bluuebunny/crossref_metadata_2025 huggingface dataset:
Accumulation of hexahydro-1,3,5-trinitro-1,3,5-triazine in channel catfish (ictalurus punctatus) and aquatic oligochaetes (lumbriculus variegatus)
Authors: Belden Jason B. , Lotufo Guilherme R. , Lydy Michael J.
Published: July 2005

The extensively used military explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) has been widely released to the environment during production, usage, and disposal operations. Toxic effects of RDX have been reported in terrestrial and aquatic receptors, but investigations regarding the bioaccumulation potential of RDX in aquatic systems are scarce. The objective of the present study was to describe the toxicokinetics of RDX during aqueous exposure for the channel catfish (Ictalurus punctatus) and aquatic oligochaetes (Lumbriculus variegatus) and to compare the amount of RDX accumulation in juvenile catfish following aqueous exposure only, dietary exposure only, and a combination of dietary and aqueous exposure. The toxicokinetics measurements included bioconcentration factors (BCFs), uptake rates, elimination rates, and biological half-lives. First-order, single-compartment models described the toxicokinetics for both species. Uptake of RDX into oligochaetes was relatively rapid (uptake clearance constant [ku] of 5.17 ml/g/h) compared to that in catfish (ku = 1.28 ml/g/h). However, elimination also was more rapid in oligochaetes, with biological half-lives of 0.28 and 1.09 h for oligochaetes and catfish, respectively. Thus, both species had very similar estimated BCFs of 2.1 ml/g for oligochaetes and 2.0 ml/g for catfish. Accumulation of RDX in fish that were fed oligochaetes exposed to an exceedingly high water concentration of RDX was minimal. The present investigation indicates that RDX uptake via the aqueous route is the expected dominant uptake pathway, with dietary uptake contributing minimally to the overall body burden in fish inhabiting RDX-contaminated sites. Because of the exceedingly low bioaccumulative potential and low reported toxicity of RDX, the presence of this explosive in aquatic systems is unlikely to pose unacceptable risks to invertebrates and fish.
Effect of soil properties and aging on the toxicity of copper for enchytraeus albidus, enchytraeus luxuriosus, and folsomia candida
Authors: de Barros Amorim Mónica João , Römbke Jorg , Schallnaß Hans-Joachim , Soares Amadeu Mortágua Velho Maia
Published: July 2005

In the present study, the effect of the heavy-metal salt copper chloride (CuCl2-2H2O) in soils freshly spiked (3 d) and aged (70 ± 10 d; mean ± SD) was studied in the test species Enchytraeus albidus, E. luxuriosus, and Folsomia Candida. Up to nine soils were used: Besides the Organisation for Economic Co-operation and Development (OECD) artificial soil and the Agricultural Testing and Research Agency (Landwirtschaftliche Untersuchungs- und Forschungsanstalt, Speyer, Germany) 2.2 natural standard soils, the others were selected based on the EURO Soil approach, taking into account the effect of different soil parameters (pH, organic matter, grain size distribution, and carbon to nitrogen ratio). Additionally, the effect of the chloride ions was studied separately. The results revealed the following: First, a soil effect was observed; for example, in F. Candida, median effective concentrations (EC50s) varied between 262 mg/kg in a sample from the same site as the original EURO Soil 5 soil and greater than 1,000 mg/kg in OECD soil. Second, an aging effect was observed, mainly in F. Candida. For example, toxicity of offspring survival was increased twofold in the OECD soil and approximately eightfold with aging in the EURO Soil 7 soil, whereas the enchytraeid species did not react differently after aging. Third, an effect of chloride ions on reproduction of the animals was found; however, this effect was independent of the aging period. Fourth, species variation was seen in terms of sensitivity (EC50), decreasing in the following order: E. luxuriosus > E. albidus » F. Candida. Differences in toxicity of offspring survival between enchytraeids and F. Candida might be explained by the different routes of uptake.
Metamorphosis of two amphibian species after chronic cadmium exposure in outdoor aquatic mesocosms
Authors: James Stacy M. , Little Edward E. , Semlitsch Raymond D.
Published: July 2005

Amphibian larvae at contaminated sites may experience an alteration of metamorphic traits and survival compared to amphibians in uncontaminated conditions. Effects of chronic cadmium (Cd) exposure on the metamorphosis of American toads (Bufo americanus) and southern leopard frogs (Rana sphenocephala) were determined. The two species were reared separately from shortly after hatching through metamorphosis in outdoor mesocosms (1,325-L polyethylene cattle tanks) that simulated natural ponds and enhanced environmental realism relative to the laboratory. Both species exhibited a decrease in survival with increasing initial nominal aqueous Cd concentration. Cadmium treatment did not influence mass at metamorphosis for either species when survival was included as a covariate, but increased the age at metamorphosis for the American toads. The whole body Cd content of metamorphs increased with aqueous Cd treatment level for both species, and the American toads tended to possess more elevated residues. Cadmium quickly partitioned out of the water column and accumulated in and altered the abundance of the tadpoles' diet. Cadmium-contaminated sites may produce fewer metamorphs, and those that survive will metamorphose later and contain Cd. Interspecific differences in the response variables illustrate the importance of testing multiple species when assessing risk.
Partition Theorems and Computability Theory
Authors: Mileti Joseph R.
Published: July 2005

