Normal specification of the extraembryonic lineage after somatic nuclear transfer

To examine the establishment and maintenance of trophectoderm (TE) lineage in somatic cloned blastocysts, the expression of Cdx2, a key molecule for specification of TE fate, was immunohistochemically examined simultaneously with Oct4 expression. Cloned mouse embryos were made by nuclear transfer using cumulus cells, tail-tip fibroblasts, and embryonic stem cells. After 96 h of culture, the rates of Oct4-expressing blastocysts were as low as 50% and 60% for cumulus and fibroblast clones, respectively. However, regardless of Oct4 expression, the majority of those cloned blastocysts (> 90%) normally expressed Cdx2. Thus, even though somatic cloned embryos have reduced potential to produce the inner cell mass lineage, the TE lineage can be established and maintained.     

红豆越桔VviDREB1基因转化烟草的研究

通过构建pVB4215植物双元表达载体,采用农杆菌介导法转化烟草,研究VviDREB1在植物体中的异源表达特性.结果显示,实验获得了25个Hyg抗性株系,经过PCR、RT-PCR和GUS组织化学染色检测及Hyg基因的PCR复检等多点验证,证实表达载体边界内序列完整地整合到2个烟草株系的基因组中.转基因烟草株系在4℃低温处理20 h后,恢复生长5 h,叶片光系统PSⅡ抗寒性分析结果表明,转基因植株的叶片快速叶绿素荧光曲线OJIP各点数值高于对照植株,VviDREB1基因能够显著提高烟草的荧光产量,最大光化学效率Fv/Fm和以吸收光能为基础的性能指数PIABS较对照高,说明VviDREB1对保护植物组织细胞内光合系统PSⅡ有明显的作用,转VviDREB1基因烟草对低温有一定的忍耐能力.     

Application of Multiplexed Kinase Inhibitor Beads to Study Kinome Adaptations in Drug-Resistant Leukemia

Protein kinases play key roles in oncogenic signaling and are a major focus in the development of targeted cancer therapies. Imatinib, a BCR-Abl tyrosine kinase inhibitor, is a successful front-line treatment for chronic myelogenous leukemia (CML). However, resistance to imatinib may be acquired by BCR-Abl mutations or hyperactivation of Src family kinases such as Lyn. We have used multiplexed kinase inhibitor beads (MIBs) and quantitative mass spectrometry (MS) to compare kinase expression and activity in an imatinib-resistant (MYL-R) and -sensitive (MYL) cell model of CML. Using MIB/MS, expression and activity changes of over 150 kinases were quantitatively measured from various protein kinase families. Statistical analysis of experimental replicates assigned significance to 35 of these kinases, referred to as the MYL-R kinome profile. MIB/MS and immunoblotting confirmed the over-expression and activation of Lyn in MYL-R cells and identified additional kinases with increased (MEK, ERK, IKKα, PKCβ, NEK9) or decreased (Abl, Kit, JNK, ATM, Yes) abundance or activity. Inhibiting Lyn with dasatinib or by shRNA-mediated knockdown reduced the phosphorylation of MEK and IKKα. Because MYL-R cells showed elevated NF-κB signaling relative to MYL cells, as demonstrated by increased IκBα and IL-6 mRNA expression, we tested the effects of an IKK inhibitor (BAY 65-1942). MIB/MS and immunoblotting revealed that BAY 65-1942 increased MEK/ERK signaling and that this increase was prevented by co-treatment with a MEK inhibitor (AZD6244). Furthermore, the combined inhibition of MEK and IKKα resulted in reduced IL-6 mRNA expression, synergistic loss of cell viability and increased apoptosis. Thus, MIB/MS analysis identified MEK and IKKα as important downstream targets of Lyn, suggesting that co-targeting these kinases may provide a unique strategy to inhibit Lyn-dependent imatinib-resistant CML. These results demonstrate the utility of MIB/MS as a tool to identify dysregulated kinases and to interrogate kinome dynamics as cells respond to targeted kinase inhibition.     

土壤水分胁迫对夏玉米植株性状整齐度的影响

利用大型防雨设施池栽,严格调控水量,研究夏玉米全程及阶段性土壤水分胁迫对植株性状整齐度的影响。结果表明,夏玉米生育全程水分胁迫,植株性状整齐度全面劣化,产量极低;苗期阶段水分胁迫导致大、小苗,壮、弱苗两极分化明显,株高整齐度显著下降,其负效应持续至生育后期,生育进程推迟,千粒重显著降低,对产量造成一定影响;穗期阶段水分胁迫对穗长、穗料数整齐度影响明显,产量降幅度较大;花粒期阶段水平胁迫对千粒重及穗     

土壤干旱条件下玉米叶片内源激素含量及光合作用的变化

利用HPLC和ELISA技术研究了土壤干旱条件下玉米叶片内源IAA、ABA、ZR、DHZR、iPA含量的变化情况 ,以及叶片光合作用过程中 ,光合速率 (Pn) ,气孔导度 (Gs) ,PSⅡ光化学效率 (Fv/Fm)的变化情况 ,结果发现 :叶片中IAA浓度 ,在整个干旱过程中变化不大 ,与对照相比下降不明显 ;IAA的浓度对干旱的反应不敏感 ;叶片中ABA浓度在干旱最初 3d里急剧升高 ,直至最大值 ,之后有所下降 ;ABA的浓度对干旱反应敏感 ,但在整个干旱过程中ABA浓度并不随土壤相对含水量的减少而逐步升高 ;叶片中ZR和DHZR的浓度在干旱过程中与对照相比变化不明显 ,iPA浓度在干旱 4d后显著下降 ;叶片Pn在干旱初始 4d里随RWC的减小而缓慢下降 ,4d之后下降迅速 ;Gs从干旱第一天起即迅速减小 ,到第 4天近乎于零 ,Fv/Fm从干旱第 5天后开始逐步减小。这些结果证实在干旱过程中叶片ABA浓度的升高对气孔导度的调节作用 ,干旱初期光合速率的下降主要是气孔关闭所致 ;干旱后期光合速率的快速下降可能是PSⅡ光反应效率降低造成的     

