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Wathes D.C.,Reproduction Group | Cheng Z.,Reproduction Group | Fenwick M.A.,Reproduction Group | Fenwick M.A.,Hammersmith Hospital | And 3 more authors.
Reproduction | Year: 2011

Postpartum dairy cows enter a period of negative energy balance (NEB) associated with low circulating IGF1, during which the uterus must undergo extensive repair following calving. This study investigated the effects of NEB on expression of IGF family members and related genes in the involuting uterus. Cows were allocated to two treatments using differential feeding and milking regimes to produce mild NEB or severe NEB (SNEB). Uterine endometrial samples collected 2 weeks post partum were analysed by quantitative PCR. The expression of IGF-binding protein 4 (IGFBP4) mRNA increased in the endometrium of SNEB cows, with trends towards increased IGFBP1 and reduced IGFBP6 expression. There were no significant differences between treatments in mRNA expression of IGF1, IGF2 or of any hormone receptor studied, but significant correlations across all cows in the expression levels of groups of receptors suggested common regulatory mechanisms: type 1 IGF receptor (IGF1R), IGF2R and insulin receptor (INSR); GHR with ESR1; and ESR2 with NR3C1. The expression of IGF1R and INSR also positively correlated with the circulating urea concentration. Matrix metalloproteinases (MMPs) are important in tissue remodelling and can affect IGF signalling via interaction with IGFBPs. The expression levels of MMP1, MMP3, MMP9 and MMP13 mRNAs all showed major upregulation in the endometrium of cows in SNEB and all except MMP9 were highly correlated with expression of IGFBP4. Alpha(2)-HS-glycoprotein (AHSG) and PDK4, two genes implicated in insulin resistance, were also highly expressed in SNEB. These results suggest that cows in SNEB experience alterations to the IGF and insulin signalling pathways in the postpartum endometrium. This may affect the rate of tissue repair with a possible negative impact on subsequent fertility. © 2011 Society for Reproduction and Fertility.


Jopcik M.,Slovak Academy of Sciences | Bauer M.,Animal Production Research Center | Bauer M.,Constantine the Philosopher University | Moravcikova J.,Slovak Academy of Sciences | And 3 more authors.
Plant Cell, Tissue and Organ Culture | Year: 2013

Plant transgenesis often requires the use of tissue-specific promoters to drive the transgene expression exclusively in targeted tissues. Although the eukaryotic promoters are expected to stay silent in Escherichia coli, when the promoter-transgene units within the plant transformation vectors are constructed and propagated, some eukaryotic promoters have been reported to be active in prokaryotes. The potential activity of plant promoter in E. coli cells should be considered in cases of expression of proteins that are toxic for host cells, environmental risk assessment or the stability in E. coli of plant vectors for specific Cre/loxP applications. In this study, DNA fragments harbouring four embryo- and/or pollen-specific Arabidopsis thaliana promoters were investigated for their ability to drive heterologous gene expression in E. coli cells. For this, they were fused to gfp:gus reporter genes in the pCAMBIA1304 vector. Although BPROM, bacterial sigma70 promoter recognition program identified several sequences with characteristics similar to bacterial promoters including -10 and -35 sequences in each of tested fragments, the experimental approach showed that only one promoter fragment was able to drive relatively strong- and one promoter fragment relatively weak-GUS expression in E. coli cells. Remaining two tested promoters did not drive any transgene expression in bacteria. Our results also showed that cloning of a shorter plant promoter sequence into vectors containing lacZ α-complementation system can increase the probability of gene expression driven by upstream located lac promoter. This should be considered when cloning of plant expression units, the expression of which is unwanted in E. coli. © 2012 Springer Science+Business Media Dordrecht.


Fadol S.R.,Animal Production Research Center | Babiker S.A.,University of Khartoum
Livestock Research for Rural Development | Year: 2010

Twenty four Sudan Baggara Zebu bulls were divided into two groups of similar body weight (245kg ± 8.08) and fed molasses based diet for a pre-experimental period of two weeks followed by a feeding period of 12 weeks At the end of this pre-experimental periods the two animals groups were randomly assigned to the feed management, which was ad libtum feeding (group A) or restricted feeding (group B). Group (A) was offered the diet ad libitum for the whole feeding period. Feeding in group (B) was offered into four interchangeable periods of three weeks each where feed was offered 50% of ad libitum for 3 weeks followed by ad libitum feeding for the next 3 weeks and so on. There were no differences between the two groups in final weight, total gain and daily gain. Live weight increased linearly in the ad libitum fed but not in the restricted fed bull groups. Daily dry matter intake and total dry matter intake were respectively greater (p<0.00119) and (p<0.0119) in ad libitum fed bulls than in restricted bulls. Slaughter and empty body weights were not different in the two groups. Hot and cold carcass weights were respectively greater (P<0.0413) and (p<0.0386) in the ad libitum fed group than in the restricted bulls. Muscle percentage was also greater (p<0.00461)) while bone and fat percentages increased slightly in ad libitum fed bulls. Muscle to bone ratio was not different between the two feeding groups. Dressing percentage did not differ between the two groups but the ad libitum fed group dressed slightly heavier than the restricted group. Longissismus dorsi muscle area was not different between the two groups. The area of the muscle was 61.07 cm2 in carcasses of the ad libitum fed group and 49.15 cm2 in the restricted bulls. The results indicated that non-carcass components showed no significant differences between the two groups except for the weight of head which was heavier (p<0.0362) in restricted bulls than in ad libitum fed bulls. The wholesale cuts showed no different between the two groups.


