Expression of hGM-CSF in silk glands of transgenic silkworms using gene targeting vector

The silk gland of the silkworm is a highly specialized organ that has the wonderful ability to synthesize and secrete silk protein. To express human granucyto-macrophage colony-stimulating factor (hGM-CSF) in the posterior silk glands of gene-targeted silkworms, a targeting vector pSK-FibL-L-A3GFP-PH-GMCSF-LPA-FibL-R was constructed, harboring a 1.2 kb portion of the left homogenous arm (FibL-L), a 0.5 kb portion of the right homogenous arm (FibL-R), fibroin H-chain-promoter-driven hGM-CSF and silkworm actin 3-promoter-driven gfp. The targeting vector was then introduced into the eggs of silkworm, and the transgenic silkworms were verified by PCR and DNA hybridization after being screened for the gfp gene. Western blotting analysis using an antibody against hGM-CSF demonstrated a specific band with a molecular weight of 22 kD in the silk glands of the G3 generation transgenic silkworms. The level of expression of hGM-CSF in the posterior silk glands of the G3 generation transgenic silkworms was approximately 2.70 ng/g of freeze-dried powdered posterior silk gland. These results showed that the heterologous gene could be introduced into the silkworm genome and expressed successfully. Further more, the exogenous genes existing in the G5 transgenic silkworm identified by PCR confirmed its integration stability. In addition, the silk glands containing expressed hGM-CSF performed the function of significantly increasing leukocyte count of CY-treated mice in a time-and-dose-dependent manner.     

MiR-221 promotes cardiac hypertrophy in vitro through the modulation of p27 expression

Cardiac hypertrophy has been known as an independent predictor for cardiovascular morbidity and mortality. Molecular mechanisms underlying the development of heart failure remain elusive. Recently, microRNAs (miRs) have been established as important regulators in cardiac hypertrophy. Here, we reported miR-221 was up-regulated in both transverse aortic constricted mice and patients with hypertrophic cardiomyopathy (HCM). Forced expression of miR-221 by transfection of miR-221 mimics increased myocyte cell size and induced the re-expression of fetal genes, which were inhibited by the knockdown of endogenous miR-221 in cardiomyocytes. The TargetScan algorithm-based prediction identified that p27, a cardiac hypertrophic suppressor, is the putative target of miR-221, which was confirmed by luciferase assay and Western blotting. In conclusion, our results demonstrated that miR-221 regulated cardiomyocyte hypertrophy probably through down-regulation of p27, suggesting that miR-221 may be a new intervention target for cardiac hypertrophy.     

Two non-allelic nuclear genes restore fertility in a gametophytic pattern and enhance abiotic stress tolerance in the hybrid rice plant

In indica rice, the HongLian (HL)-type combination of cytoplasmic male sterility (CMS) and fertility restoration (Rf) is widely used for the production of commercial hybrid seeds in China, Laos, Vietnam and other Southeast Asian countries. Generally, any member of the gametophytic fertility restoration system, 50% of the pollen in hybrid F(1) plants displays recovered sterility. In this study, however, a HL-type hybrid variety named HongLian You6 had approximately 75% normal (viable) pollen rather than the expected 50%. To resolve this discrepancy, several fertility segregation populations, including F(2) and BC(1)F(1) derived from the HL-CMS line Yuetai A crossed with the restorer line 9311, were constructed and subjected to genetic analysis. A gametophytic restoration model was discovered to involve two non-allelic nuclear restorer genes, Rf5 and Rf6. The Rf5 had been previously identified using a positional clone strategy. The Rf6 gene represents a new restorer gene locus, which was mapped to the short arm of chromosome 8. The hybrid F(1) plants containing one restorer gene, either Rf5 or Rf6, displayed 50% normal pollen grains with I(2)-KI solution; however, those with both Rf5 and Rf6 displayed 75% normal pollens. We also established that the hybrid F(1) plants including both non-allelic restorer genes exhibited an increased stable seed setting when subjected to stress versus the F(1) plants with only one restorer gene. Finally, we discuss the breeding scheme for the plant gametophytic CMS/Rf system.     