The connections between mathematical logic and combinatorics have a rich history. This paper focuses on one aspect of this relationship: understanding the strength, measured using the tools of computability theory and reverse mathematics, of various partition theorems. To set the stage, recall two of the most fundamental combinatorial principles, König's Lemma and Ramsey's Theorem. We denote the set of natural numbers by ω and the set of finite sequences of natural numbers by ω<ω. We also identify each n ∈ ω with its set of predecessors, so n = {0, 1, 2, …, n − 1}.
Strong compactness and stationary sets
Authors: Krueger John
Published: July 2005

We construct a model in which there is a strongly compact cardinal κ such thai the set S(κ, κ+) ={ a Є Pκκ+: o.t.(a) = (a⋂ κ)+}is non-stationary.
On constructing completions
Authors: Crosilla Laura , Ishihara Hajime , Schuster Peter
Published: July 2005

The Dedekind cuts in an ordered set form a set in the sense of constructive Zermelo–Fraenkel set theory. We deduce this statement from the principle of refinement, which we distill before from the axiom of fullness. Together with exponentiation, refinement is equivalent to fullness. None of the defining properties of an ordering is needed, and only refinement for two–element coverings is used.

In particular, the Dedekind reals form a set: whence we have also refined an earlier result by Aczel and Rathjen, who invoked the full form of fullness. To further generalise this, we look at Richman's method to complete an arbitrary metric space without sequences, which he designed to avoid countable choice. The completion of a separable metric space turns out to be a set even if the original space is a proper class: in particular, every complete separable metric space automatically is a set.
Numerical study on the underground coal gasification for inclined seams
Authors: Yang Lanhe
Published: July 2005

According to the characteristics for combustion and gasification reactions occurring in the gasification gallery, the mathematical functional relationship between the chemical reaction rate and every influencing factor is studied. The dynamic nonlinear coupling mathematical models on underground coal gasification of inclined seams are established. The determination methods of major model parameters are introduced. Additionally, the control volume method is adopted to find the numerical solution to the mathematical models. The patterns of development and variation for temperature field, concentration field and pressure field in gasification panel are studied. On the basis of the model test, calculation results are analyzed. From the distribution of temperature field, its calculation value is a little higher than the experimental one, with the relative error of every measuring point virtually within 17%. Research shows that, the experiment value of gas heat value and calculated value take on a good conformity; due to the influence of temperature, in the high temperature zone, the change gradient of the experiment value for concentration field of gas compositions is greater than that of the calculation value. The simulated results indicate that the relative error of the pressure field calculation is 4.13%–12.69% and 8.25%–17.47%, respectively, 7 h and 45 h after the ignition. The drop rate for the fluid pressure is 6.01% and 10.91%, respectively. Research shows that the simulated values conform with experimental values comparatively well, which demonstrates that the numerical simulation on the “three fields” in underground coal gasification is correct. © 2005 American Institute of Chemical Engineers AIChE J, 2005
Enrichment using antibody‐coated microfluidic chambers in shear flow: Model mixtures of human lymphocytes
Authors: Sin Aaron , Murthy Shashi K. , Revzin Alexander , Tompkins Ronald G. , Toner Mehmet
Published: July 2005

Isolation of phenotypically‐pure cell subpopulations from heterogeneous cell mixtures such as blood is a difficult yet fundamentally important task. Current techniques such as fluorescent activated cell sorting (FACS) and magnetic‐activated cell sorting (MACS) require pre‐incubation with antibodies which lead to processing times of at least 15–60 min. In this study, we explored the use of antibody‐coated microfluidic chambers to negative deplete undesired cell types, thus obtaining an enriched cell subpopulation at the outlet. We used human lymphocyte cell lines, MOLT‐3 and Raji, as a model system to examine the dynamic cell binding behavior on antibody coated surfaces under shear flow. Shear stress ranging between 0.75 and 1.0 dyn/cm2 was found to provide most efficient separation. Cell adhesion was shown to follow pseudo‐first order kinetics, and an anti‐CD19 coated (Raji‐depletion) device with ∼2.6 min residence time was demonstrated to produce 100% pure MOLT‐3 cells from 50‐50 MOLT‐3/Raji mixture. We have developed a mathematical model of the separation device based on the experimentally determined kinetic parameters that can be extended to design future separation modules for other cell mixtures. We conclude that we can design microfluidic devices that exploits the kinetics of dynamic cell adhesion to antibody coated surfaces to provide enriched cell subpopulations within minutes of total processing time. © 2005 Wiley Periodicals, Inc.
Continuous operation of foamed emulsion bioreactors treating toluene vapors
Authors: Kan Eunsung , Deshusses Marc A.
Published: July 2005

Continuous operation of a new bioreactor for air pollution control called the foamed emulsion bioreactor (FEBR) has been investigated. The effect of several liquid feeding strategies was explored. The FEBR exhibited high and steady toluene removal performance (removal efficiency of 89%–94%, elimination capacity of 214–226 g/m3h at toluene inlet concentration of 1 g/m3) for up to 360 h, when 20% of the culture was replaced every 24 h by a nutrient solution containing 4 g/L of potassium nitrate as a nitrogen source. This feeding mode supported a high cell activity measured as INT reduction potential and active cell growth without being subject to nitrogen limitation. In comparison, operating the FEBR with the liquid in a closed loop (i.e., batch) resulted in a significant decrease of both the removal efficiency of toluene and INT reduction activity. Operation with feeding active cells resulted in stable and effective treatment, but would require a significant effort for mass culture preparation. Therefore, the continuous process with periodically feeding nutrients was found to be the most practical and effective operating mode. It also allows for stable operation, as was shown during removal of low concentration of toluene or after pollutant starvation. Throughout the study, INT reduction measurements provided insight into the process. INT reduction activity data proved that under normal operating conditions, the FEBR performance was limited by both the kinetics and by mass transfer. Overall, the results illustrate that engineered gas‐phase bioreactors can potentially be more effective than conventional biofilters and biotrickling filters for the treatment of air pollutants such as toluene. © 2005 Wiley Periodicals, Inc.
Are Hormones from the Neuropeptide Y Family Recognized by Their Receptors from the Membrane‐Bound State?
Authors: Bader Reto , Zerbe Oliver
Published: July 2005

