Methyl β-cyclodextrin reduces accumulation of reactive oxygen species and cell death in yeast

Stabilized F-actin structures have been shown to be detrimental to both mammalian and yeast cells. In yeast, stabilization of actin caused by addition of jasplakinolide, by point mutations in the act1 gene, or by deletion of certain genes that regulate F-actin leads to cell death with hallmarks of apoptosis. In particular, there is an elevation in the levels of reactive oxygen species, and we have shown the importance of the Ras/cAMP pathway for this effect. Here we show that in yeast cells deleted for end3, which functions to regulate actin organization during endocytosis, treatment of cells with methyl β-cyclodextrin reduces levels of reactive oxygen species and inhibits cell death progression. Methyl β-cyclodextrin is widely used to disrupt lipid rafts that contain cholesterol. The mechanism through which the rescue is achieved was investigated and we demonstrate that methyl β-cyclodextrin reduces accumulation of Ras2 at the plasma membrane in Δend3 cells. We use FRAP and live cell imaging to determine the possible mechanism through which methyl β-cyclodextrin functions to elicit this effect on Ras2 localization. Finally, we demonstrate that addition of methyl β-cyclodextrin to wild-type cells can act to protect cells from acute oxidative stress caused by addition of hydrogen peroxide.     

Quantitative FRET measurement based on spectral unmixing of donor,acceptor and spontaneous excitation‐emission spectra

The spontaneous excitation‐emission (ExEm) spectrum is introduced to the quantitative mExEm‐spFRET methodology we recently developed as a spectral unmixing component for quantitative fluorescence resonance energy transfer measurement, named as SPEES‐FRET method. The spectral fingerprints of both donor and acceptor were measured in HepG2 cells with low autofluorescence separately expressing donor and acceptor, and the spontaneous spectral fingerprint of HEK293 cells with strong autofluoresence was measured from blank cells. SPEES‐FRET was performed on improved spectrometer‐microscope system to measure the FRET efficiency (E) and concentration ratio (R _C) of acceptor to donor vales of FRET tandem plasmids in HEK293 cells, and obtained stable and consistent results with the expected values. Moreover, SPEES‐FRET always obtained stable results for the bright and dim cells coexpressing Cerulean and Venus or Cyan Fluorescent Protein (CFP)‐Bax and Yellow fluorescent protein (YFP)‐Bax, and the E values between CFP‐Bax and YFP‐Bax were 0.02 for healthy cells and 0.14 for the staurosporine (STS)‐treated apoptotic cells. Collectively, SPEES‐FRET has very strong robustness against cellular autofluorescence, and thus is applicable to quantitative evaluation on the protein‐protein interaction in living cells with strong autofluoresence.      

The discovery of anthranilic acid-based MMP inhibitors. Part 3: incorporation of basic amines.

Anthranilic acid derivatives bearing basic amines were prepared and evaluated in vitro and in vivo as inhibitors of MMP-1, MMP-9, MMP-13, and TACE. Piperazine 4u has been identified as a potent, selective, orally active inhibitor of MMP-9 and MMP-13.     

PLZF-RARα融合基因的生物学功能研究

构建了8个PLZF-RARα融合基因突变体.用“滞后”胶实验证实PLZF-RARα与PML-RARα一样,亦能以同二聚体的形式结合到维甲酸反应元件(RARE)上,且PLZF的POZ结构域介导PLZF-RARα同二聚体的形成和稳定.但两者与RARE的结合类型存在差异,在DR5G,PML-RARα的结合强度大于PLZF-RARα;而在DR5T,则是PLZF-RARα强于PML-RARα.进一步工作证实PLZF-RARα能与RXR形成异二聚体,并产生4种复合物.用免疫沉淀法发现PLZF-RARα亦能与PLZF形成异二聚体,而且也是通过POZ结构域介导PLZF-RARα和PLZF异二聚体的形成.同PML-RARα一样,PLZF-RARα对RARE的结合反应亦受维甲酸调控.     

