bluuebunny/crossref_metadata_2025 huggingface dataset:
Hydrophobic salts markedly diminish viscosity of concentrated protein solutions
Authors: Du Wei , Klibanov Alexander M.
Published: October 2010

Reducing viscosities of concentrated solutions of therapeutic proteins is important for their subcutaneous and intravenous delivery. Although inorganic salts and optimizing the pH were previously reported to dramatically lower the viscosity of a monoclonal antibody solution, herein we have determined these effects not to be general. Separately, we have found that hydrophobic ionic excipients, both anionic and cationic, substantially decrease the viscosity of concentrated (300–400 mg/mL) aqueous solutions of bovine serum albumin and γ‐globulin. The more hydrophobic the excipient, the greater its viscosity‐lowering effect is. With cationic ones, the concomitant contribution of the counter‐ion broadly follows the chaotropic order. The most potent excipients lower the viscosity over fourfold to levels far below the 50 cP threshold for subcutaneous injections. The observed viscosity reductions are rationalized in terms of three‐dimensional transient protein networks formed in concentrated solutions due to hydrophobic and, to a lesser extent, ionic interactions. These reversible protein aggregates are responsible for strong resistance to flow in concentrated protein solutions and hence their high viscosity; hydrophobic ions apparently effectively compete for these interprotein interactions, thereby giving rise to less viscous solutions. Biotechnol. Bioeng. 2011; 108:632–636. © 2010 Wiley Periodicals, Inc.
Microalgal bacterial floc properties are improved by a balanced inorganic/organic carbon ratio
Authors: Van Den Hende Sofie , Vervaeren Han , Saveyn Hans , Maes Guy , Boon Nico
Published: October 2010

Microalgal bacterial floc (MaB‐floc) reactors have been suggested as a more sustainable secondary wastewater treatment. We investigated whether MaB‐flocs could be used as tertiary treatment. Tertiary influent has a high inorganic/organic carbon ratio, depending on the efficiency of the secondary treatment. In this study, the effect of this inorganic/organic carbon ratio on the MaB‐flocs performance was determined, using three sequencing batch photobioreactors. The MaB‐flocs were fed with synthetic wastewater containing 84, 42, and 0 mg L−1 C‐KHCO3 supplemented with 0, 42, 84 mg L−1 C‐sucrose, respectively, representing inorganic versus organic carbon. Bicarbonate significantly decreased the autotrophic index of the MaB‐flocs and resulted in poorly settling flocs. Moreover, sole bicarbonate addition led to a high pH of 9.5 and significant lower nitrogen removal efficiencies. Sucrose without bicarbonate resulted in good settling MaB‐flocs, high nitrogen removal efficiencies and neutral pH levels. Despite the lower chlorophyll a content of the biomass and the lower in situ oxygen concentration, 92–96% of the soluble COD‐sucrose was removed. This study shows that the inorganic/organic carbon ratio of the wastewater is of major importance and that organic carbon is requisite to guarantee a good performance of the MaB‐flocs for wastewater treatment. Biotechnol. Bioeng. 2011; 108:549–558. © 2010 Wiley Periodicals, Inc.
Differential effect of exocytic SNAREs on the production of recombinant proteins in mammalian cells
Authors: Peng Ren‐Wang , Abellan Eric , Fussenegger Martin
Published: October 2010

Mammalian cells play a dominant role in the industrial production of biopharmaceutical proteins. However, the productivity of producer cells is often hindered by a bottleneck in the saturated secretory pathway, where a sophisticated mechanism of vesicle trafficking is mediated by numerous proteins and their complexes, among which are the cross‐kingdom conserved SNAREs [soluble NSF (N‐ethylmaleimide‐sensitive factor) receptor]. The SNAREs assemble into complexes by means of four interactive α‐helices and, thus, trigger the fusion of transport vesicles with the respective target membranes. We report that the transgenic expression of exocytic SNAREs, which control the fusion of secretory vesicles to the plasma membrane, differentially impacts the secretory capacity of HEK‐293, HeLa, and CHO‐K1 cells. While other exocytic SNAREs have no effect or a negative effect, SNAP‐23 [synaptosome‐associated protein of 23 kDa] and VAMP8 [vesicle‐associated membrane protein 8] specifically increase the production of recombinant proteins when they are ectopically and stably expressed in mammalian cells. The targeted and effective intervention in the secretory capacity of SNARE proteins is a novel engineering strategy, which could lead to the development of new therapies by increasing the production of biopharmaceutical proteins or by boosting the secretion of cell implants in cell therapy initiatives. Biotechnol. Bioeng. 2011; 108:611–620. © 2010 Wiley Periodicals, Inc.
Improved galactose fermentation of Saccharomyces cerevisiae through inverse metabolic engineering
Authors: Lee Ki‐Sung , Hong Min‐Eui , Jung Suk‐Chae , Ha Suk‐Jin , Yu Byung Jo , Koo Hyun Min , Park Sung Min , Seo Jin‐Ho , Kweon Dae‐Hyuk , Park Jae Chan , Jin Yong‐Su
Published: October 2010

