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.     

Wnt3a-mediated formation of phosphatidylinositol 4,5-bisphosphate regulates LRP6 phosphorylation

The canonical Wnt-beta-catenin signaling pathway is initiated by inducing phosphorylation of one of the Wnt receptors, low-density lipoprotein receptor-related protein 6 (LRP6), at threonine residue 1479 (Thr1479) and serine residue 1490 (Ser1490). By screening a human kinase small interfering RNA library, we identified phosphatidylinositol 4-kinase type II alpha and phosphatidylinositol-4-phosphate 5-kinase type I (PIP5KI) as required for Wnt3a-induced LRP6 phosphorylation at Ser1490 in mammalian cells and confirmed that these kinases are important for Wnt signaling in Xenopus embryos. Wnt3a stimulates the formation of phosphatidylinositol 4,5-bisphosphates [PtdIns (4,5)P2] through frizzled and dishevelled, the latter of which directly interacted with and activated PIP5KI. In turn, PtdIns (4,5)P2 regulated phosphorylation of LRP6 at Thr1479 and Ser1490. Therefore, our study reveals a signaling mechanism for Wnt to regulate LRP6 phosphorylation.     

Analysis of factors affecting the electric conductivity of cow's milk--breed, locale, season

The problem was studied in a set of 678 mammary glands of 212 cows - crossbreds between the Holstein-Friesian breed with the Slovak Spotted cattle, crossbreds of the Black-Pied Lowland and Bohemian Spotted breed, and three-way crossbreds of the Black-Pied Lowland, Swedish White and Bohemian Spotted cattle. The confrontation of the confidence intervals of milk conductivity (MC) in healthy mammary glands at alpha = 0.05 showed that in the crossbreds of the Black-Pied Lowland cattle the MC was statistically significantly higher than in the breed types of the Holstein-Friesian cattle. The MC values decrease when there is a larger proportion of the blood of beef-and-dairy cattle (Slovak Spotted and Bohemian Spotted breeds). The locality where the stock was kept was found to act as an important factor underlying MC only in cases when both the geographical conditions and breed proportions were different. In a set homogenized with respect to the breeds, statistically significant differences in MC were demonstrated between the spring and summer season, spring and autumn, summer and autumn, and autumn and winter season.     

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.     

洋麻炭疽病(Colletotrichum hibisci Pollacci)的防治研究

 洋麻炭疽病早在1912年即于台湾发現,以后传到內地,按其引种的来源分析,是自印度传入中国的。     

The structure of confined oxygen in silica xerogels

The microscopic structure of oxygen confined in silica xerogels has been studied as a function of temperature. In large pores, a crystalline solid forms with a structure consistent with that of the bulk. The size of the crystallites is much larger than the pore size, indicating that cooperative effects among pores are important in freezing. As the pore size is decreased, a crossover occurs where solidification results in an amorphous phase in the pores. The resulting amorphous phase is solid but is less ordered than the liquid phase.     

The effect of bentazone manufactured in Czechoslovakia on the synthesis of bacterial proteins in the rumen of sheep]

The effect of the perorally ingested pesticide bentazone (195 mg.kg-1) of Czechoslovak origin on the amino acid composition of proteins of bacteria adhering to the ventral and dorsal ruminal wall investigated in six sheep. Proteosynthesis of adherent bacteria was studied by a modified and quantified elution method (elution of bacteria by an isotonic buffered solution at 4 degrees C). By the latter, a concentrate of undamaged bacteria a adhering to the sheep ruminal epithelium could be obtained. The yield of the method was estimated by scanning electrone microscopy and it was 93.3% (Legáth et al., 1990). The high correlation coefficient (r = 0.94, p < 0.01) suggested that in comparison with the control group of sheep, bentazone in the diet did not have a marked effect on the amount of amino acids in hydrolyzates of bacteria adhering to the dorsal and ventral ruminal epithelium. From the analysis of the single amino acids, it however follows that bentazone in the feeding ration caused significant changes in the concentrations of some amino acids in the proteins of bacteria adhering to the epithelium in the ventral and dorsal part of the rumen (p < 0.05) (Tab. I, II). In both topographico-anatomical parts of the rumen phenylalanine levels significantly increased whereas those of alanine and glycine decreased (Fig. 1). Pesticides can be one of the factors that negatively affect the biosynthetic processes in the rumen of ruminants.(ABSTRACT TRUNCATED AT 250 WORDS)