Tongel P.,Animal Production Research Center | Broucek J.,Animal Production Research Center
American Society of Agricultural and Biological Engineers Annual International Meeting 2013, ASABE 2013 | Year: 2013

SCC counter which is suitable for somatic cell count (SCC) measurement at individual cow has been developed in our Institute. It consists of metal box with electronics and servomotor. There is a mechanical arm on the front size of it, where a transparent glass tube with running metal ball is placed. SCC counter works on the basis of milk viscosity measurement. After the glass tube is filled with milk mixed with reagent, the tube declines for certain time (20 s) from the horizontal position to the angle of 25°. The metal ball moves downwards and it's velocity depend on the density of milk. The length of the ball's path is inverse proportional to the viscosity of the milk. Near the measurement tube the scale is placed, which shows the SCC. Higher viscosity means higher SCC. Three types of equipment were developed. The first has short measurement tube /20 cm/ and it is suitable for quick measurement, when the accuracy is not so important. Second one has longer measurement tube /50 cm/ and is suitable for more precise measurement. The third equipment has four long measurement tubes on the same arm and can measure four samples at once. This type of equipment can be regularly used for measurement of milk from each quarter of the udder. In an experiment we tested the equipment for its accuracy and reliability in practical conditions. Samples of milk were taken from True - test during milking for SCC measurement. Three control ones of the same milk were sent to three different certificated laboratories for SCC analysis. Total two hundred samples have been measured. The average value of SCC from these three laboratories was calculated. The correlation coefficient between SCC from SCC counter and the average value from laboratories was computed (r = 0.96, p< 0.01). The differences between data from laboratories were higher than data from SCC counter and average SCC. It was found out that the SCC counter can measure SCC in a range 150 000 to 10 000 000 with sufficient accuracy. The reliability was evaluated too and it has shown suitable for practical use. This research has been done with financial support "MLIEKO No. 26220220098" supported by the Operational Program Research and Development funded from the European Regional Development Fund and project APVV-0632-10 of the Slovak Research and Development Agency Bratislava.


Tongel P.,Animal Production Research Center | Broucek J.,Animal Production Research Center
American Society of Agricultural and Biological Engineers Annual International Meeting 2013, ASABE 2013 | Year: 2013

Rapid electronic mastitis test (REM test), which is suitable for quick mastitis monitoring of the herd of dairy cows, has been developed in our Institute. It consists of five measurement chambers, where four of them are for milk electrical conductivity measurement and the fifth, the common, is for temperature measurement. The third generation of REM test was designed and constructed in this year. It was found out in our previous experiments, that the accuracy of milk electrical conductivity measurement is crucial for mastitis indication. From this reason the measurement of accuracy and reliability has to be done. The accuracy of measurement was measured at first in laboratory conditions, and then the accuracy and reliability were evaluated in practical conditions. The results have shown the accuracy was excellent (1% from measurement range) and the reliability was suitable for practical use. The statistical value of electrical conductivity of healthy cows was 4.8 mS/cm (sd 0.4 mS/cm) and cows with mastitis was 6.4 mS/cm (sd 0.9 mS/cm). The difference between them was significant (p<0.01). In the experiment in practical conditions we have found out that absolute EC values of healthy cow can change up to 10 mS/cm between milking but the differences of EC between quarters are lesser than 0.5 mS/cm. Absolute value of milk EC of single quarter has substantially lesser sensitivity in mastitis monitoring then IQD (inter quarter difference). The differences of EC between quarters in the healthy udder were under 0.5 mS/cm. If the difference of EC of some quarter in the udder is higher than 0.5 mS/cm, than the EC of the quarter with the lowest value then this quarter with higher EC is suspicious of mastitis. When EC values of all quarters in the udder are higher than 6.0 mS/cm and differences between quarters are lesser than 0.5 mS/cm, than the cow is suspicious of another illness than mastitis. The higher absolute values the higher suspicious of illness. The correlation coefficient between absolute value of EC and SCC of single quarters was r = 0.63. The sensitivity of the mastitis detection by means of milk EC measurement was calculated as 87% and the reliability 92%. It was evaluated for IQD with threshold difference 0.5 mS/cm and for cows with mastitis which have SCC>500 000. On the basis of our findings and our previous experiments we can say the using of REM test in mastitis monitoring is suitable for daily monitoring of dairy cows especially for cows on their first lactation. This research has been done with financial support "MLIEKO No. 26220220098" supported by the Operational Program Research and Development funded from the European Regional Development Fund and project APVV-0632-10 of the Slovak Research and Development Agency, Bratislava.


