基于拉曼技术的单细胞生长检测方法

目的 单细胞生长检测可以更加科学地揭示微生物代谢变化的规律,为后期微生物工程应用提供指导。针对微生物生长应用于食品安全期和最佳食用期的精准检测问题,本文提出一种基于拉曼技术的单细胞生长检测方法。方法 首先,通过同步培养实验采集了枯草芽孢杆菌两个批次共900个单细胞拉曼光谱（SCRS）数据,其中600个用于训练和测试,另一批次300个用于模型验证。其次,基于主成分分析的特征关系矩阵,提出CP-SP特征评估方法以筛选SCRS特征用于模型检测。再基于XGBoost构建检测模型,并应用网格搜索和交叉验证对检测模型进行调优。最后,应用混淆矩阵、ROC曲线评估模型对细胞滞后期、对数期和稳定期的检测准确率、敏感性和特异性。结果 选用CP-SP筛选的第一、第二和第四主成分较特征贡献率前3个主成分的分类性能提高了3.1%,调优后的细胞生长检测模型测试准确率为96.0%,验证准确率为92.3%。结论 基于拉曼技术的单细胞生长检测方法能准确识别单细胞生长状态且具有较高的泛化能力,可为食品安全和保鲜制定精准调控机制提供科学指导。     

城市建筑屋顶光伏发电潜力评估方法和模型

建筑屋顶作为闲置的土地资源已成为光伏发电重要的潜在空间,屋顶光伏发电是脱碳电力供应的主要方式,将在实现城市碳中和进程中发挥重要作用。对建筑屋顶光伏发电潜力进行精确评估将有助于分布式光伏的科学规划和合理布局,提升土地利用效率。旨在对建筑屋顶光伏发电潜力影响因素和评估方法,以及光伏发电潜力主要评估模型进行系统性阐述,比较分析不同评估方法的优缺点,总结未来研究的重点方向。现有研究表明,建筑屋顶光伏发电潜力评估已从经验取值发展为定量空间分析,评估尺度、评估精度和评估成本已经成为不同评估方法综合权衡的重点。现有三种评估方法中,采样法计算成本和数据成本较低,但评估结果不确定性较大、精度较低;全面评估法评估精度较高,但数据获取成本和计算成本较高;机器学习法能够高效挖掘大数据潜力,且算法性能显著提升,因而相较于其他方法更适宜大尺度建筑屋顶光伏发电潜力评估。当前建筑屋顶光伏发电潜力评估仍然存在大尺度精细评估缺乏、评估结果不确定性大以及评估模型计算量大等问题。未来研究重点应关注三个方面：1)建立适宜不同区域的高精度简化模型并完善技术潜力评估模型;2)阐明建筑屋顶光伏发电潜力的影响因素,为代表性建筑分类体系...     

The chemokine interleukin-8 acutely reduces Ca(2+) currents in identified cholinergic septal neurons expressing CXCR1 and CXCR2 receptor mRNAs

The chemokine IL-8 is known to be synthesized by glial cells in the brain. It has traditionally been shown to have an important role in neuroinflammation but recent evidence indicates that it may also be involved in rapid signaling in neurons. We investigated how IL-8 participates in rapid neuronal signaling by using a combination of whole-cell recording and single-cell RT-PCR on dissociated rat septal neurons. We show that IL-8 can acutely reduce Ca(2+) currents in septal neurons, an effect that was concentration-dependent, involved the closure of L- and N-type Ca(2+) channels, and the activation of G(ialpha1) and/or G(ialpha2) subtype(s) of G-proteins. Analysis of the mRNAs from the recorded neurons revealed that the latter were all cholinergic in nature. Moreover, we found that all cholinergic neurons that responded to IL-8, expressed mRNAs for either one or both IL-8 receptors CXCR1 and CXCR2. This is the first report of a chemokine that modulates ion channels in neurons via G-proteins, and the first demonstration that mRNAs for CXCR1 are expressed in the brain. Our results suggest that IL-8 release by glial cells in vivo may activate CXCR1 and CXCR2 receptors on cholinergic septal neurons and acutely modulate their excitability by closing calcium channels.     

A multi-scaled approach for simulating chemical reaction systems

In this paper we give an overview of some very recent work, as well as presenting a new approach, on the stochastic simulation of multi-scaled systems involving chemical reactions. In many biological systems (such as genetic regulation and cellular dynamics) there is a mix between small numbers of key regulatory proteins, and medium and large numbers of molecules. In addition, it is important to be able to follow the trajectories of individual molecules by taking proper account of the randomness inherent in such a system. We describe different types of simulation techniques (including the stochastic simulation algorithm, Poisson Runge–Kutta methods and the balanced Euler method) for treating simulations in the three different reaction regimes: slow, medium and fast. We then review some recent techniques on the treatment of coupled slow and fast reactions for stochastic chemical kinetics and present a new approach which couples the three regimes mentioned above. We then apply this approach to a biologically inspired problem involving the expression and activity of LacZ and LacY proteins in E. coli, and conclude with a discussion on the significance of this work.     

