Cardiac repolarization and depolarization in people with Type 1 diabetes with normal ejection fraction and without known heart disease: a case-control study.

AIMS: To investigate depolarization and repolarization durations in people with Type 1 diabetes, including the relationship to age. METHODS: 855 persons with Type 1 diabetes without known heart disease were included and matched with 1710 participants from a general population study. Clinical examinations, questionnaires and biochemistry were assessed. A 10-second 12-lead ECG was performed and analysed digitally. RESULTS: QTc was longer in people with Type 1 diabetes compared to controls (414±16 vs. 411±19 ms, P <0.001), and particularly so in young people with Type 1 diabetes. The fully adjusted increase was 13.8 ms (95% confidence interval (CI): 8.6-19.0 ms, P <0.001) at age 20 years and 3.4 ms (CI: 1.5-5.3 ms, P<0.001) at age 40 years. The rate-corrected QRSc was increased in people with Type 1 diabetes (97±11 vs. 95±11 ms, P <0.001) and was age-independent (P =0.5). JTc was increased in the young people with Type 1 diabetes (10.7 ms (CI: 5.4-16.0 ms, P <0.001) at age 20 years), but not in older people with Type 1 diabetes (interaction age-diabetes, P <0.01). CONCLUSIONS: For people with Type 1 diabetes, cardiac depolarization is increased at all ages, whereas repolarization is increased only relatively in young people with Type 1 diabetes. Hence, young people with Type 1 diabetes may be more prone to ventricular arrhythmias. The findings contribute to the understanding of sudden cardiac death in young people with Type 1 diabetes.

Near-normalization of glycaemic control with glucagon-like peptide-1 receptor agonist treatment combined with exercise in patients with type 2 diabetes.

AIMS: To investigate the effects of exercise in combination with a glucagon-like peptide-1 receptor agonist (GLP-1RA), liraglutide, or placebo for the treatment of type 2 diabetes. METHODS: Thirty-three overweight, dysregulated and sedentary patients with type 2 diabetes were randomly allocated to 16 weeks of either exercise and liraglutide or exercise and placebo. Both groups had three supervised 60-minute training sessions per week including spinning and resistance training. RESULTS: Glycated haemoglobin (HbA1c) levels dropped by a mean ± standard deviation of 2.0% ± 1.2% (from 8.2% ± 1.4%) in the exercise plus liraglutide group vs 0.3% ± 0.9% (from 8.0% ± 1.2%) in the exercise plus placebo group ( P < .001), and body weight was reduced more with liraglutide (-3.4 ± 2.9 kg vs -1.6 ± 2.3 kg; P < .001). Compared with baseline, similar reductions were seen in body fat (exercise plus liraglutide: -2.5% ± 1.4% [ P < .001]; exercise plus placebo: -2.2% ± 1.9% [ P < .001]) and similar increases were observed in maximum oxygen uptake (exercise plus liraglutide: 0.5 ± 0.5 L O2 /min [ P < .001]; exercise plus placebo: 0.4 ± 0.4 L O2 /min [ P = .002]). Greater reductions in fasting plasma glucose (-3.4 ± 2.3 mM vs -0.3 ± 2.6 mM, P < .001) and systolic blood pressure (-5.4 ± 7.4 mm Hg vs -0.6 ± 11.1 mm Hg, P < .01) were seen with exercise plus liraglutide vs exercise plus placebo. The two groups experienced similar increases in quality of life during the intervention. CONCLUSIONS: In obese patients with type 2 diabetes, exercise combined with GLP-1RA treatment near-normalized HbA1c levels and caused a robust weight loss when compared with placebo. These results suggest that a combination of exercise and GLP-1RA treatment is effective in type 2 diabetes.

Glycated haemoglobin and the risk of cardiovascular disease, diabetes and all-cause mortality in the Copenhagen City Heart Study.

