Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Effect of porcine somatotropin on number of granulosa cell luteinizing hormone/human chorionic gonadotropin receptors, oocyte viability, and concentrations of steroids and insulin-like growth factors I and II in follicular fluid of lean and obese gilts.

Prepubertal gilts of obese (n = 24) or lean (n = 24) genetic lines were injected (s.c.) daily with 0, 2, or 4 mg of porcine somatotropin (pST) for 6 wk starting at 160 d of age to determine whether pST affects follicular function. Blood and ovaries were collected at slaughter 24 h after the last injection. Surface follicles greater than or equal to 1.0 mm in diameter were counted, and pools of follicular fluid (FFL) and granulosa cells were collected from 1.0- to 3.9-mm (small) and 4.0- to 6.9-mm (medium) follicles. Oocytes were collected from small and medium follicles and evaluated for maturational stage and viability. Porcine somatotropin increased (P less than .08) the numbers of small but not the numbers of medium follicles per gilt (P greater than .10). Oocyte maturation and viability were not affected by pST or genetic line. Porcine somatotropin increased (P less than .05) concentrations of insulin-like growth factor I (IGF-I) in serum and FFL of both obese and lean gilts; IGF-I was lower (P less than .01) in lean gilts. Treatment with pST decreased (P less than .05) IGF-II in FFL of lean but not in that of obese gilts. Dose of pST and line had no effect on concentrations of progesterone in FFL of small or medium follicles or on concentrations of estradiol in FFL of small follicles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of ventilation and air temperature on pollution levels in underground car parks

The relationship between the levels of CO and airborne Pb was evaluated against conditions of traffic flow, ambient air temperature and the ventilation rates in 3 underground car parks. During the study period significant variations were noted in the levels of the pollutants in each of the car parks which frequently exceeded the proposed Malaysian Air Quality Standard of 0.7 μg m^?3Pb and 9 μg L^?1 (8 hr average) CO for indoor environs. High correlation coefficient values were obtained between the levels of CO and traffic flow and Pb and traffic flow. High traffic flow also increased the mean air temperature inside the car parks. Reductions in the ventilation rates in the car parks caused an almost two-fold increase in the level of CO and vice versa. A simple mathematical model depicting the CO concentrations in relation to other parameters was formulated.     

Administration of porcine somatotropin by sustained-release implant: growth and endocrine responses in genetically lean and obese barrows and gilts.

Previous studies have documented the effectiveness of porcine somatotropin (pST) administered by daily injection in promoting lean tissue growth in lean and obese pigs and the influence of sex and genotype. The present study examined the accretive responses in pigs of different lines and sexes to a slow release formulation of pST (pST-SR). Implants that deliver 2.0 mg of pST/d were implanted in genetically lean and obese barrows and gilts at 65 +/- .7 kg BW (mean +/- SE). Pigs received no, one, or two implants (i.e., doses of 0, 2.0, and 4.0 mg of pST/d). Pigs (four per line x sex x dose) were housed individually and continuously supplied with fresh water and a 19% CP diet containing 1.08% lysine. Pigs were slaughtered on d 0 (four per line x sex) and at the end of the trial (approximately 42 d after implantation) for estimation of initial composition and calculation of accretion rates. Blood samples were collected at d 0, 7, 14, 28, and 42 to measure endocrine and metabolite responses to pST-SR. Sustained-release pST elevated (P < .05) circulating pST throughout the trial with peak concentrations at d 7. On d 7, serum pST concentrations in the pigs given 2.0 mg of pST-SR per day were 16-fold greater than those in control pigs, and in pigs given 4.0 mg of pST-SR per day pST concentrations were 33-fold greater than in controls. Elevated serum pST resulted in increased (P < .05) serum concentrations of insulin-like growth factor (IGF)-I, IGF-II, insulin, and glucose and in reduced (P < .05) concentrations of urea nitrogen and IGF binding protein (IGFBP)-2. Gain was not influenced by pST-SR dose; however, feed consumption was reduced (P < .05) and efficiency of gain was increased (P < .05). Accretion of all body components except cold carcass weight, cecum, and untrimmed Boston butt and ham were changed (P < .05) with pST-SR administration. Heart and stomach were the only components of the carcass and offal whose accretion was not affected by line or sex. Increases in accretion of carcass components (< 75%) induced by sustained-release pST were considerably less than those measured in the organs (liver, 157%; lungs, 748%). The pST-SR treatment resulted in elevated serum concentrations of pST and its mediators and improved efficiency and composition of gain.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of Dietary Supplementation of Barodon on Growth Performance,Innate Immunity and Disease Resistance of Juvenile Olive Flounder,Paralichthys olivaceus,Against Streptococcus iniae

This study was conducted to evaluate the effects of dietary supplementation of Barodon, an anionic alkali mineral complex, on growth, feed utilization, humoral innate immunity and disease resistance of olive flounder. A basal experimental diet was used as a control and supplemented with 0.1, 0.2, 0.3, 0.4, or 0.5% Barodon. Triplicate groups of fish (26.4 ± 0.2 g) were fed one of the diets to apparent satiation twice daily for 10 wk. The growth performance was enhanced (P < 0.05) linearly and quadratically in fish fed diets containing Barodon compared with that in fish fed the control. Feed utilization was significantly improved by Barodon supplementation. Serum lysozyme and antiprotease activities were increased quadratically in Barodon fed groups. Also, significantly higher superoxide dismutase activity was found in Barodon‐fed fish. Dietary supplementation of 0.1–0.3% Barodon resulted in significant enhancement of fish disease resistance against Streptococcus iniae. The findings in this study indicate that dietary supplementation of Barodon can enhance growth, feed utilization, innate immunity, and disease resistance of olive flounder and that the optimum level seems to be 0.1% in diets.     

Is N‐feedback involved in the regulation of nitrogenase activity in Medicago truncatula?

To determine whether senescing leaves provoke an active nitrogen (N) remobilization that results in the reduction of nitrogenase activity, 60% of Medicago truncatula lower leaves were either darkened or individually excised for two weeks. Although a considerable amount of N was remobilized, N₂ fixation activity was found to be increased to maintain the N source/sink balance, indicating an absence of the negative N‐feedback regulation of nitrogenase activity in the senescing M. truncatula.     

Surgical treatment of mammary carcinomas in dogs with or without postoperative chemotherapy

This retrospective study identified prognostic factors associated with survival; and compared survival data in 94 canine mammary carcinoma (MCA) dogs treated with surgery (n = 58), or surgery and adjunct chemotherapy (n = 36), and a subset of dogs with poor prognostic factors. On multivariate analysis independent predictors of median survival time (MST) were clinical stage, lymphatic invasion (LI; present 179 days; none 1098 days), ulceration (present 118 days; none 443 days) and surgical margins (incomplete 70 days; complete 872 days). Complete surgical margins were associated with MST in dogs with stages 1–3 MCA (incomplete 68 days; complete 1098 days) and dogs with LI (incomplete 70 days; complete 347 days). There was no statistically significant improvement in MST in dogs with advanced disease (stage 4 or LI) treated with adjunctive chemotherapy (chemotherapy 228 days; none 194 days); although five dogs with complete surgical margins that received mitoxantrone and carboplatin had a mean survival of 1139 days.