Purification and some chemical properties of 30 kDa Ginkgo biloba glycoprotein, which reacts with antiserum against beta 1-->2 xylose-containing N-glycans. (17/6126)

From the seeds of Ginkgo biloba, a glycoprotein, which is a major component that reacts with an antiserum against beta 1-->2 xylose-containing N-glycans, has been purified and characterized. The N-terminal amino acid sequence of the purified glycoprotein was H-K-A-N-X-V-T-V-A-F-V-M-T-Q-H-L-L-F-G-Q-. The molecular mass was estimated to be 17 kDa and 16 kDa by SDS-PAGE under reducing conditions, however, the molecular mass of this glycoprotein in the native state was 30,762 by MALDI-TOF MS, suggesting that this glycoprotein consists of two subunits; one is glycosylated and the other is not. The structure of N-glycan linked to this glycoprotein (designated 30 kDa GBGP) was identified as Man3Fuc1Xyl1GlcNAc2, which is the predominant N-glycan linked to the storage glycoproteins in the same seeds (Kimura, Y et al. (1998) Biosci. Biotechnol. Biochem. 62, 253-261). From the peptic digest of the carboxymethylated glycosylated subunit, one glycopeptide was purified by RP-HPLC and the amino acid sequence was identified as H-K-A-N-N(Man3Fuc1Xyl1Glc-NAc2)-V-T-V-A-F, which corresponded to the N-terminal amino acid sequence.  (+info)

Foliar modifications induced by inhibition of polar transport of auxin. (18/6126)

The effects of auxin polar transport inhibitors, 9-hydroxy-fluorene-9-carboxylic acid (HFCA); 2, 3, 5-triiodobenzoic acid (TIBA) and trans-cinnamic acid (CA) on leaf pattern formation were investigated with shoots formed from cultured leaf explants of tobacco and cultured pedicel explants of Orychophragmus violaceus, and the seedlings of tobacco and Brassica chinensis. Although the effective concentration varies with the inhibitors used, all of the inhibitors induced the formation of trumpet-shaped and/or fused leaves. The frequency of trumpet-shaped leaf formation was related to the concentration of inhibitors in the medium. Histological observation of tobacco seedlings showed that there was only one main vascular bundle and several minor vascular bundles in normal leaves of the control, but there were several vascular bundles of more or less the same size in the trumpet-shaped leaves of treated ones. These results indicated that auxin polar transport played an important role on bilateral symmetry of leaf growth.  (+info)

Effects of feeding beef females supplemental fat during gestation on cold tolerance in newborn calves. (19/6126)

Effects of prepartum fat supplementation of the dam on cold tolerance of calves were determined in two studies. In Exp. 1, 22 F1, crossbred heifers gestating F2 calves received diets containing either 1.7 or 4.7% dietary fat starting at d 230+/-2d of gestation. Safflower seeds (Carthamus tinctorius) containing 37% oil with 79% linoleic acid were the supplemental fat source in isocaloric-isonitrogenous diets. Calves were separated from their dams at birth, fed pooled dairy-cow colostrum, muzzled to prevent sucking, and returned to their dams in a heated (22 degrees C) barn for 3.5 h. At 4 h of age, a jugular catheter was inserted. At 5 h of age, calves were placed in a 0 degrees C room for 140 min and rectal temperatures and blood samples were obtained at 10- and 20-min intervals. Blood was assayed for glucose, cortisol, and cholesterol. In Exp. 2, 18 multiparous, crossbred beef cows bred to Murray Grey sires were randomly assigned to receive diets containing either 1.7 or 3.1% dietary fat starting at 235+/-2 d gestation. Safflower seeds were used as the supplemental fat source in isocaloric-isonitrogenous diets. At d 260 of gestation, premature parturition was induced in one-half of the cows from each diet group by feeding Ponderosa pine (Pinus ponderosa) needles. Experimental protocols were the same as in Exp. 1, except that cold exposure was at 9 degrees C for 200 min. Rectal temperatures were affected in Exp. 1 by time and diet x time (both P < .01) and diet x calf sex (P < .05) and in Exp. 2 by calf age (P < .05), time, and calf age x time (both P < .01). Plasma cortisol concentrations were affected by time (P < .01) and calf sex x time (P < .05) in Exp. 1 and by time ( P < .01) in Exp. 2. Cholesterol concentrations in Exp. 1 were affected by diet x time (P < .05) and in Exp. 2 by time (P < .05). Plasma glucose concentrations were affected in Exp. 1 by diet (P < .05) and in Exp. 2 by calf age, time, and calf age x time (all P < .01). We conclude from Exp. 1 that feeding heifers supplemental fat during late gestation increased glucose concentrations in the newborn calf, resulting in favorable responses in body temperature in the cold-stressed newborns. This increase in substrate availability suggests a potential positive effect on heat generation in newborns during sustained periods of cold stress. In Exp. 2, premature calves had compromised cold tolerance possibly due to impaired shivering or brown adipose tissue thermogenesis.  (+info)

Role of pfkA and general carbohydrate catabolism in seed colonization by Enterobacter cloacae. (20/6126)

