Cryopreservation of Citrus aurantifolia seeds and embryonic axes using a desiccation protocol

The desiccation and freezing tolerance of seeds, with and without testas, and embryonic axes of Citrus aurantifolia were investigated. Seeds were desiccated with silica gel, under the laminar air flow cabinet or by placing them on a laboratory bench. Whatever the desiccation method employed, survival before and after cryopreservation was higher for seeds without testas. When freezing intact seeds, the highest survival percentage (41.3 %) was achieved after desiccation to 7.3 % moisture content (fresh weight basis) on the laboratory bench. Survival of seeds cryopreserved without testas could reach up to 85 % after desiccation under the laminar air flow cabinet or on the laboratory bench, corresponding to moisture contents of 7.1 and 4.5 %, respectively. After desiccation with silica gel, survival reached a maximum of 60.0 %, for a seed moisture content of 3.3 %. Survival of control embryonic axes was high (80-100 %) whatever the sucrose concentration in the preculture medium and the duration of the desiccation period. After cryopreservation, no survival was noted with embryonic axes, which had not been precultured nor desiccated. Survival of non-desiccated embryonic axes after cryopreservation increased progressively in line with increasing sucrose concentrations in the preculture medium, from 7.5 % with 0.1 M sucrose to 77.5 % with 0.7 M sucrose. Survival of desiccated and cryopreserved embryos was always high, whatever the preculture treatment and desiccation period, ranging from 55.8 % to 92.5 %.     

Cryopreservation of chayote (Sechium edule JACQ. SW.) zygotic embryos and shoot-tips from in vitro plantlets

This paper presents the development of cryopreservation protocols for zygotic embryos and apices of chayote (Sechium edule Jacq. Sw.), a tropical plant species with recalcitrant seeds. Zygotic embryos of two cultivars, Ccocro negro (CN) and Claudio (Cl) could withstand cryopreservation, with survival percentages of 10 and 30 %, after desiccation to 23 and 19 % moisture content (fresh weight basis), respectively. Apices sampled on in vitro plantlets of cultivars Cl, 13 and JM were successfully cryopreserved using a vitrification technique. Optimal conditions included the culture of mother-plants for 22 days on medium containing 0.3 M sucrose, culture of excised apices on the same medium for 1 day, loading of apices for 20 min with 2M glycerol + 0.4M glycerol, treatment with a series of diluted PVS2 solution (60 % PVS2 followed by 80 % PVS2 solution for 15 min (cultivar Cocoro Blanco [CB]) or 30 min (cultivars CN and Cl) at each concentration), rapid freezing and thawing, washing of shoot-tips with a 1.2 M sucrose solution, followed by recovery on media with progressively decreasing sucrose concentrations until the standard concentration of 0.1 M was reached. The highest survival percentages achieved ranged between 17 and 38 %, depending on the cultivar.     

Effect of cryopreservation on the structural and functional integrity of cell membranes of sugarcane (Saccharum sp.) embryogenic calluses

In this paper, we investigated if the differences consistently noted in survival and plantlet production between cryopreserved and non-cryopreserved, control sugarcane embryogenic calluses were related to modifications induced during cryopreservation in the structural and functional integrity of cell membranes. For this, the evolution of electrolyte leakage, lipid peroxidation products and cell membrane protein contents was measured during 5 d after cryopreservation. Differences between control and frozen calluses were observed only during the first 2 (electrolyte leakage) or 3 d (lipid peroxidation products and membrane protein content) after freezing. It was not possible to link these differences with the differences noted in survival and plant production between control and cryopreserved calluses. Additional studies are thus needed to elucidate which biochemical factors, linked to survival and plantlet regeneration, are affected by cryopreservation.     

Factorization of analytic functions by means of Koenig's theorem and Toeplitz computations

Summary. By providing a matrix version of Koenig's theorem we reduce the problem of evaluating the coefficients of a monic factor r(z) of degree h of a power series f(z) to that of approximating the first h entries in the first column of the inverse of an Toeplitz matrix in block Hessenberg form for sufficiently large values of n. This matrix is reduced to a band matrix if f(z) is a polynomial. We prove that the factorization problem can be also reduced to solving a matrix equation for an matrix X, where is a matrix power series whose coefficients are Toeplitz matrices. The function is reduced to a matrix polynomial of degree 2 if f(z) is a polynomial of degreeN and . These reductions allow us to devise a suitable algorithm, based on cyclic reduction and on the concept of displacement rank, for generating a sequence of vectors that quadratically converges to the vector having as components the coefficients of the factor r(z). In the case of a polynomial f(z) of degree N, the cost of computing the entries of given is arithmetic operations, where is the cost of solving an Toeplitz-like system. In the case of analytic functions the cost depends on the numerical degree of the power series involved in the computation. From the numerical experiments performed with several test polynomials and power series, the algorithm has shown good numerical properties and promises to be a good candidate for implementing polynomial root-finders based on recursive splitting strategies. Applications to solving spectral factorization problems and Markov chains are also shown. Received September 9, 1998 / Revised version received November 14, 1999 / Published online February 5, 2001     

