ABSTRACT: BACKGROUND: Our aim was to compare the effects of a Paleolithic ('Old Stone Age') diet and a diabetes diet as generally recommended on risk factors for cardiovascular disease in patients with type 2 diabetes not treated with insulin. METHODS: In a randomized cross-over study, 13 patients with type 2 diabetes, 3 women and 10 men, were instructed to eat a Paleolithic diet based on lean meat, fish, fruits, vegetables, root vegetables, eggs and nuts; and a Diabetes diet designed in accordance with dietary guidelines during two consecutive 3-month periods. Outcome variables included changes in weight, waist circumference, serum lipids, C-reactive protein, blood pressure, glycated haemoglobin (HbA1c), and areas under the curve for plasma glucose and plasma insulin in the 75g oral glucose tolerance test. Dietary intake was evaluated by use of 4-day weighed food records. RESULTS: Study participants had on average a diabetes duration of 9 years, a mean HbA1c of 6,6 % units by Mono-S standard and were usually treated with metformin alone (3 subjects) or metformin in combination with a sulfonylurea (3 subjects) or a thiazolidinedione (3 subjects). Mean average dose of metformin was 1031 mg per day. Compared to the diabetes diet, the Paleolithic diet resulted in lower mean values of HbA1c (-0.4% units, p=0.01), triacylglycerol (-0.4 mmol/L, p=0.003), diastolic blood pressure (-4 mmHg, p=0.03), weight (-3 kg, p=0.01), BMI (-1 kg/m2, p=0.04) and waist circumference (-4 cm, p=0.02), and higher mean values of high density lipoprotein cholesterol (+0.08 mmol/L, p=0.03). The Paleolithic diet was mainly lower in cereals and dairy products, and higher in fruits, vegetables, meat and eggs, as compared with the Diabetes diet. Further, the Paleolithic diet was lower in total energy, energy density, carbohydrate, dietary glycemic load, saturated fatty acids and calcium, and higher in unsaturated fatty acids, dietary cholesterol and several vitamins. Dietary GI was slightly lower in the Paleolithic diet (GI=50) than in the Diabetic diet (GI=55). CONCLUSIONS: Over a 3-month study period, a Paleolithic diet improved glycemic control and several cardiovascular risk factors compared to a Diabetes diet in patients with type 2 diabetes. Trial registration ClinicalTrials.gov NCT00435240.

ABSTRACT: BACKGROUND: A Paleolithic diet has been suggested to be more in concordance with human evolutionary legacy than a cereal based diet. This might explain the lower incidence among hunter-gatherers of diseases of affluence such as type 2 diabetes, obesity and cardiovascular disease. The aim of this study was to experimentally study the long-term effect of a Paleolithic diet on risk factors for these diseases in domestic pigs. We examined glucose tolerance, post-challenge insulin response, plasma C-reactive protein and blood pressure after 15 months on Paleolithic diet in comparison with a cereal based swine feed. METHODS: Upon weaning twenty-four piglets were randomly allocated either to cereal based swine feed (Cereal group) or cereal free Paleolithic diet consisting of vegetables, fruit, meat and a small amount of tubers (Paleolithic group). At 17 months of age an intravenous glucose tolerance test was performed and pancreas specimens were collected for immunohistochemistry. Group comparisons of continuous variables were made by use of the t-test. P<0.05 was chosen for statistical significance. Simple and multivariate correlations were evaluated by use of linear regression analysis. RESULTS: At the end of the study the Paleolithic group weighed 22% less and had 43% lower subcutaneous fat thickness at mid sternum. No significant difference was seen in fasting glucose between groups. Dynamic insulin sensitivity was significantly higher (p=0.004) and the insulin response was significantly lower in the Paleolithic group (p=0.001). The geometric mean of C-reactive protein was 82% lower (p=0.0007) and intra-arterial diastolic blood pressure was 13% lower in the Paleolithic group (p=0.007). In evaluations of multivariate correlations, diet emerged as the strongest explanatory variable for the variations in dynamic insulin sensitivity, insulin response, C-reactive protein and diastolic blood pressure when compared to other relevant variables such as weight and subcutaneous fat thickness at mid sternum. There was no obvious immunohistochemical difference in pancreatic islets between the groups, but leukocytes were clearly more frequent in sampled pancreas from the Cereal group. CONCLUSIONS: This study in domestic pigs suggests that a Paleolithic diet conferred higher insulin sensitivity, lower C-reactive protein and lower blood pressure when compared to a cereal based diet.

