bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tissue-specific expression of estrogen receptors and their role in the regulation of neutrophil infiltration in various organs following trauma-hemorrhage
Authors: Yu Huang-Ping , Shimizu Tomoharu , Hsieh Ya-Ching , Suzuki Takao , Choudhry Mashkoor A , Schwacha Martin G , Chaudry Irshad H 
Published: February 2006

Abstract
Although 17β-estradiol (E2) administration after trauma-hemorrhage (T-H) reduces tissue neutrophil sequestration in male rodents, it remains unknown which of the estrogen receptor (ER) subtypes mediates this effect and whether the same ER subtype is involved in all the tissues. We hypothesized that the salutary effects of E2 on attenuation of neutrophil accumulation following T-H are tissue and receptor subtype-specific. Male Sprague-Dawley rats underwent sham operation or T-H (mean blood pressure, 40 mmHg for 90 min and then resuscitation). E2 (50 μg/kg), ER-α agonist propyl pyrazole triol (PPT; 5 μg/kg), ER-β agonist diarylpropiolnitrile (DPN; 5 μg/kg), or vehicle (10% dimethyl sulfoxide) was administered subcutaneously during resuscitation. Twenty-four hours thereafter, tissue myeloperoxidase (MPO) activity (a marker of neutrophil sequestration), cytokine-induced neutrophil chemoattractant (CINC)-1, CINC-3, and intercellular adhesion molecule (ICAM)-1 levels in the liver, intestine, and lung were measured (n=6 rats/group). ER-α and ER-β mRNA levels in sham-operated rats were also determined. T-H increased MPO activity, CINC-1, CINC-3, and ICAM-1 levels in the liver, intestine, and lung. These parameters were improved significantly in rats receiving E2 after T-H. Administration of the ER-α agonist PPT but not the ER-β agonist DPN improved the measured parameters in the liver. In contrast, DPN but not PPT significantly improved these parameters in the lung. In the intestine, ER subtype specificity was not observed. ER-α mRNA expression was highest in the liver, whereas ER-β mRNA expression was greatest in the lung. Thus, the salutary effects of E2 administration on tissue neutrophil sequestration following T-H are receptor subtype and tissue-specific.
Isolation and Cultivation of Human Testicular Peritubular Cells: A New Model for the Investigation of Fibrotic Processes in the Human Testis and Male Infertility
Authors: Albrecht Martin , Rämsch Romi , Köhn Frank M. , Schwarzer J. Ullrich , Mayerhofer Artur 
Published: February 2006

Context: Fibrotic remodeling, especially of the tubule wall, in testes of infertile men is common, but reasons or consequences of these striking changes are not known. Based on cell culture and ex vivo studies, we previously suggested that mast cells via their products tryptase and histamine are involved in the development of fibrosis. However, studies in a relevant human testicular model are required to further test this hypothesis and the mechanisms of testicular fibrosis in general.Objective: The objective of the study was the isolation, culture, and characterization of adult human testicular peritubular cells.Patients and Interventions: Peritubular cells were obtained from biopsies of men suffering from obstructive azoospermia (n = 8) and varicocele (n = 2) but displaying normal spermatogenesis.Results: Explant cultures were obtained from all biopsies. Immunostaining of the cultured cells and corresponding paraffin-embedded tissues with antibodies against markers of fibroblasts (CD90/Thy-1) and smooth muscle cells (α-smooth muscle actin) clearly proved their origin from the peritubular region. These cells displayed morphological features of myofibroblasts, and gene array analyses as well as immunohistochemistry revealed the predominant expression of extracellular matrix genes and genes coding for basement membrane components. The cultured cells retain receptors for the major mast cell products histamine and tryptase. The addition of histamine (100 μm) and the tryptase agonist peptide SLIGKV (10 μm) resulted in a transient increase in intracellular calcium levels, confirming the functionality of the receptors.Conclusions: We conclude that human peritubular cells are a novel model for the investigation of paracrine, including mast cell initiated, interactions in the human testis, which will allow the study of fibrotic processes underlying male idiopathic infertility.
Seasonal influence of wave action on thread production in Mytilus edulis
Authors: Moeser Gretchen M. , Leba Heather , Carrington Emily 
Published: February 2006

