Leaf and stem explants of passionfruit (Passiflora eadulis fv flavicarpa) were co-cultivated with a disarmed strain of Agrobacterium tunefaciens harbouring the co-integrate vector pMON200. Four plants of passionfruit were regenerated from leaf explants on agar-solidified Murashige and Skoog (1962) based medium containing 4.43 M 6-benzyl-aminopurine and supplemented with 86 M kanamycin sulphate. The four plants were rooted by transfer to MS based medium with 14.7 M 3-indolebutyric acid and 2.68 M -naphthyleneacetic acid for 7 d, followed by MS based medium lacking growth regulators. Both media used for rooting contained 172 M kanamycin sulphate. Rooted plants were potted and grown to maturity. Three of the plants synthesised nopaline and expressed neomycin phosphotransferase activity; DNA dot blot and polymerase chain reaction analyses confirmed the presence of the neomycin phosphotransferase gene in three plants.Abbreviations BAP
6-benzylaminopurine
- CTAB
hexadecy-Itrimethylammonium bromide
- EDTA
ethylenediaminetetraacetic acid
- IBA
3-indolebutyric acid
- MS
Murashige and Skoog (1962)
- NAA
-naphthyleneacetic acid
- NPTII
neomycin phosphotransferase II
-
nptII
neomycin phosphotransferase II gene
- PCR
polymerase chain reaction
- SDS
sodium dodecyl sulphate
- Tris
tris(hydroxymethyl)aminomethane 相似文献
A five site potential model combining Lennard–Jones plus Coulomb potential functions has been developed for chloroform molecule.
The partial charges needed for Coulombic interactions were derived using the chelpg procedure implemented in the gaussian
92 program. These calculations were performed at the MP2 level with MC-311G* basis set for Cl and 6-311G** for C and H atoms.
The parameters for the Lennard–Jones potentials were optimized to reproduce experimental values for the density and enthalpy
of vaporization of the pure liquid at 298 K and 1 atm. The statistical mechanics calculations were performed with the Monte
Carlo method in the isothermic and isobaric (NpT) ensemble. Besides the values obtained for density, ρ, and molar enthalpy
of vaporization at constant pressure, Δ HV, for liquid chloroform, results for molar volume, Vm, molar heat capacity, Cp, isobaric thermal expansivity, αp, and isothermal compressibility, κT, for this pure liquid are also in very good agreement with experimental observations. Size effects on the values of thermodynamic
properties were investigated. The potential model was also tested by computing the free energy for solvating one chloroform
molecule into its own liquid at 298 K using a statistical perturbation approach. The result obtained compares well with the
experimental value. Site–site pair correlation functions were calculated and are in good accordance with theoretical results
available in the literature. Dipole–dipole correlation functions for the present five site model were also calculated at different
carbon–carbon distances. These correlations were compared to those obtained using the four site model reported in the literature.
An investigation of the solvent dependence of the relative free energy for cis/trans conversion of a hypothetical solute in
TIP4P water and chloroform was accomplished. The results show strong interaction of water and chloroform molecules with the
gauche conformer. The value obtained for the free energy barrier for cis/trans rotation in TIP4P water is higher than that
for chloroform. This result is in agreement with the continuous theory for solvation as the conformer with higher dipole moment
is more favoured by the solvent with higher dieletric constant. The results also show an increase in entropy as the solute
goes from the cis to the trans geometry and this result is more appreciable in the aqueous solution.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
In vitro propagation of neem (Azadirachta indica A. Juss.) may offer an efficient alternative to seed propagation of this species. For optimization of in vitro propagation, different basal salt formulations, growth regulators, and culture container sealants (polytetrafluoroethylene
hydrophobic membranes [PTFE]) were evaluated. Nodal segments cultured on Murashige and Skoog (MS) medium showed the highest
shoot formation per explant (1.67). Explants cultured in flasks containing MS medium with 0.5 mg L−1 benzyladenine, 0.5 mg L−1 kinetin, and 0.05 mg L−1 naphthaleneacetic acid, and sealed with two PTFE membranes, produced the highest number of shoots (4.04). In contrast, explants
cultured in flasks without membranes showed leaf chlorosis and senescence. For plant recovery, regenerants were acclimatized
