Maize histone H2B-mCherry: a new fluorescent chromatin marker for somatic and meiotic chromosome research

Cytological studies of fluorescent proteins are rapidly yielding insights into chromatin structure and dynamics. Here we describe the production and cytological characterization of new transgenic maize lines expressing a fluorescent histone fusion protein, H2B-mCherry. The transgene is expressed under the control of the maize ubiquitin1 promoter, including its first exon and intron. Polymerase chain reaction-based genotyping and root-tip microscopy showed that most of the lines carrying the transgene also expressed it, producing bright uniform staining of nuclei. Further, plants showing expression in root tips at the seedling stage also showed expression during meiosis, late in the life cycle. Detailed high-resolution three-dimensional imaging of cells and nuclei from various somatic and meiotic cell types showed that H2B-mCherry produced remarkably clear images of chromatin and chromosome fiber morphology, as seen in somatic, male meiotic prophase, and early microgametophyte cells. H2B-mCherry also yielded distinct nucleolus staining and was shown to be compatible with fluorescence in situ hybridization. We found several instances where H2B-mCherry was superior to DAPI as a generalized chromatin stain. Our study establishes these histone H2B-mCherry lines as new biological reagents for visualizing chromatin structure, chromosome morphology, and nuclear dynamics in fixed and living cells in a model plant genetic system.     

Testing Germination of Species for Hydroseeding Degraded Mediterranean Areas

The germination performance of native species and their suitability for a rapid erosion control are uncertain. Together with their relatively low commercial availability and high costs, these are still strong reasons preventing their common use in hydroseeding for restoration of Mediterranean degraded slopes, despite the increasing number of studies recommending it. In this study, 14 non‐native (commercial) and native herb and woody species were tested. Their germination performance was evaluated under laboratory (Petri dishes) and greenhouse conditions (seeds sown in target substrate). The results obtained were compared with the seedling densities in a Mediterranean quarry slope hydroseeded with the same species. In the laboratory, commercial species had a better germination performance than most native species, but this trend was not maintained in the greenhouse. Greenhouse tests were extended beyond spring and showed that many native species germinated better, or exclusively in autumn. Germination performance and success decreased, from laboratory to greenhouse and field conditions, for many species, but not for all. Relative to field performance, the predictive value of laboratory and greenhouse tests was poor, yet sowing on the target substrate under greenhouse conditions may be a better approach for certain native species. The main drawbacks revealed by native species in the present study included: (1) relatively slow germination; (2) seasonality; and (3) seed dormancy‐breaking requirements. The results suggest that these problems may be overcome through species selection, seed pre‐treatments, hydroseeding scheduling, and/or manipulation of seeding density and relative species proportion.     

Application of adaptive DO-stat feeding control to Pichia pastoris X33 cultures expressing a single chain antibody fragment (scFv)

In this study, fed-batch cultures of a Pichia pastoris strain constitutively expressing a single chain antibody fragment (scFv) under the control of the glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter were performed in a pilot 50 L bioreactor. Due to the very high cell density achieved within the first 75?h, typically between 140 and 160?g-DCW/L of dry cell weight (DCW), most of the scFv is produced under hard oxygen transfer limitation. To improve scFv productivity, a direct adaptive dissolved oxygen (DO)-stat feeding controller that maximizes glycerol feeding under the constraint of available oxygen transfer capacity was developed and applied to this process. The developed adaptive controller enabled to maximize glycerol feeding through the regulation of DO concentration between 3 and 5?% of saturation, thereby improving process productivity. Set-point convergence dynamics are characterized by a fast response upon large perturbations to DO, followed by a slower but very robust convergence in the vicinity of the boundary with almost imperceptible overshoot. Such control performance enabled operating closer to the 0?% boundary for longer periods of time when compared to a traditional proportional–integral–derivative algorithm, which tends to destabilize with increasing cell density.     

铼-188标记生物分子在肿瘤治疗中的应用

目的：研究铼．188标记生物分子在肿瘤治疗中的应用。方法：选取小白鼠作为实验的研究对象,将荷瘤鼠的肉瘤切成小块接种到小白鼠身上,达到试验条件时使用,即将没有明显差异的小白鼠16只随机分为4组,每组4只,注射含有铼一188的药物后分别在不同的时间将其处死,之后取出重要器官进行测量分析,进而得出铼一188的应用效果。结果：瘤内注射的要去在不同时间放射性在瘤内的保持率分别为（90．5±7．7）D％（1h）,（92．2±8．6）D％（24h）,（88．3±10．9）D％（48h）和（91．5±7．6）D％（72h）,在荷瘤鼠内注入生理盐水、非放硫化铼和188Re．硫化铼混悬液,肿瘤质量分别为2885．3±1241.3、2839．9±1965．2和98．4±45．5mg。188Re-硫化铼混悬液在生理盐水、磷酸盐缓冲液和小牛血清中均可稳定72h,而且188W-188Re发生器的应用还可以降低188Re-硫化铼混悬液的价格。随着处死时间的延迟,小鼠肿瘤质量和体积逐渐减小,相临两组比较,后组测定值均明显小于前组,数据经统计学比较具有显著差异（P〈0．05）。结论：188Re-硫化铼混悬液是一种适宜的肿瘤治疗剂。     

