Dimeric inhibitors of human salivary alpha-amylase from emmer (Triticum dicoccon Schrank) seeds

The proteins belonging to the cereal trypsin/alpha-amylase inhibitor family are abundant water/salt-soluble flour proteins active against alpha-amylases from several seed parasites and pests and inactive against endogenous alpha-amylases. Three alpha-amylase inhibitor families have been described in cereals that vary in size and are differently expressed among Triticeae seeds. The present work investigates the presence of human salivary alpha-amylase inhibitors in emmer (Triticum dicoccon Schrank) flour. The isolation was obtained by a series of chromatography steps, and the purification progress was monitored through the inhibition of human salivary alpha-amylase activity. The purified fraction was subjected to protein sequencing by tandem mass spectrometry (MSMS) of the tryptic digests obtained after the sample separation on 2-DE. MSMS data indicated that the emmer alpha-amylase inhibitory fraction was composed of two newly identified proteins [emmer dimeric inhibitor 1 (EDI-1) and emmer dimeric inhibitor 2 (EDI-2)] sharing very high identity levels with related proteins from Triticum aestivum.     

Bioactive Compounds and Antioxidant Capacity of Strawberry Jams

Strawberries represent the main source of ellagic acid derivatives in the Brazilian diet. They are also good sources of flavonoids, mainly anthocyanins, and phenolic acids, to which many beneficial effects have been attributed. However, as the fruit is not available all the year, the objective of this work was to determine whether the jams could also represent a good source of bioactive compounds. In the current study, five different commercially available strawberry jams were characterized in relation to flavonoids, total phenolics, free and total ellagic acid contents, and antioxidant capacity. Anthocyanins were detected only in two jams at very low content. Kaempferol glycosides were the main flavonoids present (from 0.38 to 1.05 mg/100 g fresh weight, FW), while quercetin glycosides were present in the range 0.14–1.20 mg/100 g FW. Free and total ellagic acid content ranged from 0.4 to 2.9 mg/100 g FW, and from 17.0 to 29.5 mg/100 g FW, respectively. Total phenolics varied from 58 to 136 mg/100 g FW, and the antioxidant capacity from 0.55 to 0.76 μmol BHT (Butylhydroxytoluene) equivalents/g FW. Overall, results indicated that jams can also represent a good source of antioxidant compounds, although compared to the fruit important losses seem to occur.     

Use of s-100 and chromogranin a antibodies as immunohistochemical markers on detection of malignancy in aortic body tumors in dog

To define the characteristics of malignancy we performed routine histology and an immunohistochemical study on seventeen aortic body tumors in dogs. We essayed tumors using a panel of immunohistochemical markers: neuron specific enolase (NSE), chromogranin A (CrA) and S-100. Among 17 cases, the neoplastic cells were positive for NSE (17 cases, 100%), S-100 (9 cases, 53%), and CrA (8 cases, 47%), respectively. The sustentacular cells density and chief cell staining intensity were both inversely related to tumor grade. The most relevant data was consistent with a negative staining of S-100 correlated with absence or decreased number of sustentacular cells in tumors grade III. This report indicates that the immunohistochemical panel has utility for the diagnosis of chemodectoma and the negative staining to CrA and S-100 markers in tumors grade III expresses an indication of malignant behaviour of the tumor.     

From field to health: a simple way to increase the nutraceutical content of grape as shown by NO-dependent vascular relaxation