Although Saccharomyces cerevisiae is capable of fermenting galactose into ethanol, ethanol yield and productivity from galactose are significantly lower than those from glucose. An inverse metabolic engineering approach was undertaken to improve ethanol yield and productivity from galactose in S. cerevisiae. A genome‐wide perturbation library was introduced into S. cerevisiae, and then fast galactose‐fermenting transformants were screened using three different enrichment methods. The characterization of genetic perturbations in the isolated transformants revealed three target genes whose overexpression elicited enhanced galactose utilization. One confirmatory (SEC53 coding for phosphomannomutase) and two novel targets (SNR84 coding for a small nuclear RNA and a truncated form of TUP1 coding for a general repressor of transcription) were identified as overexpression targets that potentially improve galactose fermentation. Beneficial effects of overexpression of SEC53 may be similar to the mechanisms exerted by overexpression of PGM2 coding for phosphoglucomutase. While the mechanism is largely unknown, overexpression of SNR84, improved both growth and ethanol production from galactose. The most remarkable improvement of galactose fermentation was achieved by overexpression of the truncated TUP1 (tTUP1) gene, resulting in unrivalled galactose fermentation capability, that is 250% higher in both galactose consumption rate and ethanol productivity compared to the control strain. Moreover, the overexpression of tTUP1 significantly shortened lag periods that occurs when substrate is changed from glucose to galactose. Based on these results we proposed a hypothesis that the mutant Tup1 without C‐terminal repression domain might bring in earlier and higher expression of GAL genes through partial alleviation of glucose repression. mRNA levels of GAL genes (GAL1, GAL4, and GAL80) indeed increased upon overexpression of tTUP. The results presented in this study illustrate that alteration of global regulatory networks through overexpression of the identified targets (SNR84 and tTUP1) is as effective as overexpression of a rate limiting metabolic gene (PGM2) in the galactose assimilation pathway for efficient galactose fermentation in S. cerevisiae. In addition, these results will be industrially useful in the biofuels area as galactose is one of the abundant sugars in marine plant biomass such as red seaweed as well as cheese whey and molasses. Biotechnol. Bioeng. 2011; 108:621–631. © 2010 Wiley Periodicals, Inc.
Engineering butanol‐tolerance in escherichia coli with artificial transcription factor libraries
Authors: Lee Ju Young , Yang Kyung Seok , Jang Su A , Sung Bong Hyun , Kim Sun Chang
Published: October 2010

Escherichia coli has been explored as a host for butanol production because of its many advantages such as a fast growth and easy genetic manipulation. Butanol toxicity, however, is a major concern in the biobutanol production with E. coli. In particular, E. coli growth is severely inhibited by butanol, being almost completely stopped by 1% (vol/vol) butanol. Here we developed a new method to increase the butanol‐tolerance of E. coli with artificial transcription factor (ATF) libraries which consist of zinc finger (ZF) DNA‐binding proteins and an E. coli cyclic AMP receptor protein (CRP). Using these ATFs, we selected a butanol‐tolerant E. coli which can tolerate up to 1.5% (vol/vol) butanol, with a concomitant increase in heat resistance. We also identified genes of E. coli that are associated with the butanol‐tolerance. These results show that E. coli can be engineered as a promising host for high‐yield butanol production. Biotechnol. Bioeng. 2011; 108:742–749. © 2010 Wiley Periodicals, Inc.
Ectopic expression of human mTOR increases viability, robustness, cell size, proliferation, and antibody production of chinese hamster ovary cells
Authors: Dreesen Imke A.J. , Fussenegger Martin
Published: October 2010

Engineering of mammalian production cell lines to improve titer and quality of biopharmaceuticals is a top priority of the biopharmaceutical manufacturing industry providing protein therapeutics to patients worldwide. While many engineering strategies have been successful in the past decade they were often based on the over‐expression of a single transgene and therefore limited to addressing a single bottleneck in the cell's production capacity. We provide evidence that ectopic expression of the global metabolic sensor and processing protein mammalian target of rapamycin (mTOR), simultaneously improves key bioprocess‐relevant characteristics of Chinese hamster ovary (CHO) cell‐derived production cell lines such as cell growth (increased cell size and protein content), proliferation (increased cell‐cycle progression), viability (decreased apoptosis), robustness (decreased sensitivity to sub‐optimal growth factor and oxygen supplies) and specific productivity of secreted human glycoproteins. Cultivation of mTOR‐transgenic CHO‐derived cell lines engineered for secretion of a therapeutic IgG resulted in antibody titers of up to 50 pg/cell/day, which represents a four‐fold increase compared to the parental production cell line. mTOR‐based engineering of mammalian production cell lines may therefore have a promising future in biopharmaceutical manufacturing of human therapeutic proteins. Biotechnol. Bioeng. 2011; 108:853–866. © 2010 Wiley Periodicals, Inc.
Purification of singly PEGylated α‐lactalbumin using charged ultrafiltration membranes
Authors: Ruanjaikaen Krisada , Zydney Andrew L.
Published: October 2010

One of the challenges in producing a PEGylated therapeutic protein is that the PEGylation reaction typically generates a mixture of both singly and multiply PEGylated species. The objective of this study was to examine the feasibility of using ultrafiltration for the purification of a singly PEGylated protein from the multiply PEGylated conjugates. Data were obtained with α‐lactalbumin that was PEGylated with a 20 kDa activated PEG, with the ultrafiltration performed over a range of pH and ionic strength using both unmodified and negatively charged composite regenerated cellulose membranes. Purification of the singly PEGylated α‐lactalbumin from the multiply PEGylated species was accomplished using a diafiltration process with a negatively charged membrane at pH 5 and an ionic strength of 0.4 mM, conditions that maximized the electrostatic exclusion of the multiply PEGylated species from the charged membrane. The diafiltration process provided more than 97% yield with greater than 20‐fold purification between the singly and doubly PEGylated proteins and nearly complete removal of the more heavily PEGylated species. The singly PEGylated α‐lactalbumin was recovered as a dilute filtrate solution, although this dilution could be eliminated using a cascade filtration or the final product could be re‐concentrated in a second ultrafiltration as part of the final formulation. These results demonstrate the feasibility of using ultrafiltration for the purification of singly PEGylated protein therapeutics. Biotechnol. Bioeng. 2011; 108:822–829. © 2010 Wiley Periodicals, Inc.
Enantiomeric separation of d,l‐tryptophan and d,l‐kynurenine by HPLC using pre‐column fluorescence derivatization with R(−)‐DBD‐PyNCS
Authors: Iizuka Hideaki , Hirasa Yasushi , Kubo Kazumi , Ishii Kana , Toyo'oka Toshimasa , Fukushima Takeshi
Published: October 2010

