bluuebunny/crossref_metadata_2025_split huggingface dataset:
Linkers for improved cleavage of fusion proteins with an engineered α‐lytic protease
Authors: Lien Samantha , Milner Steven J. , Graham Lloyd D. , Wallace John C. , Francis Geoffrey L. 
Published: November 2004

AbstractAddition of an N‐terminal fusion partner can greatly aid the expression and purification of a recombinant protein in Escherichia coli. We investigated two genetically engineered proteases designed to remove the fusion partner after the protein of interest has been expressed. Recombinant human insulin‐like growth factor‐II (hIGF‐II) has been produced from E. coli‐derived fusion proteins using a novel enzymatic cleavage system that uses a mutant of α‐lytic protease. Initially, two potential fusion protein linkers were designed, Pro‐Ala‐Pro‐His (PAPH) and Pro‐Ala‐Pro‐Met (PAPM), and were tested as substrates in the form of synthetic dodecapeptides. Using mass spectrometry and reverse‐phase HPLC, the position of cleavage was confirmed and the kinetics of synthetic peptide cleavage were examined. Use of the linkers in hIGF‐II fusion proteins produced in E. coli was then evaluated. The fusion proteins constructed consist of the first 11 amino acids of porcine growth hormone linked N‐terminally to hIGF‐II by six amino acids that include the dipeptide Val‐Asn followed by a variable tetrapeptide protease cleavage motif. Mass spectrometry and N‐terminal sequencing confirmed that proteolytic cleavage of the fusion proteins had occurred at the predicted sites. Using the fusion proteins as substrates, the cleavage of the rationally designed motifs by the α‐lytic protease mutant was compared. The fusion protein containing the motif PAPM had a kcat/KM ratio indicating a 1.6‐fold preference over the PAPH fusion protein for cleavage by this enzyme. Furthermore, when hIGF‐II fusion proteins containing the designed cleavable linkers were processed with the engineered α‐lytic protease, they gave greatly improved yields of native hIGF‐II compared to an analogous fusion protein cleaved by H64A subtilisin. Comparison of the peptide and protein cleavage studies shows that the efficient proteolysis of the cleavage motifs is an inherent property of the designed sequences and is not determined by secondary or tertiary structure in the fusion proteins. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 335–343, 2001.
Abnormal dispersion of a purkinje cell subset in the mouse mutant cerebellar deficient folia (cdf)
Authors: Beierbach Elaine , Park Chankyu , Ackerman Susan L. , Goldowitz Dan , Hawkes Richard 
Published: November 2004

AbstractPurkinje cells of different molecular phenotypes subdivide the cortex of the cerebellum both rostrocaudally into parasagittal bands and mediolaterally into transverse zones. Superimposed on the Purkinje cell compartmentation, the cerebellar cortex is pleated into a reproducible array of lobes and lobules. During cerebellar development, Purkinje cell bands are formed through the rostrocaudal dispersal of embryonic clusters, triggered primarily by a Reelin‐dependent signaling pathway. In the naturally occurring mouse mutant cerebellar deficient folia (cdf), there is a failure of Purkinje cell dispersion that results in widespread Purkinje cell ectopia in the adult. The ectopia is restricted primarily to that subset of Purkinje cells that does not express zebrin II/aldolase C and that forms ectopic clusters in among the cerebellar nuclei. Most Purkinje cells that express zebrin II are located normally in a monolayer. Thus, the cerebellum of cdf mutants has a failure of Purkinje cell dispersion that is confined primarily to a zebrin II‐negative (zebrin II−) subpopulation. Despite the Purkinje cell ectopia, the parasagittal band organization of the cerebellum is still clear. The shortening of the cortex is distributed evenly over all lobules, with the result that transverse expression boundaries are relocated with respect to the lobules and fissures. The number of Purkinje cells in the cdf/cdf cerebellum is similar to the number in littermate controls. Therefore, it appears that the lesion in cdf results in the failure of a zebrin II− Purkinje cell subset to disperse either due to a cell intrinsic defect or due to an abnormal interaction between the Purkinje cells and either granule cells or afferent inputs. J. Comp. Neurol. 436:42–51, 2001. © 2001 Wiley‐Liss, Inc.
Distribution of estrogen receptor β immunoreactivity in the rat central nervous system
Authors: Shughrue Paul J. , Merchenthaler Istvan 
Published: November 2004