OBJECTIVE: Individuals with diabetes mellitus (DM) have a considerably elevated risk of developing serious health problems including cardiovascular disease (CVD). Long-term elevated levels of blood glucose in nondiabetic individuals may also be associated with increased risk of CVD. The aim of this study was to investigate the relationships between glycated haemoglobin A(1c) (HbA(1c) ) and CVD, DM and all-cause mortality. SUBJECTS AND DESIGN: The Copenhagen City Heart Study is a prospective study of individuals from the Danish general population. The cohort was followed for 10 years via national registers with respect to incident CVD, DM and all-cause mortality. Follow-up was 100% complete. RESULTS: A total of 5127 subjects were included, of whom 597 had DM. In the nondiabetic population, HbA(1c) was significantly associated with incident CVD events in both univariate [hazard ratio (HR) 1.38, 95% CI 1.11-1.71] and multivariate analyses (HR 1.31, 95% CI 1.05-1.64). In the nondiabetic population, increased levels of HbA(1c) were correlated with developing DM. There was a threefold increase in risk of incident DM per unit increase in HbA(1c) with a univariate HR of 3.83 (95% CI 1.96-7.51). This relationship was essentially unchanged after multivariate adjustments (HR 4.19, 95% CI 2.01-8.71). Furthermore, we found that net reclassification improvement for diagnosed DM and CVD was significantly improved with the addition of HbA(1c) in the analyses. Although not statistically significant, we found a strong trend towards an association between HbA(1c) and all-cause mortality (HR 1.21, 95% CI 0.99-1.47). We did not find the same associations amongst the population with DM. CONCLUSION: In the Danish general population, HbA(1c) was strongly associated with CVD in individuals without DM.

A diet high in omega-3 fatty acids does not improve or protect cognitive performance in Alzheimer's transgenic mice.

Although a number of epidemiologic studies reported that higher intake of omega-3 fatty acids (largely associated with fish consumption) is protective against Alzheimer's disease (AD), other human studies reported no such effect. Because retrospective human studies are problematic and controlled longitudinal studies over decades are impractical, the present study utilized Alzheimer's transgenic mice (Tg) in a highly controlled study to determine whether a diet high in omega-3 fatty acid, equivalent to the 13% omega-3 fatty acid diet of Greenland Eskimos, can improve cognitive performance or protect against cognitive impairment. Amyloid precursor protein (APP)-sw+PS1 double transgenic mice, as well as nontransgenic (NT) normal littermates, were given a high omega-3 supplemented diet or a standard diet from 2 through 9 months of age, with a comprehensive behavioral test battery administered during the final 6 weeks. For both Tg and NT mice, long-term n-3 supplementation resulted in cognitive performance that was no better than that of mice fed a standard diet. In NT mice, the high omega-3 diet increased cortical levels of omega-3 fatty acids while decreasing omega-6 levels. However, the high omega-3 diet had no effect on cortical fatty acid levels in Tg mice. Irrespective of diet, no correlations existed between brain omega-3 levels and cognitive performance for individual NT or Tg mice. In contrast, brain levels of omega-6 fatty acids were strongly correlated with cognitive impairment for both genotypes. Thus, elevated brain levels of omega-3 fatty acids were not relevant to cognitive function, whereas high brain levels of omega-6 were associated with impaired cognitive function. In Tg mice, the omega-3 supplemental diet did not induce significant changes in soluble/insoluble Abeta within the hippocampus, although strong correlations were evident between hippocampal Abeta(1-40) levels and cognitive impairment. While these studies involved a genetically manipulated mouse model of AD, our results suggest that diets high in omega-3 fatty acids, or use of fish oil supplements (DHA+EPA), will not protect against AD, at least in high-risk individuals. However, normal individuals conceivably could derive cognitive benefits from high omega-3 intake if it corrects an elevation in the brain level of n-6 fatty acids as a result. Alternatively, dietary fish may contain nutrients, other than DHA and EPA, that could provide some protection against AD.

Lifelong immunization with human beta-amyloid (1-42) protects Alzheimer's transgenic mice against cognitive impairment throughout aging.

