Inhibitory effect and mechanisms of an anthocyanins- and anthocyanidins-rich extract from purple-shoot tea on colorectal carcinoma cell proliferation

One newly bred variety of tea cultivar, purple-shoot tea, was selected to evaluate its antiproliferative effects on colorectal carcinoma cells, as well as normal colon cells. The phytochemicals and identified catechins of purple-shoot tea extract (PTE) were significantly higher than that of ordinary tea, especially the anthocyanins (surpassed by 135-fold) and anthocyanidins (surpassed by 3.5-fold). PTE inhibited the proliferation of COLO 320DM (IC(50) = 64.9 μg/mL) and HT-29 (IC(50) = 55.2 μg/mL) by blocking cell cycle progression during the G(0)/G(1) phase and inducing apoptotic death. Western blotting indicated that PTE induced cell cycle arrest by reducing the expression of cyclin E and cyclin D1 in COLO 320DM and the upregulation of p21 and p27 cyclin-dependent kinase inhibitors in HT-29. Two cells treated with PTE also indicated the cleavage of PARP, activation of caspase 3, and an increased Bax/Bcl-2 ratio. Our results showed that PTE is a potential novel dietary agent for colorectal cancer chemoprevention.     

Effect of routine cardiovascular catheterization on cardiac troponin I concentration in dogs

ObjectiveTo determine changes in cardiac troponin I concentration (cTnI) associated with cardiovascular catheterization in dogs.Animals, materials and methodscTnI was measured after transarterial coil embolization of patent ductus arteriosus (PDA), balloon valvuloplasty (BV), and pacemaker implantation (PACE). Dogs undergoing ovariohysterectomy (OHE) were used as a control, with 15 animals in each group. Blood for the cTnI assay was collected at baseline (T0), at 5 h (T5), 24 h (T24) and 10 days (T240) post-procedure. The effects of age, duration and difficulty of the procedure were evaluated.ResultsThere was no difference in cTnI concentration at T0 for any of the groups. There was a significant increase in cTnI concentration for BV and PACE, but not PDA at T5 and T24. PACE at T24 and T240 also had higher cTnI than control. Dogs with longer procedure times had significantly higher concentration of cTnI. There was no correlation between the difficulty of the procedure or peri-procedure complications and cTnI.ConclusioncTnI increased during some cardiovascular catheterization procedures, but returned to normal values at 24–240 h. Patients undergoing long catheterization procedures have increased risk for myocardial injury, but this was not related to short-term prognosis.     

Development of the activities of pancreatic and caecal enzymes in White Roman goslings

1. This study was to investigate the development of the activities of pancreatic and caecal enzymes in White Roman goslings from hatching to 28 d of age. 2. A total of 80 1-d-old goslings were used. At hatching, 3 and 7 d of age, 16 goslings (8 males and 8 females) were used. At 11, 14, 21 and 28 d of age, 8 goslings, 4 males and 4 females were selected. The activities of amylase, lipase, trypsin and chymotrypsin in each segment of the small intestine (duodenum, jejunum, ileum) and their contents and the activity of cellulase in the caecal contents were measured. 3. The specific activity (SA) of amylase in the duodenal mucosa and contents increased significantly both from 11 to 14 and 14 to 21 d of age and declined after 28 d of age. In the jejunum and ileum, there was a significant peak in the SA of amylase in the mucosa in goslings from 7 to 21 d of age. On average, the SA of lipase of mucosa and content in the small intestine was initially low but gradually increased from 14 to 21 and from 21 to 28 d of age. 4. The SA of trypsin in the contents of the duodenum and the jejunum increased both from 7 to 11 and 11 to 14 d of age and there was high activity in the ileal contents from 14 to 21 d of age. From 3 to 14 d of age, the SA of chymotrypsin in the duodenal mucosa significantly increased and peaked. There was a significant peak at 11 d of age in the SA of chymotrypsin in the duodenal or jejunal contents and in the ileal contents at 7 d. The SA of cellulase in the caecal contents increased linearly with age, reaching a plateau at 28 d of age. 5. The rate of activity development varied from hatching to 28 d of age. However, the mean SA of amylase and lipase of intestinal contents reached a peak at 21 d, trypsin and chymotrypsin at 11 d and cellulase at 28 d of age, respectively. The quantitative changes in SA of cellulase, amylase, lipase, trypsin and chymotrypsin increased by about 4-, 3-, 5-, 2- and 3-fold, respectively, in the intestinal content of goslings. Thus, development of proteases in the intestine of goslings peaked more rapidly than amylase, lipase and cellulase during the first 4 weeks.     

