Dim Nighttime Illumination Interacts With Parametric Effects of Bright Light to Increase the Stability of Circadian Rhythm Bifurcation in Hamsters

The endogenous circadian pacemaker of mammals is synchronized to the environmental day by the ambient cycle of relative light and dark. The present studies assessed the actions of light in a novel circadian entrainment paradigm where activity rhythms are bifurcated following exposure to a 24-h light:dark:light:dark (LDLD) cycle. Bifurcated entrainment under LDLD reflects the temporal dissociation of component oscillators that comprise the circadian system and is facilitated when daily scotophases are dimly lit rather than completely dark. Although bifurcation can be stably maintained in LDLD, it is quickly reversed under constant conditions. Here the authors examine whether dim scotophase illumination acts to maintain bifurcated entrainment under LDLD through potential interactions with the parametric actions of bright light during the two daily photophases. In three experiments, wheel-running rhythms of Syrian hamsters were bifurcated under LDLD with dimly lit scotophases, and after several weeks, dim scotophase illumination was either retained or extinguished. Additionally, “full” and “skeleton” photophases were employed under LDLD cycles with dimly lit or completely dark scotophases to distinguish parametric from nonparametric effects of bright light. Rhythm bifurcation was more stable in full versus skeleton LDLD cycles. Dim light facilitated the maintenance of bifurcated entrainment under full LDLD cycles but did not prevent the loss of rhythm bifurcation in skeleton LDLD cycles. These studies indicate that parametric actions of bright light maintain the bifurcated entrainment state; that dim scotophase illumination increases the stability of the bifurcated state; and that dim light interacts with the parametric effects of bright light to increase the stability of rhythm bifurcation under full LDLD cycles. A further understanding of the novel actions of dim light may lead to new strategies for understanding, preventing, and treating chronobiological disturbances. (Author correspondence: jevans@msm.edu)     

The affinity purification and characterization of ATP synthase complexes from mitochondria

The mitochondrial F₁-ATPase inhibitor protein, IF₁, inhibits the hydrolytic, but not the synthetic activity of the F-ATP synthase, and requires the hydrolysis of ATP to form the inhibited complex. In this complex, the α-helical inhibitory region of the bound IF₁ occupies a deep cleft in one of the three catalytic interfaces of the enzyme. Its N-terminal region penetrates into the central aqueous cavity of the enzyme and interacts with the γ-subunit in the enzyme''s rotor. The intricacy of forming this complex and the binding mode of the inhibitor endow IF₁ with high specificity. This property has been exploited in the development of a highly selective affinity procedure for purifying the intact F-ATP synthase complex from mitochondria in a single chromatographic step by using inhibitor proteins with a C-terminal affinity tag. The inhibited complex was recovered with residues 1–60 of bovine IF₁ with a C-terminal green fluorescent protein followed by a His-tag, and the active enzyme with the same inhibitor with a C-terminal glutathione-S-transferase domain. The wide applicability of the procedure has been demonstrated by purifying the enzyme complex from bovine, ovine, porcine and yeast mitochondria. The subunit compositions of these complexes have been characterized. The catalytic properties of the bovine enzyme have been studied in detail. Its hydrolytic activity is sensitive to inhibition by oligomycin, and the enzyme is capable of synthesizing ATP in vesicles in which the proton-motive force is generated from light by bacteriorhodopsin. The coupled enzyme has been compared by limited trypsinolysis with uncoupled enzyme prepared by affinity chromatography. In the uncoupled enzyme, subunits of the enzyme''s stator are degraded more rapidly than in the coupled enzyme, indicating that uncoupling involves significant structural changes in the stator region.     

Direct and inverse relationships between Riccati systems coupled with multiplicative terms

An analytical and computational framework for the derivation of solitary solutions to biological systems describing the cooperation and competition of species and expressed by the system of Riccati equations coupled with multiplicative terms is presented in this paper. It is demonstrated that relationships between these solitary solutions can be either direct or inverse. Thus, an infinitesimal perturbation of one population would lead to an infinitesimal change in the other population – if only both solitary solutions are coupled with the direct relationship. But, in general, that is not true if solitary solutions are coupled with the inverse relationship – an infinitesimal perturbation of one population may result into a non-infinitesimal change in the other population. Necessary and sufficient conditions for the existence of solitary solutions are derived in the space of the system's parameters and initial conditions.     

