Kinetic Analysis of Cerebrovascular Isoleucine Transport from Saline and Plasma

The concentration dependence of regional isoleucine transport across the blood-brain barrier was determined in anesthetized rats with the in situ brain perfusion technique of Takasato et al. [Am. J. Physiol. 247, H484-493 (1984)]. This technique allows, for the first time, accurate measurements of cerebrovascular amino acid transport in the absence of competing amino acids using saline perfusate, and in the presence of physiological concentrations of amino acids using plasma perfusate. Cerebrovascular isoleucine transport from saline perfusate followed Michaelis-Menten saturation kinetics where Vmax = 9 - 11 X 10(-4) mumol X s-1 X g-1 and Km = 0.054-0.068 mumol X ml-1 in six brain regions. A component of nonsaturable transport was not detected in any brain region even though perfusate isoleucine concentration was increased to greater than or equal to 150 times the normal plasma concentration. Isoleucine influx during plasma perfusion was only 8% of that predicted from the saline perfusion data due to transport inhibition by competing amino acids in plasma. Competitive inhibition increased the apparent Km for isoleucine transport from plasma by greater than or equal to 24-fold to 1.5-1.7 mumol X ml-1. These data provide accurate new estimates of the kinetic constants that describe amino acid transport across the blood-brain barrier. In addition, they indicate that the cerebrovascular transfer-site affinity (1/Km) for isoleucine is approximately fivefold greater than previously reported with the brain uptake index technique.     

毛细管电泳测定牛红细胞超氧化物歧化酶的巯基位点

本文介绍了毛细管电泳分析蛋白质酶解产物中含巯基多肽的方法。还原的及天然的牛红细胞超氧化物歧化酶(SOD)经4-乙烯吡啶修饰后,由TPCK-胰蛋白酶水解,在254nm检测到还原的SOD水解物中含3个巯基多肽,天然的SOD为1个疏基多肽且其毛细管电泳行为与上述3个多肽之一相一致。分析它们的氨基酸顺序,证实Cys-6为游离的巯基,Cys-55和Cys~(-144)形成二硫键。     

Vascular permeability to lanthanum in the rat incisor pulp

Summary Experiments were performed to compare the permeability of capillaries supplying the endoneurial environment, which is invested by perineurium, with vascular permeability in the pulp where perineurium is absent. Anaesthetised rats were perfused through the aorta with physiological solutions containing lanthanum nitrate at 37° C. Pieces of inferior alveolar nerve and segments of mandibular incisors were immersion-fixed and transverse sections were examined electron microscopically for the distribution of lanthanum. In the pulp the nerve fibres pass between lanthanum-impermeable arterioles and venules en route to the incisal end. In the peripheral pulp a few capillaries were permeable but the most permeable capillaries lay between the odontoblasts. Pulpal capillary permeability was attributed to the fenestrated endothelium and contrasted with the unfenestrated endoneurial capillaries which were impermeable to lanthanum. Whereas the tight junctions of endoneurial capillaries are known to prevent certain blood-borne substances from entering the endoneurium, it was not clear whether the permeability of the pulpal capillaries, which are distant from the nerve fibres, could affect the nerve fibre environment. No extravasated lanthanum reached the pulpal nerve fibres suggesting that they are not affected. Technically it was not possible to examine the incisal third of the tooth where the situation could be different because the volume of the pulp decreases and capillaries lie closer to the nerve fibres.     

A morphogram for silica‐witherite biomorphs and its application to microfossil identification in the early earth rock record

Archean hydrothermal environments formed a likely site for the origin and early evolution of life. These are also the settings, however, were complex abiologic structures can form. Low‐temperature serpentinization of ultramafic crust can generate alkaline, silica‐saturated fluids in which carbonate–silica crystalline aggregates with life‐like morphologies can self‐assemble. These “biomorphs” could have adsorbed hydrocarbons from Fischer–Tropsch type synthesis processes, leading to metamorphosed structures that resemble carbonaceous microfossils. Although this abiogenic process has been extensively cited in the literature and has generated important controversy, so far only one specific biomorph type with a filamentous shape has been discussed for the interpretation of Archean microfossils. It is therefore critical to precisely determine the full distribution in morphology and size of these biomorphs, and to study the range of plausible geochemical conditions under which these microstructures can form. Here, a set of witherite‐silica biomorph synthesis experiments in silica‐saturated solutions is presented, for a range of pH values (from 9 to 11.5) and barium ion concentrations (from 0.6 to 40 mmol/L BaCl₂). Under these varying conditions, a wide range of life‐like structures is found, from fractal dendrites to complex shapes with continuous curvature. The size, spatial concentration, and morphology of the biomorphs are strongly controlled by environmental parameters, among which pH is the most important. This potentially limits the diversity of environments in which the growth of biomorphs could have occurred on Early Earth. Given the variety of the observed biomorph morphologies, our results show that the morphology of an individual microstructure is a poor criterion for biogenicity. However, biomorphs may be distinguished from actual populations of cellular microfossils by their wide, unimodal size distribution. Biomorphs grown by diffusion in silica gel can be differentiated by their continuous gradient in size, spatial density, and morphology along the direction of diffusion.     

The sweet spot for biologics: recent advances in characterization of biotherapeutic glycoproteins

Introduction: Glycosylation is recognized as a Critical Quality Attribute for therapeutic glycoproteins such as monoclonal antibodies, fusion proteins and therapeutic replacement enzymes. Hence, efficient and quantitative glycan analysis techniques have been increasingly important for their discovery, development and quality control. The aim of this review is to highlight relevant and recent advances in analytical technologies for characterization of biotherapeutic glycoproteins.

