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.     

Improvement of Time-Driven Models of Lamina Cocksfoot Digestibility by a Process-Based Model to Take Account of Plant N Nutrition and Defoliation

For management purposes, models of lamina digestibility (D_W) that are thermal time driven (T_sum) fail to account for plant nitrogen status (0 < Ni < 1) and defoliation intensity (residual sheath height: S_r). The objective of this paper was to enrich them using a functional processed-based model, which assumes that the decline in D_W over time depends on the metabolic : structural tissue ratio and plant ageing. An experiment combining two N fertilizer rates and two defoliation regimes (differences in S_r) for two spring and two summer regrowth was done to analyse D_W of laminae (youngest fully expanded) and tiller (whole green lamina) levels. For laminae, we show that both processes contribute to explain the decline in D_W at a rate that depends on Ni but not on S_r. For tillers, consistent results were found justifying the use of a simple model: D_W = f(+Ni; −Ni × T_sum; −S_r). Two different databases (from experiment and commercial farms) were used for validation: RMSD varied from 35 to 49 g kg⁻¹. The process-based model strengthened the empirical model and improved the comprehensiveness of how management practices change D_W. We used both models to create applications for reasoning yield in function of herbage mass in complement than in function of time.     

A methodology for characterization and analysis of plant functional composition in grassland‐based farms

Understanding the relationships between farmers' land‐use and management decisions, and plant diversity is a challenge. It requires characterization of plant diversity within and between fields and investigation of land‐use allocation to fields. To analyse how grassland functional composition [mean plant trait and functional diversity index (FD)] varies according to scale (field, land‐use type: cutting, grazing, farm), grasslands were characterized according to leaf dry‐matter content (leaf DM) and FD computed from leaf DM values. A leaf DM‐αFD framework was used to analyse how leaf DM‐based plant strategies were distributed at land‐use type and farm scales (βFD). The study was conducted on eight dairy and beef farms (169 grasslands) differing in their stocking rate. At field and land‐use type scales, leaf DM was significantly decreased and increased with N fertilizer rate and field elevation respectively. It was significantly higher for grazing than for cutting. At the farm scale, the main differences between farms resulted from differences in plant strategy distribution between land‐use types within a farm and among farms for a given land‐use type in relation to management intensity. Farms that contributed the most to αFD had the highest stocking rate, and those which had the most contrasting grasslands for management intensity had the highest βFD. Management practices need to be examined at a land‐use type scale for evaluating the within‐ and between‐field plant functional compositions. By contrast, the value of the analysis was reduced if data were collected and averaged at the farm scale.     

Alteration in blood gases in cats naturally infected with Aelurostrongylus abstrusus

Four cats were presented with respiratory signs and first-stage larvae of Aelurostrongylus abstrusus were found in faecal samples. Anthelmintic treatment was given to the infected cats and venous blood gases were analysed during the treatment period. Blood gas analysis suggested hypoventilation and respiratory acidosis in infected cats. Hypoventilation may be the result of airway obstruction by adults and larvae in respiratory bronchioles and the alveolar canals. The blood gas values had returned close to the physiological range by two months after treatment. Assessment of respiratory acidosis may aid development of additional treatment methods in cats infected with A. abstrusus.     

The use of herbage nitrogen status to optimize herbage composition and intake and to minimize nitrogen excretion: an assessment of grazing management flexibility for dairy cows

A framework for managing rotationally grazed pastures for dairy cattle which enables the cows’ energy and protein requirements to be met while simultaneously limiting the amount of N excreted in order to reduce N losses is described. The first objective is achieved by ensuring that lamina mass and the N concentration of herbage do not limit herbage intake or feeding value. The second objective is achieved by limiting N fertilizer supply or increasing the interval between defoliations to reduce the N concentration of herbage. Lower and upper thresholds for the N concentration of herbage and lamina mass were estimated from published data. The method is illustrated using two vegetative regrowths (beginning and end of spring) in a cutting experiment with two fertilizer treatments, 0 or 120 kg N ha^?1 (?N and +N), and early or late cutting. Decreasing N supply led to a reduction in grazing management flexibility, i.e. the defoliation interval ranges which were compatible with the required sward characteristics (minimum lamina mass and N concentration of lamina) for herbage intake and to meet the protein requirements of dairy cows. Aiming for the upper threshold N concentration of herbage increased the minimum interval between defoliations only for the +N treatments. Nevertheless, grazing management flexibility remained the highest for the +N treatments.     

Effects of Dietary Supplementation of β‐hydroxy‐β‐methylbutyrate on Sow Performance and mRNA Expression of Myogenic Markers in Skeletal Muscle of Neonatal Piglets

The effects of dietary β‐hydroxy‐β‐methylbutyrate (HMB) supplementation during gestation on reproductive performance of sows and the mRNA expression of myogenic markers in skeletal muscle of neonatal pigs were determined. At day 35 of gestation, a total of 20 sows (Landrace × Yorkshire, at third parity) were randomly assigned to two groups, with each group receiving either a basal diet or the same diet supplemented with 4 g/day β‐hydroxy‐β‐methylbutyrate calcium (HMB‐Ca) until parturition. At parturition, the total and live litter size were not markedly different between treatments, however, the sows fed HMB diet had a decreased rate of stillborn piglets compared with the sows fed the control (CON) diets (p < 0.05). In addition, piglets from the sows fed HMB diet tended to have an increased birth weight (p = 0.08), and a reduced rate of low birth weight piglets (p = 0.05) compared with piglets from the CON sows. Nevertheless, lower feed intake during lactation was observed in the sows fed the HMB diet compared with those on the CON diet (p < 0.01). The relative weights of the longissimus dorsi (LD) and semitendinosus (ST) muscle were higher (p < 0.05) in neonatal pigs from the HMB than the CON sows. Furthermore, maternal HMB treatment increased the mRNA levels of the myogenic genes, including muscle regulatory factor‐4 (MRF4, p < 0.05), myogenic differentiation factor (MyoD) and insulin‐like growth factor‐1 (IGF‐1, p < 0.01). In conclusion, dietary HMB supplementation to sows at 4 g/day from day 35 of gestation to term significantly improves pregnancy outcomes and increases the expression of myogenic genes in skeletal muscle of neonatal piglets, but reduces feed intake of sows during lactation.     

