Lipid content and apoptosis of in vitro-produced bovine embryos as determinants of susceptibility to vitrification

The objective was to evaluate supplementation of fetal calf serum (FCS) and phenazine ethosulfate (PES), a metabolic regulator that inhibits fatty acid synthesis, in culture media during in vitro production (IVP) of bovine embryos. Taking oocyte fertilization (n = 4,320) as Day 0, four concentrations of FCS (0, 2.5, 5, and 10%) and three periods of exposure to PES (without addition—Control; after 60 h—PES Day 2.5 of embryo culture; and after 96 h—PES Day 4) were evaluated. Increasing FCS concentration in the culture media enhanced lipid accumulation (P < 0.05), increased apoptosis in fresh (2.5%: 19.1 ± 1.8 vs 10%: 28.4 ± 2.3, P < 0.05; mean ± SEM) and vitrified (2.5%: 42.8 ± 2.7 vs 10%: 69.2 ± 3.4, P < 0.05) blastocysts, and reduced blastocoele re-expansion after vitrification (2.5%: 81.6 ± 2.5 vs 10%: 67.3 ± 3.5, P < 0.05). The addition of PES in culture media, either from Days 2.5 or 4, reduced lipid accumulation (P < 0.05) and increased blastocoele re-expansion after vitrification (Control: 72.0 ± 3.0 vs PES Day 2.5: 79.9 ± 2.8 or PES Day 4: 86.2 ± 2.4, P < 0.05). However, just the use of PES from D4 reduced apoptosis in vitrified blastocysts (Control: 52.0 ± 3.0 vs PES Day 4: 39.2 ± 2.4, P < 0.05). Independent of FCS withdrawal or PES addition to culture media, the in vivo control group had lesser lipid accumulation, a lower apoptosis rate, and greater cryotolerance (P < 0.05). The increased lipid content was moderately correlated with apoptosis in vitrified blastocysts (r = 0.64, P = 0.01). In contrast, the increased apoptosis in fresh blastocysts was strongly correlated with apoptosis in vitrified blastocysts (r = 0.94, P < 0.0001). Therefore, using only 2.5% FCS and the addition of PES from Day 4, increased the survival of IVP embryos after vitrification. Moreover, embryo quality, represented by the fresh apoptosis rate, was better than lipid content for predicting embryo survival after vitrification.     

Effect of dietary fat or starch supply during gestation and/or lactation on the performance of sows, piglets' survival and on the performance of progeny after weaning

Two trials were carried out to compare the effects of fat or starch inclusion in sow's diet on sow and litter performance. In each trial, sows were assigned to one of two treatments. In trial 1, the sows were fed diets containing either soybean oil (5%, treatment GL5) or cornstarch (11.3%, GL0) from day 35 of gestation to weaning. Daily net energy and nutrient allowance were equalised during gestation. In trial 2, the same treatments were applied only after farrowing (treatments L5 and L0, respectively). Within each trial, a batch of piglets was studied until slaughter. In trial 1, adipose cell development and total lipid content were determined on some pigs at weaning (n = 6/treatment) and at slaughter in dorsal subcutaneous adipose tissue (n = 13/group at least) and in muscle (n = 46/group at least). Piglets' birth weight was not affected by treatment in trial 1. Survival rates at birth and after 24 h of life were higher in treatment GL5 (4.0% v. 7.5% stillborn piglets in GL0 treatment, P < 0.05; 8.7% v. 12.6% of piglets alive at 24 h of age died in treatment GL0, P = 0.06). Subsequently, overall survival rate until weaning was higher in treatment GL5 (81.4% v. 75.7% of total born piglets, P = 0.03), but litter size at weaning was not significantly affected (11.3). Litter growth rate before weaning was increased when a fat-enriched diet was provided during gestation and lactation (+140 g/day per litter; P < 0.01) and to a lower extent when provided only after farrowing (+90 g/day; P < 0.05). Energy supply through fat did not decrease the mobilisation of the sow's body reserve and backfat thickness loss was even higher with treatment GL5 (P < 0.05). After weaning, pigs' average daily gain, feed : gain ratio and carcass lean content were not affected by the energy source supplied before and/or after farrowing. At weaning, the number of adipose cells in the dorsal subcutaneous adipose tissue and in the Longissimus dorsi muscle was higher in the GL5 pigs. Muscle lipid content at weaning did not differ between treatments, but it was higher at slaughter, around 110 kg, in the GL5 pigs (3.46% v. 2.58%, P < 0.001).     

Shoot population dynamics of persisting clones of<Emphasis Type="Italic"> Miscanthus sinensis</Emphasis> in the warm-temperate region of Japan

The population dynamics of annual shoots in four clonal patches of Miscanthus sinensis was surveyed in terms of clone persistence. Over a study period of 3 years, a stable net shoot number was attained through a balanced replacement of old shoots by tillering 2–3 times a year. The birth rate was significantly correlated with the average monthly temperature. This suggests that a warm climate advances the date of tillerings, and shoots become taller as they emerge earlier. Five cohorts were identified, corresponding to shoots that were born in spring, early summer, summer, autumn and early winter. The autumn cohorts were more numerous and had a greater longevity than the other cohorts; they were thus were best suited for overwintering and, consequently, maintaining the population from one generation to the next. The shoots of earlier cohorts grew too tall to survive the cold of winter, which might cause freezing injury, while later cohorts shoots excelled in overwintering because of their shortness. However, if the delayed shoots are very short, they will also have difficulty in surviving the early season of growth. Therefore, M. sinensis shoots have an optimum size for survival.     

