Exenatide treatment did not affect bone mineral density despite body weight reduction in patients with type 2 diabetes.

Preclinical studies suggest that incretin-based therapies may be beneficial for the bone; however, clinical data are largely lacking. We assessed whether the differential effects of these therapies on body weight differed with respect to their effect on bone mineral density (BMD) and markers of calcium homeostasis in patients with type 2 diabetes (T2D). Sixty-nine metformin-treated patients with T2D were randomized to exenatide twice daily (n = 36) or insulin glargine once daily (n = 33). Total body BMD, measured by dual-energy X-ray absorptiometry, and serum markers of calcium homeostasis were assessed before and after 44-week treatment. Exenatide or insulin glargine treatment decreased body weight by 6%. Endpoint BMD was similar in both groups after 44-week therapy (LSmean ± s.e.m. between-group difference -0.002 ± 0.007 g/cm(2) ; p = 0.782). Fasting serum alkaline phosphatase, calcium and phosphate remained unaffected. Forty-four-week treatment with exenatide or insulin glargine had no adverse effects on bone density in patients with T2D, despite differential effects on body weight.

Exenatide twice-daily does not affect renal function or albuminuria compared to titrated insulin glargine in patients with type 2 diabetes mellitus: A post-hoc analysis of a 52-week randomised trial.

AIMS: To compare the effects of long-term treatment with the GLP-1RA exenatide twice-daily versus titrated insulin glargine (iGlar) on renal function and albuminuria in type 2 diabetes (T2DM) patients. METHODS: We post-hoc evaluated renal outcome-data of 54 overweight T2DM patients (mean  ±â€¯SD age 60 ±â€¯8 years, HbA1c 7.5 ±â€¯0.9%, eGFR 86 ±â€¯16 mL/min/1.73 m2, median [IQR] urinary albumin-to-creatinine-ratio (UACR) 0.75 [0.44-1.29] mg/mmol) randomised to exenatide 10 µg twice-daily or titrated iGlar on-top-of metformin for 52-weeks. Renal efficacy endpoints were change in creatinine clearance (CrCl) and albuminuria (urinary albumin-excretion [UAE] and UACR) based on 24-h urines, collected at baseline and Week-52. eGFR and exploratory endpoints were collected throughout the intervention-period, and after a 4-week wash-out. RESULTS: HbA1c-reductions were similar with exenatide (mean ±â€¯SEM -0.80 ±â€¯0.10%) and iGlar (-0.79 ±â€¯0.14%; treatment-difference 0.02%; 95% CI -0.31 to 0.42%). Change from baseline to Week-52 in CrCl, UAE or UACR did not statistically differ; only iGlar reduced albuminuria (P < 0.05; within-group). eGFR decreased from baseline to Week-4 with exenatide (-3.9 ±â€¯2.1 mL/min/1.73 m2; P = 0.069) and iGlar (-2.7 ±â€¯1.2 mL/min/1.73 m2; P = 0.034), without treatment-differences in ensuing trajectory. Exenatide versus iGlar reduced bodyweight (-5.4 kg; 2.9-7.9; P < 0.001), but did not affect blood pressure, lipids or plasma uric acid. CONCLUSIONS: Among T2DM patients without overt nephropathy, one-year treatment with exenatide twice-daily does not affect renal function-decline or onset/progression of albuminuria compared to titrated iGlar. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT00097500.

Crystal structure of a quinoenzyme: copper amine oxidase of Escherichia coli at 2 A resolution.

BACKGROUND: Copper amine oxidases are a ubiquitous and novel group of quinoenzymes that catalyze the oxidative deamination of primary amines to the corresponding aldehydes, with concomitant reduction of molecular oxygen to hydrogen peroxide. The enzymes are dimers of identical 70-90 kDa subunits, each of which contains a single copper ion and a covalently bound cofactor formed by the post-translational modification of a tyrosine side chain to 2,4,5-trihydroxyphenylalanine quinone (TPQ). RESULTS: The crystal structure of amine oxidase from Escherichia coli has been determined in both an active and an inactive form. The only structural differences are in the active site, where differences in copper coordination geometry and in the position and interactions of the redox cofactor, TPQ, are observed. Each subunit of the mushroom-shaped dimer comprises four domains: a 440 amino acid C-terminal beta sandwich domain, which contains the active site and provides the dimer interface, and three smaller peripheral alpha/beta domains (D1-D3), each of about 100 amino acids. D2 and D3 show remarkable structural and sequence similarity to each other and are conserved throughout the quinoenzyme family. In contrast, D1 is absent from some amine oxidases. The active sites are well buried from solvent and lie some 35 A apart, connected by a pair of beta hairpin arms. CONCLUSIONS: The crystal structure of E. coli copper amine oxidase reveals a number of unexpected features and provides a basis for investigating the intriguing similarities and differences in catalytic mechanism of members of this enzyme family. In addition to the three conserved histidines that bind the copper, our studies identify a number of other conserved residues close to the active site, including a candidate for the catalytic base and a fourth conserved histidine which is involved in an interesting intersubunit interaction.