AbstractThe discovery of estrogen receptor β (ERβ) and subsequent localization of its mRNA in the rat central nervous system (CNS) has provided new insights about estrogen action in brain. A critical step in understanding the role of ERβ is demonstrating that the mRNA is translated into functional protein. The present study used a new ERβ‐specific polyclonal antiserum (Z8P) and immunocytochemistry (ICC) to investigate the distribution of ERβ in the rat CNS. Ovariectomized female rats were perfusion fixed, and free‐floating sections were incubated with Z8P. After visualization with a standard ABC method, nuclear immunoreactivity was seen in neurons throughout the brain, including the olfactory nuclei, laminae IV–VI of the cerebral cortex, medial septum, preoptic area, bed nucleus of the stria terminalis, supraoptic nucleus, paraventricular nucleus, zona incerta, medial and cortical amygdaloid nuclei, cerebellum, nucleus of the solitary tract, ventral tegmental area, and spinal trigeminal nucleus. Moreover, the results of a double‐label ICC/ in situ hybridization study revealed that ERβ mRNA and immunoreactivity were colocalized in neurons of the brain, thus confirming the specificity of the antiserum. Through the use of Western blot analysis, Z8P was shown to recognize in vitro translated ERβ, but not ERα, as well as a 60‐kDa protein from rat granulosa cells and ovary extracts. The results of these studies have demonstrated that (1) ERβ mRNA is translated into immunoreactive protein throughout the rat brain, and (2) ERβ resides in the cell nucleus. Together, these data provide an anatomic foundation for future studies and advance our understanding of estrogen action in hypothalamic and extrahypothalamic brain regions. J. Comp. Neurol. 436:64–81, 2001. © 2001 Wiley‐Liss, Inc.
Quantitative assessment of retinoid signaling pathways in the developing eye and retina of the chicken embryo
Authors: Hoover Frank , Gundersen Thomas E. , Ulven Stine M. , Michaille Jean‐Jacques , Blanchet Sandrine , Blomhoff Rune , Glover Joel C. 
Published: November 2004

AbstractRetinoid signaling has been implicated as an important regulator of retinal development and differentiation. We have used state of the art high‐pressure liquid chromatography to identify and quantitate biologically active retinoids, immunohistochemistry to localize the retinoic acid synthetic enzyme retinaldehyde dehydrogenase 2 (RALDH2), and nucleic acid assays to quantitate and localize retinoid receptor gene transcripts in the developing eye and retina of the chicken. Our results demonstrate spatial distinctions in retinoid synthesis and signaling that may be related to laminar differentiation in the developing retina. Retinoic acids (RAs) and their precursor retinols (ROHs) are the predominant retinoids in the developing eye. All‐trans‐RA and all‐trans‐3,4‐didehydro‐RA are present in the neuroepithelium in approximately equal amounts from early stages of neurogenesis until shortly before hatching. The retinoid X receptor (RXR) ligand 9‐cis‐RA is undetectable at all stages; if present, it cannot exceed a small percentage of the total RA content. RAs are not detected in the pigment epithelium. All‐trans‐ROH is present in the neuroepithelium and pigment epithelium, whereas all‐trans‐3,4‐didehydro‐ROH is detected only in the pigment epithelium and/or the choroid and sclera. RALDH2 immunoreactivity is intense in the choroid, low or absent in the pigment epithelium, and moderate in the neuroepithelium, where it is highest in the outer layers. Transcripts of all five chicken retinoid receptor genes are present in the neural retina and eye throughout development. During the period of neurogenesis, at least three of the receptors (RARγ, RXRγ, RXRα), exhibit dynamic patterns of differential localization within the depths of the neural retina. J. Comp. Neurol. 436:324–335, 2001. © 2001 Wiley‐Liss, Inc.
Variability of position of the P2 glomerulus within a map of the mouse olfactory bulb
Authors: Schaefer Michele L. , Finger Thomas E. , Restrepo Diego 
Published: November 2004