Expression, purification, and characterization of the maltooligosyltrehalose trehalohydrolase from the thermophilic archaeon Sulfolobus solfataricus ATCC 35092

The maltooligosyltrehalose trehalohydrolase (MTHase) mainly cleaves the alpha-1,4-glucosidic linkage next to the alpha-1,1-linked terminal disaccharide of maltooligosyltrehalose to produce trehalose and the maltooligosaccharide with lower molecular mass. In this study, the treZ gene encoding MTHase was PCR-cloned from Sulfolobus solfataricus ATCC 35092 and then expressed in Escherichia coli. A high yield of the active wild-type MTHase, 13300 units/g of wet cells, was obtained in the absence of IPTG induction. Wild-type MTHase was purified sequentially using heat treatment, nucleic acid precipitation, and ion-exchange chromatography. The purified wild-type MTHase showed an apparent optimal pH of 5 and an optimal temperature at 85 degrees C. The enzyme was stable at pH values ranging from 3.5 to 11, and the activity was fully retained after a 2-h incubation at 45-85 degrees C. The k(cat) values of the enzyme for hydrolysis of maltooligosyltrehaloses with degree of polymerization (DP) 4-7 were 193, 1030, 1190, and 1230 s(-1), respectively, whereas the k(cat) values for glucose formation during hydrolysis of DP 4-7 maltooligosaccharides were 5.49, 17.7, 18.2, and 6.01 s(-1), respectively. The K(M) values of the enzyme for hydrolysis of DP 4-7 maltooligosyltrehaloses and those for maltooligosaccharides are similar at the same corresponding DPs. These results suggest that this MTHase could be used to produce trehalose at high temperatures.     

Quantitative analysis of anticancer 3-deoxyanthocyanidins in infected sorghum seedlings

3-Deoxyanthocyanidins are structurally related to the anthocyanin pigments, which are popular as health-promoting phytochemicals. Here, it is demonstrated that the 3-deoxyanthocyanidins are more cytotoxic on human cancer cells than the 3-hydroxylated anthocyanidin analogues. At 200 microM concentration, luteolinidin reduced the viability of HL-60 and HepG2 cells by 90 and 50%, respectively. Sorghum is a major source of 3-deoxyanthocyanidins, which are present as seed pigments and as phytoalexins responding to pathogen attack. On the basis of the collision-induced dissociation spectra of luteolinidin and apigeninidin, an LC-MS/MS method, operating in multiple-reaction monitoring mode, was developed for the specific detection and accurate quantification of these compounds in complex mixtures, which may be difficult to analyze using absorbance measurements. The results demonstrated that inoculated sorghum seedlings could be utilized for convenient and large-scale production of 3-deoxyanthocyanidins. A quantity of almost 270 microg/g (fresh weight) of luteolinidin was produced 72 h after fungal inoculation of 1-week-old seedlings.     

Metastatic mast cell tumour in a dog presenting with spinal pain

The clinical, advanced imaging and surgical features of a case of canine extradural mast cell tumour are presented. This rare location has only been reported once before as a primary lesion, although the present case may be a metastatic lesion. Given the frequent occurrence and treatment of cutaneous mast cell tumours in dogs, the finding of spinal metastasis is an important reminder to consider metastatic neoplasia in cases of previously treated cancers presenting with spinal pain.