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1

Biotechnological, organizational, legal and ethical aspects of xenotransplantation

100%
Smorag Z.
Biotechnologia
|
2006
|
issue 1
103-109
EN
The paper presents issues that shall be considered and elaborated before xenotransplantation reaches clinical stage. In a scope of a research project, such considerations as rules and procedures of a donor selection, including nature of its genetic modifications, must be undertaken. Another problem are regulatory requirements concerning pre-clinical tests on primates. Detailed conditions and requirements that must be fulfilled before clinical trials start are also discussed in the paper. Patient selection criteria as well as relationship with him/her before and after transplantation shall be clarified. A matter of high importance are organisational guidelines and relevant legal aspects of donor animals, breeding and xenotransplantation product harvesting, as well as ethical dimension of these actions.
2

Reproduction biotechnology of farm animals

100%
Smorag Z.
Biotechnologia
|
2002
|
issue 4
30-40
EN
This paper presents the most important research and application of biotechnology of farm animals reproduction: semen sexing, embryo in vitro production, embryonic and somatic cloning and transgenesis.
3

Achievements in biotechnology of animal reproduction over the past 20 years - case studies

100%
Smorag Z.
Biotechnologia
|
2010
|
issue 3
64-74
EN
Animal reproduction biotechnology is an area of fast development with possibilities of practical applications not only in breeding, but in pharmacy and biomedicine as well. Its growth has been achieved thanks to the results of last decades of the previous century in embryology, endocrinology, and molecular biology. The paper contains very brief description of the achievements of NRIAP Department of Animal Reproduction Biotechnology during the last 20 years in mammalian sex regulation, cryobiology of gametes and embryos, in vitro production of embryos, cloning and animal transgenesis.
4

Vitrification of mammalian oocytes and embryos

64%
Smorag Z., Gajda B.
Biotechnologia
|
1995
|
issue 3
68-83
EN
Vitrification is a new approach to oocyte and embryo cryoconservation.It consists in the solidification of a solution caused by draastic increase in viscosity during cooling and not by crystalization.The application of this approach to cryoconservation of oocytes and embryos of different species depends upon the development of proper procedures and non-toxic media.From the technical point of view, the vitrification method is simple and relatively easily applicable under field conditions.The authors review the current procedures applied to oocytes and embryos of laboratory and farm animals.
5

Monogenetic twin production technique in mammals

64%
Skrzyszowska M., Smorag Z.
Biotechnologia
|
1998
|
issue 2
137-144
EN
The development of non-surgical methods of embryo collection and transfer, primarily in cattle, has led to the commercialization of these techniques, including the production of genetically identical twins. The aim of this paper is to review the progress of the studies on the production of monogenetic progeny using microsurgical bisection of embryos, with a special emphasis put on factors affecting the efficiency of this method. We have also described our new alternative method of monogenetic twin production in cattle based on modified bisection of specifically hatching blastocysts, whose zona pellucida had been perforated.
6

Cryopreservation of porcine oocytes and embryos

64%
Gajda B., Smorag Z.
Biotechnologia
|
2003
|
issue 1
116-228
EN
This paper presents the current possibilities, state of knowledge and prospects for cryopreservation of pig oocytes and embryos. The main factors of cryopreservation efficiency, methods for the evaluation of cryopreserved embryos, and the possibilities of modifying their susceptibility to cryopreservation are discussed. In addition, the most significant results of pig embryo freezing and vitrification and the cryotechnical aspects of this method are presented.
7

Application of reproduction biotechnology methods in animal biodiversity conservation

64%
Gajda B., Smorag Z.
|
2007
|
issue 4
55-65
EN
This article presents the potential and prospects for the use of selected reproduction biotechnology methods in animal biodiversity conservation programs. The first part focuses on biotechnological methods related to male reproduction such as artificial insemination, semen cryoconservation, sexing and spermatozoa sorting. The second part discusses biotechnological methods related to female reproduction such as superovulation, in vitro production and transfer of embryos, cryoconservation of oocytes and embryos, embryo sexing, cloning and transgenesis.
8

Cryoconservation of mammalian embryos and oocytes

64%
Gajda B., Smorag Z.
Biotechnologia
|
1998
|
issue 2
10-33
EN
This paper presents current methods of embryo and oocyte cryoconservation in the following species: mice, rabbits, sheep and goats, pigs, horses and cows. Both the freezing and vitrification methods are discussed with special emphasis on the major factors affecting the efficiency of these two methods.
9

Flow cytometry-application in animal biotechnology

64%
Smorag Z., Bochenek M.
Biotechnologia
|
1995
|
issue 3
54-67
EN
Flow cytometry offers several advantages over the formerly used methodologies - hundreds or thousands of cells can be measured per second with high accuracy and reproducibility, rare cells can be detected from a large population.Some of the staining methods preserve cell viability so the reprodctive capacity of the sorted cells can be investigated.In biotechnology, the most popular application of flow cytometry is the DNA analysis e.g. cell cycle, chromosome karyotyping, gene mapping, etc.It is possible to analyse quality of semen and sorting of viable sperm cells.
10