Rethinking Technology‐Oriented Business Incubators: Developing a Robust Policy Instrument for Entrepreneurship,Innovation, and Regional Development?

ABSTRACT Today, technology‐oriented business incubators are a worldwide phenomenon, although empirical research evidence clearly suggests that they tend to fail in supporting entrepreneurship, innovation, and regional development and, therefore, do not fulfil their expected role as policy instrument. The paper focuses on this obvious antagonism. It deliberates upon political rationales, reviews evaluation literatures, and delineates suggestions for the future of the incubation industry. The main conclusion is that technology‐oriented business incubators should be run as private organisations without public funding.     

The effect of N and P fertilizer application and botanical composition on the leaf/stem ratio patterns in spring in Pyrenean meadows

The effect of botanical composition and nutrient availability on the relative allocation of biomass to stems and leaves in a permanent upland pasture in the central Pyrenees was assessed. Six short-term and medium-term fertilizer treatments (nitrogen and phosphorus) were applied to a meadow to create large differences in the proportions of the different species and a wide range of herbage nutrient status. The above-ground herbage dry-matter components (green leaves, sheaths and stems for grasses and dicotyledons, and senescent material) were measured for the first growth cycle.
The leaf mass depended mainly on the herbage N status, whereas the stem mass depended both on the botanical composition and on the herbage N and P status. During spring growth, the proportion of leaves in the above-ground dry matter decreased faster in plots that had the highest nutrient status or that were composed of species characteristic of nutrient-rich ( Festuca -poor) habitats. Application of fertilizer decreased the proportion of leaves both for grasses and dicotyledons, but to a larger extent for grasses. There was a single relationship between the proportion of leaves in the above-ground dry matter and the total mass of above-ground dry matter, irrespective of the sampling date, the botanical composition or the herbage nutrient status. This statistical relationship resulted from (i) a faster increase in stem mass than leaf mass for a given botanical composition when the herbage nutritional status increased, (ii) a greater stem mass when the sward was composed of species usually found in nutrient-rich habitats.     

Blood type A and B frequencies in Turkish Van and Angora cats in Turkey

Blood typing of domestic cats has been performed in domestic and purebred cats in various parts of the world and is important in clinical practice in order to prevent neonatal isoerythrolysis (NI) and acute haemolytic transfusion reactions. Prevalence of blood types vary greatly between breeds of cats. Turkish Van and Angora cats are different breeds that originated in geographically distinct regions of Turkey. The present survey determined the frequency of blood types in these Turkish pedigreed cats in Turkey. Ethylenediaminetetraacetic-acid anti-coagulated blood of a total of 113 Turkish Van and Angora cats were examined for blood typing using a slide and tube agglutination assay. Of the 85 Van cats surveyed, 40% had type A, and 60% had type B blood. Of the 28 Turkish Angora cats, 53.6% had type A, and 46.4% had type B blood. No type AB cats were found between both breeds. There was no significant association between blood types and gender of both Angora and Van cats or eye colours of Van cats (P > 0.05). Although these are limited surveys, the overall prevalence of type B cats in these two breeds was very high compared with the results of previous studies worldwide. It appears likely that blood type incompatibilities responsible for feline NI and transfusion reactions are occurring in these breeds. The risk of transfusion incompatibility in Turkish Angora and Van cats was 46.4 and 60%, respectively. It is therefore strongly recommended to breeders and clinicians that blood typing be performed prior to breeding and transfusing cats.     

Induction of oxidants in tomato leaves treated with <Emphasis Type="SmallCaps">DL</Emphasis>-β-Amino butyric acid (BABA) and infected with <Emphasis Type="Italic">Clavibacter michiganensis</Emphasis> ssp. <Emphasis Type="Italic">michiganensis</Emphasis>

Bacterial canker is an economically important disease of tomato. Resistance induced by DL-β-Amino butyric acid against bacterial canker caused by Clavibacter michiganensis ssp. michiganensis in tomato plants was investigated. Different doses of DL-β-Amino butyric acid (250–1000 μg ml⁻¹ doses) were tested on 3-week old plants inoculated with a 10⁸ CFU ml⁻¹ bacterial suspension, and disease development was evaluated after inoculation and treatment. Although in vitro growth of the bacteria was not affected by DL-β-Amino butyric acid treatment, foliage sprays of 500 μg ml⁻¹ DL-β-Amino butyric acid significantly suppressed disease development up to 54% by day 14 after inoculation at the four different doses tested. Bacterial populations were reduced by 84% in BABA-treated plants compared to water-treated plants by day 4 after inoculation. Inoculated BABA-treated plants showed significantly higher phenylalanine ammonia-lyase activity, peroxidase activity, and H₂O₂ concentration than inoculated water-treated plants during day 1 after treatment. These findings suggest that the DL-β-Amino butyric acid treatment resulted in an increase of these enzymes and in H₂O₂ concentration in planta, and was associated with induction of resistance to bacterial canker.