The enantiomeric separation of d,l‐tryptophan (Trp) and d,l‐kynurenine (KYN) was investigated by high‐performance liquid chromatography using pre‐column fluorescence derivatization with a chiral fluorescent labeling reagent, R(−)‐4‐(3‐isothiocyanatopyrrolidin‐1‐yl)‐7‐ (N,N‐dimethylaminosulfonyl)‐2,1,3‐benzoxadiazole [R(−)‐DBD‐PyNCS]. Using an octadecylsilica column, namely, an Inertsil ODS‐3 column (250 × 2.0 mm; i.d., 3 µm), four fluorescence peaks of D‐ and l‐Trp as well as d‐ and l‐KYN derivatized with R(−)‐DBD‐PyNCS were clearly observed, and their chemical structures were confirmed by HPLC–time‐of‐flight–mass spectrometry. Simultaneous separation was achieved under the mobile phase condition of 1.5% acetic acid in H2O–CH3CN (60:40), and the separation factors of d,l‐Trp and d,l‐KYN derivatized with R(−)‐DBD‐PyNCS were 1.22 and 1.19, respectively. Fluorescence detection was carried out by setting the emission wavelength at 565 nm, and the excitation wavelength at 440 nm, and the detection limits were approximately 0.3–0.5 pmol (signal‐to‐noise ratio of 3). Copyright © 2010 John Wiley & Sons, Ltd.
Location of the motor entry point and intramuscular motor point of the tibialis posterior muscle: For effective motor point block
Authors: Lee Je‐Hun , Lee Be‐Na , An Xiaochun , Chung Rak‐Hee , Han Seung‐Ho
Published: October 2010

The aim of this study was to elucidate the anatomical location of the motor entry point (MEP) and intramuscular motor point (IMP) of the tibialis posterior muscle for effective motor point block. Thirty‐six fresh specimens from 20 adult Korean cadavers (11 males and 9 females) were investigated. The reference line between the most proximal‐medial articular margin of the tibia (MPM) at the level of the knee joint and the most distal point of the malleolus of the tibia (MDM) on the surface were identified. The mean length of the reference line was 326.5 ± 27.1 mm. There were 82.5% of the total number of MEPs located at 10–30% and 67.9% of the total IMPs were 10–40% from the MPM. The safety zone for botulinum toxin (BTX) injections on the medial approach was 10–40% from the MPM. In addition, insertion of the needle to a depth of 3.5 cm from the surface of the skin was effective. These results may assist in determining more accurate localization of injection sites. Clin. Anat. 24:91–96, 2011. © 2011 Wiley‐Liss, Inc.
Sesamoid and accessory bones of the hand—An epidemiologic survey in a Mediterranean population
Authors: Amar Eyal , Rozenblat Yishai , Chechik Ofir
Published: October 2010

The prevalence and distribution of sesamoid and accessory bones in the hands is quite variable between different populations and ethnic groups. However, there are no published data on their distribution in Mediterranean populations. Studies on the validity of radiographic assessment for the presence of these bones are also lacking. A retrospective review of 442 radiographs of adult patients was performed in order to assess the incidence of sesamoid and accessory bones in the hands of a Mediterranean population. Additionally, two independent observers reviewed 174 radiographs and used the Kappa parameter in order to assess the validity of radiographic interpretation of radiographs for the presence of those bones. There was an incidence of 99.5% sesamoid bones (n = 440) in the 1st metacarpophalangeal (MCP) joint, 42.3% in the 2nd MCP joint (n = 187), and 41.1% (n = 182) in the 5th MCP joint. The incidence in the 1st interphalangeal joints was 26.2% (n = 116). The incidence of sesamoid bones in the 1st, 2nd, and 5th metacarpophalangeal joints in a Mediterranean population was found to be similar to that of Arab and Caucasian populations, whereas their prevalence in the 1st interphalageal joint coincided more with Arab populations. Interobserver reliability of the interpretation of radiographs for the presence of sesamoid bones in the hand was good (Kappa > 0.68) in all locations, except for the 1st MCP joint. Further research is warranted to further elucidate the genetic and/or environmental reasons for the differences between those populations. Clin. Anat. 24:183–187, 2011. © 2010 Wiley‐Liss, Inc.
Alpha‐lipoic acid does not alter stress protein response to acute exercise in diabetic brain
Authors: Lappalainen Jani , Lappalainen Zekine , Oksala Niku K. J. , Laaksonen David E. , Khanna Savita , Sen Chandan K. , Atalay Mustafa
Published: October 2010