Hormones and many other neurotransmitters, growth factors, odorant molecules, and light all present stimuli for a class of membrane‐anchored receptors called G protein‐coupled receptors (GPCRs). The GPCRs are the largest family of cell‐surface receptors involved in signal transduction. About 1 % of all known genes of Drosophila and more than 5 % of the genes of Caenorhabditis elegans encode GPCRs. In addition, more than 50 % of current therapeutic agents on the market target these receptors. When the enormous biological and pharmaceutical importance of these receptors is considered, it is surprising how little is known about the mechanism with which these receptors recognize their natural ligands. In this review we present a structural approach, utilizing techniques of high‐resolution NMR spectroscopy, to address the question of whether peptides from the neuropeptide Y family of neurohormones are recognized directly from solution or from the membrane‐bound state. In our studies we discovered that the structures of the membrane‐bound species are better correlated to the pharmacological properties of these peptides than the solution structures are. These findings are supported by the observation that many biophysical properties of these peptides seem to be optimized for membrane binding. We finally present a scenario of possible events during receptor recognition.
Probing the Binding Entropy of Ligand–Protein Interactions by NMR
Authors: Homans Steve W.
Published: July 2005

The importance of being dynamic. Protein–ligand affinities are governed not only by structural complementarity, but also by the dynamics of the interacting species, including the solvent. NMR‐relaxation methods offer important clues to understanding these recognition processes by providing per‐residue thermodynamics parameters.
Functional expression of the peptide transporter PEPT2 in the mammalian enteric nervous system
Authors: Rühl Anne , Hoppe Susanne , Frey Isabelle , Daniel Hannelore , Schemann Michael
Published: July 2005

The peptide transporter PEPT2 mediates transmembrane uptake of small peptides. So far, its expression has not been evidenced in the gastrointestinal tract. We have investigated peptide transport activity in the neuromuscular layers of the gastrointestinal tract by using the fluorescent tracer‐dipeptide β‐Ala‐Lys‐Nϵ‐7‐amino‐4‐methyl‐coumarin‐3‐acetic acid (Ala‐Lys‐AMCA). Whole‐mount preparations from mouse, rat, and guinea pig stomach and small and large intestine were incubated with Ala‐Lys‐AMCA in the presence or absence of the uptake‐inhibitors L‐histidine, D‐phenylalanyl‐L‐alanine (D‐Phe‐Ala), glycyl‐L‐sarcosine (Gly‐Sar), glycyl‐L‐glutamine (Gly‐Gln), benzylpenicillin, and cefadroxil. Fluorescence microscopy revealed that Ala‐Lys‐AMCA specifically accumulated in both ganglionic layers of the enteric nervous system (ENS) in all regions and species studied. This could be inhibited by Gly‐Sar, D‐Phe‐Ala, Gly‐Gln, and cefadroxil, but not by free histidine and benzylpenicillin, indicating uptake via PEPT2. Accordingly, dipeptide uptake was completely abolished in PEPT2‐deficient mice. Reverse transcriptase‐polymerase chain reaction analysis detected a PEPT2‐specific transcript in extracts from the ganglionic ENS layers of mouse small and large intestine, further proving that enteric dipeptide transport activity is specifically mediated via PEPT2. The cellular site of dipeptide uptake was immunohistochemically localized to enteric glial cells and tissue‐resident macrophages. In addition, dipeptide uptake occurred in a neurochemically defined subset of neurons in the guinea pig ENS. Our results constitute the first functional evidence for dipeptide transport activity in the ENS. PEPT2‐mediated dipeptide transport in enteric glia could contribute to the clearance of neuropeptides in the ENS. In addition, the fluorophore‐coupled dipeptide uptake via PEPT2 is a novel vital marker for glial cells in the ENS. J. Comp. Neurol. 490:1–11, 2005. © 2005 Wiley‐Liss, Inc.
Definition and novel connections of the entopallium in the pigeon (Columba livia)
Authors: Krützfeldt Nils O.E. , Wild J. Martin
Published: July 2005