Although both active and passive beta-amyloid (Abeta) immunotherapy have been shown to protect against or lessen cognitive impairment in various Alzheimer's transgenic mouse lines, these studies have focused on a single task and involved standard statistical analysis. Because Alzheimer's disease impacts multiple cognitive domains, the current study employed an extensive behavioral battery and multimetric analysis therein to determine the impact of Abeta immunization given throughout most of adult life (from 2-16 1/2 months of age) to APP+PS1 transgenic mice. At both adult (4 1/2-6 month) and aged (15-16 1/2 month) test points, the same 6-week behavioral battery was administered. Results indicate that Abeta immunotherapy partially or completely protected APP+PS1 mice at both test points from otherwise impaired performance in a variety of tasks spanning multiple cognitive domains (reference learning/memory, working memory, search/recognition). At both adult and aged test points, the cognitive benefits of Abeta immunotherapy were evident even when behavioral measures were analyzed collectively (as "overall" performance) through discriminant function analysis. Since behavioral protection at the 15-16 1/2 month test point occurred without a decrease in (or correlation to) Abeta deposition, the mechanism of Abeta immunotherapy's action most likely involves neutralization/removal of small Abeta oligomers from the brain. However, in factor analysis performed at this aged test point, brain Abeta deposition measures loaded heavily with key cognitive measures. Collectively, our results suggest that the entire process of Abeta deposition deleteriously impacts cognitive performance and that Abeta-based preventative strategies can provide long-term cognitive benefits extending well into older age.

A comparison of warmed-over flavour in pork by sensory analysis, GC/MS and the electronic nose.

Pork muscle samples (M. longissimus dorsi and M. psoas major) were obtained from pigs given one of 4 dietary treatments, (i) control diet, (ii) supplemental iron (300 mg iron (II) sulphate/kg feed), (iii) supplemental vitamin E (200 mg dl-α-tocopheryl acetate/kg of feed) and (iv) supplemental vitamin E+supplemental iron. Warmed-over flavour (WOF) was evaluated by a trained sensory panel (n=8) for the four treatments cooked and refrigerated at 4 °C for up to 5 days. Gas chromatography mass spectrometry (GC/MS) and Electronic nose analysis was performed on a subset of the full design which included samples of M. longissimus dorsi, treatments (ii) and (iii) and M. psoas major with treatment (i) for 0 days of WOF development. Day 5 of WOF development was included in the subset and represented by samples of M. longissimus dorsi, treatment (iv) and M. psoas major, treatments (ii) and (iii). Bi-linear modeling was used to determine the correlation of GC/MS and electronic nose data to sensory data. Also, the reproducibility and reliability of electronic nose data was evaluated by repeating the analysis of samples in a different laboratory and with a time difference of approximately 11 months. Mean-centring was used to normalise the data from these two different electronic noise data sets. GC/MS data correlated to sensory data with specific compounds (e.g., pentanal, 2-pentylfuran, octanal, nonanal, 1-octen-3-ol and hexanal), proving to be good indices of oxidation in cooked samples of M. longissimus dorsi and M. psoas major. Electronic nose data correlated to sensory data and separated the sensory variation. The reproducibility of this data was high with the second set of samples being predictive of the first set.

Effect of Brussels sprouts and inulin/rape seed cake on the sensory profile of pork M. longissimus dorsi.

The objective of the present study was to evaluate the influence of two diets containing either Brussels sprouts or inulin/rape seed cake, compared with a standard diet (control) for slaughter pigs on flavour and odour attributes and sensory profile of cooked pork. Three weeks prior to slaughter 24 female pigs were allocated to three diets: (1) a standard grower-finishing diet (control) for slaughter pigs containing barley, wheat and soy-bean meal, (2) the control diet containing 11 energy percent Brussels sprouts and (3) a diet containing 25% inulin and 55% rape seed cake. The odour and flavour of the cooked meat from inulin/rape seed cake-fed pigs differed significantly from the other two diets, showing reduced meat odour, increased pig and acrid odour, increased pig flavour, reduced fresh flavour and total impression. Meat from the Brussels sprouts-fed pigs deviated only slightly from the control-fed pigs.

Synthesis of nitrogen analogues of salacinol and their evaluation as glycosidase inhibitors.