Heat shock proteins (HSPs) are molecular chaperones which may act protective in cerebrovascular insults and peripheral diabetic neuropathy. We hypothesized that alpha‐lipoic acid (LA), a natural thiol antioxidant, may enhance brain HSP response in diabetes. Rats with or without streptozotocin‐induced diabetes were treated with LA or saline for 8 weeks. Half of the rats were subjected to exhaustive exercise to investigate HSP induction, and the brain tissue was analyzed. Diabetes increased constitutive HSC70 mRNA, and decreased HSP90 and glucose‐regulated protein 75 (GRP75) mRNA without affecting protein levels. Exercise increased HSP90 protein and mRNA, and also GRP75 and heme oxygenase‐1 (HO‐1) mRNA only in non‐diabetic animals. LA had no significant effect on brain HSPs, although LA increased HSC70 and HO‐1 mRNA in diabetic animals and decreased HSC70 mRNA in non‐diabetic animals. Eukaryotic translation elongation factor‐2, essential for protein synthesis, was decreased by diabetes and suggesting a mechanism for the impaired HSP response related to translocation of the nascent chain during protein synthesis. LA supplementation does not offset the adverse effects of diabetes on brain HSP mRNA expression. Diabetes may impair HSP translation through elongation factors related to nascent chain translocation and subsequent responses to acute stress. Copyright © 2010 John Wiley & Sons, Ltd.
A Strategy for Designing a Peptide Probe for Detection of β‐Amyloid Oligomers
Authors: Hu Yang , Su Baihao , Kim Chung‐Sei , Hernandez Michael , Rostagno Agueda , Ghiso Jorge , Kim Jin Ryoun
Published: October 2010

Aggregation of β‐amyloid (Aβ) is implicated in the pathology of Alzheimer's disease. Development of a robust strategy to detect Aβ oligomeric intermediates, which have been identified as significant toxic agents, would be highly beneficial in the screening of drug candidates as well as enhancing our understanding of Aβ oligomerization. Rapid, specific and quantitative detection, currently unavailable, would be highly preferred for accurate and reliable probing of transient Aβ oligomers. Here, we report the development of a novel peptide probe, PG46, based on the nature of Aβ self‐assembly and the conformation‐sensitive fluorescence of the biarsenical dye, FlAsH. PG46 was found to bind to Aβ oligomers and displayed an increase in FlAsH fluorescence upon binding. No such event was observed when PG46 was co‐incubated with Aβ low‐molecular‐weight species or Aβ fibrils. Aβ oligomer detection was fast, and occurred within one hour without any additional sample incubation or preparation. We anticipate that the development of a strategy for detection of amyloid oligomers described in this study will be directly relevant to a host of other amyloidogenic proteins.
Mechanism(s) of SWI/SNF‐Induced Nucleosome Mobilization
Authors: Liu Ning , Balliano Angela , Hayes Jeffrey J.
Published: October 2010

Impediments to DNA access due to assembly of the eukaryotic genome into chromatin are in part overcome by the activity of ATP‐dependent chromatin‐remodeling complexes. These complexes employ energy derived from ATP hydrolysis to destabilize histone—DNA interactions and alter nucleosome positions, thereby increasing the accessibility of DNA‐binding factors to their targets. However, the mechanism by which theses complexes accomplish this task remains unresolved. We review aspects of nucleosome alteration by the SWI/SNF complex, the archetypal remodeling enzyme. We focus on experiments that provide insights into how SWI/SNF induces nucleosome movement along DNA. Numerous biochemical activities have been characterized for this complex, all likely providing clues as to the molecular mechanism of translocation.
Controlled Synthesis of a Thin LTL Zeolitic Membrane Using Nano‐Sized Seeds: Characterization and Permeation Performance
Authors: Ghoroghchian F. , Aghabozorg H. , Farhadi F. , Kazemian H.
Published: October 2010

Industrial zeolitic membranes which offer a remarkable selectivity compared to polymeric membranes, suffer of the lower flux due of their larger thickness (e.g., 10–30 μm). This problem can be addressed by controlled synthesis of nanolayers, resulting in thinner membrane layers (e.g., 0.5–5 μm). An aluminosilicate gel with a molar composition of 20SiO2:Al2O3:10K2O:400H2O was used to prepare several membranes of zeolite L by means of a controlled hydrothermal synthesis on the surface of a porous alumina disc seeded with nanozeolite LTL crystals. Nanocrystallites of LTL zeolite with an average particle size of 80–100 nm were successfully synthesized and characterized. Using these nanoparticles as seeds, a zeolite L layer with an average thickness of 2 μm was synthesized on the alumina support at 150 °C.
Analysis of a pH‐Based Potentiometric Biosensor Using the Homotopy Perturbation Method
Authors: Meena A. , Rajendran L.
Published: October 2010

A mathematical model of steady‐state and non‐steady‐state responses of a pH‐based potentiometric biosensor immobilizing organophosphorus hydrolase was developed. The model is based on non‐stationary diffusion equations containing a nonlinear term related to the Michaelis‐Menten kinetics of an enzymatic reaction. An analytical expression for the substrate concentration was obtained for all values of parameter a (Thiele modulus) using the homotopy perturbation method. From this result, the concentrations of the deprotonation products of an organophosphodiester (PhH, ZH and AH) were obtained. Our analytical results were compared with available simulation results. A satisfactory agreement with the simulation data is noted.
Catalytic Steam Reforming of Fast Pyrolysis Bio‐Oil in Fixed Bed and Fluidized Bed Reactors
Authors: Lan P. , Xu Q. , Zhou M. , Lan L. , Zhang S. , Yan Y.
Published: October 2010

Catalytic steam reforming of bio‐oil is an economically‐feasible route which produces renewable hydrogen. The Ni/MgO‐La2O3‐Al2O3 catalyst was prepared with Ni as active agent, Al2O3 as support, and MgO and La2O3 as promoters. The experiments were conducted in fixed bed and fluidized bed reactors, respectively. Temperature, steam‐to‐carbon mole ratio (S/C), and liquid hourly space velocity (LHSV) were investigated with hydrogen yield as index. For the fluidized bed reactor, maximum hydrogen yield was obtained under temperatures 700–800 °C, S/C 15–20, LHSV 0.5–1.0 h–1, and the maximum H2 yield was 75.88 %. The carbon deposition content obtained from the fluidized bed was lower than that from the fixed bed. The maximum H2 yield obtained in the fluidized bed was 7 % higher than that of the fixed bed. The carbon deposition contents obtained from the fluidized bed was lower than that of the fixed bed at the same reaction temperature.
Ridge and PLS based rational function regression
Authors: Taavitsainen Veli‐Matti
Published: October 2010

