Heavy metal analysis in commercial Spirulina products for human consumption

For consumption of health foods of Spirulina, by the general public, health food stores are increasingly offering more exotic products. Though Spirulina consumption is growing worldwide, relatively few studies have reported on the quantities of heavy metals/minerals they contain and/or their potential effects on the population’s health. This study reveals the concentrations of six typical heavy metals/minerals (Ni, Zn, Hg, Pt, Mg, and Mn) in 25 Spirulina products commercialized worldwide for direct human consumption. Samples were ground, digested and quantified by Coupled Plasma Mass Spectroscopy (ICP–MS). The concentrations (mg/kg d.w.) were range from 0.001 to 0.012 (Pt) followed by 0.002–0.028 (Hg), 0.002–0.042 (Mg), 0.005–2.248 (Mn), 0.211–4.672 (Ni) and 0.533–6.225 (Zn). The inorganic elements of the present study were significantly lower than the recommended daily intake (RDI) level of heavy metal elements (mg/daily) Ni (0.4), Zn (13), Hg (0.01), Pt (0.002), Mg (400) and Mn (4). Based on this study the concentration of inorganic elements was not found to exceed the present regulation levels, and they can be considered as safe food.     

Regulation of Metabotropic Glutamate Receptor 7 (mGluR7) Internalization and Surface Expression by Ser/Thr Protein Phosphatase 1

The metabotropic glutamate receptor type 7 (mGluR7) is the predominant group III mGluR in the presynaptic active zone, where it serves as an autoreceptor to inhibit neurotransmitter release. Our previous studies show that PKC phosphorylation of mGluR7 on Ser-862 is a key mechanism controlling constitutive and activity-dependent surface expression of mGluR7 by regulating a competitive interaction of calmodulin and protein interacting with C kinase (PICK1). As receptor phosphorylation and dephosphorylation are tightly coordinated through the precise action of protein kinases and phosphatases, dephosphorylation by phosphatases is likely to play an active role in governing the activity-dependent or agonist-induced changes in mGluR7 receptor surface expression. In the present study, we find that the serine/threonine protein phosphatase 1 (PP1) has a crucial role in the constitutive and agonist-induced dephosphorylation of Ser-862 on mGluR7. Treatment of neurons with PP1 inhibitors leads to a robust increase in Ser-862 phosphorylation and increased surface expression of mGluR7. In addition, Ser-862 phosphorylation of both mGluR7a and mGluR7b is a target of PP1. Interestingly, agonist-induced dephosphorylation of mGluR7 is regulated by PP1, whereas NMDA-mediated activity-induced dephosphorylation is not, illustrating there are multiple signaling pathways that affect receptor phosphorylation and trafficking. Importantly, PP1γ1 regulates agonist-dependent Ser-862 dephosphorylation and surface expression of mGluR7.     

Stability study of the human G-protein coupled receptor, Smoothened

Smoothened is a member of the G-protein coupled receptor (GPCR) family responsible for the transduction of the Hedgehog signal to the intracellular effectors of the Hedgehog signaling pathway. Aberrant regulation of this receptor is implicated in many cancers but also in neurodegenerative disorders. Despite the pharmacological relevance of this receptor, very little is known about its functional mechanism and its physiological ligand. In order to characterize this receptor for basic and pharmacological interests, we developed the expression of human Smoothened in the yeast Saccharomyces cerevisiae and Smoothened was then purified. Using Surface Plasmon Resonance technology, we showed that human Smoothened was in a native conformational state and able to interact with its antagonist, the cyclopamine, both at the yeast plasma membrane and after purification. Thermostability assays on purified human Smoothened showed that this GPCR is relatively stable in the classical detergent dodecyl-β-d-maltoside (DDM). The fluorinated surfactant C₈F₁₇TAC, which has been proposed to be less aggressive towards membrane proteins than classical detergents, increased Smoothened thermostability in solution. Moreover, the replacement of a glycine by an arginine in the third intracellular loop of Smoothened coupled to the use of the fluorinated surfactant C₈F₁₇TAC during the mutant purification increased Smoothened thermostability even more. These data will be very useful for future crystallization assays and structural characterization of the human receptor Smoothened.     

Cloning and Sequence Analysis of cDNA for Diuretic Hormone Receptor from the Bombyx mori

The insect diuretic hormone (DH) binds to their receptor in malpighian tubules, and stimulates water secretion and cAMP synthesis. Complementary DNA encoding a diuretic hormone receptor was cloned from the malpighian tubules of Bombyx mori. The cloned cDNA encodes a protein consisting of 391 amino acid residues with the seven transmembrane domains. The receptor protein is homologous with that of other insects, and is structurally related to G-protein coupled receptors such as corticotropin relating factor (CRF), secretin, and vasoactive intestinal peptide.     

The coupled chemomechanics of the F1-ATPase molecular motor

The enzyme F₁-ATPase is a rotary nanomotor in which the central γ subunit rotates inside the cavity made of α₃β₃ subunits. The experiments showed that the rotation proceeds in steps of 120° and each 120° step consists of 80° and 40° substeps. Here the Author proposes a stochastic wave mechanics of the F₁-ATPase motor and combines it with the structure-based kinetics of the F₁-ATPase to form a chemomechanic coupled model. The model can reproduce quantitatively and explain the experimental observations about the F₁ motor. Using the model, several rate-limited situations about γ subunit rotation are proposed, the effects of the friction and the load on the substeps are investigated and the chemomechanic coupled time during ATP hydrolysis cycle is determined.     