Inter‐Fullerene Electronic Coupling Controls the Efficiency of Photoinduced Charge Generation in Organic Bulk Heterojunctions

Photoinduced charge generation (PCG) dynamics are notoriously difficult to correlate with specific molecular properties in device relevant polymer:fullerene organic photovoltaic blend films due to the highly complex nature of the solid state blend morphology. Here, this study uses six judiciously selected trifluoromethylfullerenes blended with the prototypical polymer poly(3‐hexylthiophene) and measure the PCG dynamics in 50 fs–500 ns time scales with time‐resolved microwave conductivity and femtosecond transient absorption spectroscopy. The isomeric purity and thorough chemical characterization of the fullerenes used in this study allow for a detailed correlation between molecular properties, driving force, local intermolecular electronic coupling and, ultimately, the efficiency of PCG yield. The findings show that the molecular design of the fullerene not only determines inter‐fullerene electronic coupling, but also influences the decay dynamics of free holes in the donor phase even when the polymer microstructure remains unchanged.     

Cx36 expression in the AII‐mediated rod pathway is activity dependent in the developing rabbit retina

Gap junctions are composed of connexin 36 (Cx36) and play a critical role in the rod photoreceptor signaling pathways of the vertebrate retina. Despite the fact that their connection and modulation in various rod pathways have been extensively studied in adult animals, little is known about the contribution and regulation of gap junctions to the development of the AII amacrine cell (AC)‐mediated rod pathway. Using immunohistochemistry and microinjection, this study demonstrates a steady increase in relative Cx36 protein expression in both plexiform layers of the rabbit retina at around the time of eye opening. However, immediately after eye opening, most Cx36 immunoreactive AII ACs show no gap junction coupling pattern to neighboring cells and it is not until the third postnatal week that AII cells begin to exhibit an adult‐like tracer‐coupling pattern. Moreover, studies using dark‐rearing and AMPA receptor blockade during postnatal development both revealed that relative levels of Cx36 immunoreactivity in AII ACs were increased when neural activity was inhibited . Our findings suggest that Cx36 expression in the AII‐mediated rod pathway is activity dependent in the developing rabbit retina . © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 473–486, 2016     

城市绿化对空气质量的影响研究——以中国27个省会城市为例

城市间绿化程度与空气污染比较及相关差异分析是提出城市环境管理措施的重要前提。选择全国27个主要省会城市,基于网络街景照片测定绿色指数差异,对比空气主要质量指标[空气质量指数(AQI)、细颗粒物(PM_2.5)、可吸入颗粒物(PM₁₀)、二氧化硫(SO_2)、二氧化氮(NO_2)、臭氧(O_3)、一氧化碳(CO)]的基础上,探讨了二者相关关系,旨在为提升环境质量、改善绿化水平提供基础数据。结果表明:(1)济南市和重庆市的绿色指数最高,分别达到是11.70%和11.55%,呼和浩特市和拉萨市城市绿色指数最低,在4%～5%。(2)海口市的空气质量最好,AQI为39.66,郑州市和济南市的空气质量最差,AQI年均值分别为117.34和113.93。但是不同空气指标城市排序间差异较大,比如PM_2.5、PM₁₀、NO_2以及SO_2年平均最低的城市都是海口,拉萨市年平均CO含量最低(0.55 mg·m^-3),哈尔滨市年平均O_3含量最低(77.08μg·m^-3)。(3)相关关系分析发现,增加城市的绿色程度,伴随着空气质量的改善,如沈阳、南宁、合肥等城市;但对于某些城市,则存在明显的正相关,如兰州、昆明、贵阳等城市,这意味着城市越绿伴随着空气污染的加重。尽管大量研究已经表明,城市绿色植被能够起到滞尘降低污染的作用,目前我国主要城市的空气污染程度,仅依靠城市绿化改善已经远远不够,甚至某一些城市绿色植被存在阻碍空气流通的作用。上述研究结果为科学规划城市绿化、提升空气质量提供基础数据支撑。     

Structural Characterization of Biomolecules through Atomistic Simulations Guided by DEER Measurements

