Intraoviductal introduction of plasmid DNA and subsequent electroporation for efficient in vivo gene transfer to murine oviductal epithelium

Various growth factors and proteins produced by oviductal cells have been demonstrated to interact with developing embryos. However, little is known concerning the function of mammalian oviducts at the molecular biological level. This may be partly due to lack of efficient gene transfer to oviductal cells. In this study, we developed an efficient method for transfection of oviductal epithelium using in vivo electroporation (EP) in mice. One microliter of solution containing enhanced green fluorescent protein (EGFP) expression plasmid (0.5 microg) and 0.05% trypan blue (TB) were directly introduced into the ampulla of the eCG-hCG-treated B6C3F1 females at embryonic day (E) 0.6 of pregnancy (corresponding to 14:00-15:00 of the day the plug was recognized). The entire oviduct was then electroporated using tweezer-type electrodes attached to a T820 electroporator (BTX Genetronics, Inc., San Diego, CA) with eight square-wave pulses, 50 V in strength and 50 msec in duration. On E 3.4, embryos at morula/early blastocyst stages were collected and their number, morphology, and EGFP-derived fluorescence recorded. Fluorescence in oviducts was also examined. In some cases, these fluorescent oviducts were subjected to cryostat sectioning. Strong fluorescence was observed in some of the oviductal epithelia, with a maximum level of 36%. Neither the number nor morphology of the collected embryos was affected by EP. Some embryos possessed fluorescence in the blastocoel, but not cytoplasm, suggesting incorporation of EGFP present in the oviductal luminal fluid. This system may enable development of new factors regulating development of preimplantation embryos and offers the prospect of a new approach to understanding oviductal function.     

Competition and introduction regime shape exotic bird communities in Hawaii

Complex combinations of historical and local-regional processes determine the assembly of ecological communities. We investigated such processes in the Hawaiian introduced avifauna, comprising 140 years of historical records of invasions and extinctions of birds. Here the particular introduction regime (i.e., colonization attempts and number of introduced species) and priority effects constitute the historical (and regional) component, and competition is the local component. These processes are theoretically supported by means of a Lotka–Volterra model of species competition, finding that changes in the specific introduction regime might result in different extinction dynamics. Both field data and model outcomes support the biotic resistance hypothesis, so that the invasibility of new incomers decrease with species richness. Finally, we found that the resistance to new invaders depends on the particular introduction regime. Thus, community assembly models built to predict the success of exotic species should consider more scenarios than random introduction regimes.     

Exclusive and complete introduction of amino groups and their N-sulfo and N-carboxymethyl groups into the 6-position of cellulose without the use of protecting groups

A new regioselective synthesis of 6-amino-6-deoxycellulose with a DS 1.0 (degree of substitution) at C-6, and its 6-N-sulfonated and its 6-N-carboxymethylated derivatives, without using protecting groups is described in this paper. The reaction conditions were optimized for preparing cellulose tosylate with full tosylation at C-6 and partial tosylation at C-2 and C-3. The nucleophilic substitution (S(N)) reaction of the tosyl group by NaN(3) at low temperature of 50 degrees C in Me(2)SO was achieved completely at C-6, whereas the tosyl groups at C-2 and C-3 were not displaced. In contrast to this, at 100 degrees C the tosyl groups at C-6, and also those at C-2 and C-3, were replaced by azido groups. This regioselective reaction that depends on temperature makes it possible to reach a selective and quantitative S(N) reaction at C-6 at low temperatures. In the subsequent reduction step with LiAlH(4), the azido group at C-6 was reduced to the amino group, and the tosyl groups at C-2 and C-3 were simultaneously completely removed. Also reported is a temperature-dependent, regioselective and complete iodination by nucleophilic substitution of the tosyl group at C-6 at 60 degrees C. At higher temperatures from 75 to 130 degrees C, substitution is also observed to occur at C-2. The selective iodination at 60 degrees C was employed to confirm the complete tosylation at C-6 of cellulose. The reaction products were identified by four different independent quantitative methods, namely 13C NMR, elemental analysis, ESCA, and fluorescence spectroscopy. 6-N-Sulfonated and 6-N-carboxymethylated cellulose derivatives were also synthesized. The new derivatives are potent candidates for structure-function studies, e.g., studies in relation to regioselectively 2-N-sulfonated and 2-N-carboxymethylated chitosan derivatives.     

What future for British red squirrels?

The geographic range of red squirrels contracted sharply in Britain during the 1940s arid 1950s, as increasingly large areas were colonized by the congeneric North American grey squirrel. Red squirrels remain common only on offshore islands, and in the large conifer forests of northern England and Scotland. The initial replacement of red squirrels was in arras dominated by oak woodland, probably because acorn crops are exploited less efficiently by red squirrels than by grey squirrels. Dirt studies have shown that acorns are digested less efficiently by the red squirrel, which occurs in conifers through most of its Eurasian range, than by the introduced grey squirrel, which is primarily a native of deciduous woodland. The red squirrel will probably be replaced in deciduous and mixed woodland throughout mainland Britain, and may eventually persist only in large areas of conifers which arc far from oak trees. The conservation of red squirrels on islands is therefore particularly important for their survival, perhaps making it worthwhile to create new island populations where they do not at present exist.     