SUMMARY
The blue mussel Mytilus edulis maintains a strong attachment to the substrate in high energy environments by producing byssal threads. On the shores of Rhode Island, USA, mussel attachment strength increases twofold in spring compared to that in the fall. While many factors could influence attachment strength (temperature, food supply, predator cues, etc.), it has been proposed that the variation observed is primarily due to increased thread production during winter and spring in response to increased wave action. This study evaluates the influence of three aspects of wave action on the thread production of M. edulis. Mussels were exposed to flow, acceleration and byssal loading stimuli and the subsequent number of byssal threads produced in the laboratory was monitored. Increased flow elicited the strongest response, significantly decreasing thread production in mussels. This result was confirmed in flume experiments exposing mussels to a range of flows, with reduced thread production above 15 cm s–1. The influence of both acceleration and byssal loading was sporadic and inconsistent across seasons. Surprisingly, overall thread production in the laboratory was lowest in winter, a time when mussels typically peak in attachment. A similar seasonal pattern was observed in field assays, with high thread production during periods of elevated temperature, reduced wave action,and high reproductive condition. These results suggest that seasonal variation in attachment strength does not reflect increased thread production in response to wave action, and that other possible factors, such as seasonal variability in both the material properties of byssal threads and thread decay rates, warrant further investigation.
Nocturnal colour vision – not as rare as we might think
Authors: Kelber Almut , Roth Lina S. V. 
Published: February 2006

SUMMARYThe dual retina of humans and most vertebrates consists of multiple types of cone for colour vision in bright light and one single type of rod, leaving these animals colour-blind at night. Instead of comparing the signals from different spectral types of photoreceptors, they use one highly sensitive receptor, thus improving the signal-to-noise ratio. However, nocturnal moths and geckos can discriminate colours at extremely dim light intensities when humans are colour-blind, by sacrificing spatial and temporal rather than spectral resolution. The advantages of colour vision are just as obvious at night as they are during the day. Colour vision is much more reliable than achromatic contrast, not only under changing light intensities, but also under the colour changes occurring during dusk and dawn. It can be expected that nocturnal animals other than moths and geckos make use of the highly reliable colour signals in dim light.
The Subfornical Organ: A Central Target for Circulating Feeding Signals
Authors: Pulman Katherine J. , Fry W. Mark , Cottrell G. Trevor , Ferguson Alastair V. 
Published: February 2006

The mechanisms through which circulating ghrelin relays hunger signals to the CNS are not yet fully understood. In this study, we have examined the potential role of the subfornical organ (SFO), a circumventricular structure that lacks the normal blood–brain barrier, as a CNS site in which ghrelin acts to influence the hypothalamic centers controlling food intake. We report that ghrelin increased intracellular calcium concentrations in 28% (12 of 43) of dissociated SFO neurons and that the SFO expresses mRNA for the growth hormone secretagogue receptor. Whole-cell patch recordings from SFO neurons demonstrated that in 29% (9 of 31) of neurons tested ghrelin induced a mean depolarization of 7.4 ± 0.69 mV, accompanied by an increase in action potential frequency. Voltage-clamp recordings revealed that ghrelin activates a putative nonselective cationic conductance.Previous reports that the satiety signal amylin exerts similar excitatory effects on SFO neurons led us to examine whether these two peptides influence different subpopulations of SFO neurons. Concentration-dependent depolarizing effects of amylin were observed in 59% (28 of 47) of SFO neurons (mean depolarization, 8.32 ± 0.60 mV). In contrast to ghrelin, voltage-clamp recordings suggest that amylin influences a voltage-dependent current activated at depolarized potentials. We tested single SFO neurons with both peptides and identified cells responsive only to ghrelin (n= 9) and only to amylin (n= 7) but no cells that responded to both peptides. These data support a role for the SFO as a center at which ghrelin and amylin may influence separate subpopulations of neurons to influence the hypothalamic regulation of feeding.
Measurement of the Auditory Brainstem Response (ABR) to Study Auditory Sensitivity in Mice
Authors: Willott James F. 
Published: February 2006

AbstractThe ABR is an electroencephalographic response measured with scalp electrodes. It provides a quick, easy, and reliable method for physiological assessment of auditory sensitivity in mice. A series of brief tone pips or clicks is presented to an anesthetized mouse at a high rate of speed; each click evokes waves of neural activity in the brainstem that are computer‐averaged so they are differentiated from non‐auditory background voltages. The intensity of the clicks is reduced in steps until an ABR can no longer be discerned, thereby defining the ABR threshold, which is closely related to the hearing threshold. Key procedural issues are: (1) accurate calibration of the acoustics (what sounds arrive at the mouse's ear), (2) anesthetization of the mouse, (3) setting up the recording electrodes, (4) the protocol for presenting acoustic stimuli and obtaining thresholds, and (5) interpretation of ABR data.
Overview of Molecular Genetic Diagnosis
Authors: Korf Bruce R. 
Published: February 2006