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黄壤稻田土壤微生物生物量碳磷对长期不同施肥的响应

本研究以进行了22年的黄壤稻田长期定位试验为依托,分析了不同施肥模式下土壤碳(C)、磷(P)与微生物生物量C(MBC)、P(MBP)的变化及其耦合特征,旨在探讨黄壤地区合理培肥模式以提高土壤磷素有效性.试验包括10个处理,分别为不施肥(CK)、单施氮肥(N)、磷钾配合(PK)、氮钾配合(NK)、氮磷配合(NP)、氮磷钾配合(NPK)、单施有机肥(M)、3种有机无机肥配施(1/4M+3/4NP、0.5MNP、MNPK).结果表明:与不施肥相比,N和NK处理土壤有机碳(TOC)、全磷(TP)、MBC、MBP均有不同程度的降低,施磷处理(PK、NP、NPK)则有不同程度的提升;与不施肥和施用无机肥相比,施用有机肥各处理TOC、MBC、MBP及MBP/TP均显著增加,其中M和MNPK处理增幅最大.MBC/MBP、TOC/MBP、MBC/TP以施用有机肥处理最低,N处理最高.土壤MBC、MBP及其耦合关系与土壤TOC和有效磷均呈极显著相关,TOC是影响MBC、MBP、MBP/TP的直接因素,而有效磷则是影响MBC/MBP、TOC/MBP、MBC/TP的直接因素.土壤MBP及碳、磷耦合关系各指标可以有效区分单施化肥和施用有机肥的不同施肥方式,可作为评价黄壤稻田磷素肥力的生物学指标.配施有机肥是增加黄壤稻田磷有效性、提高土壤供磷潜力和保持土壤生物健康的有效途径.     

Polymerization of Horseradish Peroxidase by a Laccase‐Catalyzed Tyrosine Coupling Reaction

The polymerization of proteins can create newly active and large bio‐macromolecular assemblies that exhibit unique functionalities depending on the properties of the building block proteins and the protein units in polymers. Herein, the first enzymatic polymerization of horseradish peroxidase (HRP) is reported. Recombinant HRPs fused with a tyrosine‐tag (Y‐tag) through a flexible linker at the N‐ and/or C‐termini are expressed in silkworm, Bombyx mori. Trametes sp. laccase (TL) is used to activate the tyrosine of Y‐tagged HRPs with molecular O₂ to form a tyrosyl‐free radical, which initiates the tyrosine coupling reaction between the HRP units. A covalent dityrosine linkage is also formed through a HRP‐catalyzed self‐crosslinking reaction in the presence of H₂O₂. The addition of H₂O₂ in the self‐polymerization of Y‐tagged HRPs results in lower activity of the HRP polymers, whereas TL provides site‐selectivity, mild reaction conditions and maintains the activity of the polymeric products. The cocrosslinking of Y‐tagged HRPs and HRP‐protein G (Y‐HRP‐pG) units catalyzed by TL shows a higher signal in enzyme‐linked immunosorbent assay (ELISA) than the genetically pG‐fused HRP, Y‐HRP‐pG, and its polymers. This new enzymatic polymerization of HRP promises to provide highly active and functionalized polymers for biomedical applications and diagnostics probes.     

B3LYP/6-311++G* * study of structure and spin-spin coupling constant in heparin disaccharide

Structures of heparin disaccharide have been analyzed by DFT using the B3LYP/6-311++G( * *) method. The optimized geometries of two forms of this disaccharide, differing in the conformation ((1)C(4) and (2)S(0)) of the IdoA2S residue, confirmed considerable influences of the sulfate and the carboxylate groups upon the pyranose ring geometries. The computed energies showed that disaccharide having the (1)C(4) form of the IdoA2S residue is more stable than that with the (2)S(0) form. Interatomic distances, bond and torsion angles showed that interconversion of the IdoA2S residue results in geometry changes in the GlcN,6S residue as well. Three-bond proton-proton and proton-carbon spin-spin coupling constants computed for both forms agree with the experimental data and indicate that only two chair forms contribute to the conformational equilibrium in disaccharide. Influences of the charged groups upon the magnitudes of spin-spin coupling constants are also discussed.