 52 patients with type 2 diabetes mellitus (T2DM) (average 62.3 years). Methods included the measurement of fasting glucose and immunoreactive insulin (IRI), daily profile of blood glucose and M value. Results:The obtained data were as follows: average HbA1c 8.0%, average glucose of daily profile 222mg/dL. Median data were M value 151, HOMA-R 1.07, HOMA-β 11.1. Divided into 4 groups due to M value, the levels of HOMA-R and HOMA-β in each group were 0.68, 1.08, 1.64, 1.38 and 16.9, 16.3, 10.2, 5.3, respectively. Significant correlation were observed between M value and HOMA-R (p<0.01), and between M value and HOMA-β (p<0.01). Discussion and Conclusion: As M value increases, HOMA-R increases and HOMA-β decreases. These findings suggested that diabetic patients would have insulin resistance and decreased β cell function correlated to the severity of diabetes, and that obtained results would become the basal data in this field, expecting the further development in the future research.

Biology of Human Aging and Recent Nutrition Therapy

Discussion concerning Calorie Restriction (CR) and low carbohydrate diet (LCD) has continued for years. Authors have developed research of LCD using super LCD with 12% of carbohydrate. The biology of human aging includes study between human and mammalian such as mice, rhesus monkeys. Moderate CR reduced age-related mortality, incidence of diabetes, cancer, cardiovascular disease. By very low calorie diet (VLCD), body weight can be reduced, but the diabetic pathological state remains. Besides CR and LCD, Zone, Ornish, LEARN Diets and Mediterranean style can be applicable. There are very low-carbohydrate ketogenic diet (VLCKD), Moderate-LCD, High- Carbohydrate Diet, as related with the definition of LCD. Morbus (M) value was investigated for clinical practice of LCD, in which M value was remarkably improved by LCD treatment.

The Use of Topical Compounded Analgesic Creams in Neuropathic Pain by the Primary Care Physician

The primary care physician often has to help diabetic patients suffering from localized neuropathic pain. Most of the current anti-epileptics and anti-depressants however, especially when prescribed to the elderly patients, have troublesome drawbacks: drug-interactions, side effects and tolerability issues. Topical compounded creams might be a good alternative, especially since it is easy to test in the general practice to discover responders to these creams. At our Institute for Neuropathic Pain we see many patients suffering from peripheral neuropathic pain. We have developed a number of compounded creams based on amongst others amitriptyline 10%, ketamine 10%, clonidine 0.2%, baclofen 5%, and phenytoin 10%. Many patients profit from such topical formulations and we will describe how a primary care physician can fast decide whether as specific compounded cream is useful for a patient. Patients suffering from peripheral neuropathic pain, as in diabetic neuropathy, chronic idiopathic axonal polyneuropathy (CIAP) and chemotherapy induced polyneuropathy (CIPN) as well as from complex regional pain syndrome (CRPS), seem often to be responsive to such topical approach. Meanwhile, more than 800 Dutch medical doctors have been prescribing one or more of our compounded creams, and patients and medical doctors from abroad frequently contact us to explore the therapeutic options for their patients based on these creams. We will give some practical tips how to differentiate between responders and non-responders.

Influence of Diabetes and Hemodialysis against Nerve Conduction Studies

Background: Diabetic peripheral neuropathy (DPN) has been prevalent and discussed, and nerve conduction studies (NCS) has been continued. We have checked NCS using recently introduced useful DPN-Check device. Subjects and Methods: The subjects were 66 patients (pts) classified into 4 groups according to existence of diabetes mellitus (DM) and hemodialysis (HD); Group1: DM (+), HD (+) in 15 pts, group 2: DM (-), HD (+) in 15 pts, group 3: DM (+), HD (-) in 20 pts, group 4: 16 healthy controls. Methods included measurements of sural nerve conduction velocity (SNCV) and sural nerve action potential (SNAP) using HDN-1000. Results: Average age in each group was 64.4 years to 72.6 years. SNCV value of 4 group in average was 37.1 m/sec, 46.3 m/sec, 49.3 m/sec, 53.2 m/sec, respectively, and value of group 1 was significantly lower than those of group 2,3,4 (p<0.01). Similarly, average SNAP was 4.1 μV, 8.7 μV, 8.0 μV, 21.6 μV, respectively, and group 1,2,3 were significantly lower than group 4 (p<0.01). There was significant correlation between SNCV and SNAP in all subjects (p<0.01). Significant correlations were shown between DM duration and SNCV, and DM duration and SNAP (p<0.01). Discussion and Conclusion: SNCV and SNAP were measured successfully and easily by HDN-1000, indicating clinical availability. Obtained data suggested that 1) SNCV is not significantly decreased due to only uremic neuropathy, 2) SNCV is significantly decreased in patients with both HD and DM, 3) SNAP is significantly decreased in patents with DM for years and 4) SNAP would be remarkably decreased when HD is in addition to DM. These results would become the basal data of future NCS for DM and HD.

Investigation of Elevated Ketone Bodies in Low Carbohydrate Diet (LCD)

Background: Discussion has been continued concerning Calorie Restriction (CR) and Low Carbohydrate Diet (LCD). For years, authors have investigated LCD associated with lipids, renal function and Ketone Bodies (KB) for clinical application of super LCD formular meal. In this study, 3-hydroxybutyric acid (3-OHBA) and acetoacetic acid (AcAc) was measured. Subjects and Methods: The subjects were 105 patients with type 2 diabetes mellitus (T2DM), (M/F 47/58, 62.7 years. in average) they were admitted for treatment of T2DM. The protocol consists of 3 steps. 1. Calorie Restriction (CR) diet was given on day 1 and 2 with 60% of carbohydrates. 2. Low Carbohydrate Diet (LCD) was given after day 3 with 12% of carbohydrates which is super-LCD formula meal. 3. Total ketone bodies (T-KB), 3-OHBA and AcAc were measured, and investigated the value and ratio of these markers. Results: Median T-KB was 349, 415, 486, 415, 445 μmol/L, in day 4-6, 7-9, 10-11, 12-15, 21-30, in 5 groups. Discussion and Conclusion: Hyperketonemia is due to continuation of LCD, which is physiological ketosis without clinically hazardous acidosis. As value of 3-OHBA increased, the ratio of 3-OHBA/T-KB increased. These results may become the fundamental data for clarifying the pathophysiological role of 3-OHBA and AcAc in hyperketonemia from carbohydrate restriction. respectively. There was significant correlation between value of 3-OHBA and ratio of 3-OHBA/T-KB (p<0.01, r=0.72). When 3-OHBA value was less or more than 1000 μmol/L, 3-OHBA ratio showed 65-89% or 90-94%, respectively.