AbstractThis unit describes the two approaches for molecular genetic diagnosis, direct mutation analysis and linkage studies. Direct mutation analysis is possible if the gene responsible for a disorder has been cloned and techniques exist to detect mutations. Genetic linkage analysis is used when polymorphic markers are available that are closely linked to a disease gene but the gene itself has not been cloned, or when it is difficult or impossible to detect mutations in the gene. For direct mutation analysis, the unit offers a review of the types of mutations that can be detected and the strategies that can be employed for their detection. The unit then describes the types of disorders that are best suited for linkage‐based analysis and offers guidance for interpreting the data.
Yeast as a Model for Human Disease
Authors: Smith Michael G. , Snyder Michael 
Published: February 2006

AbstractThe sequencing of the human genome promised the identification of disease‐causing genes and, subsequently, therapies for those diseases. However, when identifying the genetic basis of a disease, it is not uncommon to discover an abnormal protein whose normal function is unknown. The genetic manipulations required to assign function to genes is often extremely difficult, if not impossible, in human cells. Model organisms have been used to facilitate understanding of gene function because of the ease of genetic manipulations and because many features of eukaryotic physiology have been conserved across phyla. Yeast is a simple eukaryote with a tractable genome, a short generation time, and a large network of researchers who have generated a vast arsenal of research tools. These traits make yeast ideally suited to help reveal the function of genes implicated in human disease.
Detecting Protein–Protein Interactions with CFP‐YFP FRET by Acceptor Photobleaching
Authors: Karpova Tatiana , McNally James G. 
Published: February 2006

AbstractFRET is a light microscopy method for detecting protein‐protein interactions within intact cells. The FRET protocol presented here is for CFP‐ and YFP‐tagged proteins examined with an argon laser on a scanning confocal microscope. FRET is assayed by one of the most straightforward approaches available, namely, acceptor photobleaching. In this procedure, the YFP‐tagged protein (the FRET “acceptor”) is photobleached at a cellular site of interest, and then the intensity of the CFP‐tagged protein (the FRET “donor”) at that same site is measured. In principle, FRET is detected when the CFP intensity increases after the photobleaching of YFP. This unit describes the appropriate steps to perform this measurement, as well as the necessary controls to ensure that an increase in CFP intensity, if detected, in fact reflects bona fide FRET. Successful application of the protocol will support the conclusion that the CFP‐ and YFP‐tagged proteins directly interact at the site of the photobleaching
Multiplexed Microsphere‐Based Flow Cytometric Immunoassays
Authors: Kellar Kathryn L. , Mahmutovic Aida J. , Bandyopadhyay Kakali 
Published: February 2006

AbstractMultiplexed microsphere‐based immunoassays can be developed to simultaneously measure multiple analytes in a biologic system by flow cytometric resolution of spectrally distinct microspheres coupled with capture molecules and reporter fluorochromes bound to detection antibodies. A multiplexed sandwich immunoassay is based on an ELISA format that is transferred directly to microspheres to quantitate multiple antigens. These assays require smaller sample volumes, are less expensive, and are as reproducible, reliable, and sensitive as ELISAs. However, potential cross‐reactivities between multiplexed antibodies, antigens, and specimens need to be systematically eliminated during the validation process. Sandwich and competitive immunoassays, which require only one antigen‐specific antibody, can be combined in the same multiplexed array. Antibody‐capture immunoassays are used to detect multiple antibodies from a specimen for diagnostic or surveillance purposes. The protocols for these three multiplexed immunologic assays are accompanied by methods for coupling analytes to microspheres and biotinylation of antibodies with a water‐soluble derivative.
Detection of Gene Fusions in Acute Leukemia Using Bead Microarrays
Authors: Pihan German 
Published: February 2006