The avian entopallium (E) is the major thalamorecipient zone, within the telencephalon, of the tectofugal visual system. Because of discrepancies concerning the structure of this nuclear mass in pigeons, and in light of recent evidence concerning entopallial projections in other avian species, we here redefine and chart some novel entopallial projections in the pigeon by using a combination of cytochrome oxidase (CO) activity, calcium binding protein immunohistochemistry (CBPi), normal histology, and tract tracing. We show that 1) E is defined by the accurate overlap of CO activity and the dense terminations of thalamic (rotundal) efferents; 2) the perientopallium (Ep), E's overlying belt region, receives a relatively sparse rotundal input and is a major source of projections to wider regions of the hemisphere; and 3) E can be subdivided into internal (Ei) and external (Ex) portions on the basis of normal histology, CBPi, and differential projections. Thus, Ei, but not Ex, makes a reciprocal connection with a distinct nucleus in the ventrolateral mesopallium and is a major source of projections to the lateral striatum. These findings suggest the necessity for a revision of the original proposal of a strictly serial flow of visual information through the entopallial complex and further regions of the hemisphere and also require a modification of the long‐standing view that E is comparable to only one specific lamina (IV) of extrastriate visual cortex of mammals (Karten and Hodos [1970] J. Comp. Neurol. 140:35–51; Shimizu and Karten [1990] In: Finlay BL, et al., editors. The neocortex. New York: Plenum Press. p 75–86). Rather, E appears to be composed of a variety of neuronal types possibly equivalent to those in several neocortical laminae. J. Comp. Neurol. 490:40–56, 2005. © 2005 Wiley‐Liss, Inc.
Development of putative GABAergic neurons in the appendicularian urochordateOikopleura dioica
Authors: Søviknes Anne Mette , Chourrout Daniel , Glover Joel C.
Published: July 2005

Studying the developing brain of urochordates can increase our understanding of brain evolution in the chordate lineage. To begin addressing regional patterns of neuronal differentiation in appendicularian urochordates, we examined the development of putative GABAergic neurons inOikopleura dioicausing GABA immunohistochemistry and in situ hybridization for the GABA‐synthesizing enzyme GAD. First, we assessed the developmental dynamics of neuron number and organization in the cerebral and caudal ganglia. We then identified and mapped the positions of putative GABAergic neurons using confocal microscopy. We found GAD mRNA‐positive and GABA‐immunopositive neurons in the first brain nerves and the cerebral and caudal ganglia, but not in the caudal nerve cord. In both ganglia GAD mRNA‐positive and GABA‐immunopositive neurons are found in the same characteristic intraganglionic locations. The differentiation of these GABAergic markers occurs first in the first brain nerves and the cerebral ganglion and then with a several‐hour delay in the caudal ganglion. In all three structures GAD mRNA expression appears 2–3 hours prior to GABA expression. In general, GABA is expressed by the same number of neurons as express GAD. Several discrepancies suggest differential regulation of the GABAergic phenotype in different neurons, however. Our results show that the GABAergic phenotype has a stereotyped pattern of expression along the anteroposterior axis of the CNS. Given recent genome sequencing and developmental patterning gene studies in this species, the GABAergic neurons inO. dioicaprovide a good model for assessing, at the invertebrate–vertebrate transition, the molecular mechanisms that specify the GABAergic phenotype. J. Comp. Neurol. 490:12–28, 2005. © 2005 Wiley‐Liss, Inc.
Modified Hadamard transform microchip electrophoresis
Authors: Guchardi Renato , Schwarz Maria A.
Published: July 2005

Sensitivity is a crucial point in the development applications for medicine or environmental samples in which the analytes are present in the nanomolar range. Besides further technical development of detection systems, the multiplex sample injection technique can be applied for enhancing the signal‐to‐noise ratio. Hadamard transform is easily applied to microchip electrophoresis due to the fact that sample injection is generally achieved through cross, double‐tee, or tee injector structures. This paper reports the first demonstration of a modified Hadamard transform electrophoresis on a microchip by using an amperometric detector. Contrary to the previous Hadamard applications, the resolution (number of points per unit of time) of electropherograms obtained is independent of the number of injections.
Nanoparticle‐filled capillary electrophoresis for the separation of long DNA molecules in the presence of hydrodynamic and electrokinetic forces
Authors: Tseng Wei‐Lung , Huang Ming‐Feng , Huang Yu‐Fen , Chang Huan‐Tsung
Published: July 2005

We report the analysis of long DNA molecules by nanoparticle‐filled capillary electrophoresis (NFCE) under the influences of hydrodynamic and electrokinetic forces. The gold nanoparticle (GNP)/polymer composites (GNPPs) prepared from GNPs and poly(ethylene oxide) were filled in a capillary to act as separation matrices for DNA separation. The separations of λ‐DNA (0.12–23.1 kbp) and high‐molecular‐weight DNA markers (8.27–48.5 kbp) by NFCE, under an electric field of —140 V/cm and a hydrodynamic flow velocity of 554 μm/s, were accomplished within 5 min. To further investigate the separation mechanism, the migration of λ‐DNA was monitored in real time using a charge‐coupled device (CCD) imaging system. The GNPPs provide greater retardation than do conventional polymer media when they are encountered during the electrophoretic process. The presence of interactions between the GNPPs and the DNA molecules is further supported by the fluorescence quenching of prelabeled λ‐DNA, which occurs through an energy transfer mechanism. Based on the results presented in this study, we suggest that the electric field, hydrodynamic flow, and GNPP concentration are the three main determinants of DNA separation in NFCE.
Peak shape modeling by Haarhoff‐Van der Linde function for the determination of correct migration times: A new insight into affinity capillary electrophoresis
Authors: Le Saux Thomas , Varenne Anne , Gareil Pierre
Published: July 2005

