Behavioral, nutritional, and toxicological responses of cattle to ensiled leafy spurge.
(1/140)
Yearling cattle (n = 25; 416.1 +/- 25.9 kg) were stratified by weight and gender across five groups. Group 1 (OAT) was offered oat/rape haylage (ORH) for ad libitum consumption during two daily feeding periods. Group 2 (SPURGE) was offered leafy spurge/grass haylage (LSGH) for ad libitum consumption during the same feeding periods. Group 3 was offered ORH in an amount equal to the average amount of LSGH consumed by SPURGE at the previous feeding. Group 4 (MIX) was offered LSGH mixed with ORH for ad libitum consumption during the two feeding periods. Group 5 (PAIR) received the equivalent amount of ORH consumed by MIX at the previous feeding. The DMI for OAT, SPURGE, and MIX were similar at the first feeding (P = .52). The SPURGE group consumed very little LSGH thereafter and was removed from the trial. The OAT and MIX groups consumed similar amounts of DM daily on d 1 to 4 when the ration offered to MIX was only 7% LSGH (P = .33). When LSGH made up > or = 21% of the mixture (d 7 to 32), the OAT group consumed more daily DM than did MIX (P < .05). The spurge/oatlage ration offered to MIX was less digestible than the oatlage-only ration offered to PAIR (P < or = .01). Even though blood chemistry did not indicate that LSGH consumption caused organ damage, its intake caused minor alterations (P < or = .05) in serum albumin, calcium, gamma glutamyltransferase, P, K, and urea nitrogen. No gross or microscopic lesions, infectious agents, or significant numbers of parasites were detected in any of the carcasses or tissues examined. The MIX group had diarrhea for much of the trial. In Trial 2, five yearling cattle were adapted to a mixture of 21% LSGH and 79% ORH. Then they were simultaneously offered three mixtures of spurge and oat haylages: 1) spurge ensiled with a microbial inoculant (LSGH); 2) spurge ensiled with the same inoculant and a cellulolytic/hemicellulolytic enzyme (ENZ); and 3) spurge ensiled with the same inoculant and molasses (MOL). The mixture with ENZ was preferred over those with MOL or LSGH (P < .001), but the amounts consumed were low and similar to those for LSGH-ORH in Trial 1 when amounts of ENZ and LSGH in the mixtures were similar. The ENZ mixture may have been more palatable than LSGH and MOL because it had less (P < .05) lactic acid, but intake of ENZ indicates that it had aversive characteristics, like LSGH. Ensiling leafy spurge did little, if anything, to improve its palatability to cattle. (+info)
Isolation, characterization, and functional analysis of a novel cDNA clone encoding a small rubber particle protein from Hevea brasiliensis.
(2/140)
Biochemical evidence reported so far suggests that rubber synthesis takes place on the surface of rubber particles suspended in the latex of Hevea brasiliensis. We have isolated and characterized a cDNA clone that encodes a protein tightly bound on a small rubber particle. We named this protein small rubber particle protein (SRPP). Prior to this study, this protein was known as a latex allergen, and only its partial amino acid sequence was reported. Sequence analysis revealed that this protein is highly homologous to the rubber elongation factor and the Phaseolus vulgaris stress-related protein. Southern and Northern analyses indicate that the protein is encoded by a single gene and highly expressed in latex. An allergenicity test using the recombinant protein confirmed that the cloned cDNA encodes the known 24-kDa latex allergen. Neither ethylene stimulation nor wounding changed the transcript level of the SRPP gene in H. brasiliensis. An in vitro rubber assay showed that the protein plays a positive role in rubber biosynthesis. Therefore, it is likely that SRPP is a part of the rubber biosynthesis machinery, if not the rubber polymerase, along with the rubber elongation factor. (+info)
Seroepidemiologic survey of Orientia tsutsugamushi, Rickettsia typhi, and TT118 spotted fever group rickettsiae in rubber estate workers in Malaysia.