Many members of the forkhead genes family of transcription factors have been implicated as important regulators of metabolism, in particular glucose homeostasis e.g. Foxo1, Foxa3 and Foxc2. The purpose of this study was to exploit the possibility that yet unknown members of this gene family play a role in regulating glucose tolerance in adipocytes. We identified Foxf2 in a screen for adipose expressed forkhead genes. In vivo over-expression of Foxf2 in an adipose tissue restricted fashion demonstrated that such mice display a significantly induced insulin secretion in response to an intravenous glucose load as compared with wild type littermates. In response to increased Foxf2 expression, Irs1 mRNA and protein levels are significantly down regulated in adipocytes, however the ratio of serine versus tyrosine phosphorylation IRS1 seems to remain unaffected. Furthermore, adipocytes over-expressing FOXF2 have a significantly lower insulin mediated glucose uptake compared with wild type adipocytes. These findings argue that Foxf2 is a previously unrecognized regulator of cellular and systemic whole body glucose tolerance, at least in part, due to lower levels of IRS1. Foxf2 and its downstream target genes can provide new insights with regard to identification of novel therapeutic targets.

The objective of our study was to determine the prevalence of anti-infectious agent antibodies and autoantibodies in a unique non-Westernized population from Kitava, Papua New Guinea (PNG), compared to Western populations. We matched 120 serum samples from Kitavans with 437 samples from four healthy control groups. Sera were tested for the presence of anti-infectious agent antibodies (treponema, toxoplsmosis, Epstein-Barr virus, cytomegalovirus, rubella) and autoantiobodies [anti-double-stranded (ds)DNA, anti-chromatin, anti-ribonucleoprotein (RNP), anti-SSB, anti-SSA, anti-Scl-70, anti-Smith, anti-centromer, anti-SmRNP, anti-Jo-1, and anti-ribosomal-P] using the Bio-Rad BioPlex 2200. Antitreponemal antibodies were detected in 87% of PNG sera versus 0-6% of controls (P < 0.0001). Anti-dsDNA antibodies were detected in 31% of PNG samples, which was significantly higher than in three of the control groups (<10%). The outstanding high rate of antitreponemal antibodies detected in Kitavans possibly represents prior yaws disease. A low prevalence of cardiovascular disease was previously documented in Kitavans and has been attributed, in addition to their diet, to the high prevalence of natural cardioprotective autoantibodies (the IgM-antiphosphorylcholine antibodies) in this population. Treponemal infection has been shown to induce the appearance of antiphosphorylcholine antibodies. These protective autoantibodies may cross-react with the pathogenic anti-dsDNA antibodies. Thus, it is suggested that infection with treponema is associated with the presence of protective as well as pathogenic autoantibodies.

Abstract The hydroxyl radical ((*)OH) is believed to act as the small non-enzymatic agent involved in the brown-rot decay of wood. However, knowledge about the relation between hydroxyl radical production and the activity of wood-decomposing fungi in wood or about the significance of these radicals during interactions with other organisms is limited due to a lack of reliable methods for detecting the radicals. A sensitive and specific fluorescence method was developed in this study to detect the production of (*)OH by wood-decomposing fungal species. The method was based on the hydroxylation of coumarin-3-carboxylic acid, which produces 7-hydroxy-coumarin-3-carboxylic acid (7-OHCCA), a fluorescent, stable and specific product. Wood discs colonized by fungi were placed on water agar containing coumarin-3-carboxylic acid, where the formation of 7-OHCCA occurred and the fluorescence could be measured. The production of (*)OH was above the detection limit for eight of the 10 fungal species. The highest level, 8-25 times that of the detection limit, was produced by the brown-rot fungus Antrodia vaillantii. In interaction experiments where A. vaillantii had established contact with the antagonistic bacterium Pseudomonas fluorescens, the production of (*)OH increased, whereas contact with Bacillus subtilis did not change the amount of (*)OH generated compared with controls containing only the fungus. In contrast, the production of (*)OH increased above the control level when the fungus Coniophora puteana was in contact with any of these bacteria. The method was also tested in soil, with the result that 40% of 7-OHCCA added to the soil could be recovered with K(2)HPO(4) buffer.

