Coupling and Activation of the ß1 Adrenergic Receptor - The Role of the Third Intracellular Loop.

G protein-coupled receptors (GPCRs) belong to the most diverse group of membrane receptors with a conserved structure of seven transmembrane (TM) α-helices connected by intracellular and extracellular loops. Intracellular loop 3 (ICL3) connects TM5 and TM6, the two helices shown to play significant roles in receptor activation. Herein, we investigate the activation and signaling of the ß1 adrenergic receptor (ß1AR) using mass spectrometry (MS) with a particular focus on the ICL3 loop. First, using native MS, we measure the extent of receptor coupling to an engineered Gαs subunit (mini Gs) and show preferential coupling to ß1AR with an intact ICL3 (ß1AR_ICL3) compared to the truncated ß1AR. Next, using hydrogen-deuterium exchange (HDX)-MS, we show how helix 5 of mini Gs reports on the extent of receptor activation in the presence of a range of agonists. Then, exploring a range of solution conditions and using comparative HDX, we note additional HDX protection when ICL3 is present, implying that mini Gs helix 5 presents a different binding conformation to the surface of ß1AR_ICL3, a conclusion supported by MD simulation. Considering when this conformatonal change occurs we used time-resolved HDX and employed two functional assays to measure GDP release and cAMP production, with and without ICL3. We found that ICL3 exerts its effect on Gs through enhanced cAMP production but does not affect GDP release. Together, our study uncovers potential roles of ICL3 in fine-tuning GPCR activation through subtle changes in the binding pose of helix 5, only after nucleotide release from Gs.

Status and causes of soil compaction at apple orchards in the Weibei Dry Highland, Northwest China.

To find out the recessive factors of soil degradation threatening the healthy development of Weibei apple orchards, we examined soil compaction status and its inducing factors. This study could provide a theoretical basis for apple orchard scientific management. We evalua-ted the changes of soil bulk density and compaction with the depth of soil layer in 0-60 cm in four apple orchards with various planting period, including <10 years (4-6 years), 10-20 years (14-16 years) and ＞20 years (24-26 years). The location and degradation degree of soil compaction in orchard were investigated. Through analyzing the number of soil aggregates and the stability, soil clay and organic matter contents, we tried to find the reasons for the internal compacting of soil in Weibei orchards. The results showed that soil bulk density and compactness in the 0-60 cm soil layer significantly increased with increasing planting years and soil depth. With the 20 cm soil layer as a boundary, soil of Weibei orchards in different planting years showed obvious variation characteristics of loose in upper and compact in lower. The above indicators in soil layer above 20 cm basically met the normal requirements of apple trees, whereas soil layer below 20 cm exceeded the threshold for healthy growth of apple trees. The main reasons for soil compaction below the subsurface layer were poor soil aggregation, the lack of soil organic matter, less human disturbance during fruit planting, and the movement of scattered clay particles to the lower layer. With increasing years of fruit planting, soil compaction became more severe.

Ultrastable Luminescent Organic-Inorganic Perovskite Quantum Dots via Surface Engineering: Coordination of Methylammonium Bromide and Covalent Silica Encapsulation.

Encapsulation of luminescent perovskite quantum dots (QDs) into a solid matrix has been approved to be an efficient way to improve their stability. In this work, we reported a green encapsulation method to produce ultrastable CH3NH3PbBr3 QDs incorporated into the SiO2 matrix. Specifically, fresh-prepared CH3NH3PbBr3 QDs were covalently embedded into silica by an aqueous sol-gel method assisted with CH3NH3Br, which not only effectively inhibited the water-driven degradation of QDs through surface coordination, but also strongly stabilized the QDs in solid powder via concentration gradient. As far as we know, this silica encapsulation of perovskite QDs in aqueous environments is reported for the first time. Luminescent properties of perovskite QDs during the course of gelation as well as in resulting composite powder were investigated using steady-state and time-resolved spectroscopies, and a 2 wt % QD-doped sample treated with 11.5 mM of CH3NH3Br was demonstrated to be the optimal phosphor. The green-emissive phosphor had a PLQY of 60.3% and a full width at half maxima of ∼25 nm, exhibiting ultrahigh stability tested by cycle heating (120 °C), continuous heating (80 °C, 60 h), and light irradiation (450 nm light, 350 h). The phosphor was readily blended with polymers and applied as a color-converting layer on blue light-emitting diodes.

A fluorescent nanoprobe for real-time monitoring of intracellular singlet oxygen during photodynamic therapy.