(3/140)
The seroprevalence of Orientia tsutsugamushi, Rickettsia typhi, and TT118 spotted fever group (SFG) rickettsiae in 300 rubber estate workers in Slim River, Malaysia was determined in December 1996 and March 1997. In December, which was the wet season, 23.3%, 3.0%, and 57.3% of the population had antibodies detected against the three rickettsiae, respectively. The highest seropositive rate of 40% was detected for single infection with SFG rickettsiae, followed by a rate of 15.3% for both O. tsutsugamushi and SFG rickettsiae among the rubber estate workers. Subjects less than 21 years old had a lower seroprevalence of SFG rickettsiae compared with the other age groups. Indians had a higher seroprevalence of O. tsutsugamushi compared with other ethnic groups. Rubber tappers had a higher seroprevalence of SFG rickettsiae compared with other occupational groups. During the dry season in March 1997, there was a significant increase in the seroprevalence of R. typhi. The seroconversion rates for IgM against O. tsutsugamushi, R. typhi, and SFG rickettsiae were 5.7%, 12.3%, and 15.1%, respectively, during the four-month period. Significant variations of antibody titers towards the three rickettsiae was noted among subjects who were bled twice. This suggests a significant and continual exposure of rubber estate workers to the three rickettsiae. (+info)
Tumor promoters in commercial indoor-plant cultivars of the Euphorbiaceae.
(4/140)
Certain decorative indoor-plant cultivars are derived from toxic wild plant species. Native members of the Euphorbiaceae (spurge) contain highly irritating and tumor-promoting diterpene esters. Plant breeders and gardeners are constantly searching for less toxic cultivars of the popular Euphorbiaceae indoor plants. In this investigation, 22 commercial cultivars of Euphorbiaceae indoor plants were examined for tumor promoter contents by high-performance liquid chromatography (HPLC). Cultivars of E. milii (E. lomii hybrids), and in particular E. leuconeura, contained ingenol derivatives, whereas cultivars of E. pulcherrima and Codiaeum variegatum were devoid of these compounds. Tumor-promoting activity was assessed by induction of a luciferase reporter gene, which was placed under the control of an Epstein-Barr virus early antigen promoter. The response was closely correlated with ingenol ester content; the latex of the two E. leuconeura cultivars tested gave the strongest response. The HPLC and bioassay methods used in this study provide a basis for the development of nontoxic indoor-plant cultivars and perhaps for consumer-oriented labeling. (+info)
Three-dimensional structures of enzyme-substrate complexes of the hydroxynitrile lyase from Hevea brasiliensis.
(5/140)
The 3D structures of complexes between the hydroxynitrile lyase from Hevea brasiliensis (Hb-HNL) and several substrate and/or inhibitor molecules, including trichloracetaldehyde, hexafluoracetone, acetone, and rhodanide, were determined by X-ray crystallography. The complex with trichloracetaldehyde showed a covalent linkage between the protein and the inhibitor, which had apparently resulted from nucleophilic attack of the catalytic Ser80-Ogamma. All other complexes showed the substrate or inhibitor molecule merely hydrogen bonded to the protein. In addition, the native crystal structure of Hb-HNL was redetermined at cryo-temperature and at room temperature, eliminating previous uncertainties concerning residual electron density within the active site, and leading to the observation of two conserved water molecules. One of them was found to be conserved in all complex structures and appears to have mainly structural significance. The other water molecule is conserved in all structures except for the complex with rhodanide; it is hydrogen bonded to the imidazole of the catalytic His235 and appears to affect the Hb-HNL catalyzed reaction. The observed 3D structural data suggest implications for the enzyme mechanism. It appears that the enzyme-catalyzed cyanohydrin formation is unlikely to proceed via a hemiacetal or hemiketal intermediate covalently attached to the enzyme, despite the observation of such an intermediate for the complex with trichloracetaldehyde. Instead, the data are consistent with a mechanism where the incoming substrate is activated by hydrogen bonding with its carbonyl oxygen to the Ser80 and Thr11 hydroxy groups. A hydrogen cyanide molecule subsequently replaces a water molecule and is deprotonated presumably by the His235 base. Deprotonation is facilitated by the proximity of the positive charge of the Lys236 side chain. (+info)
Attachment forces of ants measured with a centrifuge: better 'wax-runners' have a poorer attachment to a smooth surface.
