Application of disease-associated differentially expressed genes – Mining for functional candidate genes for mastitis resistance in cattle

In this study the mRNA differential display method was applied to identify mastitis-associated expressed DNA sequences based on different expression patterns in mammary gland samples of non-infected and infected udder quarters of a cow. In total, 704 different cDNA bands were displayed in both udder samples. Five hundred-and-thirty two bands, (75.6%) were differentially displayed. Ninety prominent cDNA bands were isolated, re-amplified, cloned and sequenced resulting in 87 different sequences. Amongst the 19 expressed sequence tags showing a similarity with previously described genes, the majority of these sequences exhibited homology to protein kinase encoding genes (26.3%), to genes involved in the regulation of gene expression (26.3%), to growth and differentiation factor encoding genes (21.0%) and to immune response or inflammation marker encoding genes (21.0%). These sequences were shown to have mastitis-associated expression in the udder samples of animals with and without clinical mastitis by quantitative RT-PCR. They were mapped physically using a bovine-hamster somatic cell hybrid panel and a 5000 rad bovine whole genome radiation hybrid panel. According to their localization in QTL regions based on an established integrated marker/gene-map and their disease-associated expression, four genes (AHCY, PRKDC, HNRPU, OSTF1) were suggested as potentially involved in mastitis defense

2003b. Abnormal segregation of prion protein octapeptiderepeat alleles in cattle

Abstract. The study was conducted on full-families of Black-and-White cattle obtained as 25 AI sire families and 355 cows, as well as their progenies, mostly heifers at the age of 1-3 months. The sire group was composed by the casual qualification of 10 PRNP 6/6 and 15 PRNP 6/5 individuals on the basis of accessible young progenies. The randomly selected group of cows is characterised by a very high frequency of PRNP 6/6 (74.9%), followed by lower frequency of PRNP 6/5 (24.5%) and a very low frequency of PRNP 5/5 genotype (0.6%). The progenies represent all expected genotypes, such as: PRNP 6/6 (60.5%), PRNP 6/5 (35.8%) and PRNP 5/5 (3.7%), respectively. Taking into consideration the genotypes of parents and progenies, the segregation of PRNP 6 and PRNP 5 alleles was analysed. Results of the non-informative mating variant of % PRNP 6/6 × & PRNP 6/6 (n = 87) are affected by the PRNP 6/6 progeny genotype in all cases. Subsequently, the results of mating variants % PRNP 6/6 × &PRNP 6/5 (n = 29) and % PRNP 6/5 & PRNP 6/6 (n = 179) showed statistically non-significant differences in both above-mentioned alternations. The progeny group related from % PRNP 6/5 × & PRNP 6/5 parental mating obtained fully informative and most valuable results based on the presented research concept. In the common group of 58 calves, the genotype PRNP 6/6 is represented by 26 individuals (44.8 %), PRNP 6/5 -by 19 individuals (32.8 %) and PRNP 5/5 -by 13 individuals (22.4 %). Therefore, the theoretical genotype rate (25% : 50% : 25%) is drastically deformed and the differentiation between the observed and expected numbers of animals is statistically highly significant (c 2 = 12.72; 2 df.). These differences are affected by two times higher PRNP 6/6 homozygous (c 2 = 9.12; 1 df.) and responsively by the low number of PRNP 6/5 heterozygous animals (c 2 = 3.45; 1 df.). Further investigations are carried out to explain the genetic determination of abnormal PRNP octa-peptide repeat allele segregation, which suggests possible lethal cis-trans linkage effects

Application of disease-associated differentially expressed genes – Mining for functional candidate genes for mastitis resistance in cattle

In this study the mRNA differential display method was applied to identify mastitis-associated expressed DNA sequences based on different expression patterns in mammary gland samples of non-infected and infected udder quarters of a cow. In total, 704 different cDNA bands were displayed in both udder samples. Five hundred-and-thirty two bands, (75.6%) were differentially displayed. Ninety prominent cDNA bands were isolated, re-amplified, cloned and sequenced resulting in 87 different sequences. Amongst the 19 expressed sequence tags showing a similarity with previously described genes, the majority of these sequences exhibited homology to protein kinase encoding genes (26.3%), to genes involved in the regulation of gene expression (26.3%), to growth and differentiation factor encoding genes (21.0%) and to immune response or inflammation marker encoding genes (21.0%). These sequences were shown to have mastitis-associated expression in the udder samples of animals with and without clinical mastitis by quantitative RT-PCR. They were mapped physically using a bovine-hamster somatic cell hybrid panel and a 5000 rad bovine whole genome radiation hybrid panel. According to their localization in QTL regions based on an established integrated marker/gene-map and their disease-associated expression, four genes (AHCY, PRKDC, HNRPU, OSTF1) were suggested as potentially involved in mastitis defense

Rehabilitation after one-stage anterior cruciate reconstruction and osteochondral grafting

At least 10–20% of all ACL reconstructions require additional cartilage repair. The aim of this study was to compare the activity recovered by patients after one-stage open ACL reconstruction and osteochondral autologous grafting of articular cartilage lesions and after isolated open ACL reconstruction. The study group included 21 patients with chronic ACL deficiency and grade III or IV cartilage lesion according to the ICRS scale who were treated with combined ACL reconstruction and osteochondral grafting in one step. The control group included 32 patients with chronic ACL insufficiency and no chondral deficit higher than grade I on the ICRS scale who underwent isolated reconstruction of the ligament. For the assessment, the Lysholm and Gillquist (L&G) score and the functional Marshall score were used. Both groups displayed a statistically significant improvement in the L&G score and the Marshall score between the preoperative and 12-month assessments. The mean gain in L&G score over this period was 30.66±7.79 in the study group and 31.65±6.96 in the control group. The difference between the control group and the study group was not significant. The difference between 12 months and initial assessment was counted. The mean gain in Marshall score was 9.05±3.81 in the study group and 10.71±3.43 in the control group. The difference between the initial and the 12-month evaluation was statistically significant (p=0.49). Return to normal activity was slower and patient satisfaction was lower during the first year after operation in the study group than in the control group, however the overall advantage of the one-step operation outweighs the slightly inferior functional results at 12 months

Comparison of bioabsorbable interference screws and posts for distal fixation in anterior cruciate ligament reconstruction

Comparison of the results of bioabsorbable interference screws and posts for hamstring graft distal fixation in ACL reconstructions are presented. The results of 20 patients with bioabsorbable screws were compared to 22 patients with posts. The assessement was based on Lysholm-Gillquist and Marshall scores and the KT-1000 device. In the study group the points gained were 38.9 in the Lysholm-Gillquist and 12.89 in the Marshall scale. The average KT-1000 difference was 2.46 mm. In the control group the points gained were 32.93 in the Lysholm-Gillquist and 11.47 in the Marshall scale. The average KT-1000 difference was 2.5 mm. There were 14 patients in the study group with interference screw problems; in 2 the implants were removed. (1) There are no differences in outcome using bioabsorbable interference screws and posts for distal fixation of hamstring ACL grafts. (2) The lack of bioabsorbtion with poly L-lactide interference screws is frequent and causes problems