Agonist-regulated Cleavage of the Extracellular Domain of Parathyroid Hormone Receptor Type 1

The receptor for parathyroid hormone (PTHR) is a main regulator of calcium homeostasis and bone maintenance. As a member of class B of G protein-coupled receptors, it harbors a large extracellular domain, which is required for ligand binding. Here, we demonstrate that the PTHR extracellular domain is cleaved by a protease belonging to the family of extracellular metalloproteinases. We show that the cleavage takes place in a region of the extracellular domain that belongs to an unstructured loop connecting the ligand-binding parts and that the N-terminal 10-kDa fragment is connected to the receptor core by a disulfide bond. Cleaved receptor revealed reduced protein stability compared with noncleaved receptor, suggesting degradation of the whole receptor. In the presence of the agonistic peptides PTH(1–34), PTH(1–14), or PTH(1–31), the processing of the PTHR extracellular domain was inhibited, and receptor protein levels were stabilized. A processed form of the PTHR was also detected in human kidney. These findings suggest a new model of PTHR processing and regulation of its stability.     

Coupled oscillations of a protein microtubule immersed in cytoplasm: an orthotropic elastic shell modeling

Revealing vibration characteristics of sub-cellular structural components such as membranes and microtubules has a principal role in obtaining a deeper understanding of their biological functions. Nevertheless, limitations and challenges in biological experiments at this scale necessitates the use of mathematical and computational models as an alternative solution. As one of the three major cytoskeletal filaments, microtubules are highly anisotropic structures built from tubulin heterodimers. They are hollow cylindrical shells with a ∼ 25 nm outer diameter and are tens of microns long. In this study, a mechanical model including the effects of the viscous cytosol and surrounding filaments is developed for predicting the coupled oscillations of a single microtubule immersed in cytoplasm. The first-order shear deformation shell theory for orthotropic materials is used to model the microtubule, whereas the motion of the cytosol is analyzed by considering the Stokes flow. The viscous cytosol and the microtubule are coupled through the continuity condition across the microtubule–cytosol interface. The stress and velocity fields in the cytosol induced by vibrating microtubule are analytically determined. Finally, the influences of the dynamic viscosity of the cytosol, filament network elasticity, microtubule shear modulus, and circumferential wave-number on longitudinal, radial, and torsional modes of microtubule vibration are elucidated.     

气候变化和人类活动干扰下骆驼刺潜在分布格局变化特征

气候变化和人类活动是影响物种分布的最重要因素。骆驼刺是我国荒漠区的重要建群种,建群种的丧失将对荒漠生态系统产生严重损伤。因此预测气候变化和人类活动影响下骆驼刺适宜生境的变化特点对保护我国荒漠生态系统具有重要的意义。本研究采用61个骆驼分布点数据,和21个环境因子数据,运用最大熵模型（MaxEnt）预测骆驼刺当前在有、无人类活动干扰下的适宜生境分布,对影响其分布的环境因素进行了评估。并预测未来（2021-2040,2041-2060,2061-2080,2081-2100）四条共享社会经济路径（SSP126,SSP245,SSP370,SSP585）情景下,骆驼刺潜在分布区的变化特点。研究结果表明,在无人类活动干扰的情况下,骆驼刺适生区面积达132.29万km²,覆盖我国西北干旱区的大部分面积。年均降水量、最冷季度平均温、平均气温日较差、年均温、温度季节性、降水的季节性和高程被确定为影响骆驼刺潜在分布区的最重要因素。而在加入人类活动强度因素之后,骆驼刺适生区面积显著下降至71.31万km²,且呈现出破碎化状态。此时,年降水量、人类活动强度和高程是影响骆驼刺分布的最重要原因。在未来气候情景下,骆驼刺的适生区将产生一定的扩张。尤其在中高强迫（SSP370）和高强迫（SSP585）情景下,骆驼刺的适生面积将随时间推移产生显著的增加,且这种扩张主要向高纬度和高海拔地区发展。骆驼刺能保留大部分原有生境,仅在内蒙古中西部地区发生少量收缩。在气候变化条件下,骆驼刺能够在西北干旱区生存并扩张,故可将其作为防沙治沙的优良物种进行保护与开发。     

Amino Acid Residues Critical for Endoplasmic Reticulum Export and Trafficking of Platelet-activating Factor Receptor

Several residues are conserved in the transmembrane domains (TMs) of G-protein coupled receptors. Here we demonstrate that a conserved proline, Pro²⁴⁷, in TM6 of platelet-activating factor receptor (PAFR) is required for endoplasmic reticulum (ER) export and trafficking after agonist-induced internalization. Alanine-substituted mutants of the conserved residues of PAFRs, including P247A, were retained in the ER. Because a PAFR antagonist, Y-24180, acted as a pharmacological chaperone to rescue ER retention, this retention is due to misfolding of PAFR. Methylcarbamyl (mc)-PAF, a PAFR agonist, did not increase the cell surface expression of P247A, even though another ER-retained mutant, D63A, was effectively trafficked. Signaling and accumulation of the receptors in the early endosomes were observed in the mc-PAF-treated P247A-expressing cells, suggesting that P247A was trafficked to the cell surface by mc-PAF, and thereafter disappeared from the surface due to aberrant trafficking, e.g. enhanced internalization, deficiency in recycling, and/or accelerated degradation. The aberrant trafficking was confirmed with a sortase-A-mediated method for labeling cell surface proteins. These results demonstrate that the conserved proline in TM6 is crucial for intracellular trafficking of PAFR.