AbstractOlfactory receptor neurons (ORNs) that express a common odorant receptor molecule target specific glomeruli in the olfactory bulb. We systematically assessed the location of the olfactory glomeruli that receive input from ORNs expressing P2 receptors in the P2‐internal ribosome entry site‐tau‐lacZ mouse. We present a new mapping method that includes an Internet‐accessible computer program for generating two‐ and three‐dimensional maps of the glomerular sheet in the olfactory bulbs of mice. Cylindrical coordinates were used to define glomerular location: The coordinates were given as the anteroposterior (AP) distance parallel to the long axis of the bulb (rostrocaudal; RC) and angular measurements with origin defined by the remnant ependymal layer in the center of the granule cell layer in the bulb. Using this method, we can apply rigorous statistical methods to give objective estimates of position and variability. At the 95% confidence interval, the lateral P2 glomerulus lies at coordinates 1,008 μm ± 306 μm AP × 146° ± 12°, and the medial P2 glomerulus lies at 1,828 μm ± 196 μm AP × 204° ± 8°. We estimate that these coordinates encompass a domain containing 29 and 37 of the 1,800 glomeruli (∼2%) for the lateral and medial glomeruli, respectively. Furthermore, the data reported here demonstrate that the rostrocaudal position of small P2 glomeruli is three times more variable than that of large glomeruli. J. Comp. Neurol. 436:351–362, 2001. © 2001 Wiley‐Liss, Inc.
Peptidergic neurohemal system of an insect: Mass spectrometric morphology
Authors: Predel Reinhard 
Published: November 2004

AbstractNeuropeptides are by far the most diverse group of messenger molecules in insects. To understand cell signaling and function, it is essential to reveal the complete neuropeptide profile of a single neuron/nerve/neurohemal organ first. In this study, matrix‐assisted laser desorption ionization time of flight (MALDI‐TOF) mass spectrometry was used to analyze the peptidergic system of an insect, focussing on the neurohemal structures. Major neurohemal organs were investigated, including the retrocerebral complex, perisympathetic organs, and all nerves supplying these organs with neurosecretions. Additionally, peripheral neurohemal release sites such as the dilator muscle of the antennal circulatory organ and lateral heart nerves were studied, as well as parts of the stomatogastric nervous system. The following neuropeptide families were analyzed: kinins, allatostatins, leucomyosuppressin, corazonin, adipokinetic hormones, myoinhibitory peptide, sulfakinins, periviscerokinins, YLSamide, VEAacid, SKNacid, proctolin, the head peptide, and pyrokinins. Beyond a contribution to a map of the distribution of neuropeptides in a neurohemal system, the following conclusions can be drawn from these experiments. (1) Nearly all abundant peaks in the different mass spectra represent peptides that have already been identified. (2) Although only adult males were used in this study, variations in the peptide abundances were observed that are possibly correlated with different physiological/developmental conditions. (3) Peptides have a body‐region–specific distribution in the neurohemal system. (4) A clear compartmentalisation of the retrocerebral complex could be observed. J. Comp. Neurol. 436:363–375, 2001. © 2001 Wiley‐Liss, Inc.
Linear stability of solitary waves of the Green‐Naghdi equations
Authors: Li Yi A. 
Published: November 2004

AbstractWe investigate the eigenvalue problem obtained from linearizing the Green‐Naghdi equations about solitary wave solutions. Unlike weakly nonlinear water wave models, the physical system considered here has nonlinearity in its highest derivative term. This results in more detailed asymptotic analysis of the eigenvalue problem in the presence of a large parameter. Combining the technique of singular perturbation with the Evans function, we show that for solitary waves of small amplitude, the problem has no eigenvalues of positive real part and the Evans function is nonvanishing everywhere except the origin. This fact then leads to the linear stability of these solitary waves. © 2001 John Wiley & Sons, Inc.
Stability of one‐dimensional boundary layers by using Green's functions
Authors: Grenier Emmanuel , Rousset Frédéric 
Published: November 2004

AbstractThe aim of this paper is to investigate the stability of one‐dimensional boundary layers of parabolic systems as the viscosity goes to 0 in the noncharacteristic case and, more precisely, to prove that spectral stability implies linear and nonlinear stability of approximate solutions. In particular, we replace the smallness condition obtained by the energy method [10, 13] by a weaker spectral condition. © 2001 John Wiley & Sons, Inc.
Infants' long‐term memory for a serial list: Recognition and reactivation
Authors: Gulya Michelle , Galluccio Lissa , Wilk Amy , Rovee‐Collier Carolyn 
Published: November 2004