Sirotkin A.V.,Animal Production Research Center | Sirotkin A.V.,Constantine the Philosopher University | Laukova M.,Slovak Academy of Sciences | Ovcharenko D.,Altogen Biosystems | And 2 more authors.
Journal of Cellular Physiology | Year: 2010

Previous studies have shown that microRNAs (miRNAs) can control steroidogenesis in cultured granulosa cells. In this study we wanted to determine if miRNAs can also affect proliferation and apoptosis in human ovarian cells. The effect of transfection of cultured primary ovarian granulosa cells with 80 different constructs encoding human pre-miRNAs on the expression of the proliferation marker, PCNA, and the apoptosis marker, Bax was evaluated by immunocytochemistry. Eleven out of 80 tested miRNA constructs resulted in stimulation, and 53 miRNAs inhibited expression of PCNA. Furthermore, 11 of the 80 miRNAs tested promoted accumulation of Bax, while 46 miRNAs caused a reduction in Bax in human ovarian cells. In addition, two selected antisense constructs that block the corresponding miRNAs mir-15a and mir-188 were evaluated for their effects on expression of PCNA. An antisense construct inhibiting mir-15a (which precursor suppressed PCNA) increased PCNA, whereas an antisense construct for mir-188 (which precursor did not change PCNA) did not affect PCNA expression. Verification of effects of selected pre-mir-10a, mir-105, and mir-182 by using other markers of proliferation (cyclin B1) and apoptosis (TdT and caspase 3) confirmed specificity of miRNAs effects on these processes. This is the first direct demonstration of the involvement of miRNAs in controlling both proliferation and apoptosis by ovarian granulose cells, as well as the identification of miRNAs promoting and suppressing these processes utilizing a genome-wide miRNA screen. © 2009 Wiley-Liss, Inc.


Krupova Z.,Animal Production Research Center | Krupa E.,Institute of Animal Science | Wolfova M.,Institute of Animal Science
Czech Journal of Animal Science | Year: 2013

The impact of variation in economic conditions on the economic values of fourteen production and functional traits was examined for the Improved Valachian breed using a bio-economic model implemented in the ECOWEIGHT software. The following economic parameters were investigated: market prices of lambs, milk, and cheese (variation ± 40%), costs for roughage, concentrates, and total feeding rations , costs for labour and veterinary care, fixed costs (variation ± 20% for all costs), and discount rate of revenues and costs (0 and 3%). Results of the analyses were presented in detail for the marginal and relative economic values of the four most important traits: milk yield in the 150-day milking period, conception rate of ewes, litter size per lambed ewe, and productive lifetime of ewes. Furthermore, cumulative relative economic values of the four trait complexes - milk production, growth, functional, and wool traits - were presented. Prices for sheep products were found to be the most important factor for both the marginal and the relative economic values of the evaluated traits. The four traits with the highest relative economic values in the base calculation stayed the most important for all investigated economic parameters ranges. The relative economic values of the remaining 10 traits did not exceed 6.1%. The relative economic values of milk yield and litter size were the most sensitive to the variation in economic circumstances. For the investigated range of economic parameters, the relative economic value for the complex of milk production traits ranged 30.6-48.1%, for growth traits 6.3-9.4%, and that for functional traits 45.4-59.7%. The relative economic value for the wool trait did not exceed 0.3%. © 2011 Czech Academy of Agricultural Sciences.


Keady T.W.J.,Animal Production Research Center | Hanrahan J.P.,Animal Production Research Center
Grass and Forage Science | Year: 2010