The utilisation of arginine by oral streptococci grown glucose-limited in a chemostat

Abstract Legionella pneumophila occurring in drinking water was subjected to environmental stress through holding tests at ambient and elevated temperatures and by chemical disinfection. The bacterium in its native environment was more resistant to adverse conditions, as compared with laboratory-grown organisms. Of the several chemical disinfectants acceptable to drinking water treatment and tested for Legionella inactivation, ozonation was the most efficient method. The C · t ' products indicated that free chlorine was superior to mono- and dichloramines.     

Avian pluripotent stem cells

Pluripotent embryonic stem cells are undifferentiated cells capable of proliferation and self-renewal and have the capacity to differentiate into all somatic cell types and the germ line. They provide an in vitro model of early embryonic differentiation and are a useful means for targeted manipulation of the genome. Pluripotent stem cells in the chick have been derived from stage X blastoderms and 5.5 day gonadal primordial germ cells (PGCs). Blastoderm-derived embryonic stem cells (ESCs) have the capacity for in vitro differentiation into embryoid bodies and derivatives of the three primary germ layers. When grafted onto the chorioallantoic membrane, the ESCs formed a variety of differentiated cell types and attempted to organize into complex structures. In addition, when injected into the unincubated stage X blastoderm, the ESCs can be found in numerous somatic tissues and the germ line. The potential give rise to somatic and germ line chimeras is highly dependent upon the culture conditions and decreases with passage. Likewise, PGC-derived embryonic germ cells (EGCs) can give rise to simple embryoid bodies and can undergo some differentiation in vitro. Interestingly, chicken EG cells contribute to somatic lineages when injected into the stage X blastoderm, but only germ line chimeras have resulted from EGCs injected into the vasculature of the stage 16 embryo. To date, no lines of transgenic chickens have been generated using ESCs or EGCs. Nevertheless, progress towards the culture of avian pluripotent stem cells has been significant. In the future, the answers to fundamental questions regarding segregation of the avian germ line and the molecular basis of pluripotency should foster the full use of avian pluripotent stem cells.     

Grassland birds and rotational-grazing in the northeast: Breeding ecology,survival and management opportunities

We studied long-term (2002–2009) demographics of bobolinks (Dolichonyx oryzivorus) and savannah sparrows (Passerculus sandwichensis) breeding in rotationally grazed systems in the northeastern United States. Both nest success (6–44% bobolink; 7–48% savannah sparrow) and annual productivity (0.43–2.83 bobolink; 0.70–2.35 savannah sparrow) varied annually. Predation (48%) and trampling by cows (32%) accounted for most failed nests. Annual adult apparent survival ranged from 23% to 85% and showed substantial variation by species and sex. Although grazing resulted in substantial loss of vegetative cover, savannah sparrows responded similarly to predation and grazing-induced nest failure (time to renest, distance moved between nests) and generally remained in the same paddock to renest. Minimum paddock size to accommodate one female within this rotation was 70 m × 70 m (0.49 ha), allowing a female to stay within a paddock and move (approx. 50 m) between nesting attempts. To balance pasturing objectives and birds' needs in paddocks first grazed before 25 June, managers in the Northeast should rest that paddock 42–50 days before the second grazing. © 2011 The Wildlife Society.     

Silencing of the ADNP-family member, ADNP2, results in changes in cellular viability under oxidative stress

Activity-dependent neuroprotective protein (ADNP) 2 (KIAA0863; ZNF508) gene, a homeobox-profile containing gene, was identified in a screen for homologous proteins to ADNP. The human ADNP2 contains 1131 amino acid residues with a molecular weight of 122.8 KDa. In silico analysis indicated that ortholgs to ADNP2 exist in different phyla, suggesting that ADNP2 might be evolutionary conserved. Here, we began to explore the molecular and functional characterization of ADNP2. Results showed that the mouse ADNP2 mRNA is ubiquitously expressed in distinct normal tissues with increased expression in the brain, particularly in the cerebral cortex. During development, a relatively high level of ADNP2 gene expression was found in the embryonic mouse brain and was sustained throughout embryogenesis and adulthood. An increase in the mRNA was detected in differentiated P19 neuronal/glial-like cells as compared with the non-differentiated cells. To gain insight into ADNP2 function, ADNP2-deficient cell lines were established by the RNA silencing (small interfering RNA) technology. ADNP2 deficiency significantly changed the toxicity induced by hydrogen peroxide in P19 embryonic carcinoma cells, similar to what would be predicted for ADNP deficiency. These findings represent an initial characterization of ADNP2 and suggest that this gene product may have an important function in brain by playing a role in cellular survival pathways.     

Oncogenic potential of Nrf2 and its principal target protein heme oxygenase-1

Nuclear factor erythroid 2-related factor 2 (Nrf2) is an essential component of cellular defense against a vast variety of endogenous and exogenous insults, including oxidative stress. Nrf2 acts as a master switch in the circuits upregulating the expression of various stress-response proteins, especially heme oxygenase-1 (HO-1). Paradoxically, however, recent studies have demonstrated oncogenic functions of Nrf2 and its major target protein HO-1. Levels of Nrf2 and HO-1 are elevated in many different types of human malignancies, which may facilitate the remodeling of the tumor microenvironment making it advantageous for the autonomic growth of cancer cells, metastasis, angiogenesis, and tolerance to chemotherapeutic agents and radiation and photodynamic therapy. In this context, the cellular stress response or cytoprotective signaling mediated via the Nrf2–HO-1 axis is hijacked by cancer cells for their growth advantage and survival of anticancer treatment. Therefore, Nrf2 and HO-1 may represent potential therapeutic targets in the management of cancer. This review highlights the roles of Nrf2 and HO-1 in proliferation of cancer cells, their tolerance/resistance to anticancer treatments, and metastasis or angiogenesis in tumor progression.