First case of highly pathogenic H5N1 avian influenza virus in Spain

Background  

In August 2006 a major epidemic of bluetongue virus serotype 8 (BTV8) started off in North-West Europe. In the course of 2007 it became evident that BTV8 had survived the winter in North-West Europe, re-emerged and spread exponentially. Recently, the European Union decided to start vaccination against BTV8. In order to improve the understanding of the epidemiological situation, it was necessary to execute a cross-sectional serological study at the end of the BT vector season. Cattle were the target species for cross-sectional serological studies in Europe at the end of 2006 and 2007. However, there was no information on the BTV8-seroprevalence in sheep and goats.     

CO2 and O2 dynamics in human-impacted watersheds in the state of São Paulo, Brazil

We studied the spatial and temporal variation in O₂ and dissolved inorganic carbon (DIC) forms concentrations in ten subtropical watersheds located in the state of São Paulo, Brazil, with different degrees of impact by urbanization and land-use changes. Additionally, we used stable carbon isotopic composition of DIC to explain observed patterns. We found that land-cover changes and watershed geology are the main drivers of DIC distribution. Land-cover/use changes influence the riverine DIC in two ways: by replacing the original Cerrado 3 (C3)-type forest vegetation by C4-type vegetation composed of grasses (pasture), and by sugarcane. Most domestic sewage is dumped untreated into rivers in the state of São Paulo. Consequently, in the most densely populated watersheds, sewage is an important source of labile carbon and consequently of DIC to rivers. In terms of geology, although silicate weathering that produces kaolinite is the main type of weathering in the watersheds, the weathering of carbonate cements present in the geological formations of the western portion of the state of São Paulo are also an important source of DIC to rivers.     

Helophyte germination in a Mediterranean salt marsh: Gut‐passage by ducks changes seed response to salinity

Question: In seeds which are regularly consumed by waterbirds in the field, how does gut‐passage modify their response to salinity gradients? Location: Doñana National Park salt marsh, south‐west of Spain. Methods: Seeds of Scirpus litoralis and Scirpus maritimus were collected and force fed to mallards (Anas platyrhynchos). Both the ingested seeds (passage) and non‐ingested seeds (controls) were exposed, in germination chambers, to a salinity range similar to that observed in the field (0–32 dS/m). After 30 days, the total percentage germination, the duration of the dormancy period and the germination speed were computed. The response of the different germination parameters to ingestion and salinity was analyzed using generalized lineal models. Recovery tests on seeds that did not germinate in the various treatments and tests of the effect of ingestion on the intrinsic variability in seed response were also performed. Results: An increase in salinity reduced germinability and increased the length of dormancy, while gut pas sage increased the intrinsic variability of the temporal seed response in both species. In S. litoralis there was a significant interaction between the effects of salinity and passage on germination rate. Passage increased germination rate at low salinities (≤2 dS/m) but decreased it at high salinities (≥4 dS/m). Conclusion: Gut‐passage by ducks significantly changes seed response to salinity. The outcome of plant‐animal interactions can be influenced by environmental gradients. Studies of germination in response to gut passage that do not take such gradients into account may produce misleading results.     

Characterization of NADPH-diaphorase-positive glial cells of the tench optic nerve after axotomy

NADPH-diaphorase (ND) positive cell types were characterized throughout the optic nerve of the tench in normal conditions and after optic nerve transection with survival periods of 1, 3, 7, 14, 30, 60, 120 and 180 days. Astrocytic markers (S100 and glutamine synthetase) and the microglial marker tomato lectin were employed. In the control prechiasmatic optic nerve two types (types I and II) of ND-positive glial cells appeared. All type I cells showed S100 immunoreactivity, whereas only a subpopulation of them were positive to glutamine synthetase. Type II cells only presented S100 immunoreactivity. In the control anterior optic tract, all ND-positive glial cells (type III) presented immunolabeling to S100 and glutamine synthetase. After transection, types I and II did not show any changes in the staining patterns for the glial markers when observed. Two new types of ND-positive glial cells (types IV and V) were observed after axotomy. All type IV cells were S100-immunopositive, and a subpopulation presented glutamine synthetase immunolabeling. Only a subpopulation of type V cells showed glutamine synthetase immunostaining. The presence of type IV or V cells in the lesioned optic nerve occurred simultaneously with significant decreases or absence of type I cells. These data suggest that type I and III cells are astrocytes and type II cells are oligodendrocytes. Types IV and V cells are the result of the activation of type I cells after optic nerve section. The polymorphism observed in ND-positive cells may reflect different cell functions during degenerative and regenerative processes.