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1

Production of paclitaxel and other taxanes in in vitro cultures

100%
Olszowska O., Oledzka H.
|
2003
|
issue 3
62-74
EN
Paclitaxel is an effective important anticancer drug for advanced breast, ovarian and other cancers. It was first isolated from the bark of the Pacific yew (Taxus brevifolia). Paclitaxel belongs to a group of secondary metabolites collectively indicated with the name taxanes. Paclitaxel is found in most yews, but the content is very (0-0,05%) variable. Because of the scarcity of the slow growing trees and the relative low content of paclitaxel, alternative sources are needed to meet the increasing demand. The literature concerning the tissue culture of Taxus sp. as an alternative source for taxanes production in reviewed.
2

Effect of cyclosporin A used alone and in combination with either 2-chlorodeoxyadenosine or fludarabine on normal and chronic myelogenous leukemia progenitors in vitro

100%
Korycka A., Robak T.
Archivum Immunologiae et Therapiae Experimentalis
|
2003
|
vol. 51
|
issue 1
61-67
EN
We evaluated the influence of cyclosporin A (CsA) used alone or together with new purine nucleoside analogues (PNAs): 2-chlorodeoxyadenosine (2-CdA) and fludarabine (F-ara-A) on the colony growth of normal and chronic myelogenous leukemia (CML) granulocyte-macrophage progenitor cells (CFU-GM) in cultures in vitro. The assay was based on the method described by Iscove et al. in our modification. Specimens of bone marrow were collected from 15 patients with CML in the chronic phase and from 10 hematologically normal patients. CsA at the concentrations of 1, 2 and 4 mug/ml was used alone and at the concentration of 4 mug/ml it was preincubated with MNCs and after 30 minutes PNAs were added to the culture medium. 2-CdA at the concentrations of 5, 10, 20 nM and 0.4, 0.8, 1.6 muM of F-ara-A were used. After 14 days of incubation the colonies were scored under inverted microscope. We observed that CsA used alone at all three concentrations in a statistically significant degree inhibited the colony growth of CML CFU-GM, as compared to the control (p<0.02) and it did not significantly influence the normal colony growth. IC50 for CsA was 3.9 mug/ml in case of normal CFU-GM and 2.7 mug/ml in case of CML CFU-GM. After the use of CsA in combination either with the highest concentrations of 2-CdA or F-ara-A, statistically significant differences, as compared to CsA used alone were observed (p=0.008; p=0.03 for CsA with 2-CdA; and p=0.0007; p=0.005 for CsA with F-ara-A; respectively for normal and CML CFU-GM). However, there were no significant differences between the combinations of drugs and PNAs used alone. In case of the combination of CsA with the highest concentrations of both PNAs significant differences in the colony growth inhibition between normal and CML CFU-GM were observed (p=0.002 and p=0.005, respectively for 2-CdA and F-ara-A). In conclusion, at the used concentrations of the drugs a subadditive action was observed either between CsA and 2-CdA or between CsA and F-ara-A.
3

An attempt at the transformation of embryogenic genotypes of alfalfa using gene pPR97/GUS/Intron-LLpr10.1a promoter

100%
Broda Z., Morawska J.
Biotechnologia
|
2000
|
issue 4
106-111
EN
The transformation was led by pPR97/GUS/Intron LLpr10.1a promoter gene, which codes the acid protein. Two different media were used in the experiment. On Blayds medium, the highest efficiency of soamatic embryogenesis (25%) was observed in the Kometa variety. The culture media developed by Guelph University appeared to be the best for the Kama variety (18%).
4

Factors affecting production of transgenic farm animals: cattle and rabbits

75%
Jura J., Smorag Z., Katska L., Skrzyszowska M., Gajda B., Rynska B.
Biotechnologia
|
1998
|
issue 2
101-109
EN
There are many factors affecting transgenic farm animals production. One of them is the effectiveness of the transfer of zygotes and embryos obtained after DNA microinjection. Low effectiveness of the transfer of potentially transgenic blastocysts in cattle is due to their decreased developmental potential in comparison to the blastocysts developed from not microinjected zygotes. A simple short term in vitro culture used for rabbit zygotes after microinjection increased twice the number of produced potentially transgenic rabbits.
5

The use of new strategies in the cloning studies on somatic cell cloning in pigs

75%
Skrzyszowska M., Samiec M.
Biotechnologia
|
2006
|
issue 1
68-81
EN
Somatic cell nuclear transfer (SCNT) technique in pigs remains relatively low (2% to 5% of produced piglets), that is why further efforts have to be made to optimize both a multi-step cloning procedure and to improve a structuro-functional quality of recipient oocytes and nuclear donor cells. Pre- and postimplantation developmental potential of porcine SCNT-derived embryos depends to a high degree on not only coordination of mitotic cycle stage with phenotype of nuclear donor cell, but also proper combination of the methods of maternal chromosome elimination (enucleation), oocyte reconstruction techniques, the systems of artificial activation of generated nuclear-cytoplasmic hybrids (clonal cybrids) and in vitro culture of reconstructed embryos. Generally, it can result in increasing the competences of both somatic nuclear and mitochondrial genome for epigenetic remodeling/reprogramming in developing cloned embryos.
6