(6/140)
The symbiotic ant partners of glaucous Macaranga ant-plants show an exceptional capacity to run on the slippery epicuticular wax crystals covering the plant stem without any difficulty. We test the hypothesis that these specialised 'wax-runners' have a general, superior attachment capacity. We compared attachment on a smooth surface for 11 ant species with different wax-running capacities. The maximum force that could be withstood before an ant became detached was quantified using a centrifuge recorded by a high-speed video camera. This technique has the advantage of causing minimum disruption and allows measurements in very small animals. When strong centrifugal forces were applied, the ants showed a conspicuous 'freezing reflex' advantageous to attachment. Attachment forces differed strongly among the ant species investigated. This variation could not be explained by different surface area/weight ratios of smaller and larger ants. Within species, however, detachment force per body weight (F/W) scaled with the predicted value of W(-)(0.33), where W is body weight in newtons. Surprisingly, our results not only disprove the hypothesis that 'wax-runners' generally attach better but also provide evidence for the reverse effect. Superior 'wax-runners' (genera Technomyrmex and Crematogaster) did not cling better to smooth Perspex, but performed significantly worse than closely related congeners that are unable to climb up waxy stems. This suggests an inverse relationship between adaptations to run on wax and to attach to a smooth surface. (+info)
Identification of natural rubber and characterization of rubber biosynthetic activity in fig tree.
(7/140)
Natural rubber was extracted from the fig tree (Ficus carica) cultivated in Korea as part of a survey of rubber producing plants. Fourier transform infrared and (13)C nuclear magnetic resonance analysis of samples prepared by successive extraction with acetone and benzene confirmed that the benzene-soluble residues are natural rubber, cis-1,4-polyisoprene. The rubber content in the latex of fig tree was about 4%, whereas the rubber content in the bark, leaf, and fruit was 0.3%, 0.1%, and 0.1%, respectively. Gel-permeation chromatography revealed that the molecular size of the natural rubber from fig tree is about 190 kD. Similar to rubber tree (Hevea brasiliensis) and guayule (Parthenium argentatum Gray), rubber biosynthesis in fig tree is tightly associated with rubber particles. The rubber transferase in rubber particles exhibited a higher affinity for farnesyl pyrophosphate than for isopentenyl pyrophosphate, with apparent K(m) values of 2.8 and 228 microM, respectively. Examination of latex serum from fig tree by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed major proteins of 25 and 48 kD in size, and several proteins with molecular mass below 20 and above 100 kD. Partial N-terminal amino acid sequencing and immunochemical analyses revealed that the 25- and 48-kD proteins were novel and not related to any other suggested rubber transferases. The effect of EDTA and Mg(2+) ion on in vitro rubber biosynthesis in fig tree and rubber tree suggested that divalent metal ion present in the latex serum is an important factor in determining the different rubber biosynthetic activities in fig tree and rubber tree. (+info)
Enzyme kinetics of hevamine, a chitinase from the rubber tree Hevea brasiliensis.
(8/140)
The enzyme kinetics of hevamine, a chitinase from the rubber tree Hevea brasiliensis, were studied in detail with a new enzyme assay. In this assay, the enzyme reaction products were derivatized by reductive coupling to a chromophore. Products were separated by HPLC and the amount of product was calculated by peak integration. Penta-N-acetylglucosamine (penta-nag) and hexa-N-acetylglucosamine (hexa-nag) were used as substrates. Hexa-nag was more efficiently converted than penta-nag, which is an indication that hevamine has at least six sugar binding sites in the active site. Tetra-N-acetylglucosamine (tetra-nag) and allosamidin were tested as inhibitors. Allosamidin was found to be a competitive inhibitor with a K(i) of 3.1 microM. Under the conditions tested, tetra-nag did not inhibit hevamine. (+info)