This study introduces both ridge and partial least squares (PLS) regression based rational function regression techniques. The results of four different cases are compared to those obtained using other regression techniques. The results are mostly favorable, and the proposed method should be considered as a noteworthy alternative in nonlinear modeling. Copyright © 2010 John Wiley & Sons, Ltd.
New‐Phased Metastable V2O3 Porous Urchinlike Micronanostructures: Facile Synthesis and Application in Aqueous Lithium Ion Batteries
Authors: Xu Yang , Zheng Lei , Wu Changzheng , Qi Fei , Xie Yi
Published: October 2010

New‐phased metastable V2O3 porous urchinlike micronanostructures were first fabricated on a large scale by a simple top‐down strategy of pyrolyzing a vanadyl ethylene glycolated precursor in the absence of any templates or matrices. The pyrolysis mechanism was clearly revealed by synchrotron vacuum ultraviolet (VUV) photoionization mass spectra for the first time. The new‐phased metastable V2O3 exhibits a body‐centered cubic bixbyite structure and shows structural evolution from metastable cubic symmetry to thermodynamically stable rhombohedral symmetry V2O3 (R) above 510 °C. Furthermore, the prepared V2O3 porous urchinlike micronanostructures, as anode materials in aqueous lithium ion batteries, exhibit improved electrochemical properties with relatively high first discharge capacity and better cycle retention relative to thermodynamically stable V2O3 (R), which is derived from its unique microscopic crystal structure and macroscopic 3D framework with rigid morphology, porous structure, and high specific surface area.
A Photoelectrochemical Immunosensor Based on Au‐Doped TiO2 Nanotube Arrays for the Detection of α‐Synuclein
Authors: An Yarui , Tang Linlin , Jiang Xiaoli , Chen Hua , Yang Meicheng , Jin Litong , Zhang Shengping , Wang Chuangui , Zhang Wen
Published: October 2010

α‐Synuclein (α‐SYN) is a very important neuronal protein that is associated with Parkinson’s disease. In this paper, we utilized Au‐doped TiO2 nanotube arrays to design a photoelectrochemical immunosensor for the detection of α‐SYN. The highly ordered TiO2 nanotubes were fabricated by using an electrochemical anodization technique on pure Ti foil. After that, a photoelectrochemical deposition method was exploited to modify the resulting nanotubes with Au nanoparticles, which have been demonstrated to facilitate the improvement of photocurrent responses. Moreover, the Au‐doped TiO2 nanotubes formed effective antibody immobilization arrays and immobilized primary antibodies (Ab1) with high stability and bioactivity to bind target α‐SYN. The enhanced sensitivity was obtained by using {Ab2‐Au‐GOx} bioconjugates, which featured secondary antibody (Ab2) and glucose oxidase (GOx) labels linked to Au nanoparticles for signal amplification. The GOx enzyme immobilized on the prepared immunosensor could catalyze glucose in the detection solution to produce H2O2, which acted as a sacrificial electron donor to scavenge the photogenerated holes in the valence band of TiO2 nanotubes upon irradiation of the other side of the Ti foil and led to a prompt photocurrent. The photocurrents were proportional to the α‐SYN concentrations, and the linear range of the developed immunosensor was from 50 pg mL−1 to 100 ng mL−1 with a detection limit of 34 pg mL−1. The proposed method showed high sensitivity, stability, reproducibility, and could become a promising technique for protein detection.
Near‐Infrared Cell‐Permeable Hg2+‐Selective Ratiometric Fluorescent Chemodosimeters and Fast Indicator Paper for MeHg+ Based on Tricarbocyanines
Authors: Guo Zhiqian , Zhu Weihong , Zhu Mingming , Wu Xumeng , Tian He
Published: October 2010

Three tricarbocyanine dyes (IR‐897, IR‐877, and IR‐925) with different thiourea substituents that function as dosimeter units through specific Hg2+‐induced desulfurization have been demonstrated in a fast indicator paper for Hg2+ and MeHg+ ions. In comparison with available Hg2+‐selective chemodosimeters, IR‐897 and IR‐877 show several advantages, such as convenient synthesis, very long wavelengths falling in the near‐infrared (NIR) region (650–900 nm) with high molar extinction coefficients, a ratiometric response, and quite low disturbance with Ag+ and Cu2+ ions. They exhibit large redshifts, which result in a clear color change from deep blue to pea green that can be easily monitored by the naked eye for a convenient indicator paper. In emission spectra, they display a characteristic turn‐off mode at 780 nm and turn‐on mode at 830 nm with titration of Hg2+ ions. Remarkably, the signal/noise (S/N) ratio with other thiophilic metal ions (Ag+ and Cu2+) is greatly enhanced with ratiometric measurement of two channels: excitation spectra mode (I810 nm/I670 nm, monitored at 830 nm) and emission spectra mode (I830 nm/I780 nm, isosbestic absorption point at 730 nm as excitation). The distinct response is dependent upon the electron‐donating effect of the thiourea substituents; that is, the stronger the electron‐donating capability of the thiourea substituents, the faster the Hg2+‐promoted cyclization. Additionally, experiments with living SW1116 cells show that these three tricarbocyanine dyes with low toxicity can exhibit special characteristics that are favorable for visualizing intracellular Hg2+ and MeHg+ ions in biological systems, including excellent membrane permeability, minimal interfering absorption and fluorescence from biological samples, low scattering, and deep penetration into tissues.
A Neutral Zwitterionic Molecular Solid
Authors: El‐Ghayoury Abdelkrim , Mézière Cécile , Simonov Sergey , Zorina Leokadiya , Cobián Manuel , Canadell Enric , Rovira Carme , Náfrádi Bálint , Sipos Balázs , Forró László , Batail Patrick
Published: October 2010

