Modulation of coupling in the Escherichia coli ATP synthase by ADP and Pi: Role of the ε subunit C-terminal domain

The ε-subunit of ATP-synthase is an endogenous inhibitor of the hydrolysis activity of the complex and its α-helical C-terminal domain (εCTD) undergoes drastic changes among at least two different conformations. Even though this domain is not essential for ATP synthesis activity, there is evidence for its involvement in the coupling mechanism of the pump. Recently, it was proposed that coupling of the ATP synthase can vary as a function of ADP and P_i concentration. In the present work, we have explored the possible role of the εCTD in this ADP- and P_i-dependent coupling, by examining an εCTD-lacking mutant of Escherichia coli. We show that the loss of P_i-dependent coupling can be observed also in the εCTD-less mutant, but the effects of P_i on both proton pumping and ATP hydrolysis were much weaker in the mutant than in the wild-type. We also show that the εCTD strongly influences the binding of ADP to a very tight binding site (half-maximal effect ≈ 1 nM); binding at this site induces higher coupling in EF_OF₁ and increases responses to P_i. It is proposed that one physiological role of the εCTD is to regulate the kinetics and affinity of ADP/P_i binding, promoting ADP/P_i-dependent coupling.     

Surface N balances and reactive N loss to the environment from global intensive agricultural production systems for the period 1970-2030

Data for the historical years 1970 and 1995 and the FAO-Agriculture Towards 2030 projection are used to calculate N inputs (N fertilizer, animal manure, biological N fixation and atmospheric deposition) and the N export from the field in harvested crops and grass and grass consumption by grazing animals. In most industrialized countries we see a gradual increase of the overall N recovery of the intensive agricultural production systems over the whole 1970-2030 period. In contrast, low N input systems in many developing countries sustained low crop yields for many years but at the cost of soil fertility by depleting soil nutrient pools. In most developing countries the N recovery will increase in the coming decades by increasing efficiencies of N use in both crop and livestock production systems. The surface balance surplus of N is lost from the agricultural system via different pathways, including NH3 volatilization, denitrification, N2O and NO emissions, and nitrate leaching from the root zone. Global NH3-N emissions from fertilizer and animal manure application and stored manure increased from 18 to 34 Tg·yr-1 between 1970 and 1995, and will further increase to 44 Tg·yr-1 in 2030. Similar developments are seen for N2O-N (2.0 Tg·yr-1 in 1970, 2.7 Tg·yr-1 in 1995 and 3.5 Tg·yr-1 in 2030) and NO-N emissions (1.1 Tg·yr-1 in 1970, 1.5Tg·yr-1 in 1995 and 2.0 Tg·yr-1 in 2030).     

Isocitrate dehydrogenase 1 R132C mutation occurs exclusively in microsatellite stable colorectal cancers with the CpG island methylator phenotype

The CpG Island Methylator Phenotype (CIMP) is fundamental to an important subset of colorectal cancer; however, its cause is unknown. CIMP is associated with microsatellite instability but is also found in BRAF mutant microsatellite stable cancers that are associated with poor prognosis. The isocitrate dehydrogenase 1 (IDH1) gene causes CIMP in glioma due to an activating mutation that produces the 2-hydroxyglutarate oncometabolite. We therefore examined IDH1 alteration as a potential cause of CIMP in colorectal cancer. The IDH1 mutational hotspot was screened in 86 CIMP-positive and 80 CIMP-negative cancers. The entire coding sequence was examined in 81 CIMP-positive colorectal cancers. Forty-seven cancers varying by CIMP-status and IDH1 mutation status were examined using Illumina 450K DNA methylation microarrays. The R132C IDH1 mutation was detected in 4/166 cancers. All IDH1 mutations were in CIMP cancers that were BRAF mutant and microsatellite stable (4/45, 8.9%). Unsupervised hierarchical cluster analysis identified an IDH1 mutation-like methylation signature in approximately half of the CIMP-positive cancers. IDH1 mutation appears to cause CIMP in a small proportion of BRAF mutant, microsatellite stable colorectal cancers. This study provides a precedent that a single gene mutation may cause CIMP in colorectal cancer, and that this will be associated with a specific epigenetic signature and clinicopathological features.     

双源CT冠状动脉血管成像诊断心肌桥的价值探讨

目的：研究双源CT冠状动脉血管成像诊断心肌桥的临床价值。方法：选择260例具有典型心前区不适的患者进行双源CT冠脉血管成像检查,观察其发生部位,测量其长度和深度并进行分析。结果：260例受检患者中,62例共70段存在心肌桥,检出率达20.76%,高于文献报道的检出率18.2%。所有心肌桥均发生于左前降支,其中近段17段（24.4%）,中段43段（61.4%）,远段10段（14.2%）。心肌桥平均长度为15.8±6.4mm,深度为1.4±0.85mm。结论：双源CT冠状动脉血管成像因其便捷无创,不受心率严格限制且价格低廉可作为心肌桥筛查的理想检查手段。     