Polyphenolic grapevine components involved in plant resistance against pathogens possess various pharmacological properties that include nitric oxide (NO)-dependent vasodilation and anti-inflammatory and free radical scavenging activities, which may explain the protective effect of moderate red wine consumption against cardiovascular disease. The aim of this work was (a) to verify the possibility that preharvest treatments of grapevine with a plant activator, benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), could lead to an enriched nutraceutical potential of wine and (b) to characterize the profile of metabolites responsible for pharmacological activity. Plant spraying at the end of veraison, with a water suspension of BTH (0.3 mM), led to increased whole anthocyanin content as confirmed by HPLC comparative analysis. Extracts from berry skins of BTH-treated grapevines caused NO-dependent vasorelaxation, with a concentration-response curve that was significantly shifted to the left of the control non-BTH-treated curve. Moreover, 1:1000 dilutions of berry extracts from BTH-treated plants significantly increased basal production of guanosine 3',5'-cyclic monophosphate (cGMP) in human vascular endothelial cells when compared to the corresponding extracts of untreated plants. These results show that BTH treatment increases anthocyanin content of grape extracts, as well as their ability to induce NO-mediated vasoprotection. No increase of anthocyanin content was observed in the wine extracts from BTH-treated vines. It is concluded that BTH treatment could be exploited to increase the nutraceutical potential of grapes.     

New constituents of sweet Capsicum annuum L. fruits and evaluation of their biological activity

Four new acyclic diterpene glycosides named capsianosides (1-4), together with 12 known compounds, were isolated from the fresh sweet pepper fruits of Capsicum annuum L., a plant used as a vegetable food, spice, and external medicine. The chemical structures of new natural compounds, as well as their absolute configurations, were established by means of spectroscopic data including infrared, high-resolution mass spectrometry, and one- and two-dimensional nuclear magnetic resonance and by chemical derivatization. The known capsidiol (11) showed bacteriostatic properties in vitro against Helicobacter pylori with a minimum inhibitory concentration (MIC) of 200 microg/mL when compared with the commercial drug metronidazole (MIC, 250 microg/mL). Some purified components were also tested for their antioxidant activities.     

Fibrosarcoma with typical features of postinjection sarcoma at site of microchip implant in a dog: histologic and immunohistochemical study

A 9-year-old, male French Bulldog was examined for a subcutaneous mass located at the site of a microchip implant. Cytologic examination of the mass was suggestive of a malignant mesenchymal neoplasm. Histologically, the mass was confirmed as a high-grade infiltrative fibrosarcoma, with multifocal necrosis and peripheral lymphoid aggregates. By immunohistochemistry, the sample was investigated for vimentin, smooth-muscle actin (SMA), CD3, CD79alpha, and CD18. All the neoplastic cells were positive for vimentin. Scattered cells at the periphery of the lesion were also positive for SMA, highlighting a myofibroblastic phenotype. The lymphoid cells were positive for CD18 and CD3. No aluminum deposits were detected by the aurintricarboxylic acid method. A diagnosis of fibrosarcoma morphologically similar to feline postinjection sarcomas was made. Fibrosarcomas at the site of injections have been reported in dogs and ferrets. Furthermore, neoplastic growth at the site of microchip implant in dog and laboratory rodents has been described.     

Activated platelet-derived growth factor β receptor and Ras-mitogen-activated protein kinase pathway in natural bovine urinary bladder carcinomas

Bovine papillomavirus types 1 or 2 (BPV-1/2) are involved in the aetiopathogenesis of bovine urinary bladder cancer. BPV-1/2 E5 activates the platelet-derived growth factor β receptor (PDGFβR). The aim of this study was to analyse the Ras/mitogen-activated protein kinase (MAPK) pathway in relation to activation of PDGFβR in natural bovine urinary bladder carcinomas. Co-immunoprecipitation and Western blot analysis demonstrated that recruitment of growth factor receptor bound protein 2 (GRB-2) and Sos-1 to the activated PDGFβR was increased in carcinomas compared to normal tissues. Higher grade bovine urinary bladder carcinomas were associated with activation of Ras, but not with activation of downstream mitogen-activated protein kinase/extracellular signal-regulated kinase (Mek 1/2) or extracellular signal-regulated kinase (Erk 1/2).     