We report on the acid ethylenedithiotetrathiafulvaleneamidoglycine (EDT‐TTF‐CO‐NH‐CH2‐CO2H; 1; EDT‐TTF=ethylenedithiotetrathiafulvalene) and the 1:1 adduct [(EDT‐TTF).+‐CO‐NH‐CH2‐(CO2)−][(EDT‐TTF)‐CO‐NH‐CH2‐(CO2H)]⋅CH3OH (2), a new type of hydrogen‐bonded, 1:1 acid/zwitterion hybrid embrace of redox peptidics into a two‐dimensional architecture, an example of a system deliberately fashioned so that oxidation of π‐conjugated cores toward the radical‐cation form would interfere with the activity of the appended ionizable residues in the presence of a templating base during crystal growth. First‐principles calculations demonstrate that, notwithstanding preconceived ideas, a metallic state is more stable than the hole‐localized alternatives for a neat 1:1 neutral acid/zwitterion hybrid. The inhomogeneous Coulomb field associated with proton‐shared, interstacks OH⋅⋅⋅O hydrogen bonds between the ionizable residues distributed on both sides of the two‐dimensional π‐conjugated framework leads, however, to a weak hole localization responsible for the activated but high conductivity of 1 S cm−1. This situation is reminiscent of the role of the environment on electron transfer in tetraheme cytochrome c, in which the protonation state of a heme propionate becomes paramount, or ion‐gated transport phenomena in biology. These observations open rather intriguing opportunities for the construction of electronic systems at the interface of chemistry and biology.
Efficient Access to New Chemical Space Through Flow—Construction of Druglike Macrocycles Through Copper‐Surface‐Catalyzed Azide–Alkyne Cycloaddition Reactions
Authors: Bogdan Andrew R. , James Keith
Published: October 2010

A series of 12‐ to 22‐membered macrocycles, with druglike functionality and properties, have been generated by using a simple and efficient copper‐catalyzed azide–acetylene cycloaddition reaction, conducted in flow in high‐temperature copper tubing, under environmentally friendly conditions. The triazole‐containing macrocycles have been generated in up to 90 % yield in a 5 min reaction, without resorting to the high‐dilution conditions typical of macrocyclization reactions. This approach represents a very efficient method for constructing this important class of molecules, in terms of yield, concentration, and environmental considerations.
Measuring circular dichroism in a capillary cell using the b23 synchrotron radiation CD beamline at diamond light source
Authors: Jávorfi Tamás , Hussain Rohanah , Myatt Daniel , Siligardi Giuliano
Published: October 2010

Synchrotron radiation circular dichroism (SRCD) is a well‐established method in structural biology. The first UV‐VIS beamline dedicated to circular dichroism at Diamond Light Source, a third generation synchrotron facility in South Oxfordshire, has recently become operational and it is now available for the user community. Herein we present an important application of SRCD: the CD measurement of protein solutions in fused silica rectangular capillary cells. This was achieved without the use of any lens between the photoelastic modulator and the photomultiplier tube detectors by exploiting the high photon flux of the collimated beam that can be as little as half a millimeter squared. Measures to minimize or eliminate vacuum‐UV protein denaturation effects are discussed. The CD spectra measured in capillaries is a proof of principle to address CD measurements in microdevice systems using the new B23 SRCD beamline. Chirality 22:E149–E153, 2010. © 2010 Wiley‐Liss, Inc.
Behavior of Aluminum Adsorption in Different Compost‐Derived Humic Acids
Authors: Lin Yong‐Hong , Su Po‐Hsin
Published: October 2010

Humic acid plays an important role in the distribution of heavy metals in the environment. The aims of this study were conducted to evaluate the spectroscopic characteristics and aluminum (Al) adsorption of humic acids which were extracted from four composts. The functional groups were determined by Fourier transform infrared spectroscopy (FTIR) and solid‐state 13C nuclear magnetic resonance spectroscopy (13C‐NMR). The results showed that the aromatic groups were all found in the humic acids of the four composts, and the surface of humic acids included carboxylic group, hydroxylic group, and amino group. The experiment of Al adsorption was described by Freundlich equation. It showed that the adsorption of Al by humic acid from compost of cattle manure was higher, but that from the compost of pig manure, lemon manure, tea manure in equal preparation was lower. The adsorptive behavior was different due to the interaction by functional groups (OH and COOH) with Al. These results can describe the fate and transportation of Al in the soil of different organic fertilizer.
Differential bulbar and extrabulbar projections of diverse olfactory receptor neuron populations in the adult zebrafish (Danio rerio)
Authors: Gayoso José Ángel , Castro Antonio , Anadón Ramón , Manso María Jesús
Published: October 2010