MR弥散加权成像对鉴别诊断良恶性椎体压缩性骨折的临床价值研究

目的：探讨MR弥散加权成像（DWI）鉴别诊断良恶性椎体压缩性骨折的临床价值。方法：对57例经临床或病理证实的椎体良恶性压缩性骨折患者行矢状位T1M、T2WI、T2WI／FS及DWI扫描,研究其在常规序列和DWI序列上的表现,将常规MR序列和DWI序列检出率进行比较,测量正常椎体及病变椎体的表观弥散系数（ADC）值,并进行统计学分析。结果：（1）MR常规序列和DWI序列（b=500s／mm2）表现：良性椎体压缩性骨折呈长T1长或等T2改变,T2WI／FS呈高信号,DWI可以呈高信号、等信号及低信号;恶性椎体压缩性骨折呈长T1长T2信号,大部分病灶T2WUFS及DWI呈高信号,少数变现为低信号;（2）MR常规序列和DWI序列（b=500s／mm2）病灶检出率的比较：T1WI、T2WI／FS及DWI序列病灶检出率均高于T2WI序列,其间的差别有显著性意义（P〈0．01）,T1WI、T2WI／FS及DWI序列病灶检出率之间无显著性差异（P〉0．01）;（3）ADC值比较：在DWI（b=500s／mm2）上,良性组ADC值为（2．03±0．83）×10^3mm^2/s,恶性组ADC值为（1．37±0．75）×10^-3mm^2/s,正常组ADC值为（0．36±0．21）×10^-3mm^2／s,成像条件相同时,良性组高于恶性组,两组间有明显的统计学意义（P〈0．05）。结论：DWI可较好的反映椎体的弥散特征,ADC值作为量化指标可对良恶性椎体压缩性骨折进行可靠鉴别。     

Frog brain expresses a 60 KDa Ca2+ binding protein similar to mammalian calreticulin

The present report was undertaken in an effort to characterize the nature of Ca2+ binding protein(s) in the central nervous system of less evolved vertebrates. In particular we investigated whether the brain microsomal fraction of Rana esculenta expresses calsequestrin, calreticulin and/or other related Ca2+ binding protein(s). We found that a 60 KDa protein having an NH2-terminal amino acid sequence similar to mammalian calreticulin is the major microsomal Ca2(+)-binding protein.     

3种香薷属植物染色体数目与核型分析

参照植物根尖细胞学研究的方法标准,对香薷属3种(5个居群)植物进行核形态学分析。结果表明:(1)从染色体数目看,密花香薷2居群染色体数目2n=16;野苏子2居群染色体数目2n=20,染色体数目和倍性与前人报道的一致;毛穗香薷染色体数目2n=10为首次报道。(2)聚类分析结果显示,3种(5居群)植物中野苏子和密花香薷亲缘关系较近;结合现有报道数据分析表明,该属植物仅有2种倍性(二倍体和四倍体),且二倍体占主导地位。(3)核型参数分析表明:密花香薷的稻城无名山居群1核型公式为2n=2x=16=14m+2sm,居群2为2n=2x=16=16m,着丝粒指数(CI)分别为39.57和42.32,不对称系数AI值分别为2.75和2.87,核型不对称性都为1A型;毛穗香薷的核型公式为2n=2x=10=10m,着丝粒指数(CI)为41.76,不对称系数AI值为5.25,核型不对称性为1B型;野苏子的昆明西山居群核型公式为2n=2x=20=14m+6sm,聂拉木樟木沟居群为2n=2x=20=16m+4sm,着丝粒指数(CI)分别为38.49和40.97,不对称系数AI值为4.20和4.30,核型不对称性为1B型和2B型。     

Tissue Characterization after a New Disaggregation Method for Skin Micro-Grafts Generation

Several new methods have been developed in the field of biotechnology to obtain autologous cellular suspensions during surgery, in order to provide one step treatments for acute and chronic skin lesions. Moreover, the management of chronic but also acute wounds resulting from trauma, diabetes, infections and other causes, remains challenging. In this study we describe a new method to create autologous micro-grafts from cutaneous tissue of a single patient and their clinical application. Moreover, in vitro biological characterization of cutaneous tissue derived from skin, de-epidermized dermis (Ded) and dermis of multi-organ and/or multi-tissue donors was also performed. All tissues were disaggregated by this new protocol, allowing us to obtain viable micro-grafts. In particular, we reported that this innovative protocol is able to create bio-complexes composed by autologous micro-grafts and collagen sponges ready to be applied on skin lesions. The clinical application of autologous bio-complexes on a leg lesion was also reported, showing an improvement of both re-epitalization process and softness of the lesion. Additionally, our in vitro model showed that cell viability after mechanical disaggregation with this system is maintained over time for up to seven (7) days of culture. We also observed, by flow cytometry analysis, that the pool of cells obtained from disaggregation is composed of several cell types, including mesenchymal stem cells, that exert a key role in the processes of tissue regeneration and repair, for their high regenerative potential. Finally, we demonstrated in vitro that this procedure maintains the sterility of micro-grafts when cultured in Agar dishes. In summary, we conclude that this new regenerative approach can be a promising tool for clinicians to obtain in one step viable, sterile and ready to use micro-grafts that can be applied alone or in combination with most common biological scaffolds.