Evaluation of the risk of transmission of Trichinella in pork production systems in Argentina

Recently, there has been interest in programs that certify pork production practices that minimize the risk of exposure of pigs to Trichinella spiralis. Certification might be useful for reducing the risk of human trichinellosis from pork in Argentina, but more information is needed on pig production practices and sources of Trichinella infection in Argentinian pigs. In this study, 21 pig farms were assessed for Trichinella infection including some farms using total and partial confinement management, and others with pigs raised exclusively outdoors. A total of 3224 muscle samples were collected from pigs raised on these farms and tested to determine the presence of T. spiralis larvae by artificial digestion. Serum samples from the same 3224 pigs were tested for antibodies to T. spiralis by ELISA. For each farm, a questionnaire was completed summarizing information about management factors and this information was used to assess risk factors for exposure of T. spiralis. Based on the results, pigs raised outdoors were more likely to be infected than pigs raised in total or partial confinement (p ≤ 0.05). Pigs fed waste products containing meat were 12.5 times more likely to be infected than pigs not fed waste containing meat (p < 0.01). The role played by rats in transmission of Trichinella is unclear; however, on farms with evidence of wild animals and access of pigs to wildlife carcasses, the prevalence of Trichinella infection was significantly higher. All pigs raised under good hygienic and sanitary conditions were negative for Trichinella infection by both artificial digestion and ELISA.     

How Do Horses Appraise Humans' Actions? A Brief Note over a Practical Way to Assess Stimulus Perception

Heart rate (HR) is considered to be an effective tool for assessing animals' emotional response to a stimulus. We investigated changes in HR during a series of handling procedures (grooming test) in horses that had different experiences of human interaction. We used four groups of horses: grazing horses (group A), school horses (group B), six ponies, traditionally trained (group C), and six ponies, trained with modulated stimulus intensity (group D). An HR monitor was applied to each horse. The operator began the grooming test, standardized at the following points: (1) trainer's entry into the enclosure/paddock, (2) halter fastened, (3) start of grooming, (4) end of grooming, (5) saddling, and (6) inserting the bit. Group A repeated the grooming test twice: group A1 with a known operator and group A2 with an unknown operator. The data were compared by using analysis of variance (ANOVA) tests. The results showed a significant increase in HR (P ≤ .05) at point 2 compared with baseline in all groups except group D. Other significant differences were found at all points between groups A1 and A2, groups A2 and B, groups A1 and C, groups B and C, and at points 5 and 6 between groups A2 and C, as well as between groups C and D. Even when a scientific assessment of the increase in the HR is of primary importance, the use of this physiologic parameter may be helpful in assessing the horse's perceptions of stimuli presented by people during handling. Our results show that it is not the action “per se” that is important, but the manner in which horses perceive and appraise such actions in relation to the environment and their subject's experiences.     

Prevalence and concentration of Verotoxigenic Escherichia coli O157:H7 in adult sheep at slaughter from Italy

A 1-year study on the animal-level prevalence and concentration of Escherichia coli O157 in adult sheep at slaughter was performed, to collect qualitative and quantitative information on the diffusion of the pathogen in adult sheep from Italy. A total 533 samples were collected, with a similar distribution in the four seasons. For prevalence estimates, a simple random sampling technique was used. An immuno-magnetic separation technique was used for sample screening, with enumeration of the pathogen in positive samples, along with molecular and serological identification of isolates. An overall prevalence of 7.1% (38/ 533, 95% CI 4.9-9.3%) was observed for fully virulent E. coli O157. A wide interval of VTEC O157 per gram was observed (< 100 to 6 x 10(5) CFU g(-1)), with 28.9% (11/38) of positive samples > or = 1 x 10(3) CFU g(-1), set as the threshold for those animals defined 'active shedders' for the purpose of the study. Eight per cent (3/38) of animals shed > 1 x 10(4) g(-1) VTEC O157, which represents > 96% of the total VTEC O157 bacteria cultured from all animals tested. The prevalence estimate of active shedders was therefore 2.1% (95% CI 0.9-3.3%). Most (34/38, 89.5%) of the positive animals were found in summer (July-September). Prevalence and concentrations of virulent VTEC O157 obtained in this study contribute to the demonstration that adult sheep represent a relevant source of environmental contamination from virulent VTEC O157, as well as a source of VTEC O157 contamination for food of ovine origin (meat and dairy products), especially during warm months.