Ethical aspects of animal biotechnology

63%
Smorag Z.
Biotechnologia
|
1995
|
issue 3
7-12
11

Effect of protein source, witamin E and phenazine ethosulfate on developmental competence and quality of porcine embryos cultured in vitro

51%
Gajda B., Baryla M., Smorag Z.
Folia Biologica
|
2008
|
vol. 56
|
issue 1-2
57-63
EN
Effects of fetal calf serum (FCS) or bovine serum albumin (BSA), with or without vitamin E (vit. E) or phenazine ethosulfate (PES) supplementation on developmental competence and quality of cultured porcine embryos were examined. The experiment was done on zygotes and 2-cell embryos obtained from superovulated gilts. Morphologically normal zygotes were cultured in vitro in NCSU-23 medium supplemented with: experiment 1-0.004 g/ml BSA, 10% FCS, protein-free (control); experiment 2-0 (control), 25, 50 or 100 M vit. E; experiment 3-0 (control), 0.025, 0.05 or 0.075 M PES. Embryo quality criteria were developmental competence (cleavage, morula and blastocyst rates), total cell number per blastocyst and degree of apoptosis as assessed by TUNEL staining. Presence of BSA in culture medium increased significantly morula and blastocysts production as compared to FCS (P<0.001) and protein-free group (P<0.05 and P<0.001, respectively). The blastocysts cultured in protein-free medium had higher average number of apoptotic nuclei and DNA fragmented nucleus index as compared to the BSA (P<0.05 and P<0.01, respectively) and FCS (P<0.5) group. Supplementation in culture medium of 100 M vit. E increased blastocyst production as compared to control and 50 M vit-E (P<0.05). Both the number of cells per and percentage of TUNEL positive nuclei per blastocyst were slightly lower in PES treated than control groups.
12

Transvaginal method of bovine oocytes collection: aspiration efficiency, developmental competence of oocytes and application in breeding

51%
Smorag Z., Gogol P., Kuznetsov V.
Biotechnologia
|
1998
|
issue 2
153-160
EN
This paper describes the ultrasound-guided transvaginal method of oocyte collection in cattle. Both technical and physiological aspects affecting the efficiency of the method are presented. The results of the author's experiments on oocytes collection and their developmental competence is also discussed.
13

Influence of DNA concentration on the efficiency of bovine blastocyst production after WAP-bGH microinjection into germinal vesicle of immature oocytes, zygotes and embryos obtained in vivo or in vitro

39%
Jura J., Smorag Z., Katska L., Skrzyszowska M., Houdebine L.-M., Rynska B.
Biotechnologia
|
1997
|
issue 4
71-81
EN
Microinjection is one of the most successfully used methods to produce transgenic farm animals. But the effectiveness of transgenesis with the use of microinjection, specially in cattle is still low. Many steps of the transgenesis has been found to influence its effectiveness; DNA purity, the site of its injection, the culture system. There are no reports on the influence of a DNA vector concentration influence on the developmental ability of transformed bovine eggs to the blastocyst stage in the in vitro culture. In presented experiments we investigated the influence of different DNA concentrations on the developmental rate of microinjected immature bovine oocytes, zygotes and 2-cell embryos to the blastocyst stage.
14

Transgenic pigs: production and genetic modification

39%
Smorag Z., Jura J., Kopchick J. J., Gajda B., Skrzyszowska M., Rozycki M., Pasieka J.
Biotechnologia
|
1998
|
issue 2
145-152
EN
This paper presents the methods of transgenic pigs production and the results based on the long experience of the authors in this area. Moreover, the trends and current issues of transgenic modification in pigs are discussed.
15

Factors affecting production of transgenic farm animals: cattle and rabbits

39%
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.
16

The expression of WAP-Fuc gene in genotypes of transgenic farm animals ? effectiveness of transgenesis with the use of DNA microinjection

39%
Jura J., Smorag Z., Gajda B., Kareta W., Kopchick J. J., Kelder B., Prieto P. A.
Biotechnologia
|
2003
|
issue 1
216-222
EN
There are two principal applications of transgenic animals. Best known and most advanced application is to use transgenic animals (called bioreactors or molecular farms) for the production of various proteins or biopolymers of medical significance. The second application concerns efforts to improve the productivity traits of breeding animals. We have worked out methods which allow somatic cloning of mammals and gene knockout methods. These methods have been developing very rapidly in recent years and their efficiency will soon be improved to the extent that they will become profitable. For the time being, DNA microinjection with all its disadvantages, remains the principal method of producing transgenic bioreactors. In this paper, the effectiveness of production of transgenic rabbits, goats and pigs with the use of WAP-Fuc gene construct is presented.
17