Testing a short-term feeding trial to assess compositional and histopathological changes in hearts of rats fed vegetable oils.

Male, female and castrated rats, three wk of age, were fed a low-fat diet for 14 wk followed by high-fat diets (20% by weight) for one wk containing graded levels of erucic acid from 1 to 50%, to evaluate the effect of short-term feeding and interaction of male sex hormones on formation of heart lesions. Some rats within each group were returned to the low-fat diet for one wk after the test period. For comparison, one group of three-wk-old male rats was fed the high fat 50% erucic acid diet for 15 wk. Erucic acid depressed growth rate and food consumption and increased cardiac lipidosis and triglycerides proportional to the erucic acid content of the diet. There were no sex differences, and the effects disappeared once rats were returned to the low-fat diet for one week. There was a significance (P less than 0.05) in the incidence of myocardial necrosis among male rats fed increased levels of erucic acid for one week, but the response was not linear to the increase in dietary erucic acid. Furthermore, the response was much less than in males fed the 50% erucic acid diet continually for 15 weeks. These results suggest that the short-term model is not a suitable substitute for the long-term feeding trial to test the cardiopathogenicity of a vegetable oil. The significantly lower incidence in myocardial lesions in female and castrated male rats compared with male rats suggests involvement of sex hormones.(ABSTRACT TRUNCATED AT 250 WORDS)

Reduction of myocardial necrosis in male albino rats by manipulation of dietary fatty acid levels.

A comprehensive statistical analysis had shown a significant correlation between the incidence of myocardial lesions in male albino rats and the concentration of certain dietary fatty acids. To test this result under controlled conditions, male rats were fed for 16 weeks diets containing 20% by weight soybean oil or a low erucic acid rapeseed (LEAR) oil. Both dietary oils contained substantial amounts of linolenic acid, and both groups developed a high incidence of myocardial necrosis. The addition of dietary saturated fatty acids to the oil in the form of cocoa butter significantly lowered the incidence of heart lesions in both groups. The addition of cocoa butter resulted in increased absorption of saturates and increased growth. Replacement of the cocoa butter by at least an equal amount of synthetic triolein resulted in no significant changes in the cardiopathogenic response compared to the original oils, thus ensuring that the reduction in heart lesions associated with the addition of cocoa butter was not due to dilution of cardiopathogenic compounds in the original vegetable oils. These results support the hypothesis that myocardial lesions in male rats are related to the balance of dietary fatty acids and not to cardiotoxic contaminants in the oils. Changes in the dietary fatty acids did not appear to influence the proportion of the cardiac phospholipids, but their fatty acid composition was markedly influenced. Dietary linolenic acid affected the C22 polyunsaturated fatty acids (PUFA) and dietary saturates increased the level of saturates in cardiac phospholipids. The level of arachidonic acid and total C22 PUFA did not appear to be affected by diet.

Effect of cold stress on rapeseed oil fed rats.

No mortality was observed in 6 week old male Sprague-Dawley rats subjected to cold at 4 C for 3 weeks and fed either a control diet (Chow) or a semisynthetic diet containing 20% by wt rapeseed oil high in erucic acid (23.6%). All rats fed the Chow diet and 17 of 20 rats fed the rapeseed oil-containing diet survived 4 weeks in the same environment. Three rats on the latter diet died of self-mutilation. Marked myocardial lipidosis as well as a large acumulation of 20:1 and 22:1 was observed in the hearts of rats fed the rapeseed oil-containing diet. Five of 20 rats on the Chow diet and 2 of 20 rats on the rapeseed oil-containing diet had focal necrotic areas in the myocardium.

Cardiopathogenicity of soybean oil and tower rapeseed oil triglycerides when fed to male rats.

The triglycerides of soybean oil were purified by molecular distillation and those of Tower rapeseed oil by molecular distillation and adsorption chromatography. The original oils and the purified triglycerides were incorporated in semisynthetic diets at 20% by weight and fed for 16 weeks to weanling male Sprague-Dawley rats to compare the nutritional and pathological effects of the oils and their triglyceride fractions on rats. The study was carried out at two independent laboratories. No significant differences were observed between the results of the two establishments. The incidence of myocardial lesions was significantly higher in rats fed Tower rapeseed oil than in those fed soybean oil. Purification of the triglycerides by molecular distillation and adsorption chromatography appeared to have no major effect on the incidence of myocardial lesions. This supports our previous findings that the cardiopathogenicity appeared to have no major effect on the incidence of myocardial lesions. This supports our previous findings that the cardiopathogenicity of the test oils to rats resides in the triglycerides of these oils.

