Method of clonal micropropagation of different willow species and hybrids

Conditions of cultivation and micropropagation of selected biotypes of five willow species (Salyx dasyclados Wimm., S. caspica Pall., S. triandra L., S. purpurea L., and S. viminalis L.) and two hybrids (×S. acuminata S. and ×S. palustris Host.) were optimized. Data on in vitro propagation of S. caspica, S. triandra, S. purpurea together with hybrids S. acuminata and S. palustris were obtained for the first time. It has been demonstrated that the outcome of cultivation and propagation of willows strongly depends on genotypic peculiarities of initial plants. The optimal terms of isolation and sterilization of single-node segments for obtaining 50–75% of aseptic viable developing cultures were estimated. The nutritive media were selected providing induction of stem development (to 67%), their rooting (to 91%), elongation (to 3–6 cm), and multiplication (propagation coefficient of 4). The designed method (adopted to different genotypes) can be applied for obtaining aseptic in vitro cultures serving as initial plant material for genetic transformation and mass propagation of plants with new agriculturally valuable characteristics which are of interest for construction of bioenergetic plantations and for needs of the paper industry.     

Antibody-targeted photolysis: in vitro immunological, photophysical, and cytotoxic properties of monoclonal antibody-dextran-Sn(IV) chlorin e6 immunoconjugates.

A set of anti-melanoma immunoconjugates were prepared which contained chlorin e6: antibody molar ratios of 18.9:1, 11.2:1, 6.8:1, and 1.7:1. All immunoconjugates retained antigen binding activity regardless of the chromophore:antibody substitution ratio that was attained. In contrast, the ground-state absorption spectra of the immunoconjugates showed features which appeared to be dependent on the chromophore:antibody molar ratio. In addition, the quantum yield of singlet oxygen generated by the conjugated chromophores was observed to be significantly less than that observed with the unbound dye. Time-resolved absorbance spectroscopy of the chromophore excited triplet state indicated that the loss of singlet oxygen quantum yield resulted from diminished chromophore triplet yield. Analysis of data obtained from in vitro photolysis of target melanoma cells, in combination with that obtained from the immunochemical and photochemical studies, indicates that the observed immunoconjugate phototoxicity can be reasonably quantitatively represented by (1) the ability of the immunoconjugate to bind SK-MEL-2 cell surface antigen, (2) the amount of chromophore localized at the target cells by immunoconjugate binding, (3) the delivered dose of light at 634 nm, and (4) the singlet oxygen quantum yield of the antibody-bound photosensitizer. Though these data argue strongly for photolysis by the cumulative dosage of singlet oxygen at the cell membrane, nonetheless, the concurrent photoinduced release of other cytotoxic agents should not be ruled out.     

Electrophoretic analysis of the estrogen receptor. Molybdate stabilization and identification of the classical estrogen receptor

Conditions are defined which permit analysis of estrogen receptors from the mammalian uterus by polyacrylamide gel electrophoresis, thereby solving a longstanding problem encountered in previous attempts at such analysis, namely the failure of a large portion of the receptor population to enter such gels. A paramount requirement for entry of the estrogen-receptor complex into polyacrylamide gels is its maintenance in an untransformed state which does not form aggregates that are excluded from these gels. Of the multiple estrogen-binding proteins separated, only one (relative mobility of 0.5-0.6) possessed the definitive characteristics of the classical estrogen receptor. The inclusion of molybdate in extraction buffers selectively enhanced receptor recovery and facilitated its separation. Moreover, the estrogen-receptor complex so resolved is separated from other types of estrogen-binding proteins present in the uterine cytosol. These findings show that the molybdate-stabilized estrogen receptor exists in a single discrete form, but otherwise exhibits multiple forms that are probably artifactual. Electrophoresis in discontinuous buffers, but not in a continuous buffer system, promoted aggregate formation. This finding has implications concerning the subunit structure of the untransformed receptor.     