Sensing of intracellular singlet oxygen (1O2) is required in order to optimize photodynamic therapy (PDT). An optical nanoprobe is reported here for the optical determination of intracellular 1O2. The probe consists of a porous particle core doped with the commercial 1O2 probe 1,3-diphenylisobenzofuran (DPBF) and a layer of poly-L-lysine. The nanoparticle probes have a particle size of ~80 nm in diameter, exhibit good biocompatibility, improved photostability and high sensitivity for 1O2 in both absorbance (peak at 420 nm) and fluorescence (with excitation/emission peaks at 405/458 nm). Nanoprobes doped with 20% of DPBF are best suited even though they suffer from concentration quenching of fluorescence. In comparison with the commercial fluorescent 1O2 probe SOSG, 20%-doped DPBF-NPs (aged) shows higher sensitivity for 1O2 generated at an early stage. The best nanoprobes were used to real-time monitor the PDT-triggered generation of 1O2 inside live cells, and the generation rate is found to depend on the supply of intracellular oxygen. Graphical abstract A fluorescent nanoprobe featured with refined selectivity and improved sensitivity towards 1O2 was prepared from the absorption-based probe DBPF and used to real-time monitoring of the generation of intracellular 1O2 produced during PDT.

In situ silica coating-directed synthesis of orthorhombic methylammonium lead bromide perovskite quantum dots with high stability.

Luminescent perovskite quantum dots (QDs) had attracted great attention by virtue of the merits of color-tunable and narrow-band emissions. However, sofar reported perovskite QDs suffered from instability more or less. In this work, a type of silica-coated orthorhombic CH3NH3PbBr3 QDs (SiO2-QDs) with greatly improved stability was reported. The SiO2-QDs were one-pot synthesized using a reprecipitation-encapsulation method assisted with an amine functional silane, which not conly controlled the crystallization of QDs, but also encapsulated QDs with a silica layer simultaneously. More interestingly, the in situ encapsulation of silica shell induced the presence of orthorhombic perovskite that was thought to be unstable at room temperature. This is the first report of orthorhombic CH3NH3PbBr3 QDs, as far as we are concerned. The orthorhombic SiO2-QDs exhibited narrow-band green luminescence with a quantum yield of 78%, and a high production yield of ∼70wt%. Moreover, stability of SiO2-QDs was considerably improved due to silica-coating. White-LEDs were also successfully fabricated with the green SiO2-QDs and a red commercial phosphors using a noncontact configuration. These results demonstrated that the orthorhombic SiO2-QDs held great promise for high-performance display or lighting technology.

Selective impairments of alerting and executive control in HIV-infected patients: evidence from attention network test.

BACKGROUND: Attention ability can be subdivided into three functionally independent networks, i.e., alerting network, orienting network, and executive network. Previous literature has documented that deficits in attention are a common consequence of HIV infection. However, the precise nature of deficits of attention in HIV-infected patients is poorly understood. Accordingly, the aim of the study was to identify whether the HIV-infected patients showed a specific attention network deficit or a general attentional impairment. METHODS: We investigated 27 HIV-infected patients and 31 normal controls with the Attention Network Test (ANT). RESULTS: The patients exhibited less efficient alerting network and executive network than controls. No significant difference was found in orienting network effect between groups. Our results also indicate a tendency for poorer efficiency on alerting attention and executive attention in patients with CD4 ≤ 200. CONCLUSIONS: Our findings suggest that HIV-infected patients exhibited selective impairments of attention network of alerting and executive control. The link between lower CD4 T cell count and poorer attention network function imply the importance of starting antiretroviral therapy earlier to avoid irreversible neurocognitive impairment.

Family history of suicide and high motor impulsivity distinguish suicide attempters from suicide ideators among college students.

Suicide in college students has become an important public health issue in China. The aim of this study was to identify the differences between suicide attempters and suicide ideators based on a cross-sectional survey. Our results indicate that although female gender, positive screening for psychiatric illness, positive family history of suicide, elevated overall impulsivity, and elevated motor impulsivity were correlated with suicidal ideation, only positive family history of suicide and high motor impulsivity could differentiate suicide attempters from suicidal ideators. Future research with a longitudinal and prospective study design should be conducted to confirm these findings.

[Effects of fertilizing regime and planting age on soil calcium decline in Luochuan apple orchards]. / æ–½è‚¥æ–¹å¼å’Œå›­é¾„å¯¹æ´›å·è‹¹æžœå›­åœŸå£¤é’™ç´ é€€åŒ–çš„å½±å“.