Among the different experimental strategies available in capillary electrophoresis (CE) to determine binding parameters, affinity capillary electrophoresis (ACE) has been the most widely embraced due to its easiness of implementation and of data handling. Ligand‐substrate binding constants are thus directly derived from the substrate migration time shifts resulting from the variation of ligand concentration introduced in a background electrolyte. Classically, the substrate migration time is measured on top of the electrophoretic peak, assuming symmetrical peak shape. Depending on both substrate and ligand concentrations that may be required to meet detection sensitivity or complexation conditions, zonal migrations in ACE may, however, produce triangular peak shape, most often due to pronounced electromigration dispersion (EMD), and this may result in positively or negatively erroneous migration time assessments. In this work, EMD distorted triangular peak shapes obtained in the course of host‐guest complexation studies were fitted with the Haarhoff‐Van der Linde function, allowing better estimation of migration time. The model systems studied were those of β‐cyclodextrin and naproxen, 2‐naphthalenesulfonate, or 1‐adamantanecarboxylate. The impact of this correction on binding isotherms and binding constant evaluation was exemplified. Furthermore, in situations where the substrate concentration injected by far overtakes that of the ligand in the electrolyte, the interest in this peak shape correction was discussed in connection with the question of whether the free ligand concentration can be still considered equal to the ligand concentration introduced, a question that still remains under debate nowadays.
Retention behavior of proteins in size‐exclusion electrochromatography with a low‐voltage electric field perpendicular to the liquid phase streamline
Authors: Tan Guomin , Shi Qinghong , Sun Yan
Published: July 2005

A novel preparative size‐exclusion electrochromatography with an oscillatory low‐voltage electric field perpendicular to the liquid phase streamline (pSEEC) was proposed with a column design of rectangular cross‐section. The column of 12 cm length was packed with Sephadex G‐75, and the retention behavior of bovine serum albumin (BSA) and myoglobin (Myo) was extensively investigated under various conditions. The results indicated that the partition coefficient of a charged protein increased significantly on increasing the current strength as well as the difference between its pI and pH. The partition coefficient also increased on decreasing the mobile phase conductivity. For the gel‐excluded protein like BSA, the concentration polarization (CP) on the gel surface induced by the protein electromigration was the main reason for the increased retention. For a gel‐permeable protein like Myo, both the CP and electrophoretic migration in the solid phase contributed to its increased retention. Further results exhibited that the polarization would be offset by diffusion, because the accumulated protein would flux back to the bulk liquid phase. Therefore, when the electrically induced mass flux was equal to the diffusion flux, the partition coefficient did not change with a further increase of the oscillatory current cycle. Finally, pSEEC was compared with SEC in the separation of protein mixtures of BSA/Myo as well as BSA/Myo/lysozyme. The results showed much better resolutions of the protein mixtures in pSEEC with the column as short as 12 cm. The potential of pSEEC for preparative protein separation was therefore demonstrated.
Zygospore formation of desmids from Kollam district, Kerala (India) 1. Closterium
Authors: Panikkar M. V. N. , Krishnan Sreeja
Published: July 2005

The present paper deals with the zygospore formation in ten taxa of the genus Closterium: Cl. calosporum Wittr., Cl. dianae Ehrenb., Cl. dianae var. brevius (Wittr.) Petkoff, Cl. intermedium Ralfs, Cl. kuetzingii Bréb., Cl. malmei Borge f. minor Thérézien, Cl. nematodes Joshua, Cl. ralfsii Bréb., Cl. setaceum Ehrenb. and Cl. striolatum Ehrenb., collected from different parts of the Kollam district of Kerala, India. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Sexual or clonal origin? A morpho‐ecological and molecular analysis in a patch of Ajuga reptans L. (Lamiaceae)
Authors: Pfeiffer T.
Published: July 2005

Spatial clonal structure and patch colonisation in Ajuga reptans L. (Lamiaceae), a common stoloniferous semi‐rosette plant, were studied using a combined morpho‐ecological and molecular genetic approach. Within a natural patch from a forest near Diedorf (Thuringia, Germany), the spatial clonal structure was analysed, correlating both datasets: Morphologically, characteristics of clonal growth and clonal reproduction were studied, the spatial distribution of modules was mapped and merigenet relationships were reconstructed.

Samples from the patch and its surroundings, and an additional sample from Berlin were then analysed by AFLP fingerprinting using four different primer combinations to identify genets. Most divergence in banding patterns was already obtained for samples from the Diedorf forest. Within the patch, however, most samples had very similar fingerprints, indicating their belonging to the same genet and hence a clonal origin, although they are morphologically separated into three “plants”. Based on AFLP data, the relationships of one sample remained ambiguous; but the correlation with morphological data helped to interpret the pattern and indicated that the sample is probably a dividual of the clone, too. The relevance of the observed vegetative multiplication (clonal growth and subsequent clonal reproduction) for patch colonisation and maintenance are discussed. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Karpologische Untersuchung der Axyris‐Arten (Chenopodiaceae) im Zusammenhang mit ihrer Diagnostik und Taxonomie
Authors: Suchorukow A. P.
Published: July 2005

Vergleichend karpologische Untersuchungen an Arten der Gattung Axyris wurden durchgeführt. Die Früchte aller betrachteten Sippen erscheinen dimorph. Die morphologischen Merkmale der Früchte sind artspezifisch; sie werden in einem originalen diagnostischen Schlüssel zusammengefasst. Die anatomische Struktur eines jeden Fruchttyps variiert nicht innerhalb einer Art. Die flügelartigen Anhängsel werden von Perikarpzellen gebildet, die von darunterliegenden Schichten im apikalen Teil der Frucht ausgegliedert werden. Die Anhängsel entstehen aus nur einer Perikarpschicht bei den Früchten des einen Typs und aus der äußeren Schicht bei den Früchten des zweiten Typs. Eine kurze taxonomische Arten‐Liste wird beigefügt und, sofern erforderlich, wurde eine Lectotypisierung vorgenommen. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Echium boissieriSteud. (Boraginaceae), an Ibero‐Mauretanian roadside plant
Authors: Müller J. V. , Deil U. , Galán de Mera A. , Vicente Orellana J. A.
Published: July 2005