Phenylurea herbicides are used worldwide, and often pollute surface- and groundwater in concentrations exceeding the limit value for drinking water (0.1 mug l(-1)). Bacteria degrade phenylurea herbicides by successive N-dealkylation to substituted aniline products. Little is known about the corresponding fungal pathways, however. We here report degradation of chlorotoluron, diuron, isoproturon and linuron by the soil fungus Mortierella sp. Gr4. Degradation was fastest with linuron and resulted in successively dealkylated metabolites and 3,4-dichloroaniline. A major new metabolite was detected that has not yet been fully identified. Thin layer chromatography and nuclear magnetic resonance spectroscopy indicate that it is a non-aromatic diol. Degradation of isoproturon, chlorotoluron and diuron involved successive N-demethylation and, in the case of isoproturon and chlorotoluron, additional hydroxylation. A new hydroxylated isoproturon metabolite was detected. The study thus shows that the fungal pathways differ from the bacterial pathways and yield new metabolites of possible environmental concern.

Mice are increasingly used in studies where measuring insulin sensitivity (IS) is a common procedure. Glucose clamp is labor intensive, cannot be used in large numbers of animals, cannot be repeated in the same mouse and has been questioned as a valid tool for IS in mice; thus, the minimal model with 50min-intravenous glucose (IVGTT) data was adapted in mice. However, specific software and particular ability is needed. Aim of this study was to establish a simple procedure for evaluating IS during IVGTT in mice (CSI). IVGTT (n=520) were performed in NMRI and C57BL/6J mice (20-25g). After glucose injection (1 g/kg), 7 samples were collected until 50 min for glucose and insulin measurements, analyzed with minimal model which provided the validated reference IS (SI). By using the regression CSI=alpha1+alpha2xKG/AUCD, IS was calculated in 134 control animals randomly selected (regression CSI vs. SI: r=0.66, p<0.0001) and yielded alpha1=1.93 and alpha2=0.24. KG is the slope of log(glucose5-20) and AUCD is the mean dynamic area under insulin curve in the IVGTT. By keeping fixed alpha1 and alpha2, CSI was validated in 143 control mice (4.7+/-0.2 min(-1)microU(-1)ml, virtually identical to SI: 4.7+/-0.3, r=0.89, p<0.0001; and in 123 mice in different conditions: transgenic, addition of neuropeptides, incretins, insulin (CSI: 6.0+/-0.4 vs. SI: 6.1+/-0.4, r=0.94, p<0.0001). In other 120 animals, CSI revealed its ability to segregate different categories, as SI does. This easily usable formula for calculating CSI overcomes many experimental obstacles and may be a simple alternative to more complex procedures when large numbers of mice or repeated experiments in the same animals are required. Key words: glucose tolerance, insulin resistance, glucose effectiveness, animal model.

Glucagon-like 1 peptide (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) regulate islet function after carbohydrate ingestion. Whether incretin hormones are of importance for islet function after ingestion of non-carbohydrate macronutrients is not known. This study therefore examined the integrated incretin and islet hormone responses to ingestion of pure fat (oleic acid; 0.88 g/kg) or protein (milk and egg protein; 2 g/kg) over 5 hours in healthy men, aged 20-25 years (n=12); plain water ingestion served as control. Both intact (active) and total GLP-1 and GIP levels were determined as was plasma activity of dipeptidyl peptidase-4 (DPP-4). Following water ingestion, glucose, insulin, glucagon, GLP-1 and GIP levels and DPP-4 activity were stable during the 5 hour study period. Both fat and protein ingestion increased insulin, glucagon, GIP and GLP-1 levels without affecting glucose levels or DPP-4 activity. The GLP-1 responses were similar after protein and fat, whereas the early (30 min) GIP response was higher after protein than after fat ingestion (P<0.001). This was associated with seven-fold higher insulin and glucagon responses compared to fat ingestion (both P<0.001). After protein, the early GIP, but not GLP-1, responses correlated to insulin (r(2)=0.86; P=0.0001) but not glucagon responses. In contrast, after fat ingestion, GLP-1 and GIP did not correlate to islet hormones. We conclude that whereas protein and fat release both incretin and islet hormones, the early GIP secretion after protein ingestion may be of primary importance to islet hormone secretion. Key words: insulin, glucagon, GLP-1, GIP.