Shift in pike, Esox lucius L., predation pressure following the introduction of ruffe, Gymnocephalus cernuus (L.) to Loch Lomond

Rapid and dramatic change in the fish community of Loch Lomond has resulted from a series of fish introductions in recent years. A comparison of the diet of pike, Esox lucius L., in 1989–1990 with data from 1955–1967, prior to recent introductions, demonstrates a shift in prey choice. In 1955–1967 powan, Coregonus lavaretus (L.), dominated in the diet (57% of prey by number) by 1989–1990 the introduced ruffe, Gymnocephalus cernuus (L.), was the commonest prey species (44% by number). This shift in pike predation to an abundant population of introduced ruffe has consequences for native species. Assuming that the pike population has not increased in response to increased food availability due to the introduction of ruffe, pike predation pressure on native species will be relieved. This is likely to have the greatest effect on powan. Comparison of the predation rate in 1955–1967 with 1989–1990 supports the hypothesis that the rate of predation on powan has declined, although the effect that this may have on the powan population is unclear, as the role of predation in the regulation of population size is unknown for this species.     

Amplified fragment length polymorphism data provide a poor solution to the Littorina littorea puzzle

The status of Littorina littorea as an exotic species in North America has been debated in the biological literature for over a century. In recent decades, molecular data have been used to supplement historical and archaeological evidence that has suggested that the snail may have persisted in North America well before European settlers arrived. However, these earlier molecular studies have since been questioned due to incomplete sampling of both the geographical range of L. littorea and the genome of the species itself. Here we consider an amplified fragment length polymorphism screen of the nuclear genome, which provides a large number of independent dominant markers that can be used in resolving this historical puzzle. Although these data appear to refute earlier allozyme studies of L. littorea as indicating long-term divergence between European and American populations, the data are also problematic in that they include clear artefacts. We discuss these artefacts and suggest future approaches to definitively resolving the debate over L. littorea's introduction to North America.     

Convergent evolution of coloration in experimental introductions of the guppy (Poecilia reticulata)

Despite the multitude of examples of evolution in action, relatively fewer studies have taken a replicated approach to understand the repeatability of evolution. Here, we examine the convergent evolution of adaptive coloration in experimental introductions of guppies from a high‐predation (HP) environment into four low‐predation (LP) environments. LP introductions were replicated across 2 years and in two different forest canopy cover types. We take a complementary approach by examining both phenotypes and genetics. For phenotypes, we categorize the whole color pattern on the tail fin of male guppies and analyze evolution using a correspondence analysis. We find that coloration in the introduction sites diverged from the founding Guanapo HP site. Sites group together based on canopy cover, indicating convergence in response to light environment. However, the axis that explains the most variation indicates a lack of convergence. Therefore, evolution may proceed along similar phenotypic trajectories, but still maintain unique variation within sites. For the genetics underlying the divergent phenotypes, we examine expression levels of color genes. We find no evidence for differential expression, indicating that the genetic basis for the color changes remains undetermined.     

药用植物引种驯化原理研究

药用植物作为一种重要的资源,其价值主要体现在具有防治疾病的功能,这种功能的物质基础在于植物内部合成的有效成分。药用植物引种驯化的主要目的是为了扩大其资源量,能更好地开发药用植物和可持续利用。因此,药用植物引种驯化不仅要保证药用植物在引种地的生长发育和繁殖,还需保证药效成分的维持与提高,其引种驯化成功的标志是从"药效"到"药效"。为此,该研究在对近年来药用植物药效成分的形成与稳定、药用植物的发源以及药用植物引种与驯化等方面进行综述整理的基础上,提出了药用植物引种驯化的原理,对于目前大量药效成分复杂不清的药用植物,不提倡复杂引种驯化。针对这类药用植物提出了药用植物发源中心假设,即先确定其发源中心,在此中心内收集引种群体和生态各因子的信息,以此为基础,在引种地上保证与药效形成和稳定相关因子不变,确保引种驯化后药用植物的药用功效不减弱或丧失,并通过多种方法综合评价药用植物的药效,从而达到药用植物引种驯化的目的。     

Ecological traits and environmental affinity explain Red Sea fish introduction into the Mediterranean

Alien species are considered one of the prime threats to biodiversity, driving major changes in ecosystem structure and function. Identifying the traits associated with alien introduction has been largely restricted to comparing indigenous and alien species or comparing alien species that differ in abundance or impact. However, a more complete understanding may emerge when the entire pool of potential alien species is used as a control, information that is rarely available. In the eastern Mediterranean, the marine environment is undergoing an unparalleled species composition transformation, as a flood of aliens have entered from the Red Sea following the opening of the Suez Canal in 1869. In this study, we compile data on species traits, geographical distribution, and environmental affinity of the entire pool of reef‐associated fish species in the Red Sea and more generally across the Indo‐Pacific. We use this extensive data to identify the prime characteristics separating Red Sea species that have become alien in the Mediterranean from those that have not. We find that alien species occupy a larger range of environments in their native ranges, explaining their ability to colonize the seasonal Mediterranean. Red Sea species that naturally experience high maximum temperatures in their native range have a high probability of becoming alien. Thus, contrary to predictions of an accelerating number of aliens following increased water temperatures, hotter summers in this region may prevent the establishment of many alien species. We further find that ecological trait diversity of alien species is substantially more evenly spaced and more divergent than random samples from the pool of Red Sea species, pointing at additional processes, such as competition, promoting ecological diversity among alien species. We use these results to provide a first quantitative ranking of the potential of Red Sea species to become established in the eastern Mediterranean.