in a substrate of coconut fiber and eucalyptus bark (1:1) and showed 80% survival. Our results indicated that the use of flasks
with vents was beneficial for in vitro propagation of this important plant. 相似文献
Lecythis pisonis Cambess, popularly known as sapucaia, has great economic and socio-environmental potential. The objective of this study was to evaluate the establishment and in vitro morphogenesis of L. pisonis under the effect of disinfecting agents, plant growth regulators, and thermal stress. The study was divided into three experiments: (i) development of the disinfection protocol by testing different concentrations and times of exposure to sodium hypochlorite (NaOCl) and different concentrations and methods of amoxicillin application, (ii) in vitro budding induction by testing different concentrations of 6-benzylaminopurine (BAP) or kinetin (KIN) supplemented to Woody Plant Medium (WPM) and Murashige and Skoog (MS) culture media, and (iii) in vitro formation from plantlets by analyzing different concentrations of indole-3-butyric acid (IBA) with different exposure times to a thermal stress of 40°C. The disinfection of stem segments was effective using 3% NaOCl and 3.0 g L−1 amoxicillin solution. MS culture medium supplemented with 0.25 mg L−1 BAP induced more shoots in vitro. One milligram per liter IBA promoted greater rooting in vitro, and it is not necessary for thermal stress tolerance.
Plant and Soil - Root-released carboxylates enhance the availability of manganese (Mn), which enters roots through transporters with low substrate specificity. Leaf Mn concentration ([Mn]) has been... 相似文献
Plant Cell, Tissue and Organ Culture (PCTOC) - Somaclonal variation during in vitro culture is often an undesirable phenomenon but may also be a source of genetic variation useful for breeders. The... 相似文献
In his Commentary (Manger PR. 2009. Subglacial cetaceans and other mathematical mysteries: a Commentary on “A quantitative test of the thermogenesis hypothesis of cetacean brain evolution, using phylogenetic comparative methods” by C. Maximino. Mar Fresh Behav Physiol. 42: 359–362) on my paper (Maximino C. 2009. A quantitative test of the thermogenesis hypothesis of cetacean brain evolution, using phylogenetic comparative methods. Mar Freshwater Behav Physiol. 42:1–17), Dr Paul Manger noted four errors in the quantitative analysis of the relationship between cetacean encephalization quotients (EQs) and water temperatures, which I suggested was a test of his thermogenesis hypothesis (Manger PR. 2006Manger, PR. 2006. An examination of cetacean brain structure with a novel hypothesis correlating thermogenesis to the evolution of a big brain. Biol Rev Camb Philos Soc, 81: 293–338. [Google Scholar]. An examination of cetacean brain structure with a novel hypothesis correlating thermogenesis to the evolution of a big brain. Biol Rev Camb Philos Soc. 81:293–338). These referred to incorrect raw data on water temperatures for two species, odd use of midpoint temperatures as independent variable, lack of inclusion of data on Mysticeti and the use of a differently derived EQ and midpoints instead of the EQs proposed by Manger and temperature ranges; Dr Manger proposed that these errors invalidate the analysis, with special emphasis in an observation that, since my paper did not address the relationship between EQs and temperature range, it did not actually test the thermogenesis hypothesis. In this Reply, I apologize for the mistakes which were made, and show that re-analysis using all the proposed alterations do not qualitatively or quantitatively alter the final result. I also argue that the relationship between phylogenetically correct EQs and midpoint temperatures is a better test of the thermogenesis hypothesis than the relationship between non-phylogenetic EQs and temperature ranges. 相似文献
The soluble lipase from Pseudomonas fluorescens (PFL) forms bimolecular aggregates in which the hydrophobic active centers of the enzyme monomers are in close contact. This bimolecular aggregate could be immobilized by multipoint covalent linkages on glyoxyl supports at pH 8.5. The monomer of PFL obtained by incubation of the soluble enzyme in the presence of detergent (0.5% TRITON X-100) could not be immobilized under these conditions. The bimolecular aggregate has two amino terminal residues in the same plane. A further incubation of the immobilized derivative under more alkaline conditions (e.g., pH 10.5) allows a further multipoint attachment of lysine (Lys) residues located in