CD and NMR investigation of collagen peptides mimicking a pathological Gly–Ser mutation and a natural interruption in a similar highly charged sequence context

Even a single Gly substitution in the triple helix domain of collagen leads to pathological conditions while natural interruptions are suggested to play important functional roles. Two peptides—one mimicking a pathological Gly–Ser substitution (ERSEQ) and the other one modeling a similar natural interruption sequence (DRSER)—are designed to facilitate the comparison for elucidating the molecular basis of their different biological roles. CD and NMR investigation of peptide ERSEQ indicates a reduction of the thermal stability and disruption of hydrogen bonding at the Ser mutation site, providing a structural basis of the OI disease resulting from the Gly–Ser mutation in the highly charged RGE environment. Both CD and NMR real‐time folding results indicate that peptide ERSEQ displays a comparatively slower folding rate than peptide DRSER, suggesting that the Gly–Ser mutation may lead to a larger interference in folding than the natural interruption in a similar RSE context. Our studies suggest that unlike the rigid GPO environment, the abundant R(K)GE(D) motif may provide a more flexible sequence environment that better accommodates mutations as well as interruptions, while the electrostatic interactions contribute to its stability. These results shed insight into the molecular features of the highly charged motif and may aid the design of collagen biomimetic peptides containing important biological sites.     

The Greenhouse Gas Footprint of China's Food System: An Analysis of Recent Trends and Future Scenarios

Food chain systems (FCSs), which begin in agricultural production and end in consumption and waste disposal, play a significant role in China's rising greenhouse gas (GHG) emissions. This article uses scenario analysis to show China's potential trajectories to a low‐carbon FCS. Between 1996 and 2010, the GHG footprint of China's FCSs increased from 1,308 to 1,618 megatonnes of carbon dioxide equivalent (Mt CO₂‐eq), although the emissions intensity of all food categories, except for aquatic food, recorded steep declines. We project three scenarios to 2050 based on historical trends and plausible shifts in policies and environmental conditions: reference scenario; technology improvement scenario; and low GHG emissions scenario. The reference scenario is based on existing trends and exhibits a large growth in GHG emissions, increasing from 1,585 Mt CO₂‐eq in 2010 to 2,505 Mt CO₂‐eq in 2050. In the technology improvement scenario, emissions growth is driven by rising food demand, but that growth will be counterbalanced by gains in agricultural technology, causing GHG emissions to fall to 1,413 Mt CO₂‐eq by 2050. Combining technology improvement with the shift to healthier dietary patterns, GHG emissions in the low GHG emissions scenario will decline to 946 Mt CO₂‐eq in 2050, a drop of 41.5% compared with the level in 2010. We argue that these are realistic projections and are indeed indicative of China's overall strategy for low‐carbon development. Improving agricultural technology and shifting to a more balanced diet could significantly reduce the GHG footprint of China's FCSs. Furthermore, the transition to a low‐carbon FCS has potential cobenefits for land sustainability and public health.     

Doxycycline enhances the Ras‐MAPK signaling and proliferation of mouse thymic epithelial cells

Depletion of T‐cell‐dependent immunity is a major consideration for patients suffering from infections of human immunodeficiency virus (HIV), those undergoing organ transplantation, and those receiving anti‐cancer chemotherapy and/or radiotherapy. In general, T‐cell regeneration occurs in the thymus through thymopoiesis. We have found that doxycycline (Dox), a tetracycline derivative, enhances the proliferation of mouse thymic epithelial cells, which are unique in their capacity to support positive selection and are essential throughout the development of thymocytes. Cell cycle analysis indicates that the increased cell proliferation is due to a shortened G₀/G₁ phase. To reveal the underlying mechanisms, we examined the expression of an array of molecules that regulate the cell cycle. The results show that in mouse thymic medullary‐type epithelial cell line 1 (MTEC1) Dox leads to elevated levels of H‐Ras, phosphorylated extracellular signal‐regulated kinase 1/2 (p‐ERK1/2), cyclin E, cyclin dependent kinase 4/2 (CDK4/CDK2), E2F3, and c‐myc. These data, and the observation that the proliferation‐enhancing effect is largely abolished following treatment with an ERK inhibitor support an active role of the Ras‐ERK/mitogen‐activated protein kinase (MAPK) signaling pathway. In conclusion, the present study reveals a new activity of an old family of antibiotics. The in vivo effect of Dox on immune reconstitution warrants further exploration. J. Cell. Biochem. 107: 494–503, 2009. © 2009 Wiley‐Liss, Inc.     