Roadsides are frequently colonized by eurychorous species with a wide ecological and coenological amplitude. Stenoic and stenochorous plant species are often missing in synanthropic vegetation. An exception is Echium boissieri Steud., occurring in Central and Southern Spain, Portugal, Northern Morocco and Algeria. To determine whether this species is a “homeless roadside plant”, we analysed herbarium specimens, performed a detailed mapping of current populations on both sides of the Straits of Gibraltar, sampled phytosociological relevés and gathered the available coenological data from literature.

Echium boissieri is a hapaxanthous perennial herb and belongs to section Eleuterolepis. The analysis of herbarium specimens indicates a recent apophytization that implies a trend from primary (rocks, screes, landslides) and secondary habitats (extensive pastures, fallow lands) to roadsides during the last two centuries, particularly in Spain. This habitat change coincides with land‐use changes. The Chamaeleo gummiferi‐Echietum boissieri is described as a new association. This is a multi‐layered herbaceous vegetation type with an enormous species‐ and lifeform‐diversity. It is characterized by E. boissieri and Chamaeleon gummifer, and by the transgression of grassland (Phalaridetalia) and weed species (Ridolfion). The association grows on steep slopes of road embankments and on grazed pastures. The role of roadside habitats for nature conservation in intensively used landscapes is discussed. (© 2005 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Biocompatibility and degradation of poly(DL‐lactic‐co‐glycolic acid)/calcium phosphate cement composites
Authors: Ruhé P. Quinten , Hedberg Elizabeth L. , Padron Nestor Torio , Spauwen Paul H. M. , Jansen John A. , Mikos Antonios G.
Published: July 2005

Injectable calcium phosphate (Ca‐P) cement materials exhibit favorable osteocompatible behavior but are resorbed slowly because of a lack of a bone ingrowth–enabling macroporosity. In this study, poly(DL‐lactic‐co‐glycolic acid) (PLGA) microparticles (average size 66 ± 25 μm) were incorporated into Ca‐P cement to obtain a macroporous Ca‐P cement scaffold after PLGA hydrolysis in vivo. Preset PLGA/Ca‐P cement composite discs of various weight ratios (0/100, 15/85, 30/70, and 50/50) were implanted subcutaneously and in cranial defects in rats for 12 weeks. Histological analysis revealed that all macropores in the PLGA‐containing composites (average pore size 73 ± 27 μm) were filled with fibrous tissue and blood vessels (subcutaneous implants) and/or bone (cranial implants). Histologically, bone formation appeared most abundant and most consistent in the 30/70 PLGA/Ca‐P cement composites. Histomorphometrical evaluation revealed a significant increase in defect fill in the 15/85 and 30/70 PLGA/Ca‐P cement composites. Finally, subcutaneous and cranial 50/50 PLGA/Ca‐P cement composites had degraded to a large extent, without adequate replacement by bone in the cranial implants. Therefore, we conclude that PLGA/Ca‐P cement composites enable tissue ingrowth and show excellent osteocompatibility in weight ratios of 15/85 and 30/70 PLGA/Ca‐P cement. In this model, 30/70 PLGA/Ca‐P cement composites showed the most favorable biological response. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005
Endothelial cell reaction on a biological material
Authors: Breithaupt‐Faloppa Ana Cristina , Kleinheinz Johannes , Crivello Oswaldo
Published: July 2005

The successful clinical application of materials should involve detailed investigations on interaction between them and tissue with which they will contact. We examined herein the behavior of endothelial cells (ECs) on a collagen material, using histological and immunohistochemical methods. We used isolated human umbilical cord vein cells (HUVECs) identified by means of endothelial‐specific antibodies. Cells were seeded in a standard density on a collagen membrane (Lycoll®, Resorba, Nuernberg, Germany) and on gelatin‐coated, control plastic surfaces, after two passages. These were then maintained for periods of 1, 7, or 14 days. The cells adhered, spread, and proliferated, and within 24 h started forming a subconfluent monolayer. We observed that the cultured cells expressed integrins (α5β1 and α(v)β3) and synthesized fibronectin. After 14 days, we could observe a confluent layer of ECs. We could conclude that the collagen material supported growth and attachment of endothelial cells. In addition, the attachment seemed to be most related to the fibronectin synthesized by the cells and to its highly expressed receptor (the α5β1 integrin); even though this is not the only protein related to this adhesion, we observed that our cultured HUVECs did not synthesize vitronectin. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006
Paul Everett Meehl: The cumulative record
Authors: Waller Niels G. , Lilienfeld Scott O.
Published: July 2005

In this brief biographical sketch of Paul Meehl, the authors examine the “cumulative record” of his scholarship and mentorship. This record sheds light on why Meehl is widely regarded as one of the most influential clinical psychologists of the 20th century, as well as on Meehl's remarkable intellectual life. Time has proven that Meehl's writings are exceptional in their quality, influence, breadth, and depth. In addition, Meehl's cumulative record raises important questions regarding the reinforcement contingencies in major research‐oriented psychology departments. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1209–1229, 2005.
Paul E. Meehl's influence on experimental psychopathology: Fruits of the nexus of schizotypy and schizophrenia, neurology, and methodology
Authors: Lenzenweger Mark F. , Maher Brendan A. , Manschreck Theo C.
Published: July 2005