the same plane as the amino terminal residues. Monomeric PFL was immobilized at pH 10.5 in the presence of 0.5% TRITON X-100. The properties of both PFL derivatives were compared. In general, the bimolecular derivatives were more active, more selective and more stable both in water and in organic solvents than the monomolecular ones. The bimolecular derivative showed twice the activity and a much higher selectivity (100 versus 20) for the hydrolysis of R,S-2-hydroxy-4-phenylbutyric acid ethyl ester (HPBEt) in aqueous media at pH 5.0 compared to the monomeric derivative. In experiments measuring thermal inactivation at 75 °C, the bimolecular derivative was 5-fold more stable than the monomeric derivative (and 50-fold more stable than a one-point covalently immobilized PFL derivative), and it had a half-life greater than 4 h. In organic solvents (cyclohexane and tert-amyl alcohol), the bimolecular derivative was much more stable and more active than the monomeric derivative in catalyzing the transesterification of olive oil with benzyl alcohol. 相似文献
Severe asthma is associated with T helper (TH) 2 and 17 cell activation, airway neutrophilia and phosphoinositide-3-kinase (PI3K) activation. Asthma exacerbations are commonly caused by rhinovirus (RV) and also associated with PI3K-driven inflammation. Anthraquinone derivatives have been shown to reduce PI3K-mediated AKT phosphorylation in-vitro.
Objective
To determine the anti-inflammatory potential of anthraquinones in-vivo.
Methods
BALB/c mice were sensitized and challenged with crude house dust mite extract to induce allergic airways disease and treated with mitoxantrone and a novel non-cytotoxic anthraquinone derivative. Allergic mice were also infected with RV1B to induce an exacerbation.
Results
Anthraquinone treatment reduced AKT phosphorylation, hypoxia-inducible factor-1α and vascular endothelial growth factor expression, and ameliorated allergen- and RV-induced airways hyprereactivity, neutrophilic and eosinophilic inflammation, cytokine/chemokine expression, mucus hypersecretion, and expression of TH2 proteins in the airways. Anthraquinones also boosted type 1 interferon responses and limited RV replication in the lung.
Conclusion
Non-cytotoxic anthraquinone derivatives may be of therapeutic benefit for the treatment of severe and RV-induced asthma by blocking pro-inflammatory pathways regulated by PI3K/AKT. 相似文献
The ability to apply standard and interoperable solutions for implementing and managing medical registries as well as aggregate, reproduce, and access data sets from legacy formats and platforms to advanced standard formats and operating systems are crucial for both clinical healthcare and biomedical research settings.
Purpose
Our study describes a reproducible, highly scalable, standard framework for a device registry implementation addressing both local data quality components and global linking problems.
Methods and Results
We developed a device registry framework involving the following steps: (1) Data standards definition and representation of the research workflow, (2) Development of electronic case report forms using REDCap (Research Electronic Data Capture), (3) Data collection according to the clinical research workflow and, (4) Data augmentation by enriching the registry database with local electronic health records, governmental database and linked open data collections, (5) Data quality control and (6) Data dissemination through the registry Web site. Our registry adopted all applicable standardized data elements proposed by American College Cardiology / American Heart Association Clinical Data Standards, as well as variables derived from cardiac devices randomized trials and Clinical Data Interchange Standards Consortium. Local interoperability was performed between REDCap and data derived from Electronic Health Record system. The original data set was also augmented by incorporating the reimbursed values paid by the Brazilian government during a hospitalization for pacemaker implantation. By linking our registry to the open data collection repository Linked Clinical Trials (LinkedCT) we found 130 clinical trials which are potentially correlated with our pacemaker registry.
Conclusion
This study demonstrates how standard and reproducible solutions can be applied in the implementation of medical registries to constitute a re-usable framework. Such approach has the potential to facilitate data integration between healthcare and research settings, also being a useful framework to be used in other biomedical registries. 相似文献