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Biomedical subjects

B Gajda

Publications and source records attributed to B Gajda.

16 recordsLinked to original sources

Generation of transgenic rabbits by the novel technique of chimeric somatic cell cloning.

A novel technique of chimeric somatic cell cloning was applied to produce a transgenic rabbit (NT20). Karyoplasts of transgenic adult skin fibroblasts with Tg(Wap-GH1) gene construct as a marker were microsurgically transferred into one, previously enucleated, blastomere of 2-cell non-transgenic embryos, while the second one remained intact. The reconstructed embryos either were cultured in vitro up to the blastocyst stage (Experiment I) or were transferred into recipient-females immediately after the cloning procedure (Experiment II). In Experiment I, 25/102 (24.5%) embryos formed blastocysts from whole embryos and 46/102 (44.12%) embryos developed to the blastocyst stage from single non-operated blastomeres, while the reconstructed blastomeres were damaged and degenerated. Thirteen (12.7%) embryos did not exceed 3- to 4-cell stages and 18 (17.7%) embryos were inhibited at the initial 2-cell stage. Out of 14 blastocysts which were subjected to molecular analysis, the transgene was detected in the cells of 4 blastocysts. In Experiment II, 163/217 (75.0%) embryos were transferred into 9 pseudopregnant recipient-rabbits (an average of 18 embryos per recipient). Four recipient-females (44.4%) became pregnant and delivered a total of 24 (14.7%) pups. Molecular analysis confirmed that two pups (1.2%), one live and one stillborn, showed a positive transgene signal. Live transgenic rabbit NT20 appeared healthy and anatomically as well as physiologically normal. The results of our experiments showed that transgenic adult skin fibroblast cell nuclei, which have been introduced into the cytoplasmic microenvironment of single enucleated blastomeres from 2-cell stage rabbit embryos, are able to direct the development of chimeric embryos not only to the blastocyst stage but also up to term.

Animals↗

[Potter sequence in a neonate with bilateral renal dysplasia with genetic aspects].

The authors describe a neonate with Potter sequence due to bilateral renal dysplasia and discuss its genetic implications. Although this congenital anomaly of the kidneys may occur sporadically, it is now accepted that this condition has an autosomal dominant inheritance pattern with a penetrance of 50-90% and variable expression. Potter sequence is not compatible with extrauterine life; therefore, in case of maternal oligohydramnios, ultrasound should be performed in the second trimester in order to make a prenatal diagnosis, as well as to permit genetic counseling. As Potter sequence is frequently associated with clinically silent anomalies of the kidneys, ultrasound examination of the kidneys and urinary tract should also be performed in all family members.

Abnormalities, Multiple↗

[Criminal contamination by metallic mercury of cow's silage].

This paper presents an unsuccessful homicidal attempt on 23 persons by means of metallic mercury added in large amount to cow's silage. Thirteen adults and twelve children used milk from these cows for 4.5 months. The mercury concentration in the milk of these cows as well as in serum and daily urine of consumers of the milk were examined by atomic absorption spectrophotometry. Despite the increased level of mercury in the milk (average 0.023 mg/kg), the concentrations of mercury in daily urine of only three persons were raised about 50%. These people had been working in a cowshed without ventilation every day for many months and thus had been exposed to vaporised mercury.

Adult↗

Cryopreservation of mammalian ova and embryos by vitrification.

Vitrification is a new approach to oocyte and embryo cryoconservation. It consists in the solidification of a solution caused not by crystallization, but by a drastic increase in viscosity during cooling. 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.

Journal Article↗

Factors affecting the survival of one- and two-cell rabbit embryos cryopreserved by vitrification.

