A synthetic wheat with 56 chromosomes derived fromTriticum turgidum andAegilops tauschii

By colchicine treatment of hybrids between Triticum turgidum and Aegilops tauschii (as seedlings), a fertile wheat plant (SHW-L2) carrying 56 chromosomes was artificially synthesized. At metaphase I of 50 pollen mother cells, the 56 chromosomes of the new wheat SHW-L2 showed a mean pairing configuration of 2.82 univalents, 6.18 rod bivalents, 19.39 ring bivalents, 0.5 trivalents, and 0.14 quadrivalents. Cytological analyses suggested that SHW-L2 had additional 7 pairs of chromosomes from the A and D genome besides the 42 chromosomes of common wheat. The special chromosome constitution of SHW-L2 may be derived from the chromosome doubling by the colchicine treatment of seedlings and then spontaneous doubling of gametes.     

Glial cell line-derived neurotrophic factor (GDNF) is a neurotrophic factor for sensory neurons: Comparison with the effects of the neurotrophins

We compared the effects of glial cell line-derived neurotrophic factor (GDNF) on dorsal root ganglion (DRG) sensory neurons to that of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin 3 (NT-3). All of these factors were retrogradely transported to sub-populations of sensory neuron cell bodies in the L4/L5 DRG of neonatal rats. The size distribution of ¹²⁵I-GDNF-labeled neurons was variable and consisted of both small and large DRG neurons (mean of 506.60 μm²). ¹²⁵I-NGF was preferentially taken up by small neurons with a mean cross-sectional area of 383.03 μm². Iodinated BDNF and NT-3 were transported by medium to large neurons with mean sizes of 501.48 and 529.27 μm², respectively. A neonatal, sciatic nerve axotomy-induced cell death model was used to determine whether any of these factors could influence DRG neuron survival in vivo. GDNF and NGF rescued nearly 100% of the sensory neurons. BDNF and NT-3 did not promote any detectable level of neuronal survival despite the fact that they underwent retrograde transport. We examined the in vitro survival-promoting ability of these factors on neonatal DRG neuronal cultures derived from neonatal rats. GDNF, NGF, and NT-3 were effective in vitro, while BDNF was not. The range of effects seen in the models described here underscores the importance of testing neuronal responsiveness in more than one model. The biological responsiveness of DRG neurons to GDNF in multiple models suggests that this factor may play a role in the development and maintenance of sensory neurons. © 1997 John Wiley & Sons, Inc. J Neurobiol 32: 22–32, 1997.     

圈养大熊猫野化培训期的生境选择特征

大熊猫(Ailuropoda melanoleuca)是我国特有的珍稀物种,也是世界上最濒危的野生动物之一。为了将人工繁育的部分大熊猫个体重引入其历史分布区或复壮野生种群,中国保护大熊猫研究中心从2003年开始进行圈养大熊猫的野外放归工作,通过野化培训以提高圈养大熊猫适应和选择野外环境的能力。对野化培训大熊猫"淘淘"的生境选择研究表明:该野化培训大熊猫幼仔经常活动于新笋密度较大的区域[生境与对照:(2.68±1.14)对(1.58±0.66)],却避开成竹密度过大[(9.91±2.51)对(12.18±4.68)]、竹子较高[(4.57±1.09) m对(4.98±0.66) m]以及枯死竹过多[(2.52±0.86)对(3.39±1.33)]的区域;喜欢活动于离水源[(1.59±0.67)对(2.19±0.87)]和隐蔽场所较近[(5.37±2.14) m对(8.35±7.76)m],以及距离乔木较远[(3.09±0.69) m对(2.70±0.42) m]和郁闭度较低[(1.85±0.57)对(2.10±0.47)]的区域(P < 0.05),新笋密度大小是该栖息地在整个野化培训期间是否被利用的最重要因素。该野化培训大熊猫幼仔保持着与带仔母兽相近的生境选择特征,对竹子环境的选择也与卧龙野生大熊猫相似,野化培训对该大熊猫幼仔产生了积极的作用。野化培训大熊猫幼仔形成的家域和核域面积分别为9.21 hm² 和1.93 hm²,占野化培训圈面积的51.95%和10.89%,其中家域面积仅有卧龙野生大熊猫的1.4%-2.4%,所以在以后的野化培训过程中需要采取增加野化培训圈中环境丰富度等方式,促进野化培训大熊猫形成较大的家域面积。     

Heterologous expression of an aspartic protease gene from biocontrol fungus Trichoderma asperellum in Pichia pastoris

Trichoderma asperellum parasitizes a large variety of phytopathogenic fungi. The mycoparasitic activity of T. asperellum depends on the secretion of complex mixtures of hydrolytic enzymes able to degrade the host cell wall and proteases which are a group of enzymes capable of degrading proteins from host. In this study, a full-length cDNA clone of aspartic protease gene, TaAsp, from T. asperellum was obtained and sequenced. The 1,185 bp long cDNA sequence was predicted to encode a 395 amino acid polypeptide with molecular mass of 42.3 kDa. The cDNA of TaAsp was inserted into the pPIC9K vector and transformed into yeast Pichia pastoris GS115 for heterologous expression. A clearly visible band with molecular mass about 42 kDa in the SDS-PAGE gel indicated that the transformant harboring the gene TaAsp had been successfully translated in P. pastoris and produced a recombinant protein. Enzyme characterization test showed that the optimum fermentation time for P. pastoris GS115 transformant was 72 h. Enzyme activity of the recombinant aspartic proteinase remained relatively stable at 25–60 °C and pH 3.0–9.0, which indicated its good prospect of application in biocontrol. The optimal pH value and temperature of the enzyme activity were pH 4.0 and 40 °C, and under this condition, with casein as the substrate, the recombinant protease activity was 18.5 U mL^?1. In order to evaluate antagonistic activity of the recombinant protease against pathogenic fungi, five pathogenic fungi, Fusarium oxysporum, Alternaria alternata, Cytospora chrysosperma, Sclerotinia sclerotiorum and Rhizoctonia solani, were applied to the test of in vitro inhibition of their mycelial growth by culture supernatant of P. pastoris GS115 transformant.     