Immunohistochemical methods were used to characterize the expression of two calcium‐binding proteins, calretinin (CR) and S100, in the olfactory rosette of the adult zebrafish. These proteins are expressed in different sets of sensory neurons, and together represent a large proportion of these cells. Double immunofluorescence for CR and Gαolf protein, and CR immunoelectron microscopy, indicated that most CR‐immunoreactive (ir) cells were ciliary neurons. Differential S100‐ and CR‐ir projections to glomerular fields of the olfactory bulb were also observed, although these projections overlap in some glomeruli. Application of the carbocyanine dye DiI to either S100‐ir or CR‐ir glomerular regions led to labeling of cells mostly similar to S100‐ir and CR‐ir neurons, respectively. Instead, these bulbar regions project to similar telencephalic targets. On the other hand, antibodies against keyhole limpet hemocyanin (KLH)‐stained numerous sensory cells in the olfactory rosette, including cells that were CR‐ and S100‐negative. This antiserum also stained most primary bulbar projections and revealed extrabulbar olfactory primary projections coursing to the ventral area of the telencephalon through the medial olfactory tract. This extrabulbar projection was confirmed by tract‐tracing with DiI. A loose association of this extrabulbar primary olfactory projection and the catecholaminergic populations of the ventral area was also observed with double tyrosine hydroxylase/KLH‐like immunohistochemistry. Comparison between KLH‐like‐ir pathways and the structures revealed by FMRFamide immunohistochemistry (a marker of terminal ganglion cells and fibers) indicated that the KLH‐like‐ir extrabulbar projection was different from the terminal nerve system. The significance of the extrabulbar olfactory projection of zebrafish is discussed. J. Comp. Neurol. 519:247‐276, 2011. © 2010 Wiley‐Liss, Inc.
Spatiotemporal nuclear factor interleukin‐6 expression in the rat brain during lipopolysaccharide‐induced fever is linked to sustained hypothalamic inflammatory target gene induction
Authors: Damm Jelena , Luheshi Giamal N. , Gerstberger Rüdiger , Roth Joachim , Rummel Christoph
Published: October 2010

Rats injected with lipopolysaccharide (LPS) show brain‐controlled sickness symptoms, including fever. In these animals, early genomic activation of brain cells was previously monitored by immunohistochemical detection of transcription factors such as nuclear factor (NF)‐κB or signal transducer and activator of transcription (STAT)3 and was linked to the initiation or maintenance of the febrile response. To investigate whether NF‐IL6 might be another important transcription factor implicated in this kind of immune‐to‐brain signaling, rats were injected with LPS (100 μg/kg, intraperitoneally) or phosphate‐buffered saline, and brains were analyzed by immunohistochemistry, real‐time PCR, or Western blot 4, 6, 8, and 10 hours later. Moderate to strong LPS‐induced nuclear NF‐IL6 immunoreactivity (IR) occurred in a time‐dependent manner within circumventricular organs, namely, the vascular organ of the lamina terminalis, the subfornical organ, the area postrema, and the median eminence, brain structures with a leaky blood–brain barrier. Furthermore, nuclear NF‐IL6‐IR was observed in the pituitary gland, the choroid plexus, and the meninges as well as blood vessels throughout the entire brain. Endothelial, microglial, and ependymal cells, astrocytes, perivascular macrophages, and neurons exhibited LPS‐induced nuclear NF‐IL6‐IR; mRNA levels of NF‐IL6, responsive inflammatory genes, and NF‐IL6 protein levels were significantly elevated. As opposed to observations on STAT3 or NFκB, the percentage of NF‐IL6‐reactive cells increased in parallel to late phases of the febrile response. In conclusion, these results suggest a potential role for NF‐IL6 in the maintenance or possibly the termination of LPS‐induced fever. Moreover, we propose NF‐IL6 to be a delayed brain cell activation marker. J. Comp. Neurol. 519:480–505, 2011. © 2010 Wiley‐Liss, Inc.
A novel form of pigment‐dispersing hormone in the central nervous system of the intertidal marine isopod, Eurydice pulchra (leach)
Authors: Wilcockson David C. , Zhang Lin , Hastings Michael H. , Kyriacou Charalambos P. , Webster Simon G.
Published: October 2010

Pigment‐dispersing factor (PDF) is well known as a circadian clock output factor, which drives daily activity rhythms in many insects. The role of its homologue, pigment‐dispersing hormone (PDH), in the regulation of circadian and/or circatidal rhythmicity in crustaceans is, however, poorly understood. The intertidal isopod crustacean, Eurydice pulchra has well‐defined circatidal (12.4‐hour) activity rhythms. In this study we show that this runs parallel to a circadian (24‐hour) cycle of chromatophore dispersion. As a first step in determining the potential role of PDH in these rhythms, we have identified a novel form of PDH expressed in this species. Because conventional homology cloning was unsuccessful, we employed immuno‐identification and Edman microsequencing to determine the primary structure of this peptide. From this, cDNA cloning identified the nucleotide encoding sequence and thus facilitated description of PDH neurons by in situ hybridization and immunohistochemistry. We show them to be morphologically similar to those that co‐ordinate circadian activity rhythms in insects. In animals expressing both tidal (activity) and circadian (chromatophore) rhythms, however, there was no evidence for a corresponding periodicity in the expression of pdh transcript, as determined by quantitative reverse transcriptase polymerase chain reaction (qRT‐PCR) in Eurydice heads. It is therefore suggested that any role for PDH in daily/tidal timing in Eurydice is not mediated at the transcriptional level, rather rhythms in neurohemal release may be important in such co‐ordination. J. Comp. Neurol. 519:562–575, 2011. © 2010 Wiley‐Liss, Inc.
Composition of the migratory mass during development of the olfactory nerve
Authors: Miller Alexandra M. , Treloar Helen B. , Greer Charles A.
Published: October 2010