Modeling Impacts of Alternative Practices on Net Global Warming Potential and Greenhouse Gas Intensity from Rice–Wheat Annual Rotation in China

Background

Evaluating the net exchange of greenhouse gas (GHG) emissions in conjunction with soil carbon sequestration may give a comprehensive insight on the role of agricultural production in global warming.

Materials and Methods

Measured data of methane (CH₄) and nitrous oxide (N₂O) were utilized to test the applicability of the Denitrification and Decomposition (DNDC) model to a winter wheat – single rice rotation system in southern China. Six alternative scenarios were simulated against the baseline scenario to evaluate their long-term (45-year) impacts on net global warming potential (GWP) and greenhouse gas intensity (GHGI).

Principal Results

The simulated cumulative CH₄ emissions fell within the statistical deviation ranges of the field data, with the exception of N₂O emissions during rice-growing season and both gases from the control treatment. Sensitivity tests showed that both CH₄ and N₂O emissions were significantly affected by changes in both environmental factors and management practices. Compared with the baseline scenario, the long-term simulation had the following results: (1) high straw return and manure amendment scenarios greatly increased CH₄ emissions, while other scenarios had similar CH₄ emissions, (2) high inorganic N fertilizer increased N₂O emissions while manure amendment and reduced inorganic N fertilizer scenarios decreased N₂O emissions, (3) the mean annual soil organic carbon sequestration rates (SOCSR) under manure amendment, high straw return, and no-tillage scenarios averaged 0.20 t C ha⁻¹ yr⁻¹, being greater than other scenarios, and (4) the reduced inorganic N fertilizer scenario produced the least N loss from the system, while all the scenarios produced comparable grain yields.

Conclusions

In terms of net GWP and GHGI for the comprehensive assessment of climate change and crop production, reduced inorganic N fertilizer scenario followed by no-tillage scenario would be advocated for this specified cropping system.     

The HIV-1 passage from cytoplasm to nucleus: the process involving a complex exchange between the components of HIV-1 and cellular machinery to access nucleus and successful integration

The human immunodeficiency virus 1 (HIV-1) synthesizes its genomic DNA in cytoplasm as soon as it enters the cell. The newly synthesized DNA remains associated with viral/cellular proteins as a high molecular weight pre-integration complex (PIC), which precludes passive diffusion across intact nuclear membrane. However, HIV-1 successfully overcomes nuclear membrane barrier by actively delivering its DNA into nucleus with the help of host nuclear import machinery. Such ability allows HIV-1 to productively infect non-dividing cells as well as dividing cells at interphase. Further, HIV-1 nuclear import is also found important for the proper integration of viral DNA. Thus, nuclear import plays a crucial role in establishment of infection and disease progression. While several viral components, including matrix, viral protein R, integrase, capsid, and central DNA flap are implicated in HIV-1 nuclear import, their molecular mechanism remains poorly understood. In this review, we will elaborate the role of individual viral factors and some of current insights on their molecular mechanism(s) associated with HIV-1 nuclear import. In addition, we will discuss the importance of nuclear import for subsequent step of viral DNA integration. Hereby we aim to further our understanding on molecular mechanism of HIV-1 nuclear import and its potential usefulness for anti-HIV-1 strategies.     

The deubiquitinating enzyme complex BRISC is required for proper mitotic spindle assembly in mammalian cells

Deubiquitinating enzymes (DUBs) negatively regulate protein ubiquitination and play an important role in diverse physiological processes, including mitotic division. The BRCC36 isopeptidase complex (BRISC) is a DUB that is specific for lysine 63–linked ubiquitin hydrolysis; however, its biological function remains largely undefined. Here, we identify a critical role for BRISC in the control of mitotic spindle assembly in cultured mammalian cells. BRISC is a microtubule (MT)-associated protein complex that predominantly localizes to the minus ends of K-fibers and spindle poles and directly binds to MTs; importantly, BRISC promotes the assembly of functional bipolar spindle by deubiquitinating the essential spindle assembly factor nuclear mitotic apparatus (NuMA). The deubiquitination of NuMA regulates its interaction with dynein and importin-β, which are required for its function in spindle assembly. Collectively, these results uncover BRISC as an important regulator of the mitotic spindle assembly and cell division, and have important implications for the development of anticancer drugs targeting BRISC.     