Paul E. Meehl made numerous contributions to clinical science and a hallmark of many of these contributions was their integrative nature. Meehl's positions on complex topics, especially one such as schizophrenia, were reflective of input from a variety of disciplines and levels of analysis. In this essay the authors focus on Meehl's uniquely rich contribution to our understanding of schizophrenia through his theoretical model of schizotypy, his abiding interest in exploring neurologically based indicators of schizophrenia liability and encouragement to others to pursue such indicators, and his passion for rigorous research methodology. Meehl's contributions in each of these areas continue to influence the direction and research strategies used in experimental psychopathology to illuminate the fundamental nature of schizophrenia. These contributions have also shaped inquiry into many other psychopathological entities. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1295–1315, 2005.
Meehl on metatheory
Authors: Rozeboom Wm. W.
Published: July 2005

This article proffers a critical survey of Paul E. Meehl's thoughts on the philosophy of science, as well as constructive criticisms of his views on corroboration of scientific theories. Using examples from clinical psychology and allied domains, six major topics are addressed: (a) the nature of theoretical constructs, (b) statistical diagnosis of clinical categories, (c) detection of hypothesized taxa, (d) null‐hypothesis significance testing, (e) complexified hypothesis testing, and (f) cliometric theory appraisal. Through numerous examples, it is shown that Paul Meehl had a superb ability to recognize, articulate, and clarify hidden complexities, and other underappreciated obscurities in important working concepts. © 2005 Wiley Periodicals, Inc. J Clin Psychol 61: 1317–1354, 2005.
Sustained elevation of neutrophils in rats induced by lentivirus‐mediated G‐CSF delivery
Authors: Barry Simon , Brzezinski Margaret , Yanay Ofer , Seppen Jurgen E. , Osborne William R. A.
Published: July 2005

Background

Patients with severe chronic and cyclic neutropenia, characterized by neutrophil numbers <500 cells/µl, are treated daily with recombinant granulocyte colony‐stimulating factor (G‐CSF). As an alternative delivery approach we investigated the ability of lentivirus vectors to provide sustained G‐CSF expression.

Methods

Fischer rats were injected intramuscularly (IM) with vesicular stomatitis virus G (VSV‐G)‐pseudotyped lentivirus pRRL‐CMV‐G‐CSF‐SIN that encoded rat G‐CSF cDNA regulated by the human cytomegalovirus (CMV) promoter and incorporated a self‐inactivating (SIN) construct in the 3′ long terminal repeat (LTR). Control rats received normal saline or lentivirus encoding the enhanced green fluorescent protein (eGFP). Rats were serially monitored for blood cell production and tissues assayed for provirus distribution.

Results

Rats receiving a single IM injection of lentivirus exhibited elevated neutrophil counts for 14 months. Virus administration of 6 × 107 infectious units induced sustained levels of neutrophil production having a mean ± standard deviation (SD) of 5650 ± 900 cells/µl and rats that received a 10‐fold lower dose of virus showed mean neutrophil counts of 3340 ± 740 cells/µl. These were significantly higher than the mean of control animals receiving saline or control lentivirus (1, 760 ± 540 cells/µl, P < 0.0001). White blood cell (WBC) counts were significantly elevated in treated over control animals (P < 0.0001). Hematocrits (P > 0.3), lymphocytes (P > 0.2) and platelets (P > 0.1) were not significantly different between control and treated animals. Genomic DNA from muscle at the injection sites was positive for provirus, whereas lung, spleen, liver, kidney and non‐injected muscle samples were all negative, suggesting lack of virus spread.

Conclusions

These studies indicate that lentivirus vectors administered IM provide sustained, therapeutic levels of neutrophils and suggest this approach to treat patients with severe and cyclic neutropenia. Copyright © 2005 John Wiley & Sons, Ltd.
Site‐specific detection of S‐nitrosylated PKB α/Akt1 from rat soleus muscle using CapLC‐Q‐TOFmicro mass spectrometry
Authors: Lu Xiao‐Ming , Lu Mary , Tompkins Ronald G. , Fischman Alan J.
Published: July 2005

Protein Kinase Bα(PKBα, or Akt1) is believed to play a crucial role in programmed cell death, cancer progression and the insulin‐signaling cascade. The protein is activated by phosphorylation at multiple sites and subsequently phosphorylates and activates eNOS. Free cysteine residues of the protein may capture reactive, endogenously produced nitric oxide (NO) as S‐nitrosothiols. Site‐specific detection of S‐nitrosylated cysteine residues, usually at low stoichiometry, has been a major challenge in proteomic research largely due to the lack of mass marker for S‐nitrosothiols that are very labile under physiologic conditions. In this report we describe a sensitive and specific MS method for detection of S‐nitrosothiols in PKB α/Akt1 in rat soleus muscle.