Putting the plants back into plant ecology: six pragmatic models for understanding and conserving plant diversity

BACKGROUND: There is a compelling need to protect natural plant communities and restore them in degraded landscapes. Activities must be guided by sound scientific principles, practical conservation tools, and clear priorities. With perhaps one-third of the world's flora facing extinction, scientists and conservation managers will need to work rapidly and collaboratively, recognizing each other's strengths and limitations. As a guide to assist managers in maintaining plant diversity, six pragmatic models are introduced that are already available. Although theoretical models continue to receive far more space and headlines in scientific journals, more managers need to understand that pragmatic, rather than theoretical, models have the most promise for yielding results that can be applied immediately to plant communities. SIX PRAGMATIC MODELS: For each model, key citations and an array of examples are provided, with particular emphasis on wetlands, since "wet and wild" was my assigned theme for the Botanical Society of America in 2003. My own work may seem rather prominent, but the application and refinement of these models has been a theme for me and my many students over decades. The following models are reviewed: (1) species-area: larger areas usually contain more species; (2) species-biomass: plant diversity is maximized at intermediate levels of biomass; (3) centrifugal organization: multiple intersecting environmental gradients maintain regional landscape biodiversity; (4) species-frequency: a few species are frequent while most are infrequent; (5) competitive hierarchies: in the absence of constraints, large canopy-forming species dominate patches of landscape, reducing biological diversity; and (6) intermediate disturbance: perturbations such as water level fluctuations, fire and grazing are essential for maintaining plant diversity. CONCLUSIONS: The good news is that managers faced with protecting or restoring landscapes already have this arsenal of tools at their disposal. The bad news is that far too few of these models are appreciated.     

Conditional knockout mice to study alternative splicing in vivo

Analysis of genomes has revealed that the total number of human genes is comparable to those of simpler organisms, and thus, the number of genes does not correlate with the complexity and functional diversity of different organisms. Multiple mechanisms, including alternative splicing, are believed to contribute to the molecular complexity in higher eukaryotes. Given the fact that more than half of human genes undergo alternative splicing, however, little is known about the biological relevance of most alternative splicing events and their regulatory mechanisms. Recent work has highlighted the power of reverse genetic approaches in addressing regulated splicing in animal models. Here, we focus on the conditional knockout approach adapted for splicing research with the intention to provide a general guide to the generation of mouse models to study regulated splicing in development and disease.     

Study of Tissue-Specific CpG Methylation of DNA in Extended Genomic Loci

Modern approaches for studies on genome functioning include investigation of its epigenetic regulation. Methylation of cytosines in CpG dinucleotides is an inherited epigenetic modification that is responsible for both functional activity of certain genomic loci and total chromosomal stability. This review describes the main approaches for studies on DNA methylation. Under consideration are site-specific approaches based on bisulfite sequencing and methyl-sensitive PCR, whole-genome approaches aimed at searching for new methylation hot spots, and also mapping of unmethylated CpG sites in extended genomic loci.     

Osmotic sucrose enhancement of single-cell embryogenesis and transformation efficiency in <Emphasis Type="Italic">Oncidium</Emphasis>

Traditional breeding processes to genetically modify the long reproductive cycle and slow seed maturation of orchids have limits. We developed a more efficient protocol using particle bombardment to produce transgenic plants of Oncidium Sharry Baby OM8 (Orchidaceae). Pretreating protocorm-like bodies (PLBs) with 0.5 M sucrose for 2 h increased single-cell embryogenesis 3- to 4-fold; however, shoot formation was suppressed. In addition, new PLBs were regenerated from the entire sucrose-pretreated PLBs, whereas in untreated PLBs, this occurred only from the bases. Pretreated PLBs were bombarded with pSPFLP containing genes encoding a sweet pepper ferredoxin-like protein (pflp), hygromycin phosphotransferase (hpt) and -glucuronidase (GUS) driven by the cauliflower mosaic virus 35S promoter. Pretreated PLBs showed a 14.8-fold increase in GUS expression over the untreated PLBs 40days after bombardment. The presence of pflp and hpt transgenes in the 40 putatively stably transformed lines that produced 113 clones was confirmed by PCR analysis. Six lines (eight clones) were positive for both pflp and hpt transgenes. In addition, clones derived from these lines were either all positive or all negative for the two transgenes, which suggests homogeneity in pretreated PLBs with more single-cell embryogenesis. Thus, sucrose pretreatment enhanced the regeneration of PLBs, single-cell embryogenesis and efficiency of transformation.