Enterobacter cloacae A-11 is a transposon mutant of strain 501R3 that was deficient in cucumber spermosphere colonization and in the utilization of certain carbohydrates (D. P. Roberts, C. J. Sheets, and J. S. Hartung, Can. J. Microbiol. 38:1128-1134, 1992). In vitro growth of strain A-11 was reduced or deficient on most carbohydrates that supported growth of strain 501R3 but was unaffected on fructose, glycerol, and all amino acids and organic acids tested. Colonization by strain A-11 was significantly reduced (P +info)

A mutational analysis of leaf morphogenesis in Arabidopsis thaliana. (21/6126)

As a contribution to a better understanding of the developmental processes that are specific to plants, we have begun a genetic analysis of leaf ontogeny in the model system Arabidopsis thaliana by performing a large-scale screening for mutants with abnormal leaves. After screening 46,159 M2 individuals, arising from 5770 M1 parental seeds exposed to EMS, we isolated 1926 M2 putative leaf mutants, 853 of which yielded viable M3 inbred progeny. Mutant phenotypes were transmitted with complete penetrance and small variations in expressivity in 255 lines. Most of them were inherited as recessive monogenic traits, belonging to 94 complementation groups, which suggests that we did not reach saturation of the genome. We discuss the nature of the processes presumably perturbed in the phenotypic classes defined among our mutants.  (+info)

Characterization of molecular mobility in seed tissues: an electron paramagnetic resonance spin probe study. (22/6126)

The relationship between molecular mobility (tauR) of the polar spin probe 3-carboxy-proxyl and water content and temperature was established in pea axes by electron paramagnetic resonance (EPR) and saturation transfer EPR. At room temperature, tauR increased during drying from 10(-11) s at 2.0 g water/g dry weight to 10(-4) s in the dry state. At water contents below 0.07 g water/g dry weight, tauR remained constant upon further drying. At the glass transition temperature, tauR was constant at approximately 10(-4) s for all water contents studied. Above Tg, isomobility lines were found that were approximately parallel to the Tg curve. The temperature dependence of tauR at all water contents studied followed Arrhenius behavior, with a break at Tg. Above Tg the activation energy for rotational motion was approximately 25 kJ/mol compared to 10 kJ/mol below Tg. The temperature dependence of tauR could also be described by the WLF equation, using constants deviating considerably from the universal constants. The temperature effect on tauR above Tg was much smaller in pea axes, as found previously for sugar and polymer glasses. Thus, although glasses are present in seeds, the melting of the glass by raising the temperature will cause only a moderate increase in molecular mobility in the cytoplasm as compared to a huge increase in amorphous sugars.  (+info)

Epidemic of self-poisoning with seeds of the yellow oleander tree (Thevetia peruviana) in northern Sri Lanka. (23/6126)

Deliberate self-harm is an important problem in the developing world. Ingestion of yellow oleander seeds (Thevetia peruviana) has recently become a popular method of self-harm in northern Sri Lanka -- there are now thousands of cases each year. These seeds contain cardiac glycosides that cause vomiting, dizziness, and cardiac dysrhythmias such as conduction block affecting the sinus and AV nodes. This paper reports a study of the condition's mortality and morbidity conducted in 1995 in Anuradhapura General Hospital, a secondary referral centre serving 750 000 people in Sri Lanka's north central province. 415 cases were admitted to the hospital during 11 months; 61% were women and 46% were less than 21 years old. A prospective study of 79 patients showed that 6% died soon after admission. 43% presented with marked cardiac dysrhythmias which necessitated ther transfer to the coronary care unit in Colombo for prophylactic temporary cardiac pacing. The reasons for the acts of self-harm were often relatively trivial, particularly in children; most denied that they wished to die. Unfortunately, the case fatality rate for oleander poisoning in Sri Lanka is at least 10%. This epidemic is not only causing many unnecessary deaths, it is also putting immense stress on the already stretched Sri Lankan health services. There is an urgent need for an intervention which could be used in rural hospitals, thus preventing the hazardous and expensive emergency transfer of patients to the capital.  (+info)

Insecticidal activity of an alpha-amylase inhibitor-like protein resembling a putative precursor of alpha-amylase inhibitor in the common bean, Phaseolus vulgaris L. (24/6126)

alpha-Amylase inhibitor (alphaAI) in the common bean, Phaseolus vulgaris L., protects seeds from insect pests such as the cowpea weevil (Callosobruchus maculatus) and the azuki bean weevil (C. chinensis). Cultivars which lack alphaAI still show resistance to both bruchids. These cultivars have a glycoprotein that reacts with anti-alphaAI-1 antibodies. The glycoprotein with a molecular mass of 29 kDa (Gp29) was purified and the encoding gene was isolated. The primary structure of Gp29 is the same as alpha-amylase inhibitor-like protein (AIL) from which the encoding gene has already been isolated. AIL resembles a putative precursor of alphaAI, even though it does not form the active inhibitor. However, AIL has some inhibitory effect on the growth of C. maculatus but not C. chinensis. The presence of AIL alone is insufficient to explain the bruchid resistance of common bean cultivars lacking alpha-AI. Common bean seeds appear to contain several factors responsible for the bruchid resistance.  (+info)