In the course of the "Neolithic Revolution", while the warming after the glacial period about 13,000 years ago was starting, the crucial evolution of mankind took place, which has consequences to this day. With the beginning of agriculture and metallurgy a transition from an accidental outlook on life to a deliberate shaping of life set in. A chance from almost egalitarian tribal societies oriented solely towards consumption into production societies organized in a hierarchically complex way was not solely the result of an adaptation to climatic conditions, but also of a positively selecting evolution of the intellect. Following the socially relevant upheavals connected with this, the prerequisites for starting modest prosperity only for a privileged few across millenniums were created. Problems and future prospects of today's global distribution of affluence are discussed.

The obesity is a multicausal disease that includes exogenous and endogenous factors. We emphasized the leptin, a hormone produced by adipose tissue, whose receiver is in the hypothalamus. The leptin has been considered important in the development of the obesity, because it influences in the energy intake and expenditure. In the revision one sets out to evaluate the leptin like factor related to the development of the obesity, as well as the influence of diet in the secretion and action of leptin. In the human obesity, the deficiency of functional leptin is rare, nevertheless, can exist leptin resistance. After the loss weight it happens reduction in the leptin levels, which can be in the recovery of body weight. The leptin is regulated by food intake and their circulating levels and potential of action are associated with content of macronutrients of the diet and the body composition individually. Nevertheless, studies demonstrated the high and the low food intake can influence in the leptin secretion, independent of the effect on adiposity. In addition, the fat of diet presents important effect in secretion and action of leptin. Also the zinc deficiency has been associated with the reduced levels of leptin. Several experimental studies have been made to evaluate the effect of the administration of leptin in treatment of obesity, by means of injections, inhalation or more active forms of leptin. Nevertheless, we need similar studies with humans. Is important the development of studies that associate the environmental factors, particularly the diet, with regulation of serum levels and action of leptin in humans.

In this investigation shown the influence of traditional hypocaloric diet on concentration of leptin level and leptin receptor in type 2 diabetic patients. Our results show that diet therapy improves metabolic control, and as a result of a diet were normoglucaemia and decrease of leptin and leptin receptor level in type 2 diabetic patients.

ABSTRACT: BACKGROUND: A Paleolithic diet has been suggested to be more in concordance with human evolutionary legacy than a cereal based diet. This might explain the lower incidence among hunter-gatherers of diseases of affluence such as type 2 diabetes, obesity and cardiovascular disease. The aim of this study was to experimentally study the long-term effect of a Paleolithic diet on risk factors for these diseases in domestic pigs. We examined glucose tolerance, post-challenge insulin response, plasma C-reactive protein and blood pressure after 15 months on Paleolithic diet in comparison with a cereal based swine feed. METHODS: Upon weaning twenty-four piglets were randomly allocated either to cereal based swine feed (Cereal group) or cereal free Paleolithic diet consisting of vegetables, fruit, meat and a small amount of tubers (Paleolithic group). At 17 months of age an intravenous glucose tolerance test was performed and pancreas specimens were collected for immunohistochemistry. Group comparisons of continuous variables were made by use of the t-test. P<0.05 was chosen for statistical significance. Simple and multivariate correlations were evaluated by use of linear regression analysis. RESULTS: At the end of the study the Paleolithic group weighed 22% less and had 43% lower subcutaneous fat thickness at mid sternum. No significant difference was seen in fasting glucose between groups. Dynamic insulin sensitivity was significantly higher (p=0.004) and the insulin response was significantly lower in the Paleolithic group (p=0.001). The geometric mean of C-reactive protein was 82% lower (p=0.0007) and intra-arterial diastolic blood pressure was 13% lower in the Paleolithic group (p=0.007). In evaluations of multivariate correlations, diet emerged as the strongest explanatory variable for the variations in dynamic insulin sensitivity, insulin response, C-reactive protein and diastolic blood pressure when compared to other relevant variables such as weight and subcutaneous fat thickness at mid sternum. There was no obvious immunohistochemical difference in pancreatic islets between the groups, but leukocytes were clearly more frequent in sampled pancreas from the Cereal group. CONCLUSIONS: This study in domestic pigs suggests that a Paleolithic diet conferred higher insulin sensitivity, lower C-reactive protein and lower blood pressure when compared to a cereal based diet.