逐日气象要素空间插值方法的比较

采用距离反比权重法(IDW)、协克里格法(CK)和薄盘样条法(TPS) 3种不同的空间插值方法,对我国1951-2005年气象数据完整的559个气象站点逐月第15日的平均基本气象要素(最高气温、最低气温、日照时数和降水量)进行了插值分析与评价.结果表明：3种插值方法中,TPS法对最高气温和最低气温插值的根均方差（RMSE）最小(1.02 ℃和1.12 ℃)、R²最大(0.9916和0.9913);不同季节中,TPS法对秋季最高气温、夏季最低气温进行插值的RMSE均最小(0.83℃、0.86 ℃),R²均为秋季最高.对于日照时数和降水量而言,TPS法的RMSE最小(0.59 h和1.01 mm)、R²最大(0.9118和0.8135);不同季节中,TPS法对冬季日照时数进行插值的RMSE最小(0.49 h)、R²最大(0.9293),TPS法对冬季降水量进行插值的RMSE最小(0.33 mm),IDW法对夏季降水量进行插值的RMSE最小(2.01 mm),CK法对春季降水量进行插值的R²最大(0.8781).TPS法可作为我国大量逐日基本气象要素的最优空间插值方法.     

Understanding the genetic and epigenetic architecture in complex network of rice flowering pathways

Although the molecular basis of flowering time control is well dissected in the long day (LD) plant Arabidopsis, it is still largely unknown in the short day (SD) plant rice. Rice flowering time (heading date) is an important agronomic trait for season adaption and grain yield, which is affected by both genetic and environmental factors. During the last decade, as the nature of florigen was identified, notable progress has been made on exploration how florigen gene expression is genetically controlled. In Arabidopsis expression of certain key flowering integrators such as FLOWERING LOCUS C (FLC) and FLOWERING LOCUS T (FT) are also epigenetically regulated by various chromatin modifications, however, very little is known in rice on this aspect until very recently. This review summarized the advances of both genetic networks and chromatin modifications in rice flowering time control, attempting to give a complete view of the genetic and epigenetic architecture in complex network of rice flowering pathways.     

Evaluating the effectiveness of landscape configuration metrics from landscape composition metrics

Although landscape configuration and landscape composition metrics are correlated theoretically and empirically, the effectiveness of configuration metrics from composition metrics has not been explicitly investigated. This study explored to what extent substantial information of configuration metrics increases from certain easily calculated and extensively used composition metrics and how strongly the effectiveness is influenced by different factors. The effectiveness of 12 landscape configuration metrics from the percentage of landscape (PLAND) of each land-use class and patch density (PD) was evaluated through the coefficient of determination (R ²) of multivariate stepwise linear regression analysis of 150 town-based landscape samples from three regions. The different landscape configuration metrics from PLAND and PD presented significantly different performances in terms of effectiveness [the contagion index and aggregation index possess minimal information, and the effective mesh size (MESH) and area-weighted mean patch fractal dimension possess abundant information]. Furthermore, the effectiveness of configuration metrics showed different responses to changing cell sizes and different land-use categorization in different regions (interspersion and juxtaposition index, patch cohesion index, and MESH exhibited large variations in R ² among the different regions). No single, uniform, consistent characteristic of effectiveness was determined across different factors. This new approach to understanding the effectiveness of configuration metrics helps clarify landscape metrics and is fundamental to landscape metric assessment.     

Asp97 is a crucial residue involved in the ligation of the [Fe4S4] cluster of IscA from Acidithiobacillus ferrooxidans

IscA was proposed to be involved in the ironsulfur cluster assembly encoded by the iscSUA operon, but the role of IscA in the iron-sulfur cluster assembly still remains controversial. In our previous study, the IscA from A. ferrooxidans was successfully expressed in Escherichia coli, and purified to be a [Fe4S4]-cluster-containing protein. Cys35, Cys99, and Cys101 were important residues in ligating with the [Fe4S4] cluster. In this study, Asp97 was found to be another ligand for the iron-sulfur cluster binding according to sitedirected mutagenesis results. Molecular modeling for the IscA also showed that Asp97 was a strong ligand with the [Fe4S4] cluster, which was in good agreement with the experimental results. Thus, the [Fe4S4] cluster in IscA from A. ferrooxidans was ligated by three cysteine residues and one aspartic acid.