Covalent docking of selected boron-based serine beta-lactamase inhibitors

Journal of Computer-Aided Molecular Design - AmpC β-lactamase is a hydrolytic enzyme conferring resistance to β-lactam antibiotics in multiple Gram-negative bacteria. Therefore,...     

Supramolecular Assemblies of Phosphorylated Cavitands

The recent results in the chemistry of cavitands have proved that they are very efficient molecular receptors and potential precursors of molecular devices. In this context, we have investigated the synthesis and binding properties of phosphorylated cavitands. The stereoselective synthesis and the structural studies of the new compounds showed that these bowl-shaped molecules possess a well defined aromatic cavity surrounded by four phosphoryl groups (P=O or P=S). They are very efficient ligands for metal and organic cations. They are able to encapsulate cationic species by cooperative effect of the preorganized aromatic cavity and the four phosphorylated groups. Moreover, the upper and lower rim functionalitics can lead to the formation of molecular capsules and supramolecular assemblies whose properties and structures have been investigated by X-ray diffraction and NMR studies in solution.     

Mechanisms of Ceftazidime and Ciprofloxacin Transport through Porins in Multidrug-Resistance Developed by Extended-Spectrum Beta-Lactamase <Emphasis Type="Italic">E.coli</Emphasis> Strains

Resistance towards antibiotics stands out today as a major issue in the clinical act of treatment of bacterial-generated infections. This process was characterized in proteoliposomes reconstituted from an E.coli strain isolated from invasive infections (blood culture) occurred in patients with a cardio-vascular device admitted for surgery. Fluorescence spectroscopy and patch-clamp technique have been used. Two types of antibiotics have been targeted: ceftazidime and ciprofloxacin. Antibiotics addition in proteoliposomes suspension undergoes a quenching in tryptophan residues from outer membrane porins structure, probably due to the formation of a transient non-fluorescent porin-antibiotic complex. Patch-clamp recordings revealed strong ion current blockages for both antibiotics, reflecting antibiotic–channel interactions but with varying strength of interaction. The present study puts forward the mechanism of multidrug-resistance in extended-spectrum beta-lactamase E.coli strains, as being caused by alterations of the antibiotics transport across the porins of the outer bacterial membrane.     

Geneticallly enginereed trehalose accumulation improves cryopreservation tolerance of chrysanthemum (Dendranthema grandiflorum kitam.) shoot-tips

Shoot-tips isolated from two transgenic lines of chrysanthemum (Dendranthema grandiflorum Kitam.) var. Indianapolis in vitro plantlets with induced capacity to biosynthesize trehalose, and from a non-transformed line, were subjected to cryopreservation using a vitrification procedure. After dissection, apices were precultured on semi-solid MS medium with 0.3 M sucrose for 4 days, loaded in a 0.4 M sucrose + 2 M glycerol solution for 20-30 min and exposed to PVS2 or PVS3 vitrification solutions for 0, 20, 40 or 60 min at room temperature prior to rapid immersion in liquid nitrogen. The highest shoot regeneration after cryopreservation was obtained with exposure to either PVS solution for 40 min. Plant regeneration from cryopreserved shoot-tips ranged between 48 percent and 67 percent for transgenic lines and between 33 percent and 36 percent for non-transgenic lines. No polymorphic loci were detected in plantlets regenerated from cryopreserved and non-cryopreserved shoot-tips with RAPD techniques using eight primers that amplified 101 monomorphic loci.     

SF and SC Activation of SF6 and SF5 Derivatives at Rhodium: Conversion of SF6 into H2S

The degradation of SF₆ and SF₅ organyls by S? F and S? C bond‐activation reactions at [{Rh(μ‐H)(dippp)}₂] under mild conditions is reported. Fluorido and thiolato species were identified as products or intermediates, and were characterized by X‐ray diffraction analysis and multinuclear NMR spectroscopy. An unprecedented cyclic process for the conversion of the potent greenhouse gas SF₆ into H₂S was developed.