Brassica oleracea varieties (red cabbage, broccoli, Savoy cabbage and cauliflower) were tested for their ability to regenerate shoots in vitro. Cotyledon, hypocotyl and root explants of 7 day-old seedlings were incubated on Murashige and Skoog's (MS) medium supplemented with 1 mg l-1 6-benzyladenine (BA) or 6-furfurylaminopurine (KIN) in combination with 0, 0.1, and 0.2 mg l-1 indole-3-butyric acid (IBA). Hypocotyls showed the best explants in almost all varieties tested with a minimum regeneration potential of 75% and producing 3.5-7.4 shoots per explant. The BA-supplemented media were optimal for both shoot regeneration and multiplication. Shoots rooted maximally (100%) on plant growth regulator-free MS medium containing 2% or 4% sucrose. Increased sucrose content improved plant acclimation in the greenhouse
A method for the simultaneous preparation of highly enriched human placental trophoblast populations (villous and extravillous) from first-trimester placental villi (5 to 12 weeks) by using sequential trypsinization, percoll gradient centrifugation, and negative selection with anti-CD9 immunomagnetic separation is described. The purification method resulted in the isolation of four distinct trophoblast populations identified on the basis of morphology and phenotyping: (i) mononuclear villous cytotrophoblast cells which, through differentiation, become committed to syncytium formation; (ii) an extravillous trophoblast population which appeared as a ''crazy pavement'' and, with subsequent subculturing, differentiated morphologically to mononuclear cells; (iii) an extravillous trophoblast fraction which fused to form multinucleated trophoblast giant cells; and (iv) floating intermediate extravillous trophoblast cells which fused together to form cell clumps and which further differentiated to a mononuclear anchoring intermediate extravillous trophoblast. Short-term cultures of the freshly isolated cell fractions consisted of heterogeneous trophoblasts at different differentiation stages as determined by their varied biochemical and morphological properties. All the isolated trophoblast populations expressed the cytokeratin intermediate filament and the epithelium-specific cell-cell adhesion molecule Ecadherin. The isolated villous trophoblasts in culture expressed integrins ␣6 and 4 and reduced levels of 1 subunits, whereas the proliferating extravillous trophoblast cultures expressed ␣1, ␣3, and ␣5 and high levels of 1 integrin subunits, vitronectin receptor (␣V3/5), and major histocompatibility complex class 1 molecules. Furthermore, the isolated trophoblast populations secreted metalloproteases (such as type IV collagenases [mainly 72-and 92-kDa enzymes, i.e., gelatinases A and B]) and urokinase plasminogen activator, as evaluated by substrate gel zymography. This method of isolation should facilitate in vitro studies of trophoblast proliferation, differentiation, invasion, virus interactions, cytokenesis, and immunology.
Investigation comprised 41 tomato genotypes originating from the population
of domestic and domesticated genotypes collected in Serbia and belonging to
the tomato collection of the Institute of Vegetable Crop Science, Smederevska
Palanka. The aim of collection screening was to choose the genotypes tolerant
to drought during plant intensive growth stage, whereby the process of
selection would set out to obtain the recombinant genotypes for this abiotic
factor. The screening criteria were established for genotype divergence in
plant height and shoot-root ratio under conditions of optimal irrigation
regime and drought. Divergence was estimated using cluster analysis with
Euclidean distance as a measure of distance, with a complete gene attachment
to grouping. Drought tolerance is expressed by the stress susceptibility
index (SSI). Various results were obtained based on the screening of
genotypes grown under optimal and dry conditions. As a measure of stress
susceptibility, based on SSI, genotypes having different drought tolerance
level were determined. On the grounds of the analyses carried out, 10
genotypes were segregated (G102, G104, G107, G109, G110, G119, G125, G126,
G128 and G141) to represent a basis to obtain the recombinant genotypes and
to initiate the selection for drought resistance. [Projekat Ministarstva
nauke Republike Srbije, br. TR 31005 i br. TR 31059]
Interferons (IFN) are produced by the placenta during pregnancy, and they can be detected in the maternal and fetal blood. Although the antiviral potential of IFNs is well established, it remains unclear whether the IFNs associated with pregnancy can prevent transplacental spread of viral infection. The present study was undertaken in order to determine the possible protective effect of placentally produced IFN-alpha on fetal acquisition of herpes simplex virus (HSV). Nine mothers with a known history of genital HSV infection were studied. In five cases IFN-alpha was detected in the placenta, maternal, and fetal blood, whereas in three cases IFN-alpha could not be detected. in the remaining case, IFN-alpha was found only in the maternal blood. As corroborated by the serological evidence of early HSV infection in the cord blood, the single case of vertical HSV transmission was observed in the group of IFN nonproducers. Furthermore, virus transmission did not occur in cases where IFN-alpha was present in the placenta and simultaneously in the maternal and fetal circulations. Thus, the present data indicate that high levels of IFN during pregnancy may protect the fetus from acquiring a possibly fatal intrauterine HSV infection.
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