Areas covered: The review gives an overview of the glycosylation trends of biotherapeutics approved in 2016 and 2017 by FDA. It describes current and novel analytical technologies for characterization of therapeutic glycoproteins and is explored in the context of released glycan, glycopeptide or intact glycoprotein analysis. Ultra performance liquid chromatography, mass spectrometry and capillary electrophoresis technologies are explored in this context.

Expert commentary: There is a need for the biopharmaceutical industry to incorporate novel state of the art analytical technologies into existing and new therapeutic glycoprotein workflows for safer and more efficient biotherapeutics and for the improvement of future biotherapeutic design. Additionally, at present, there is no ‘gold-standard’ approach to address all the regulatory requirements and as such this will involve the use of orthogonal glycoanalytical technologies with a view to gain diagnostic information about the therapeutic glycoprotein.     

Silica‐modified luminescent LaPO4:Eu@LaPO4@SiO2 core/shell nanorods: Synthesis,structural and luminescent properties

Monoclinic‐type tetragonal LaPO₄:Eu (core) and LaPO₄:Eu@LaPO₄ (core/shell) nanorods (NRs) were successfully prepared using a urea‐based co‐precipitation process under ambient conditions. An amorphous silica layer was coated around the luminescent core/shell NRs via the sol–gel process to improve their solubility and colloidal stability in aqueous and non‐aqueous media. The prepared nano‐products were systematically characterized by X‐ray diffraction pattern, transmission electron microscopy, energy dispersive X‐ray analysis, and FTIR, UV/Vis, and photoluminescence spectroscopy to examine their phase purity, crystal phase, surface chemistry, solubility and luminescence characteristics. The length and diameter of the nano‐products were in the range 80–120 nm and 10–15 nm, respectively. High solubility of the silica‐modified core/shell/Si NRs was found for the aqueous medium. The luminescent core NRs exhibited characteristic excitation and emission transitions in the visible region that were greatly affected by surface growth of insulating LaPO₄ and silica layers due to the multiphonon relaxation rate. Our luminescence spectral results clearly show a distinct difference in intensities for core, core/shell, and core/shell/Si NRs. Highly luminescent NRs with good solubility could be useful candidates for a variety of photonic‐based biomedical applications.     

Sequential reactors for the removal of endocrine disrupting chemicals by laccase immobilized onto fumed silica microparticles

Abstract

The main objective of this study is the evaluation of the capability of laccase from Myceliophthora thermophila immobilized on fumed silica microparticles (fsMP) for the removal of endocrine disrupting chemicals (EDCs) in two enzymatic reactor configurations. This type of support can also be magnetized to allow the straightforward separation of the biocatalyst under a magnetic field. The support exhibited excellent biocompatibility with the enzyme, superior tolerance to pH and temperature as well as improved stability in comparison with the free enzyme, even in the presence of organic solvents and enzyme inhibitors. The technical feasibility of the removal of EDCs by immobilized laccase was assessed in two types of enzymatic reactors operated in sequential mode: a membrane reactor using fsMP-laccase and a reactor with magnetic separation using magnetized fsMP-laccase. The extent of transformation for the target compounds: bisphenol A (BPA) and 17β-estradiol (E2) was high and comparable to free laccase in both systems (up to 80%). The possibility of reusing the immobilized enzyme, especially for magnetized supports, offers an interesting approach in the development of enzyme based processes for the biotransformation of emerging pollutants.     

Mesoporous Hybrid Electrolyte for Simultaneously Inhibiting Lithium Dendrites and Polysulfide Shuttle in Li–S Batteries

A bifunctional hybrid electrolyte composed of mesoporous silica nanosheets and liquid electrolyte is achieved for lithium–sulfur (Li–S) batteries. This hybrid electrolyte possesses abundant mesopores (2.8 nm), thin feature (20 µm), and high ionic conductivity (1.17 × 10^?1 mS cm^?1) as well as a low interfacial resistance with electrodes. Such unique features not only enable the efficient inhibition of the growth of lithium dendrites, but also significantly prevents polysulfide shuttling. Consequently, a Li–S battery with this hybrid electrolyte exhibits a relatively high reversible capacity and good capacity retention.     

Comparison of solutes, nutrients, and bacteria inputs from two types of groundwater to the Rhône river during an artificial drought

Solute, nutrient and bacterial inputs to the River Rhône from the interstitial habitat of a gravel bar and the floodplain aquifer were investigated during an artificial drought. Eight springs were investigated: four groundwater-fed springs in the floodplain, located at the bottom of the bank; and four interstitial-fed springs located at the downstream end of a gravel bar. During this period, the inflows of groundwater to the river represented an average input of 0.77 mg l^–1 of nitrogen (of which 93.3% were nitrates), 0.0187 mg l^–1 of total phosphorus (of which 42.2% was orthophosphate), 3.56 mg l^–1 of silica, 2.315 ± 0.703 mg l^–1 of dissolved organic carbon (DOC, of which 47% was biodegradable) and 7.3 × 10⁴ ± 3.7 × 10⁴ bacteria per ml (of which 8.8% were active). Silica, DOC, biodegradable DOC, and bacteria concentrations displayed temporal variations during the study, which seem to be linked to the biological activity of the groundwater biofilm. There was a strong heterogeneity between the two types of groundwater that flow to the river: concentrations of calcium and alkalinity were higher in bank springs than in gravel bars springs. In these latters, sulfate, sodium, nitrogen, phosphorus were significantly higher.