Performance of transgenic pigs produced with the use of two different growth hormone gene constructs

33%
Rozycki M., Smorag Z., Kopchick J., Chen W. Y., Jura J., Pasieka J., Orzechowska B., Gajda B., Skrzyszowska M.
Journal of Applied Genetics
|
1999
|
vol. 40
|
issue 1
29-37
EN
Four transgenic pigs, produced with the use of two different gene constructs containing the bovine growth hormone coding gene - Mt-bGH-10D6 (wild type) and Mt-bGH-M8 (mutated), were used to produce the F1 generation to investigate their performance traits. No differences were observed in fattening and slaughter traits between transgenic and non-transgenic pigs. It was found that the weight of ham proper and loin eye area was significantly higher in carriers of Mt-bGH-10D6 gene constructs.
18

Production of pigs used in xenotransplantation

33%
Jura J., Slomski R., Smorag Z., Gajda B., Wieczorek J., Lipinski D., Kalak R., Juzwa W., Zbyland J.
Biotechnologia
|
2006
|
issue 1
151-158
EN
There are four main trends in the use of transgenic animals. One of the most interesting is the use of transgenic animals as the donors of tissue or organs suitable for transplantation in human ? xenotransplantation. This field of research has been undergoing intensive and increasing study during the past few years, and some encouraging progress is being made. A pig has been identified as the most suitable donor animal. The aim of the presented experiment was to produce transgenic pigs which tissues and organs could be used for the xenotransplantation needs. To target the goal, a competitive gene construct coding the same substrate as the endogenous enzyme of organ donor was introduced. In effect, transgenic boar was produced with confirmed integration of human a1,2 fucosyltransferase gene. Also, F1 generation of transgenic pigs was generated to preserve ongoing needs of preliminary research of xenotransplantation project.
19

Transgenic rabbit producing human growth hormone in milk

27%
Lipinski D., Jura J., Kalak R., Plawski A., Kala M., Szalata M., Jarmuz M., Korcz A., Slomska K., Gronek P., Smorag Z., Pienkowski M., Slomski R.
Journal of Applied Genetics
|
2003
|
vol. 44
|
issue 2
165-174
EN
The gene construct WAP(6xHisThr):hGH containing the entire human growth hormone gene (hGH) under the rat whey acidic protein (WAP) promoter regulating the expression in mammary glands of mammals was prepared. The 5? end of the gene was modified by the addition of a sequence encoding six histidine residues and a sequence recognized by thrombin. The gene construct was introduced by microinjection into the male pronucleus of a fertilized oocyte. The founder male rabbit was obtained with the transgene mapping to chromosome 7. The presence of the growth hormone was confirmed in samples of milk collected during the lactation of F1 generation females. The growth hormone can be easily purified by affinity chromatography and cleavage by thrombin to an active form.
20

Expressive gene structures of proper and modified genes

27%
Lipinski D., Szalata M., Kalak R., Plawski A., Nuc K., Kala M., Juzwa W., Slomska K., Gronek P., Jura J., Smorag Z., Pienkowski M., Slomski R.
Biotechnologia
|
2003
|
issue 1
48-73
EN
The genetic construct WAP 6xHisHGH containing the gene encoding human growth hormone (hGH) and WAP promoter expressed in mammary gland of animals was prepared. The 5? end of the gene was modified by the addition of sequence encoding six histidine residues and the sequence recognized by thrombin. In this way, the growth hormone can be easily purified by affinity chromatography and cleaved with thrombin to an active form. In the next step, the genetic construct was introduced by microinjection into male pronuclei of fertilized oocytes. Transgene was detected in male rabbit of F0 generation (number 61). Twelve offspring of founder rabbit of generation F1 indicated transgene sequences. The presence of growth hormone was revealed in the samples of milk accumulated during the lactation of females of F1 generation. The genetic constructs containing chain 1 and chain 2 of Feld1, and the major allergen produced by cat (Fedlis domesticus) were prepared. Both genes were inactivated by introduction into the sequences a positive selectable marker aminoglycoside phosphotransferase (resistant to neomycin). Outside the region of homology to Feld1 chain 1 and chain 2 genes, the negative selectable marker ? thymidine kinase gene was introduced. The genetic constructs pNTKFd1 and pNTKFd2 can be used in further experiments involving the inactivation of Feld1 genes in cat cells. Both genes were modified by site-directed mutagenesis using megastarter with Stop codon for premature termination of translation. The presence of mutation was confirmed by sequencing. The genetic constructs with human hGH gene and cat Feld1 gene were introduced into the bovine and cat fetal fibroblasts respectively in co-transfection with plasmid pGT-N29 containing positive selectable marker by lipofection, precipitation and electroinjection methods. After the selection, surviving cells were subjected to further molecular analysis. The stabile incorporation of the genetic constructs WAP 6xHisHGH and WAPHGH into the genome were observed.
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