The syntheses of two nitrogen analogues (11 and 12) of the naturally occurring sulfonium ion, salacinol (7) are described. The latter compound is one of the active principles in the aqueous extracts of Salacia reticulata that are traditionally used in Sri Lanka and India for the treatment of diabetes. The synthetic strategy relies on the nucleophilic attack of a 1,4-dideoxy-1,4-imino-D- or L-arabinitol at the least hindered carbon of 2,4-O-benzylidene D- or L-erythritol-1,3-cyclic sulfate. The nitrogen analogues bear a permanent positive charge and serve as mimics of the sulfonium ion. We reasoned that these ammonium derivatives should function in a manner similar to that of known glycosidase inhibitors of the alkaloid class such as castanospermine (4) and deoxynojirimycin (5). Enzyme inhibition assays indicate that salacinol (7) is a weak (K(i) = 1.7 mM) inhibitor of glucoamylase, whereas compounds 11 and 12 inhibit glucoamylase with K(i) values in the range approximately 10-fold higher. The nitrogen analogues 11 and 12 showed no significant inhibitory effect of either barley alpha-amylase (AMY1) or porcine pancreatic alpha-amylase (PPA) at concentrations of 5 mM. In contrast, salacinol (7) inhibited AMY1 and PPA in the micromolar range, with K(i) values of 15 +/- 1 and 10 +/- 2 microM, respectively.

3-Methylindole (skatole) and indole production by mixed populations of pig fecal bacteria.

Pig fecal slurries converted added L-tryptophan either to indole without detectable intermediates or to 3-methylindole (skatole) via indole-3-acetate. The initial rate of production of 3-methylindole was greatest at pH 6.5 and less at pH 5.0 and 8.0; the initial rates of indole production were similar at pH 6.5 and 8.0. More than 80% of the tryptophan added was converted to 3-methylindole at pH 5.0; at pH 8.0 85% was converted to indole. Both pathways had similar Km values for tryptophan and similar maximum rates. Indole-3-carbinol and indole-3-acetonitrile completely inhibited the production of 3-methylindole from indole-3-acetate but had no effect on the reactions involving L-tryptophan.

Gas chromatographic determination of indole and 3-methylindole (skatole) in bacterial culture media, intestinal contents and faeces.

A simple, rapid and inexpensive gas chromatographic method was developed for the determination of indole and 3-methylindole (skatole) in faeces, intestinal contents and bacterial cultures. It involves a simple homogenization and extraction with chloroform. The extract is injected onto a gas chromatograph equipped with a 12.5-m fused-silica capillary column coated with BP20 and a film thickness of 0.5 micron. To simplify the chromatograms and to get a higher sensitivity a nitrogen-phosphorus-sensitive detector is applied. The detection limit for indole and 3-methylindole under the conditions employed is 20 micrograms/kg, which is well below the values typically found in intestinal contents (up to 100 mg/kg). Recovery for both compounds was close to 100%, and the mean coefficients of variation were 3.5% for indole and 3.0% for 3-methylindole. The method has demonstrated its practical value in the analysis of more than 50,000 samples in our laboratory. More than 100 samples can be analyzed per day.

Serum antibody responses of neonatal and young adult pigs to transmissible gastroenteritis coronavirus.

Serum titers of virus-neutralizing (VN) antibody were 10 to 16 times higher in neonatal pigs than in young adult pigs, after single oral doses of virulent transmissible gastroenteritis virus (TGEV). To determine the reason for this higher response, sera from neonatal and young adult pigs, 18 to 21 days after exposure to TGEV, were collected and assayed for VN antibody by plaque reduction. In addition, sera of VN-positive and VN-negative neonatal pigs were analyzed for immunoglobulin classes by radial immunodiffusion technique. The competence of neonatal pigs to produce VN antibody with increased IgG levels was demonstrated. The higher antibody response seen in neonatal pigs, when compared to sera of young adult pigs, may be attributed to the increased replication of TGEV in the intestinal tracts of neonatal pigs or to the lack of other immunogens that may interfere or compete with the production of specific antibody.

Partial characterization of procomplementary activity of porcine serum and its relationship to the fifth component of complement.

The procomplementary factor (PCF) of porcine serum, a component that enhances the hemolytic activity of guinea pig complement, was purified by precipitation with methanol and then by diethylaminoethyl-cellulose chromatography. The PCF substituted for the 5th complement component (C5) in the complement cascade in tests with functionally purified guinea pig complement components. In contrast to human C5, PCF is heat stable at 56 C.

Direct immunoelectron microscopy of transmissible gastroenteritis virus with immunoglobulins G and A and guinea pig complement.