AbstractThis unit describes a method for the rapid and simultaneous detection of hybrid mRNAs (hRNA; also known as gene fusions, RNA fusions, or chimeric RNA), resulting from recurrent chromosome translocations or inversions in acute leukemia. The principal components of the assay are a multiplex RT‐PCR, which simultaneously amplifies any of a number of possible hRNA targets, and a liquid bead microarray (LBMA), which is capable of unambiguously identifying the amplified hRNA. The entire procedure from RNA extraction to display of results can be broken into four steps: (1) conjugated‐microsphere and LBMA manufacture and quality control, (2) cDNA synthesis, (3) multiplex PCR, and (4) hybridization of multiplex PCR products to the hRNA‐detecting LBMA and assay read‐out on a flow cytometer. The assay is designed to detect the most common risk‐stratifying translocation in pediatric acute lymphoblastic leukemia, satisfying the demand for inclusion of genetic data in the diagnosis of acute leukemia as predicated by the current WHO classification of hematopoietic and lymphoid neoplasms.
Mathematical modeling of tin‐free chemically‐active antifouling paint behavior
Authors: Yebra Diego Meseguer , Kiil Søren , Dam‐Johansen Kim , Weinell Claus E. 
Published: February 2006

AbstractMathematical modeling has been used to characterize and validate the working mechanisms of tin‐free, chemically‐active antifouling (AF) paints. The model‐based analysis of performance data from lab‐scale rotary experiments has shown significant differences between antifouling technologies as regards the biocide leaching and the surface polishing processes. Hence, the modeling framework developed in this work is built so as to describe any generic, chemically‐active AF paint through model parameters, the values of which can be obtained or adjusted from relatively fast measurements. The detailed quantitative information on reacting AF paint systems obtained can be used for accelerated product optimization purposes, thus facilitating the transition to cleaner antifouling technologies using, for example, fast‐degrading natural or synthetic bioactive components. © 2006 American Institute of Chemical Engineers AIChE J, 2006
Guest Encapsulation and Self‐Assembly of a Cavitand‐Based Coordination Capsule
Authors: Haino Takeharu , Kobayashi Mutsumi , Fukazawa Yoshimasa 
Published: February 2006

AbstractThe synthesis and spectroscopic characterization of a cavitand‐based coordination capsule 1⋅4 BF4 of nanometer dimensions is described. Encapsulation studies of large aromatic guests as well as aliphatic guests were performed by using 1H NMR spectroscopy in [D1]chloroform. In addition to the computational analysis of the shape and geometry of the capsule, an experimental approach to estimate the interior size of the cavity is discussed. The cavity provides a highly rigid binding space in which molecules with lengths of approximately 14 Å can be selectively accommodated. The rigid cavity distinguished slight structural differences in the flexible alkyl‐chain guests as well as the rigid aromatic guests. The detailed thermodynamic studies revealed that not only CH–π interactions between the methyl groups on the guest termini and the aromatic cavity walls, but also desolvation of the inner cavity play a key role in the guest encapsulation. The cavity preferentially selected the hydrogen‐bonded heterodimers of a mixture of two or three carboxylic acids 18–20. The chiral capsule encapsulated a chiral guest to show diastereoselection.
A flow cytometric screening test for detergent‐resistant surface antigens in monocytes
Authors: Wolf Zsuzsanna , Orsó Evelyn , Werner Tobias , Boettcher Alfred , Schmitz Gerd 
Published: February 2006

AbstractRafts resemble cholesterol‐ and glycosphingolipid‐enriched, liquid‐ordered plasma membrane microdomains, showing resistance to nonionic detergents, and are involved in various cellular processes. In the present study, we have tested surface antigens on resting and lipopolysaccharide (LPS)‐stimulated human peripheral blood monocytes for their detergent resistance (i.e. raft‐association), by flow cytometry. Constitutive (CD14, CD32, CD55), or LPS‐induced (CD81) raft‐association, and detergent solubility (i.e. exclusion of rafts) (CD71) of monocyte antigens in the presence of 0.01% Triton X‐100 are clearly demonstrated. Flow cytometric detergent insolubility is a powerful tool for rapid screening the raft‐association of monocyte antigens in a whole‐blood assay. © 2006 International Society for Analytical Cytology
Divergent effects of IL‐12 and IL‐23 on the production of IL‐17 by human T cells
Authors: Hoeve Marieke A. , Savage Nigel D. L. , de Boer Tjitske , Langenberg Dennis M. L. , de Waal Malefyt René , Ottenhoff Tom H. M. , Verreck Frank A. W. 
Published: February 2006