The effects of equilibration time, glycerol (GLY), and 1,2-propanediol (PROH) concentration, and of vitrification and sucrose solution on the viability of 1- and 2-cell rabbit embryos were investigated. After collection, the embryos were equilibrated for 5 or 10 minutes in phosphate buffered saline (PBS) containing 10% GLY-20% PROH and were exposed for 30 seconds at 4 degrees C or were exposed and vitrified in one of two vitrification solutions 35% GLY-35% PROH or 20% GLY-50% PROH. The in vitro survival rates of 1-cell embryos equilibrated for both 5 and 10 minutes were lower (34.0 and 48.0%, respectively) than those of 2-cell embryos (78.8 and 68.5%, respectively; P<0.01). No differences were noted in the viability of embryos exposed to the 2 vitrification solutions. Following vitrification in a mixture of 35% GLY-35% PROH, the survival rates of 1- and 2-cell embryos were 18.3 and 13.7% and 19.6 and 10.4% for 5 and 10 minutes of equilibration, respectively. The survival rates of 1- and 2-cell embryos vitrified in a solution of 20% GLY-50% PROH were 25.7 and 35.4% and 26.2 and 21.3% for 5 and 10 minutes of equilibration, respectively. The survival rates of 1-and 2-cell embryos stored in 1M sucrose solution were 63.8 and 84.0%, respectively. In conclusion, the viability of vitrified 1- and 2-cell rabbit embryos was reduced as a consequence of their equilibration before vitrification, the exposure to vitrification solution and the dilution in a sucrose solution rather than of the vitrification process itself.

Journal Article↗

The effect of sucrose and trehalose on viability of one- and two-cell rabbit embryos.

The effect of sucrose and trehalose on the viability of one- and two-cell rabbit embryos was investigated. A significant decrease in the viability of one- and two-cell embryos exposed for 30 min. at 20 degrees C was observed. At 38 degrees C none of the two-cell embryos in a sucrose solution survived after 30 min exposure, while approximately 50% of the embryos survived in a trehalose solution. The cleveage rate in culture of two-cell embryos exposed both to 2.0 M or 1.45 M trehalose was significantly lower in comparison with the control group. However the survival rate after transfer of two-cell embryos exposed to 1.45 M trehalose solution at 20 degrees C remained the same as that of the control group.

Journal Article↗

Stage-dependent viability of vitrified rabbit embryos.

The aim of the work was to determine the susceptibility of rabbit embryos to vitrification at different developmental stages. The experiment was carried out on 676 embryos at 1-, 2- and 8-to 16-cell stages as well as the morula and blastocyst stages. As a vitrification medium, a mixture of 30% 1,2-propanediol + 30% glycerol (Solution I), or 35% 1,2-propanediol + 35% glycerol (Solution II), was used. The embryos were frozen in glass ampules placed in nitrogen vapour for 5 min before being plunged into liquid nitrogen. Dilution after rapid thawing was done in one step in a 1-M sucrose solution. After vitrification in Solution I, none of the 1- or 2-cell embryos survived, whereas the survival rate of 8-to 16-cell embryos, morula and blastocysts, was 23.0, 82.7 and 78.5%, respectively. After vitrification in Solution II, the survival rate of 1-, 2- and 8-to 16-cell embryos was 20.0, 43.8 and 92.9%, respectively. The proportion of live offspring on the Day 28 after transfer of 68 vitrified morula was 26.5% compared with 24.0% in the control group. Thus, the proposed vitrification procedures can be useful in the cryopreservation of rabbit embryos.

Journal Article↗

Effect of dexamethasone and hydrocortisone on the course of superovulation in cattle.

The effect of hydrocortisone and dexamethasone on superovulation was examined in 12 cows. On the day PMSG was given, each animal received either the first of five daily doses of 250 mg succinate hydrocortisone or one injection of 30 mg dexamethasone. In the 48-hr interval between the injection of PMSG and PGF(2)alpha, the concentration of progesterone rose from 6.97 to 10.22 ng/ml in the experimental groups and only to about 2.8 ng/ml in the control group. In the following days progesterone increased even more, from 15.7 to 26.0 ng/ml seven days after estrus in the experimental group and to 19.25 ng/ml in the control group. The group which received dexamethasone had an average of 4.7 corpora lutea and one embryo flushed per animal. The hydrocortisone group had an average of 2.5 corpora lutea and one cow had two embryos. The control group had 6.2 corpora lutea and 5.2 embryos per animal.

Journal Article↗

Survival of pig morula and blastocyst after exposure to vitrification media or vitrification.

The aim of this experiment was to examine the survival of porcine embryos following exposure to vitrification solutions and vitrification. The work was carried out on non-cultured and cultured morulae and blastocysts. The viability of treated embryos was assessed by their ability to develop in in vitro culture. The results showed that the most detrimental step in the vitrification of pig embryos is exposing them specifically to a vitrification medium rather than the vitrification process itself. Moreover, we demonstrated the beneficial effect of culture on the viability of pig embryos after vitrification

Journal Article↗