This study was conducted to assess the effects of fertilizing regime and orchard planting age on soil calcium contents and stocks in the apple orchards on the Loess Plateau. The apple orchards in Luochuan County, one of the best regions for apple plantation in the world, were selec-ted in this study. The contents of calcium carbonate,water-soluble calcium and exchangeable cal-cium at 0-100 cm soil layer under different fertilizing regimes and various planting ages were mea-sured, their stocks were calculated and their variation features were analyzed. The results showed that soil in the apple orchards in the study region was characterized by the decline in calcium contents. The decline was more serious in apple orchards with long-term application of chemical fertili-zer than in those with combined application of chemical fertilizer and farmyard manure. The average contents of calcium carbonate, water-soluble calcium and exchangeable calcium at 0-100 cm soil layer in apple orchards with long-term application of chemical fertilizer decreased by 38.8%, 25.4% and 5.6% respectively than those in the apple orchards with long-term application of both chemical fertilizer and farmyard manure. The stocks of calcium carbonate, water-soluble calcium and exchangeable calcium decreased by 36.4%, 26.0% and 4.3%, respectively. The decline of soil cal-cium was aggravated with the increase of orchard planting age. The contents of calcium carbonate, water-soluble calcium and exchangeable calcium at 0-100 cm soil layer in orchards of more than 25 years of planting age decreased by 48.8%, 69.4% and 39.5% respectively, compared with orchards of less than 10 years of planting age, and the stocks decreased by 40.8%, 64.1% and 33.0%, respectively. These results indicated that either long-term application of chemical fertilizer or long-term plantation of apple trees obviously depleted soil calcium carbonate, water-soluble calcium and exchangeable calcium. Therefore, it was recommended that application of chemical fertilizer and farmyard manure should be combined to mitigate soil calcium decline, and calcium management should be strengthened in apple orchards of more than 25 years of planting age. The fertilizing regime was a driving factor of soil calcium decline which had a significant temporal (orchard planting age) and spatial (soil depth) effect.

Expression analysis of eight amphioxus genes involved in the Wnt/ß-catenin signaling pathway.

The Wnt/ß-catenin signaling pathway plays a crucial role in the embryonic development of metazoans. Although the pathway has been studied extensively in many model animals, its function in amphioxus, the most primitive chordate, remains largely uncharacterized. To obtain basic data for functional analysis, we identified and isolated seven genes (Lrp5/6, Dvl, APC, CkIα, CkIδ, Gsk3ß, and Gro) of the Wnt/ß-catenin signaling pathway from the amphioxus (Branchiostoma floridae) genome. Phylogenetic analysis revealed that amphioxus had fewer members of each gene family than that found in vertebrates. Whole-mount in situ hybridization showed that the genes were maternally expressed and broadly distributed throughout the whole embryo at the cleavage and blastula stages. Among them, Dvl was expressed asymmetrically towards the animal pole, while the others were evenly distributed in all blastomeres. At the mid-gastrula stage, the genes were specifically expressed in the primitive endomesoderm, but displayed different patterns. When the embryo developed into the neurula stage, the gene expressions were mainly detected in either paraxial somites or the tail bud. With the development of the embryo, the expression levels further decreased gradually and remained only in some pharyngeal regions or the tail bud at the larva stage. Our results suggest that the Wnt/ß-catenin pathway might be involved in amphioxus somite formation and posterior growth, but not in endomesoderm specification.

Synthesis and optimization of ZnPc-loaded biocompatible nanoparticles for efficient photodynamic therapy.

Zinc(ii) phthalocyanine (ZnPc) is a promising photosensitizer for PDT but suffers from aggregation in a physiological aqueous environment. In this paper, a class of biocompatible polymeric nanoparticles (NPs) was prepared to encapsulate ZnPc molecules. Mostly because of the planar structure, ZnPc molecules were difficult to be encapsulated into the polymeric NPs unless further coated with a thick poly-l-lysine (PLL) layer. The PLL shell endowed the NPs with good biocompatibility, efficient cellular uptake, and potential bioconjugation. The degree of aggregation (DOA) of ZnPc molecules in PLL-NPs was thoroughly investigated based on self-defined relative DOA, and a loading capacity of 4 wt% was deduced as the turning point for aggravating aggregation. Similarly, the optimal loading capacity of ZnPc was determined to be 4% according to the 1O2 generation rate, demonstrating the feasibility of the DOA approach. Polymers with large rigid units (PVK and PFO) were also utilized to relieve the aggregation of ZnPc in NPs. Taking advantage of the optimized ZnPc-loaded NPs, high PDT efficacy was demonstrated in HepG2 cells and in tumor-bearing mice as well. Both high in vitro and in vivo PDT efficacy and biocompatibility are demonstrated. Aside from affording a class of efficient biocompatible nanophotosensitizers, this work is also instructive to design other types of ZnPc-based nanocarriers, in which aggregation should be well considered.