Effective pulmonary delivery of an aerosolized plasmid DNA vaccine via surface acoustic wave nebulization

Background

Pulmonary-delivered gene therapy promises to mitigate vaccine safety issues and reduce the need for needles and skilled personnel to use them. While plasmid DNA (pDNA) offers a rapid route to vaccine production without side effects or reliance on cold chain storage, its delivery to the lung has proved challenging. Conventional methods, including jet and ultrasonic nebulizers, fail to deliver large biomolecules like pDNA intact due to the shear and cavitational stresses present during nebulization.

Methods

In vitro structural analysis followed by in vivo protein expression studies served in assessing the integrity of the pDNA subjected to surface acoustic wave (SAW) nebulisation. In vivo immunization trials were then carried out in rats using SAW nebulized pDNA (influenza A, human hemagglutinin H1N1) condensate delivered via intratracheal instillation. Finally, in vivo pulmonary vaccinations using pDNA for influenza was nebulized and delivered via a respirator to sheep.

Results

The SAW nebulizer was effective at generating pDNA aerosols with sizes optimal for deep lung delivery. Successful gene expression was observed in mouse lung epithelial cells, when SAW-nebulized pDNA was delivered to male Swiss mice via intratracheal instillation. Effective systemic and mucosal antibody responses were found in rats via post-nebulized, condensed fluid instillation. Significantly, we demonstrated the suitability of the SAW nebulizer to administer unprotected pDNA encoding an influenza A virus surface glycoprotein to respirated sheep via aerosolized inhalation.

Conclusion

Given the difficulty of inducing functional antibody responses for DNA vaccination in large animals, we report here the first instance of successful aerosolized inhalation delivery of a pDNA vaccine in a large animal model relevant to human lung development, structure, physiology, and disease, using a novel, low-power (<1 W) surface acoustic wave (SAW) hand-held nebulizer to produce droplets of pDNA with a size range suitable for delivery to the lower respiratory airways.     

中国海域团水虱属一新种(甲壳纲:等足目:团水虱科)

记述采自我国厦门及广东海门的团水虱科Sphaeromatidae一新种 ,中华团水虱Sphaeromasinensis。本种与采自海南岛的三口团水虱SphaeromatristeHeller相似 ,主要区别特征如下 :本种腹部前缘有 1个大突起 ,后者无 ;本种腹节具多个瘤状小突起 ,后者腹尾节则在近中部有 3对明显的大突起。正模标本♂ ,82F 83C ,1982年 4月 10日采自厦门何厝。模式标本保存在中国科学院海洋研究所标本馆。     

Tagging and mapping the thermo-sensitive genic male-sterile gene in rice (Oryza sativa L.) with molecular markers

The thermo-sensititve genic male-sterile (TGMS) gene in rice can alter fertility in response to temperature and is useful in the two-line system of hybrid rice production. However, little is known about the TGMS gene at the molecular level. The objective of this study was to identify molecular markers tightly linked with the TGMS gene and to map the gene onto a specific rice chromosome. Bulked segregant analysis of an F₂ population from 5460s (a TGMS mutant line) x Hong Wan 52 was used to identify RAPD markers linked to the rice TGMS gene. Four hundred RAPD primers were screened for polymorphisms between the parents and between two bulks representing fertile and sterile plants; of these, 4 primers produced polymorphic products. Most of the polymorphic fragments contained repetitive sequences. Only one singlecopy sequence fragment was found, a 1.2-kb fragment amplified by primer OPB-19 and subsequently named TGMS1.2. TGMS1.2 was mapped on chromosome 8 with a RIL population and confirmed by remapping with a DHL population. Segregation analysis using TGMS1.2 as a probe indicated that TGMS1.2 both consegregated and was lined with the TGMS gene in this population. It is located about 6.7 cM from the TGMS gene. As TGMS1.2 is linked to the TGMS gene, the TGMS gene must be located on chromosome 8.This research was supported by the Rockefeller Foundation and China National High-Tech Research and Development Program. The first author is a Rockefeller Career Fellow at Texas Tech University     

Automated measurement and statistical modelling of elastic laminae in arteries

Structural features of elastic laminae within arteries can provide vital information for both the mechanobiology and the biomechanics of the wall. In this paper, we propose, test and illustrate a new computer-based scheme for automated analysis of regional distributions of elastic laminae thickness, inter-lamellar distances and fragmentation furcation points (FPs) from standard histological images. Our scheme eliminates potential artefacts produced by tissue cutting, automatically aligns tissue according to physiologic orientations and performs cross-sectional measurements along radial directions. A statistical randomised complete block design and F test were used to assess the potential (non)-uniformity of lamellar thicknesses and separations along both radial and circumferential directions. Illustrative results for both normotensive and hypertensive thoracic porcine aorta revealed marked heterogeneity along the radial direction in nearly stress-free samples. Clearly, regional measurements can provide more detailed information about morphologic changes that cannot be gained by globally averaged evaluations alone. We also found that quantifying FP densities offers new information about potential elastin fragmentation, particularly in response to increased loading due to hypertension.