Antiviral flavonoids from the root bark of Morus alba L

A prenylated flavonoid, moralbanone, along with seven known compounds kuwanon S, mulberroside C, cyclomorusin, eudraflavone B hydroperoxide, oxydihydromorusin, leachianone G and alpha-acetyl-amyrin were isolated from the root bark of Morus alba L. Leachianone G showed potent antiviral activity (IC(50) = 1.6 microg/ml), whereas mulberroside C showed weak activity (IC(50) = 75.4 microg/ml) against herpes simplex type 1 virus (HSV-1). Their structures were elucidated by spectroscopic methods.     

涡度相关观测的能量闭合状况及其对农田蒸散测定的影响

涡度相关法被认为是测定农田蒸散量的标准方法。然而,能量不闭合现象在涡度相关测量中普遍存在。分析能量不闭合对涡度相关观测的影响,对于提高涡度相关观测精度具有重要意义。以蒸渗仪法为参照,探讨涡度相关观测的能量闭合状况对农田蒸散测定的影响,在导致涡度相关观测能量不闭合的诸多因素中,寻找对蒸散测定有影响的因素。结果表明:涡度相关观测的白天能量平衡比率(EBR)呈秋冬高、春夏低的变化特征,麦季日均EBR范围在0.26—2.84之间,平均1.15;玉米季日均EBR范围在0.19—2.59之间,平均0.78。无论麦季或玉米季,涡度相关法测定的平均蒸散量(ETec)均明显低于蒸渗仪法观测值(ETL),但两者显著相关(P<0.01),并有相似的季节变化。平均蒸散比(ETec/ETL)麦季约为0.61,玉米季约为0.50。在冬小麦田和夏玉米田,ETec/ETL均与EBR显著相关(P<0.01)。麦田种植密度大,下垫面较均匀,蒸散比与EBR成正比(P<0.01),且不受叶面积指数(LAI)大小影响;反之,玉米田种植密度小,只有当LAI>1,下垫面变得较均匀后,蒸散比与EBR的关系才变得显著(P<0.01)。风速小时ETec/ETL与EBR显著相关,风速增加时二者相关性减弱。尤其在玉米田,当摩擦风速(u*)大于0.3 m/s时,ETec/ETL与EBR的相关性不再显著。风速小时,大气湍流微弱,湍流的涡旋较大。在有限的观测时段(0.5h)内,涡度相关仪的传感器难以捕捉足够的湍涡能量,所测湍流能量偏低,导致能量不闭合。以上结果为应用能量平衡比率校正农田蒸散提供了可能途径。     

Colonization of endophyte Acremonium sp. D212 in Panax notoginseng and rice mediated by auxin and jasmonic acid

Endophytic fungi can be beneficial to plant growth. However, the molecular mechanisms underlying colonization of Acremonium spp. remain unclear.In this study, a novel endophytic Acremonium strain was isolated from the buds of Panax notoginseng and named Acremonium sp. D212. The Acremonium sp. D212 could colonize the roots of P. notoginseng,enhance the resistance of P. notoginseng to root rot disease, and promote root growth and saponin biosynthesis in P. notoginseng. Acremonium sp. D212 could secrete indole-3-acetic acid(IAA) and jasmonic acid(JA), and inoculation with the fungus increased the endogenous levels of IAA and JA in P. notoginseng. Colonization of the Acremonium sp. D212 in the roots of the rice line Nipponbare was dependent on the concentration of methyl jasmonate(Me JA)(2–15 μmol/L) and 1-naphthalenacetic acid(NAA)(10–20 μmol/L). Moreover, the roots of the JA signaling-defective coi1-18 mutant were colonized by Acremonium sp. D212 to a lesser degree than those of the wild-type Nipponbare and mi R393 boverexpressing lines, and the colonization was rescued by Me JA but not by NAA. It suggests that the cross-talk between JA signaling and the auxin biosynthetic pathway plays a crucial role in the colonization of Acremonium sp. D212 in host plants.