The embryonic development of the olfactory nerve includes the differentiation of cells within the olfactory placode, migration of cells into the mesenchyme from the placode, and extension of axons by the olfactory sensory neurons (OSNs). The coalition of both placode‐derived migratory cells and OSN axons within the mesenchyme is collectively termed the “migratory mass.” Here we address the sequence and coordination of the events that give rise to the migratory mass. Using neuronal and developmental markers, we show subpopulations of neurons emerging from the placode by embryonic day (E)10, a time at which the migratory mass is largely cellular and only a few isolated OSN axons are seen, prior to the first appearance of OSN axon fascicles at E11. These neurons also precede the emergence of the gonadotropin‐releasing hormone neurons and ensheathing glia which are also resident in the mesenchyme as part of the migratory mass beginning at about E11. The data reported here begin to establish a spatiotemporal framework for the migration of molecularly heterogeneous placode‐derived cells in the mesenchyme. The precocious emigration of the early arriving neurons in the mesenchyme suggests they may serve as “guidepost cells” that contribute to the establishment of a scaffold for the extension and coalescence of the OSN axons. J. Comp. Neurol. 518:4825–4811, 2010. © 2010 Wiley‐Liss, Inc.
Heavy metal contaminants can eliminate quantum dot fluorescence
Authors: Zarkowsky David , Lamoreaux Laurie , Chattopadhyay Pratip , Koup Richard A. , Perfetto Stephen P. , Roederer Mario
Published: October 2010

Quantum dots (QD) are fluorescent nanocrystals that are highly useful in imaging and flow cytometric analyses. During routine use of monoclonal antibody conjugates of QD, we have occasionally seen partial or total loss of fluorescence when using certain lots of fixative solutions. We hypothesized that a low level contamination with heavy metal cations was responsible, since low level metal contaminants are not uncommon in formalin solutions. By titrating known concentrations of heavy metal cations into staining solutions, we found that millimolar concentrations of ferrous and zinc ions, and as low as 50 nanomolar cupric ions, completely eliminated QD fluorescence. By mass spectroscopic quantification of metals in commercial fixative solutions previously shown to perform poorly or well with regard to QD fluorescence, we confirmed that the presence of copper in solution was correlated with poor performance. Notably, prior addition of EDTA to chelate the divalent cations in these solutions prevented the inhibition of QD fluorescence. Finally, the copper‐induced loss of QD fluorescence is irreversible: cells labeled with QD are highly fluorescent and can be rendered nonfluorescent by the addition of cupric sulfate, even after washing extensively. Indeed, these cells can then be successfully stained with other QD reagents, providing a method for immunofluorescence restaining of cells without contaminating fluorescence from the first stain. Published 2010 Wiley‐Liss, Inc.
Cortisol levels in relation to maternal interaction and child internalizing behavior in preterm and full‐term children at 18 months corrected age
Authors: Brummelte Susanne , Grunau Ruth E. , Zaidman‐Zait Anat , Weinberg Joanne , Nordstokke David , Cepeda Ivan L.
Published: October 2010

Cortisol levels were compared in children born preterm at extremely low gestational age (ELGA; 24–28 weeks), very low gestational age (VGLA; 29–32 weeks), and full‐term in response to cognitive assessment at 18 months corrected age (CA). Further, we investigated the relationship between maternal interactive behaviors and child internalizing behaviors (rated by the mother) in relation to child cortisol levels. EGLA children had higher “pretest” cortisol levels and a different pattern of cortisol response to cognitive assessment compared to VGLA and full‐terms. Higher cortisol levels in ELGA, but not full‐term, children were associated with less optimal mother interactive behavior. Moreover, the pattern of cortisol change was related to internalizing behaviors among ELGA, and to a lesser degree VLGA children. In conclusion, our findings suggest altered programming of the hypothalamic‐pituitary‐adrenal (HPA) axis in preterm children, as well as their greater sensitivity to environmental context such as maternal interactive behavior. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 53:184–195, 2011
Motor development in 9‐month‐old infants in relation to cultural differences and iron status
Authors: Angulo‐Barroso Rosa M. , Schapiro Lauren , Liang Weilang , Rodrigues Onike , Shafir Tal , Kaciroti Niko , Jacobson Sandra W. , Lozoff Betsy
Published: October 2010

Motor development, which allows infants to explore their environment, promoting cognitive, social, and perceptual development, can be influenced by cultural practices and nutritional factors, such as iron deficiency. This study compared fine and gross motor development in 209 9‐month‐old infants from urban areas of China, Ghana, and USA (African‐Americans) and considered effects of iron status. Iron deficiency anemia was most common in the Ghana sample (55%) followed by USA and China samples. Controlling for iron status, Ghanaian infants displayed precocity in gross motor development and most fine‐motor reach‐and‐grasp tasks. US African‐Americans performed the poorest in all tasks except bimanual coordination and the large ball. Controlling for cultural site, iron status showed linear trends for gross motor milestones and fine motor skills with small objects. Our findings add to the sparse literature on infant fine motor development across cultures. The results also indicate the need to consider nutritional factors when examining cultural differences in infant development. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 53:196–210, 2011
Current literature in diabetes
Authors:   John Wiley & Sons, Ltd.
Published: October 2010

In order to keep subscribers up‐to‐date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of diabetes/metabolism. Each bibliography is divided into 26 sections: 1 Reviews; 2 General; 3 Genetics; 4 Epidemiology; 5 Immunology; 6 Obesity; 7 Prediction and Prevention; 8 Intervention: a) General; b) Care; c) Drug Therapy; d)Economics; e) Gene therapy; f) Nursing; g) Nutrition; h) Surgery; i) Transplantation; 9 Pathology and Complications: a) General; b) Cardiovascular; c) Eye disease; d) Gestational and fetal; e) Neurological; f) Podiatrical; g) Renal; 10 Endocrinology & Metabolism; 11 Experimental Studies; 12 Diagnosis and Techniques. Within each section, articles are listed in alphabetical order with respect to author