The effects of allowance of extended (deferred) grazed herbage (AEGH) and herbage allocation management (HAM) were studied in ewe lambs (248) and late-gestation ewes (152), respectively, on commercial farms in south-east Ireland in 2005-06. In Experiment 1, which consisted of four treatments, the effects of AEGH (0·75, 1·25 and 1·75 kg DM per head daily) and concentrate supplementation (0·5 kg per head daily with the 0·75 kg DM herbage allowance) on lamb performance during the extended grazing (16 December to 3 March) and subsequent grazing (4 March to 11 August) seasons were evaluated. Increasing AEGH increased herbage intake linearly (P < 0·001) and live weight (P < 0·001) at the end of extended grazing and the subsequent grazing season. In Experiment 2, single- and twin-bearing ewes were allocated to either a conventional (single- and twin-bearing ewes grazed separately) or leader-follower system (twin- and single-bearing ewes, as leaders and followers respectively) of HAM from 30 January to 24 March. The same quantities of herbage were offered daily for each system. System of HAM affected ewe condition score at lambing but did not alter (P > 0·05) subsequent lamb birth or weaning weights. It is concluded that increasing AEGH to ewe lambs increased liveweight gain during extended grazing and resulted in heavier animals in mid August of the subsequent grazing season. For ewe lambs each 1 kg concentrate DM had the same feed value as 2·4 kg DM AEGH. Use of a leader-follower system for ewes in late pregnancy did not alter lamb birth weight or subsequent performance. © 2010 Blackwell Publishing Ltd.


The aim of the present study was to understand the interrelationships between stress, hormones and basic ovarian functions in the ovary. For this purpose, we compared the expression of markers of proliferation (PCNA, cyclin B1), of apoptosis (Bax, caspase-3) and secretory activity (release of progesterone, P4, and insulin-like growth factor, IGF-I) in whole ovarian follicles and granulosa cells cultured in conditions of normal temperature (37.5°C) and feeding (with serum), high temperature (41.5°C, with serum) and malnutrition (37.5°C, without serum), with and without hormones [IGF-I, leptin and follicle-stimulating hormone (FSH)]. The expression of proliferation and apoptosis markers was evaluated by SDS PAGE-western blotting whereas radioimmunoassay (RIA) measured the release of hormones. High temperature dramatically induced a reduction in both proliferation and apoptosis markers in both ovarian follicles and granulosa cells and induced a significant increase in P4 and IGF-I release by ovarian granulosa cells but not in P4 secretion by ovarian follicles. Serum deprivation increased accumulation of cyclin B1 but not other markers of proliferation (PCNA) and apoptosis (Bax, caspase-3) or P4 release in ovarian follicles. On the contrary, it inhibited the expression of apoptotic marker (Bax), release of both P4 and IGF-I but it did not affect proliferation marker (PCNA) in granulosa cells. Adding IGF-I, leptin and FSH affected proliferation, apoptosis and secretory activity of ovarian cell functions but also prevented an inhibitory effect of high temperature on the expression of Bax and PCNA and an inhibitory action of serum deprivation on PCNA in ovarian follicles. Furthermore, treatment with these hormones prevented an inhibitory action of thermal stress on Bax, PCNA, P4 and IGF-I in ovarian granulosa cells. The present observations (1) confirm the involvement of hormones (IGF-I, leptin and FSH) in the control of proliferation, apoptosis and secretory activity of ovarian cells, (2) demonstrate for the first time that heat stress/increased temperature can induce a reduction in ovarian cell proliferation and apoptosis and an oversecretion of ovarian hormones, (3) show that malnutrition/serum deprivation can reduce both apoptosis and secretory activity of ovarian cells, (4) demonstrate the differences in the response of granulosa and other ovarian follicular cells to stresses, and (5) are the first demonstration that hormones (IGF-I, leptin and FSH) could be used for preventing the effect of stresses on ovarian cell functions. © 2010. Published by The Company of Biologists Ltd.


Uhrin P.,Medical University of Vienna | Zaujec J.,Medical University of Vienna | Breuss J.M.,Medical University of Vienna | Olcaydu D.,Medical University of Vienna | And 10 more authors.
Blood | Year: 2010

During embryonic development, lymph sacs form from the cardinal vein, and sprout centrifugally to form mature lymphatic networks. Separation of the lymphatic from the blood circulation by a hitherto unknown mechanism is essential for the homeostatic function of the lymphatic system. O-glycans on the lymphatic endothelium have recently been suggested to be required for establishment and maintenance of distinct blood and lymphatic systems, primarily by mediating proper function of podoplanin. Here, we show that this separation process critically involves platelet activation by podoplanin. We found that platelet aggregates build up in wild-type embryos at the separation zone of podoplanin+ lymph sacs and cardinal veins, but not in podoplanin -/- embryos. Thus, podoplanin -/- mice develop a "nonseparation" phenotype, characterized by a blood-filled lymphatic network after approximately embryonic day 13.5, which, however, partially resolves in postnatal mice. The same embryonic phenotype is also induced by treatment of pregnant mice with acetyl salicylic acid, podoplanin-blocking antibodies, or by inactivation of the kindlin-3 gene required for platelet aggregation. Therefore, interaction of endothelial podoplanin of the developing lymph sac with circulating platelets from the cardinal vein is critical for separating the lymphatic from the blood vascular system. © 2010 by The American Society of Hematology.

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