Porcine colostral immunoglobulin (Ig)G and IgA, isolated from transmissible gastroenteritis virus-infected sows, were compared by direct immunoelectron microscopy. It was estimated, using antibodies with a less than a twofold difference in virus-neutralizing activity, that IgG was 500 times more efficient than was IgA for coating transmissible gastroenteritis virions. Guinea pig complement enhanced the antibody coating with IgG, but did not increase virus-neutralizing activity of IgG or IgA.

Methanol precipitation of transmissible gastroenteritis virus.

Methanol precipitation of transmissible gastroenteritis virus was tested at Ph 4.0, 5.0, 6.0, and 7.0 and at methanol concentrations of 15%, 25%, and 30%. Supernatant and precipitate fractions were tested for complement-fixing and agar-diffusion soluble antigens and plaque-forming units, and were examined by electron microscopy. Virus could be obtained free of detectable agar-diffusion antigens and most of the complement-fixing antigens. Most of the virions were without peplomers after methanol treatment but they retained infectivity.

Efficacy of isolated colostral IgA, IgG, and IgM(A) to protect neonatal pigs against the coronavirus of transmissible gastroenteritis.

To ascertain what class of immunoglobulin (Ig; IgA, IgG, or IgM) is most efficacious in protection, a large quantity of colostrum from sows immunized with virulent transmissible gastroenteritis (TGE) virus was fractionated by chromatographic and gel filtration methods. The isolated IgG, IgA, and IgM(A) had specific virus-neutralizing activities of 1:7.6, 1:342, and 1:302 per milligram of protein, respectively. Each Ig was fed to groups of hysterectomy-derived colostrum-deprived neonatal pigs before and after exposure (challenge) with virulent TGE virus. The 7 pigs fed IgG survived the challenge exposure, but 2 of 7 fed IgA and 1 of 7 fed IgM(A) died of TGE. Three of the survivor pigs that had been fed IgG and 2 of the survivor pigs that had been fed IgA had increased serum antibody titers between 8 and 19 days after challenge exposure, but none of the survivor pigs fed IgM(A) had TGE antibody. In contrast, 12 of 14 virus-control pigs died of TGE and the 2 survivors had antibody conversion. The data show that all 3 Ig classes in immune colostrum will protect neonatal pigs against exposure with virulent TGE virus.

Partial characterization of the principal soluble antigens associated with the coronavirus of transmissible gastroenteritis by complement fixation and immunodiffusion.

A microtiter complement fixation (CF) test to detect transmissible gastroenteritis (TGE) viral antigen was developed, using TGE hyperimmune pig serum as an antibody source. Sera from TGE covalescent pigs did not fix complement by this test. Maximal virus and soluble antigen (SA) titers were obtained 36 to 48 h after inoculation of swine testes cells. Cell-associated virus and SA titers were higher than those in the culture fluid, which had to be concentrated 20X before use as antigen in agar immunodiffusion tests (ID). By sucrose density-gradient centrifugation, the SA had a buoyant density of 1.10 g/ml and could be separated from the virus that banded in the 1.19-g/ml region. Virus and SA from three different isolates of TGE had the same buoyant densities. Heating and proteolytic enzyme digestion established the protein nature of the SA. As assayed by CF and ID, there were stability differences between crude and purified preparations of SA. Antibody prepared in rabbits against the SA neutralized the TGE virus.

Chromatographic separation of gram quantities of immunoglobulins from porcine colostrium against transmissible gastroenteritis virus.

Similar immunoglobulin (Ig) classes were obtained from porcine colodtral whey by either column or batch chromatographic procedures; a stepwise buffer elution technique was used. Specific transmissible gastroenteritis virus neutralizing antibody was found in the 4 major fractions eluted comprising of IgG1, IgG2, IgA, and IgM. The IgG1, and IgG2 were essentially homogeneous, and the IgA- AND IgM-rich fractions had to be recycled several times through Sephadex G-200 to obtain pure IgA and IgM that had specific virus neutralizing activities per mg of protein of 342.1 and 302. 4, compared with 7.6 for IgG. By a combination of the batch chromatographic procedures and gel filtration, gram amounts of specific Ig could be fractionated from the same colostrum.