AbstractIL‐23 is regarded as a major pro‐inflammatory mediator in autoimmune disease, a role which until recently was ascribed to its related cytokine IL‐12. IL‐23, an IL‐12p40/p19 heterodimeric protein, binds to IL‐12Rβ1/IL‐23R receptor complexes. Mice deficient for p19, p40 or IL‐12Rβ1 are resistant to experimental autoimmune encephalomyelitis or collagen‐induced arthritis. Paradoxically, however, IL‐12Rβ2‐ and IL‐12p35‐deficient mice show remarkable increases in disease susceptibility, suggesting divergent roles of IL‐23 and IL‐12 in modulating inflammatory processes. IL‐23 induces IL‐17, which mediates inflammation and tissue remodeling, but the role of IL‐12 in this respect remains unidentified. We investigated the roles of exogenous (recombinant) and endogenous (macrophage‐derived) IL‐12 and IL‐23, on IL‐17‐induction in human T‐cells. IL‐23 enhanced IL‐17 secretion, as did IL‐2, IL‐15, IL‐18 and IL‐21. In contrast, IL‐12 mediated specific inhibition of IL‐17 production. These data support the role of IL‐23 in inflammation through stimulating IL‐17 production by T lymphocytes, and importantly indicate a novel regulatory function for IL‐12 by specifically suppressing IL‐17 secretion. These data therefore extend previous reports that had indicated unique functions for IL‐23 and IL‐12 due to distinct receptor expression and signal transduction complexes, and provide novel insights into the regulation of immunity, inflammation and immunopathology.
Critical role for c‐kit (CD117) in T cell lineage commitment and early thymocyte development in vitro
Authors: Massa Steffen , Balciunaite Gina , Ceredig Rhodri , Rolink Antonius G. 
Published: February 2006

AbstractThe precise roles played by the transmembrane receptor tyrosine kinase c‐kit and its ligand stem cell factor in early T cell development are difficult to study. Using cloned Pax5‐deficient progenitor B cells, we show that following Notch signaling, which induces their commitment to the T cell developmental pathway, c‐kit expression is rapidly up‐regulated at both the transcriptional and cell surface level. Using either an anti‐c‐kit monoclonal antibody or Gleevec, a pharmacological inhibitor of c‐kit signaling, we show that the Notch‐induced T cell differentiation of either Pax5‐deficient progenitor B cells, or the equivalent cell from the bone marrow of normal mice, is strictly dependent on c‐kit signaling, whereas the differentiation of normal progenitors into the B cell lineage is not. Moreover, we show that the Notch and IL‐7 signaling‐induced proliferation and differentiation of CD44+CD25–c‐kithigh and CD44+CD25+c‐kithigh thymocytes along the T cell, but not natural killer cell or macrophage, pathway also requires c‐kit signaling, whereas the Notch‐induced proliferation and differentiation of CD44–CD25+c‐kitint cells along the T cell pathway is independent of c‐kit. These results further highlight the complex inter‐relationships existing between c‐kit, Notch and IL‐7 receptor signaling that control the proliferation and differentiation of early T cell progenitors.See accompanying commentary: http://dx.doi.org/10.1002/eji.200635950
Phase Relations Between β‐Tricalcium Phosphate and Hydroxyapatite with Manganese(II): Structural and Spectroscopic Properties
Authors: Mayer Isaac , Cuisinier Frederic J. G. , Popov Ina , Schleich Yechezkel , Gdalya Sarit , Burghaus Olaf , Reinen Dirk 
Published: February 2006

AbstractThe preparation of Mn‐containing β‐tricalcium phosphate (β‐TCP) samples was achieved in two ways: a) transformation of precipitated Mn‐containing calcium hydroxyapatite (HA) to β‐TCP by heating at 1100 °C, and b) preparation by solid‐state reaction of a mixture of CaCO3, (NH4)2HPO4, and Mn(NO3)2 at 1100 °C. Powder X‐ray diffraction (XRD) analyses of the samples, obtained by both methods, show well‐defined patterns with structural data of the rhombohedral R3c, β‐TCP phase. The calculated lattice constants are smaller than those known for β‐Ca3(PO4)2 because of substitution of Ca2+ by Mn2+. EPR spectroscopy indeed reveals that manganese is divalent in the samples. Apparently, the Ca(5) site in the β‐TCP structure is occupied by Mn2+. The distribution of Mn2+ between the β‐TCP and the HA phase in the case of preparation (b) was studied by EPR spectroscopy, and a pronounced preference for the former lattice was found. Micron‐ and submicron‐sized crystals with visible faces were observed by TEM in the case of β‐TCP prepared by solid‐state reaction, and large micron‐sized, droplike‐shaped crystals, sensitive to beam radiation, were found in the case of samples prepared by heating HA at elevated temperatures. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006)