Interaction of hydrophobic bis (D-mannose) derivatives with adipocyte and erythrocyte sugar transport systems

The inhibition of sugar uptake by a series of hydrophobic bis(D-mannose) derivatives has been measured in rat adipocytes. When the D-mannose moieties of the bis compounds are separated by a hexane bridge the transport inhibition constant (Ki) is greater than for a decane-bridged molecule. This is probably due to the increased hydrophobicity of the bridge of the decane-bridged compound. The enhancement in affinity due to the second sugar in the bis(D-mannose) derivatives is probably only 2-fold, since half reduction of the bis(D-mannosyloxy)hexane increases Ki approx. 2-3-fold. N'-DNP-1,3-bis(D-mannos-4'-yloxy)propyl-2-amine has very high affinity in insulin-treated cells. The affinity is approx. 1000-fold higher than for D-mannose. This enhancement is probably due to the hydrophobicity of the DNP group. The distance from the sugar to the hydrophobic group is important because an increase in Ki occurs if an aminocaproyl spacer is introduced between the DNP group and 1,3-bis(D-mannos-4'-yloxy)propyl-2-amine. Aminocaproyl and glycyl spacers also increase the Ki for NAP derivatives of 1,3-bis(D-mannos-4'-yloxy)propyl-2-amine. Each of the hydrophobic bis(D-mannose) derivatives has a lower Ki in insulin-treated cells. This may be due to an insulin responsive hydrophobic interaction between the hydrophobic portion of the sugar and a hydrophobic domain in the transport system. The inhibition constants for the hydrophobic bis(D-mannose) compounds have also been measured in human erythrocytes.     

Pollen-based biome reconstruction for southern Europe and Africa 18,000 yr bp

Pollen data from 18,000 ¹⁴C yr bp were compiled in order to reconstruct biome distributions at the last glacial maximum in southern Europe and Africa. Biome reconstructions were made using the objective biomization method applied to pollen counts using a complete list of dryland taxa wherever possible. Consistent and major differences from present‐day biomes are shown. Forest and xerophytic woods/scrub were replaced by steppe, both in the Mediterranean region and in southern Africa, except in south‐western Cape Province where fynbos (xerophytic scrub) persisted. Sites in the tropical highlands, characterized today by evergreen forest, were dominated by steppe and/or xerophytic vegetation (cf. today’s Ericaceous belt and Afroalpine grassland) at the last glacial maximum. Available data from the tropical lowlands are sparse but suggest that the modern tropical rain forest was largely replaced by tropical seasonal forest while the modern seasonal or dry forests were encroached on by savanna or steppe. Montane forest elements descended to lower elevations than today.     

Genetics and Mechanism of Resistance to Meloidogyne arenaria in Peanut Germplasm

Segregation of resistance to Meloidogyne arenaria in six BC₅F₂ peanut breeding populations was examined in greenhouse tests. Chi-square analysis indicated that segregation of resistance was consistent with resistance being conditioned by a single gene in three breeding populations (TP259-3, TP262-3, and TP271-2), whereas two resistance genes may be present in the breeding populations TP259-2, TP263-2, and TP268-3. Nematode development in clonally propagated lines of resistant individuals of TP262-3 and TP263-2 was compared to that of the susceptible cultivar Florunner. Juvenile nematodes readily penetrated roots of all peanut genotypes, but rate of development was slower (P = 0.05) in the resistant genotypes than in Florunner. Host cell necrosis indicative of a hypersensitive response was not consistently observed in resistant genotypes of either population. Three RFLP loci linked to resistance at distances of 4.2 to 11.0 centiMorgans were identified. Resistant and susceptible alleles for RFLP loci R2430E and R2545E were quite distinct and are useful for identifying individuals homozygous for resistance in segregating populations.     

Implicit associations with social status: the effects of relationship involvement

The study tested the proposition that relationship involvement influences the implicit responses of women to high- and low-status professions. It was hypothesized that when a high-involvement context was primed, women would have more positive implicit associations with high-status occupations than when a low-involvement context was primed. In contrast, when a high-involvement context was primed, women would have more negative associations with low-status occupations than when a low-involvement context was primed. To test the hypothesis, 123 female participants received a high or low relationship involvement prime. Then the participants completed a single category implicit associations test designed to measure the participants' associations with either high- or low-status occupations. As predicted, the relationship involvement prime influenced the positivity of associations made with high- and low-status occupations. The study pointed to the possibility that persons possess evolutionary-based implicit associations.