AbstractSerial lists contain information about item identity and item order. Using a task designed for nonverbal animals, we previously found that 3‐ and 6‐month‐olds exhibited a primacy effect after 24 hr, remembering both item identity and item order. Presently, we examined their memory of list information after longer delays. In Experiment 1, the serial‐position curve reverted to a U‐shape after 1 week at both ages, revealing that the common practice of attributing primacy and recency effects to long‐ and short‐term memory, respectively, is flawed. In Experiment 2, a precuing procedure confirmed that 6‐month‐olds' memory still contained order information after 1 week, but 3‐month‐olds' reactivated memory contained none. Experiments 3A and 3B confirmed that increasing the complexity of information that was learned shortened the delay after which it could be retrieved. Testing infants after delays longer than have previously been used with animals or human adults sheds new light on an old phenomenon. © 2001 John Wiley & Sons, Inc. Dev Psychobiol 38: 174–185, 2001
Estimation of multiple talker locations using randomly positioned microphones (method of subarray selection)
Authors: Kobayashi Kazunori , Hokari Haruhide , Shimada Shoji 
Published: November 2004

AbstractTechnologies for estimating talker location are applicable to teleconference systems with the sense of presence and security systems. Our proposed method estimates multiple talker locations by using synchronous multipliers; it was assumed that the microphones surrounded the talkers. This paper considers the situation in which the talkers surround the microphones. In this arrangement, the main problem is that the capacity for detecting talker location is decreased. Therefore, we propose a new method of subarray selection. Experiments show that the proposed method of subarray selection is effective in estimating talker location. © 2001 Scripta Technica, Electron Comm Jpn Pt 3, 84(9): 42–48, 2001
Rollback recovery for a communication system with two failure states
Authors: Kimura Mitsutaka , Yasui Kazumi , Nakagawa Toshio 
Published: November 2004

AbstractAs computer communication technology has developed, system control mechanisms have been realized by a number of processors, and, hence, it has become necessary to carry out processing distributively and rapidly. Also, as the system mechanisms have become increasingly complicated, the problems of improving reliability and increasing capabilities have become important. In this paper, we consider a control system which is roughly divided into one mark processor and a group of other processors, and formulate a stochastic model by taking notice of only communications between two processors. We perform scheduled checkpointing and execute rollback recovery to restore the consistency of a whole system when two faults of processor failures and communication errors have occurred. The expected cost is obtained and an optimal checkpoint interval which minimizes it is discussed. © 2001 Scripta Technica, Electron Comm Jpn Pt 3, 84(9): 49–55, 2001
The RCRA Toxicity Characteristic Leaching Procedure (TCLP): A Concept for a New Method
Authors: Kimmell Todd A. , Williams Llewellyn R. , Sorini Susan S. 
Published: November 2004

AbstractThe U.S. Environmental Protection Agency has committed to reexamining its use of the Toxicity Characteristic
Leaching Procedure (TCLP). The TCLP was developed to support the Resource Conservation and Recovery
Act (RCRA) Toxicity Characteristic (40 CFR 261.24) and to help predict whether toxic
constituents of solid wastes would be mobilized upon their contact with municipal waste leachate. The method
involves a batch extraction test in which wastes are exposed to an aqueous liquid designed to simulate the
solvent properties of municipal waste leachates. The resulting extract (i.e., TCLP leachate) is
analyzed for the presence of various organic and inorganic contaminants. This article presents a concept for a
new method that addresses a number of critical design criteria. The concept is based on a preliminary method
developed by the authors as part of a work group chaired by the American Society for Testing and Materials. 2001
© John Wiley & Sons, Inc.
Greening the Government and Environmental Management Compliance Using Interactive Web Technologies
Authors: Wowak Kristin , Cleet Christopher 
Published: November 2004