Comparative proteomic analysis of mesenchymal stem cells derived from human bone marrow,umbilical cord,and placenta: Implication in the migration

Umbilical cord (UC) and placenta (P) have been suggested as alternatives to bone marrow (BM) as sources of mesenchymal stem cells (MSC) for cell therapy, with both UC‐ and P‐MSC possess immunophenotypic and functional characteristics similar to BM‐MSC. However, their migration capacity, which is indispensable during tissue regeneration process, is unclear. Under defined conditions, the migration capacity of BM‐ and P‐MSC was found 5.9‐ and 3.2‐folds higher than that of UC‐MSC, respectively. By the use of 2‐DE and combined MS and MS/MS analysis, six differentially expressed proteins were identified among these MSC samples, with five of them known to be involved in cell migration as migration enhancing or inhibiting proteins. Consistent with their migration capacity, the levels of migration enhancing proteins including cathepsin B, cathepsin D and prohibitin,were significantly lower in UC‐MSC when compared with those in BM‐ and P‐MSC. For the migration inhibiting proteins such as plasminogen activator inhibitor‐1 (PAI‐1) and manganese superoxide dismutase, higher expression was found in the UC‐MSC. We also showed that the overexpression of the PAI‐1 impaired the migration capacity of BM‐ and P‐MSC while silencing of PAI‐1 enhanced the migration capacity of UC‐MSC. Our study indicates that PAI‐1 and other migration‐related proteins are pivotal in governing the migration capacity of MSC.     

Distribution of heme oxygenase-2 and NADPH-diaphorase in the spinal trigeminal nucleus of the rat

Heme oxygenase (HO)/carbon monoxide (CO) and nitric oxide synthase (NOS)/nitric oxide (NO) systems are involved in sensory information processing. The present study was undertaken to examine the distribution of HO-2 and NOS in the spinal trigeminal nucleus (STN) of the rat, using histochemistry and immunohistochemistry. Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) staining was found that NADPH-d activity was more prominent in the nucleus caudalis (Vc) and the dorsomedial subdivision of the nucleus oralis (Vo) than in other spinal trigeminal regions. Immunohistochemistry for HO-2 revealed that HO-2 staining neurons distributed extensively, which intensity was higher in the rostral than caudal part of the STN. The colocalization of NADPH-d and HO-2 was mainly confined in the Vc. The expression and distribution of NADPH-d and HO-2 suggest that NO and CO are likely neurotransmitters and might function in the processing orofacial signal in the STN together.     

Effects of elevated CO2 concentration, supplemental irrigation and nitrogenous fertilizer application on rain-fed spring wheat yield