Pneumonia Associated with Inhaled Plant Material in Swine.

A high incidence of foreign-body pneumonia was found in an institutional swine herd. Foreign material of plant origin was found in 40 of 68 lungs. Most of the foreign matter was of feed origin but four particles of wood were demonstrated. Numerous particles up to 50 µm wide were found in alveoli, and particles as large as 300 µm were found in bronchioles. Lungs of four control animals were used to study the histological reaction to the inhaled foreign bodies. Polymorphonuclear leucocytes, macrophages, and giant cells were the principal cell types associated with the plant particles. Hyperplasia and destruction of respiratory epithelium and lymphoreticular proliferation were also noted.

Evaluation of the serological response of sheep in one flock to Mycobacterium paratuberculosis by crossed immunoelectrophoresis.

Sera from 74 sheep culled from one flock on the basis of performance and response to immunological tests for paratuberculosis or maedi visna were used to evaluate the serological response to a sonicated antigen of Mycobacterium paratuberculosis by crossed immunoelectrophoresis. A total of seven precipitating components was demonstrated. Four components (A,C,V,W) were detected in low frequency only with sera from animals with paratuberculosis while two components (X,Y) were detected in high frequency with sera from animals with or without paratuberculosis. One component (D) was observed in high frequency with sera from animals with paratuberculosis. The magnitude of the serological response to the D component as measured by crossed immunoelectrophoresis correlated well with bacterial load and generally agreed with the quantitative assessment by agar gel immunodiffusion. A development time for crossed immunoelectrophoresis of 24-72 hours after electrophoresis was required to achieve correlation with agar gel immunodiffusion.

Serodiagnosis of ovine paratuberculosis, using lipoarabinomannan in an enzyme-linked immunosorbent assay.

The use of lipoarabinomannan (LAM; obtained from Mycobacterium paratuberculosis) in and ELISA (LAM-ELISA) to test 75 sheep sera from a paratuberculosis-infected flock resulted in an approximate threefold increase in sensitivity (from 23.5% to 70.6%), compared with the use of Annau's polysaccharide in a complement fixation test (P-CFT). Even after manipulation of the LAM-ELISA cut-off value to produce a specificity of 100% to match that of the P-CFT, the sensitivity still was approximately twofold greater than that of the P-CFT. Anti-bovine monoclonal antiglobulin-enzyme conjugates matched commercially available anti-ovine polyclonal antiglobulin-enzyme conjugates with respect to sensitivity and specificity. False-positive results were found to be less frequent after combining 2 serodiagnostic tests, LAM-ELISA and D antigenagar gel immunodiffusion, resulting in an increase in specificity from 88.1% to 95.2%. The repeatability of true seropositive and seronegative results was found to be 89.5% and 91.1%, respectively, for sera obtained less than or equal to 1 month prior to slaughter and 91.7% and 95.5%, respectively, for reanalysis of sera obtained at the time of slaughter.

Some aspects of the composition of avian ascitic fluid.

Ascitic fluid from male Single Comb White Leghorn chickens fed a diet containing 20% by weight rapeseed oil rich (51.6 weight percent) in erucic (22:1n9) acid contained 2.84 +/- .09 g/dl protein and 2.55 +/- .22 micrograms/ml DNA. A characteristic feature of ascitic fluid as compared to liver and plasma was its high polyunsaturates, low saturates, and low saturate to unsaturate ratio. In general, the fatty acid profile of ascitic fluid is more comparable to the fatty acid profile of plasma lipids than to that of liver lipids. These data suggest that there is no direct absorption of fat and transfer to the ascitic fluid.

Growth, heart weight, cardiac lipid, and pathology of chickens fed soybean oil or oil extracted from different rapeseed cultivars.

A total of 384 male Single Comb White Leghorn chicks of the Hyline strain were randomized in 24 Petersime battery units of 16 birds per unit. Four replicate pens were fed either a basal (control) diet containing no added oil or a diet supplemented with 20% by weight of either soybean oil or rapeseed oil (RSO) from cultivars Tower, Candle, Regular, or R-500. Levels of crucic acid (22:1) in the RSO's ranged from .1 to 51.6%. Four birds from each unit were killed at 4, 8, 12, and 16 weeks for cardiac lipid analysis and pathology. Feeding RSO had no consistent effect on body weights, heart weights, or the heart-to-body weight ratio. Only the diet containing R-500 (51.6% 22:1) consistently increased cardiac lipid levels as measured gravimetrically. Maximum deposition of the long-chain monoenes (22:1) and eicosenoic acids (20:1) occurred in the cardiac lipids by 8 weeks, and the levels remained high throughout the 16-week trial. The relative concentration of these long-chain monoenes in the cardiac lipids was positively correlated to the dietary levels of these acids. The incidence of lesions found in heart, liver, and skeletal muscle was significantly higher in chickens fed RSO's high in 22:1 than in chickens fed control or soybean oil, and the incidence significantly correlated to the level of 22:1 in the diet. The incidence of heart and muscle lesions and sinusoidal distention in chickens fed RSO's low in 22:1 was not significantly different from that of chickens fed the control diet or soybean oil, but the incidence of hepatic necrosis was significantly higher for chickens fed RSO's low in 22:1 compared to those fed the control diet or soybean oil.