Comparison of Expression of Inflammatory Cytokines in the Spinal Cord Between Young Adult and Aged Beagle Dogs

Aging is an inevitable process that occurs in the whole body system accompanying with many functional and morphological changes. Inflammation is known as one of age-related factors, and inflammatory changes could enhance mortality risk. In this study, we compared immunoreactivities of inflammatory cytokines, such as interleukin (IL)-2 (a pro-inflammatory cytokine), its receptor (IL-2R), IL-4 (an anti-inflammatory cytokine), and its receptor (IL-4R) in the cervical and lumbar spinal cord of young adult (2–3 years old) and aged (10–12 years old) beagle dogs using immunohistochemistry and western blotting. IL-2 and IL-2R-immunoreactive nerve cells were found throughout the gray matter of the cervical and lumbar spinal cord of young adult and aged dogs. In the spinal cord neurons of the aged dog, immunoreactivity and protein levels were apparently increased compared with those in the young adult dog. Change patterns of IL-4- and IL-4R-immunoreactive cells and their protein levels were also similar to those in IL-2 and IL-2R; however, IL-4 and IL-4R immunoreactivity in the periphery of the neuronal cytoplasm in the aged dog was much stronger than that in the young adult dog. These results indicate that the increase of inflammatory cytokines and their receptors in the aged spinal cord might be related to maintaining a balance of inflammatory reaction in the spinal cord during normal aging.     

小叶锦鸡儿灌丛下土壤水分对降雨的响应

以科尔沁沙地主要固沙灌木小叶锦鸡儿为研究对象,在其生长季次降雨21.5 mm后180 h内,利用TDR和微渗仪测量小叶锦鸡儿灌丛下不同部位的土壤含水量和土壤蒸发,并计算该灌丛下不同部位储水量和水量平衡.结果表明:降雨结束后初期,灌丛枝干的茎流作用使其根部的土壤含水量明显高于其他部位;灌丛根部水分的入渗速率大于灌丛中部和灌丛外缘.因冠幅的庇荫作用,灌丛下蒸发量小于灌丛外裸露沙地.水量平衡表明:小叶锦鸡儿灌丛下降雨后前期蒸散量明显高于灌丛外裸露沙地,与灌丛下根系的分布有直接关系.     

Tumor necrosis factor regulates intestinal epithelial cell migration by receptor-dependent mechanisms

Altered mucosal integrity andincreased cytokine production, including tumor necrosis factor (TNF),are the hallmarks of inflammatory bowel disease (IBD). In this study,we addressed the role of TNF receptors (TNFR) on intestinal epithelialcell migration in an in vitro wound closure model. With mouse TNFR1 orTNFR2 knockout intestinal epithelial cells, gene transfection, andpharmacological inhibitors, we show a concentration-dependentreceptor-mediated regulation of intestinal cell migration by TNF. Aphysiological TNF level (1 ng/ml) enhances migration through TNFR2,whereas a pathological level (100 ng/ml) inhibits wound closure through TNFR1. Increased rate of wound closure by TNFR2 or inhibition by TNFR1cannot be explained by either increased proliferation orapoptosis, respectively. Furthermore, inhibiting Src tyrosine kinase decreases TNF-induced focal adhesion kinase (FAK) tyrosine phosphorylation and cellular migration. We therefore conclude thatTNFR2 activates a novel Src-regulated pathway involving FAK tyrosinephosphorylation that enhances migration of intestinal epithelial cells.

     

Cytosolic prion protein is not toxic and protects against Bax-mediated cell death in human primary neurons

Recently, it was observed that reverse-translocated cytosolic PrP and PrP expressed in the cytosol induce rapid death in neurons (Ma, J., Wollmann, R., and Lindquist, S. (2002) Science 298, 1781-1785). In this study, we investigated whether accumulation of prion protein (PrP) in the cytosol is toxic to human neurons in primary culture. We show that in these neurons, a single PrP isoform lacking signal peptide accumulates in the cytosol of neurons treated with epoxomicin, a specific proteasome inhibitor. Therefore, endogenously expressed PrP is subject to the endoplasmic reticulum-associated degradation (ERAD) pathway and is degraded by the proteasome in human primary neurons. In contrast to its toxicity in N2a cells, reverse-translocated PrP (ERAD-PrP) is not toxic even when neurons are microinjected with cDNA constructs to overexpress either wild-type PrP or mutant PrPD178N. We found that ERAD-PrP in human neurons remains detergent-soluble and proteinase K-sensitive, in contrast to its detergent-insoluble and proteinase K-resistant state in N2a cells. Furthermore, not only is microinjection of a cDNA construct expressing CyPrP not toxic, it protects these neurons against Bax-mediated cell death. We conclude that in human neurons, ERAD-PrP is not converted naturally into a form reminiscent of scrapie PrP and that PrP located in the cytosol retains its protective function against Bax. Thus, it is unlikely that simple accumulation of PrP in the cytosol can cause neurodegeneration in prion diseases.