Bacteria in plant tissue cultures

75%
Orlikowska T., Zawadzka M.
|
EN
Bacterial contamination is quite frequent in plant tissue cultures although, theoretically, cultures have to be axenic. It is their ubiquity and adaptability to different conditions that enable bacteria to colonize also tissue cultures. Among them there are not only the typical endo- and egzophytic species connected with the plant kingdom, but also those which are common among people. The bacteria species isolated from plant tissue cultures were found to be vitropathogenic (pathogens facultative to in vitro explants), latent (pathogens not virulent in vitro), and cryptic (present in tissues but invisible). Some bacteria produce growth regulators, which can modify the morphogenetic mode of explants. They are all undesirable ones in cultures, but the explants contaminated with pathogenic species should be eliminated obligatorily. Various groups of bacteria, as well as the techniques of detecting, identifying and eliminating them, are briefly described.
7

Stimulation of anthocyanin production by two-phase growth system, in callus of Rudbeckia hirta

75%
Luczkiewicz M., Cisowski W.
Biotechnologia
|
1999
|
issue 4
199-210
EN
Biotechnological research to achieve and then increase anthocyanin production in callus tissue of R. hirta L., involved testing a number of growth media, modified with growth regulators. The evaluation of the growth of propagated tissues, their water contents and in particular their ability to produce anthocyanins, led to the development of an original two-phase growth system. This new system has the advantages of both growth-stimulating media and pigment production media. As a result of the research, callus tissue was obtained on a two-phase growth medium, which was made of modified Schenk-Hildebrandt medium (growth phase) and Miller's medium (production phase). The callus synthesised a 12-component anthocyanin compound complex, which constituted 2,18% of dry mass. This is a considerable amount compared to 0,28% in the natural plant. Phytochemical analysis (TLC, HPLC, PC, UV, 1H-NMR) of the anthocyanin complex isolated from callus produced with the two-phase system proved that the dominating compounds in the pigment complex were: cyanidin-3-0-(6-0-malonyl-b -D-glucopyranoside) and cyanidin-3-0-(b-D-glucopyranoside).
8

Regeneration of Arabidopsis thaliana (L.) Heynh. plants via direct somatic embryogenesis

75%
Gaj M. D.
Biotechnologia
|
2001
|
issue 2
90-98
EN
In Arabidopsis biotechnology plants are regenerated in vitro via shoot organogenesis induced in callus derived from different somatic tissues. An alternative way of in vitro plant regeneration via somatic embryogenesis has not been applied in Arabidopsis so far. Recently, it was found that development of Arabidopsis somatic embryos can be induced in the culture of immature zygotic embryos and that the callus phase is not nocessary for the initiation of embryogenesis. The aim of the presented research was to determinate the in vitro culture conditions enabling high efficiency of somatic embryo induction and their conversion into plants. The influence of induction medium composition including liquid or agar medium, type and concentration of auxin, carbohydrates and ammonium sources as well as duration of auxin treatment of explants on DSE efficiency were evaluated. Advantages of described regeneration system via DSE are as follows: short time needed to induce somatic embryos (10-15 days), high efficiency of the process (up to 90% explants responded), numerous embryos produced per explant (on average 17) and high percentage of embryo conversion into fertile plants (70-80%).
9

Densytometric analysis of diosgenin in extracts from callus tissue of Polygonatum verticillatum (L.) All.

75%
Szybka-Hrynkiewicz P., Janeczko Z.
Biotechnologia
|
2004
|
issue 2
93-99
EN
In this study, we examined the effect of plant growth regulators (2, 4-D, 6-BAP) and steroidal hormone (17-beta-estradiol) on a quantity of diosgenin in callus tissue of Polygonatum verticillatum (L.) All. The callus grown in in vitro conditions on MS medium with: 1/1, 2/2, 5/5, 10/10 ([mg/dm3] 2,4-D/[mg/dm3] 6-BAP) and 1/1/1, 2/2/2, 5/5/5, 10/10/10 ([mg/dm3] 2,4-D/[M/dm3] 17beta-estradiol/[mg/dm3] 6-BAP) was used for the densytometric analysis to determine the presence of diosgenin. The content of diosgenin ranged from 0,0642 to 1,1458%. An increase in diosgenin content due to addition of 6-BAP and 2,4-D was observed. The developed procedure was estimated by statistical analysis which indicates that it can be used for routine analysis.
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