Pathological changes associated with the feeding of soybean oil or oil extracted from different rapeseed cultivars to single comb white Leghorn cockerels.

A total of 384 Single Comb White Leghorn cockerels of the Hyline strain were fed either a basal (control) diet containing no added oil or a diet supplemented with 20% by weight of soybean oil, rapeseed oil from cultivars Tower, Candle, a mixture of Echo and Arlo high erucic acid rapeseed (HEAR), or R-500. Levels of erucic acid (22:1) in the rapeseed diets varied from .03 to 10.31%. Three birds from each unit were killed at 28, 56, 84, and 112 days and a wide range of tissues were examined histologically. A number of birds in all dietary groups had healed lesions of avian encephalomalacia. Two cockerels fed HEAR oil and 12 fed R-500 developed marked ascites, firm shrunken livers, hydropericardium, and cachectic muscular atrophy. Marked periacinar necrosis was present in birds dying with ascites. A significantly higher number of birds fed the rapeseed oils developed hepatic sinusoidal distention than birds fed the basal or soybean oil diets. Degenerative myocardial changes were seen only in R-500 and HEAR oil-fed birds. This, coupled with hepatic changes, producing shrunken firm livers, led to development of ascites, cachectic muscular atrophy, and periacinar hepatic necrosis. These changes were probably produced by the high erucic acid content of the HEAR and R-500 oils and the resultant grossly imbalanced diet.

Major Sperm Protein Genes from Globodera rostochiensis.

Three genes in the major sperm protein (MSP) gene family from the potato cyst nematode Globodera rostochiensis were cloned and sequenced. In contrast to the absence of introns in Caenorhabditis elegans MSP genes, these genes in G. rostochiensis contained a 57 nucleotide intron, with normal exon-intron boundaries, in the same relative location as the intron in Onchocerca volvulus. The MSP genes of G. rostochiensis had putative CAAT, TATA, and polyadenylation signals. The predicted G. rostochiensis MSP gene product is 126 amino acids long, one residue shorter than the products in the other species. The comparison of MSP amino acid sequences from four diverse nematode species suggests that O. volvulus, Ascaris suum, and C. elegans may be more closely related to each other than they are to G. rostochiensis.

Bovine neoplasms encountered in Canadian slaughterhouses: a summary.

During the past five and one half years veterinarians of the Food Production and Inspection Branch of Agriculture Canada submitted tissues from 1535 cases of suspected bovine neoplasia for diagnosis to the Animal Diseases Research Institute, Nepean. Neoplasia was confirmed in 1370 cases. Lymphosarcoma was diagnosed in 738 (53.9%) of these cases, but it was not possible to accurately assess the prevalence of this condition at slaughter. Other commonly found neoplasms in order of frequency were neurofibroma, squamous cell carcinoma, uterine carcinoma, adrenal tumors, ovarian tumors and primary liver tumors. Many metastatic anaplastic carcinomas involving lymph nodes were not specifically identified as to tissue of origin. Nonneoplastic lesions were found in 165 submissions.

Clinical study of a C31G containing mouthrinse: effect on salivary microorganisms.

In vitro studies have demonstrated the antiplaque properties of C31G, a potent broad spectrum antimicrobial agent consisting of an equimolar mixture of alkyl dimethyl glycine and alkyl dimethyl amine oxide, buffered with citric acid. In this initial clinical study, C31G at concentrations of 0.05%, 0.1%, 0.2% and 0.5%. Listerine, and placebo were tested in a complete crossover design. Twelve subjects were evaluated, with a minimum of 2 days between treatments. Parameters monitored were salivary bacterial counts and saliva glycolysis. The 0.5% and 0.2% C31G mouthrinses significantly reduced total bacterial counts in saliva samples obtained up to and including three hours after rinsing, compared with counts obtained prerinsing or after placebo rinsing. Both 0.5%, and 0.2% C31G significantly inhibited glycolysis of salivary bacteria for up to 6 hours postrinsing, compared with pH values obtained prerinsing. 0.1% and 0.05% C31G exhibited little or no effect in either assay. Listerine showed a significant reduction in bacterial counts for up to 1 hour postrinsing, compared with prerinse counts, but the effect was less sustained. Listerine showed no significant inhibition of glycolysis at any time point. No tooth staining or altered taste sensation was noted with either product.