PKB α/Akt1 was isolated by immunoprecipitation and 2D‐gel electrophoresis, subjected to in‐gel tryptic digestion, and cysteinyl nitrosothiols were reacted with iodoacetic acids [2‐C12/C13 = 50/50] under ascorbate reduction conditions. This resulted in the production of relatively stable carboxymethylcysteine (CMC) immonium ions (m/z 134.019 and m/z 135.019) within a narrow argon collision energy (CE = 30 ± 5 V) in the high MS noise region. In addition, free and disulfide‐linked cysteine residues were converted to carboxyamidomethylcysteines (CAM). Tryptic S‐nitrosylated parent ion was detected with a mass accuracy of 50 mDa for the two CMC immonium ions at the triggered elution time during capillary liquid chromatography (LC) separation. A peptide containing Cys296 was discriminated from four co‐eluting tryptic peptides under lock mass conditions (m/z 785.8426). S‐nitrosothiol in the tryptic peptide, ITDFGLBKEGIK (B: CAM, [M + 2H]2+ = 690.86, Found: 690.83), is believed to be present at a very low level, since the threshold for the CMC immonium trigger ions was set at 3 counts/s in the MS survey.

The high levels of NO that are produced under stress conditions may result in increased S‐nitrosylation of Cys296 which blocks disulfide bond formation between Cys296 and Cys310 and suppresses the biological effects of PKB α/Akt1. With the procedures developed here, this process can be studied under physiological and pathological conditions. Copyright © 2005 John Wiley & Sons, Ltd.
Optimized sample preparation for isotopic analyses of CO2 in air: systematic study of precision and accuracy dependence on driving variables during CO2 purification process
Authors: Bertolini T. , Rubino M. , Lubritto C. , D'Onofrio A. , Marzaioli F. , Passariello I. , Terrasi F.
Published: July 2005

A systematic analysis of efficiency, reproducibility and accuracy of cryogenic purification of CO2 from air samples for isotopic analyses is presented. The technical characteristics of the cryogenic line are given in detail. To study the cryogenic process, three different operating parameters are considered: flow rate of the gas entering the line, pressure of the gas in the line, and CO2‐trap shape. Experimental results demonstrate that efficiency, reproducibility and accuracy strongly depend on the CO2trap shape. Moreover, a dependence of reproducibility and accuracy on the flow rate of the gas is found, but not on its pressure. High precision (≤0.02‰ for δ13C and ≤0.05‰ for δ18O) and good accuracy (<0.09‰ for δ13C and <0.14‰ for δ18O) is achieved after applying the N2O correction. Copyright © 2005 John Wiley & Sons, Ltd.
Loss of dopaminergic neurons by the induction of inducible nitric oxide synthase and cyclooxygenase‐2 via CD40: Relevance to Parkinson's disease
Authors: Okuno Tatsusada , Nakatsuji Yuji , Kumanogoh Atsushi , Moriya Masayuki , Ichinose Hiroshi , Sumi Hisae , Fujimura Harutoshi , Kikutani Hitoshi , Sakoda Saburo
Published: July 2005

A glial reaction associated with up‐regulation of inflammatory molecules has been suggested to play an important role in dopaminergic neuron loss in Parkinson's disease (PD). Among inflammatory molecules, inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) have been focused upon as key factors in the pathogenesis. However, the mechanism of how these molecules are induced in PD brains is not clearly understood. We focused on CD40, which is expressed on neural cells and could be implicated in the neuroinflammation by inducing inflammatory molecules. We showed that both iNOS and COX‐2 were up‐regulated in microglia and astrocytes by CD40 stimulation in association with a low dose of interferon‐γ (IFN‐γ) in vitro. Selective loss of dopaminergic neurons was induced by costimulation with CD40 and IFN‐γ in mesencephalic cultures, which was protected by selective inhibitors of iNOS and/or COX‐2. We also found in CD40‐stimulated astrocytes an increase of a low‐affinity IgE receptor CD23, which is known to induce iNOS expression. Together these data suggest that up‐regulated iNOS and COX‐2 via the CD40 pathway may lead to dopaminergic neuron loss and may participate in the neuroinflammaory pathway of PD. © 2005 Wiley‐Liss, Inc.
Variations in extracellular levels of dopamine, noradrenaline, glutamate, and aspartate across the sleep–wake cycle in the medial prefrontal cortex and nucleus accumbens of freely moving rats
Authors: Léna I. , Parrot S. , Deschaux O. , Muffat‐Joly S. , Sauvinet V. , Renaud B. , Suaud‐Chagny M.‐F. , Gottesmann C.
Published: July 2005

We used intracerebral microdialysis coupled with electrophysiologic recordings to determine relative changes in the concentrations of several neurotransmitters in the medial prefrontal cortex and nucleus accumbens of freely moving rats during waking, slow‐wave sleep, and rapid eye movement (REM) sleep. The concentrations of noradrenaline, dopamine, glutamate, and aspartate in 2‐min dialysate samples were analyzed by capillary electrophoresis combined with laser‐induced fluorescence detection. Changes in glutamate and aspartate concentrations were found only in the nucleus accumbens, in which a decrease was obtained during both slow‐wave sleep and REM sleep compared to waking. A progressive reduction in the release of noradrenaline was observed from waking to REM sleep in both structures. In contrast, dopamine concentrations were higher during waking and REM sleep compared to that during slow‐wave sleep. The latter results demonstrate that contrary to the findings of earlier electrophysiologic studies carried out on ventral tegmental area dopaminergic neurons, changes in the release of dopamine in projection areas occur across the sleep–wake cycle. The elevated levels of dopamine during waking and REM sleep in the medial prefrontal cortex and the nucleus accumbens could result from changes during these two states in afferent modulation at the level of cell bodies or at the level of dopaminergic terminals. © 2005 Wiley‐Liss, Inc.