Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Understanding the site of action of photosystem II inhibitors

It is believed that the different classes of compound which interfere with photosynthesis system II (PS 11) must act by binding at different sites, because they appear to be structurally so dissimilar. However, it can be argued that some of the classes of compound of diverse structural type share very similar external surface features. Now that details about the binding oj PSII inhibitors are becoming available, it will be possible to see how many of the different compound classes actually bind similarly. In future, increasingly detailed knowledge of the structures of potential sites of action will be available. QSAR methods will need to be used together with molecular modelling to be effective tools for agrochemical design.     

An economically based evaluation index for perennial and short‐term ryegrasses in New Zealand dairy farm systems

Grazed pastures based on ryegrass species provide most of the feed for dairy cattle in New Zealand. There are many cultivars of perennial (Lolium perenne), annual and Italian (L. multiflorum), and hybrid (L. boucheanum) ryegrasses available for dairy farmers to use in pasture renewal. This study describes an index which ranks ryegrass cultivars relative to a genetic base according to the estimated economic value (EV) of seasonal dry matter (DM) traits. A farm system model was used to derive EVs (Grazed pastures based on ryegrass species provide most of the feed for dairy cattle in New Zealand. There are many cultivars of perennial (Lolium perenne), annual and Italian (L. multiflorum), and hybrid (L. boucheanum) ryegrasses available for dairy farmers to use in pasture renewal. This study describes an index which ranks ryegrass cultivars relative to a genetic base according to the estimated economic value (EV) of seasonal dry matter (DM) traits. A farm system model was used to derive EVs ($ ha^?1 calculated as change in operating profit divided by unit change of the trait) for additional DM produced in different seasons of the year in four regions. The EV of early spring DM was consistently high across all regions, whereas EV for late spring DM was moderate to low. Genotype × environment analysis revealed significant reranking of DM yield among ryegrass cultivars across regions. Hence, separate performance values (PVs) were calculated for two mega‐environments and then combined with the corresponding season and region EV to calculate the overall EV for twenty‐three perennial ryegrass and fifteen short‐term ryegrass cultivars. The difference in operating profit between the highest ranked and lowest ranked perennial ryegrass cultivar ranged from $556 ha^?1 to $863 ha^?1 year^?1 depending on region. For short‐term ryegrasses used for winter feed, the corresponding range was $394 to $478 ha^?1 year^?1. Using PV for DM yield, it was estimated that plant improvement in perennial ryegrass has added $12–$18 ha^?1 year^?1 (depending on region) operating profit on dairy farms since the mid‐1960s.     

Erosion rates near rome, Italy

Before man's intensive use of the land the rates of erosion near Rome, Italy, were approximately 2 to 3 centimeters per thousand years. With intensive occupation by man the rates have increased by an order of magnitude.     

Monensin controlled-release intraruminal capsule for control of bloat in pastured dairy cows

Monensin, a polyether ionophore antibiotic, is potentially an important agent for bloat relief in dairy cows grazing temperate legume-based pasture. A series of studies was undertaken to determine the effect of monensin, when delivered continuously in the rumen of lactating dairy cows by means of controlled-release capsules (monensin CRC). Such devices release approximately 300 mg/head/day for 100 d. A short-term pilot study made at Ruakura, New Zealand, tested monensin CRC in cows selected for high susceptibility to bloat and grazing lucerne (Medicago sativa) or red clover (Trifolium pratense). Treatment significantly reduced the incidence of bloat, while milk yield and protein yield were increased. There was no effect on fat yield. Following the pilot study, 6 large-scale field experiments involving a total of 368 lactating dairy cows, were made in Australia and New Zealand to confirm the effectiveness of monensin CRC for bloat control and to measure the effect of such treatment on milk production and composition. A severe bloat problem occurred in 2 experiments, mild bloat occurred in 2 others, while no visual signs of bloat were observed in the remaining 2 experiments. Bloat was significantly (P less than 0.05) reduced by monensin CRC treatment when data was pooled over the 4 experiments in which bloat occurred. Daily milk yield was increased in all experiments from a mean of 17.7 in untreated groups to 18.8 kg/head/day (P less than 0.05) in monensin CRC-treated cows. Protein percentage was not affected by treatment, while there was a decrease from 4.29 to 4.10% fat, although total fat yield was not affected.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of Hormonal Supplementation on Nuclear Maturation and Cortical Granules Distribution of Canine Oocytes During Various Reproductive Stages

The aim of this study was to evaluate the effects of hCG, progesterone and oestradiol supplementation on nuclear and cytoplasmic maturation of canine oocytes cultured for 24, 48, 72 and 96 h. Oocytes obtained from 18 healthy bitches were divided into three groups according to their reproductive status (follicular, luteal and anoestrus stages) and cultured in TCM 199 + 25 UI/ml of hCG + 1 μg/ml of progesterone + 1 μg/ml of 17‐β oestradiol or without hormonal supplementation (control) for different periods. Then, they were stained with FITC‐LCA‐Hoescht for chromatin configuration and cortical granules distribution and evaluated under an epifluorescence microscope. Culture time and the influence of different stages of the oestrous cycle were also evaluated. The present study demonstrated that there was no significant difference among the reproductive stages. With regards to culture medium, only oocytes from the supplemented medium were able to complete meiosis; however, significant difference was only noticed in the percentage of MI stage oocytes (p < 0.05) in the follicular and luteal group at 72 h of culture. Most oocytes in germinal vesicle, germinal vesicle breakdown and metaphase I stage had cortical granules distributed throughout the cytoplasm (immature pattern), irrespective of the culture period (p < 0.05). Cortical granules distributed immediately beneath the plasma membrane (mature) was only observed in metaphase II stage oocytes, but not all of them presented matured cytoplasm. Our results reveal that cortical granules distribution in canine oocytes matured in vitro did not progressed in correspondence with nuclear stage changes and are in accordance with those from other species.