AbstractThe Internet is a powerful tool to inform, communicate, and implement Federal pollution prevention
(P2) programs. In 1996, TRW envisioned the creation of a forum that would enable the tracking of P2
compliance quicker and more accurately than conventional methods allowed. Towards this objective, TRW worked
closely with the Department of Energy's (DOE) Pollution Prevention Team to design and create an
interactive, agency‐wide Web site to track P2 compliance as required by Executive Order (EO)
13101.Interactive Web technologies have direct future applications for a myriad of P2 compliance initiatives,
activities, and areas, including other “Greening the Government” executive orders. Use of
interactive Web technologies can enhance existing information sources by allowing P2 coordinators to track and
report compliance both in real‐time and online, and to provide agency‐specific P2 information and
training. TRW has developed a knowledge base and a set of lessons learned to assist federal P2 managers in
identifying areas of application for enhanced P2 compliance through the use of next‐generation
interactive Web technologies. Copyright TRW Inc. 2001. All rights reserved. TRW has granted permission for
reprint of this article in this journal. No part of this copyrighted work may be reproduced, modified, or
distributed in any form or by any means or stored in any database or retrieval system, without prior written
permission of TRW Inc. © 2001 John Wiley & Sons, Inc.
Making Legitimate Decisions: Scientific and Democratic Ideals in Federal Facility Management
Authors: Mercer Doug 
Published: November 2004

AbstractEnvironmental decision making is conventionally conceived as involving a scientific technical analysis and a
political process of public participation. The ideal of the technical process is that it be an unbiased analysis
of the best available scientific information. The ideal political process would involve all of the affected
stakeholders. The actual decision usually entails a compromise between these two ideals, frustrating decision
makers with high standards for both. This article reports the results of a survey of public attitudes about the
role of science and public participation in decisions at the Idaho National Engineering and Environmental
Laboratory (INEEL), an 890‐square‐mile site managed by the Department of Energy
(DOE). INEEL is cleaning up land contaminated with radioactive and conventional wastes resulting from
historical operations. Its approach to cleanup has been both technically and politically controversial. The
article concludes with the suggestion that decision makers think of technical and political aspects of a
decision as a single process, rather than two processes with different ideals. © 2001 John
Wiley & Sons, Inc.
Coordination of Response Activities with Restoration of Natural‐Resource Injuries under CERCLA at DOD Sites
Authors: Hyatt Diane , Bernhard Teresa S. 
Published: November 2004

AbstractThe Department of Defense (DOD) is proactively acting as a natural resource trustee to address
injuries to natural resources during response under the Comprehensive Environmental Response, Compensation and
Liability Act (CERCLA). Taking the leadership role as lead agent and natural resource trustee improves
environmental protectiveness standards in remedies while realizing more efficient, cost‐effective data
collection to meet interrelated goals. Coordinating timely stakeholder input can promote consistent decisions
from all federal agencies and reduce potential for liabilities resulting from natural‐resource damage
claims from state or tribal trustees. This article also provides a case study of a Maryland facility that is
successfully implementing the principles described herein. The Department of the Navy and stakeholder
participants have successfully utilized this approach for the CERCLA assessment phase to fulfill the
responsibilities of CERCLA response and trusteeship. © 2001 John Wiley & Sons, Inc.
Next Steps in Navigating the DOD Environmental Cleanup Negotiation Process
Authors: Pohlman Teresa 
Published: November 2004

AbstractMany objectives influence the Department of Defense (DOD( environmental cleanup program, including
those from the regulatory, legal, political, public, temporal, economic, technical, and public health
(social) perspectives. Although all of these objectives are integral to the environmental cleanup
process, preliminary research by the author suggests that the two with the greatest influence (accepting
the legal and social as implicitly incorporated in the baseline process) are regulatory and public. These
objectives have grown to greatly influence the DOD environmental program over the past 30 years, as evidenced by
the evolution of the program from being mildly regulated with little public input to being heavily regulated
with increasing public participation and scrutiny. The evolution of the DOD environmental program has resulted
in an urgent need to more effectively and efficiently pursue negotiations with regulatory agencies while dealing
equitably with public participation and influence. This article will attempt to address this need by
(1) presenting the historical context of the DOD environmental program in terms of major pieces of
legislation, (2) examining emerging issues catalyzed by this legislation and by the objectives
influencing the program, and (3) suggesting options for exploring new negotiating approaches and
procedures to address the emerging issues. © 2001 John Wiley & Sons, Inc.