It is expected that the CO₂ concentration of the Earth’s atmosphere will reach 600–1000 ppm by the end of the 21st century. Therefore, in this study, we evaluated the effects of elevated CO₂ concentrations on the development of rain-fed spring wheat in an attempt to identify a practical pathway to increase crop production. To accomplish this, a field experiment was conducted at Guyuan Experimental Station in a semiarid region of China during 2005–2007. During this experiment, the CO₂ concentration was increased to 40.0 ppm and supplemental irrigation and nitrogenous fertilizer (N fertilizer) were applied. The experimental results showed that the elevated CO₂ concentration significantly improved the thousand-grain weight and the grain number per spike. Furthermore, supplemental irrigation and N fertilizer application during the elongation and booting stage of rain-fed spring wheat in conjunction with an elevated CO₂ concentration improved the water use efficiency (WUE), nitrogen use efficiency (NUE), thousand-grain weight, and the yield by 14.6%, 39.6%, 9.3%, and 14.7%, respectively, when compared to groups subjected to the same treatment but not grown under elevated CO₂ concentrations. Furthermore, the spring wheat yield was improved by 81.8% in response to an elevated CO₂ concentration, 60 mm of supplemental irrigation and applied N fertilizer (37.5 g m^?2 NH₄NO₃). However, the presence of an elevated CO₂ concentration without supplemental irrigation and N fertilizer only resulted in an increase in the wheat yield of 7.8%. Consequently, the combination of elevated CO₂ concentration, supplemental irrigation and N fertilizer application played an important role in the improvement of WUE, NUE, thousand-grain weight, and grain yield of rain-fed spring wheat in this region.     

CD95 (Fas) ligand-expressing vesicles display antibody-mediated, FcR-dependent enhancement of cytotoxicity

Bioactive Fas ligand (FasL)-expressing vesicles were generated (vesicle preparation, VP) from two cell lines overexpressing FasL. The effect of NOK-1 anti-FasL mAb (mouse IgG1) on the cytotoxicity of FasL VP against various targets was determined. At high concentrations (1-10 microg/ml), NOK-1 inhibited the cytotoxicity. By contrast, NOK-1 in the dose range of 1-100 ng/ml significantly enhanced cytotoxicity against the FcR(+) LB27.4, M59, and LF(+) targets, but not the FcR(-) Jurkat and K31H28 hybridoma T cell targets. The ability to enhance FasL VP-mediated cytotoxicity could be blocked by the FcR-specific mAb 2.4G2. Enhancement was also observed with FcR(+) A20 B lymphoma but not with the FcR(-) A20 variant. Enhancement of FasL VP cytotoxicity was observed with five IgG anti-FasL mAbs, but not with an IgM anti-FasL mAb. Inhibition was observed with high doses of all mAb except the IgG anti-FasL mAb G247-4, which is specific to a segment outside the FasL binding site. Interestingly, under identical conditions but in the presence of 2.4G2, G247-4 inhibited the cytotoxicity of FasL VP. In addition, G247-4 inhibited the FasL VP-mediated killing of FcR(-) Jurkat. The data demonstrate that FasL-expressing bioactive vesicles display a property heretofore unknown in bioactive agents that express FasL-mediated cytotoxicity. The mechanism of the Ab-mediated, FcR-dependent enhancement of cytotoxicity of bioactive vesicles and its physiological significance are discussed.     

基于地统计学方法的稻田灰飞虱与蜘蛛时空动态分析

通过田间调查,用地统计学方法对灰飞虱Laodelphax striatellus (Fallén)及其重要天敌蜘蛛的时空动态进行了比较分析。通过变差函数模型的拟合及空间格局的分析,得出灰飞虱长翅成虫、若虫与蜘蛛的变程范围相近,结构性强度的范围相近,但同一时间灰飞虱与蜘蛛的变程和结构性强度则均不尽相同;灰飞虱若虫与蜘蛛自始至终为聚集分布,但灰飞虱长翅成虫与蜘蛛的分布型则有差异,在某些时间为均匀分布。用普通克立格方法作出空间分布图,在时间序列上进行比较分析,得出稻田蜘蛛与灰飞虱成若虫的空间分布均有相似性,尤其与若虫相似性更强,表现出比较明显的空间跟随现象,且空间跟随现象与灰飞虱虫量的消长有密切的关系,但是由于天气、稻田环境、灰飞虱其他天敌以及蜘蛛其他猎物的存在等因素的影响使得其空间跟随又有着相当的复杂性。