bluuebunny/crossref_metadata_2025 huggingface dataset:
Activity-Dependent Induction of Multitransmitter Signaling Onto Pyramidal Cells and Interneurons of Hippocampal Area CA3
Authors: Romo-Parra Héctor , Vivar Carmen , Maqueda Jasmín , Morales Miguel A. , Gutiérrez Rafael
Published: May 2006

The granule cells of the dentate gyrus (DG) are considered to be glutamatergic, but they contain glutamic acid decarboxylase, γ-amino butyric acid (GABA), and the vesicular GABA transporter mRNA. Their expression is regulated in an activity-dependent manner and coincides with the appearance of GABAergic transmission from the mossy fibers (MF) to pyramidal cells in area CA3. These data support the hypothesis that MF are able to release glutamate and GABA. Following the principle that a given neuron releases the same neurotransmitter(s) onto all its targets, we here demonstrate the emergence, after a generalized convulsive seizure, of MF GABAergic signaling sensitive to activation mGluR-III onto pyramidal cells and interneurons of CA3. Despite this, excitation overrides inhibition in interneurons, preventing disinhibition. Furthermore, on blockade of GABA and glutamate ionotropic receptors, an M1-cholinergic depolarizing signal is also revealed in both targets, which postsynaptically modulates the glutamatergic and GABAergic fast neurotransmission. The emergence of these nonglutamatergic signals depends on protein synthesis. In contrast to cholinergic responses evoked by associational/commissural fibers activation, cholinergic transmission evoked by DG stimulation is only observed after seizures and is strongly depressed by the activation of mGluR-II, whereas both are depressed by M2-AChR activation. With immunohistological experiments, we show that this cholinergic pathway runs parallel to the MF. Thus seizures compromise a delicate balance of excitation and inhibition, on which a complex interaction of different neurotransmitters emerges to counteract excitation at pre- and postsynaptic sites. Particularly, MF GABAergic inhibition emerges to exert an overall inhibitory action on CA3.
GABAergic Regulation of REM Sleep in Reticularis Pontis Oralis and Caudalis in Rats
Authors: Sanford Larry D. , Tang Xiangdong , Xiao Jihua , Ross Richard J. , Morrison Adrian R.
Published: May 2006

The nucleus reticularis pontis oralis (RPO) and nucleus reticularis pontis caudalis (RPC) are implicated in the generation of rapid eye movement sleep (REM). Work in cats has indicated that GABA in RPO plays a role in the regulation of REM. We assessed REM after local microinjections into RPO and RPC of the γ-aminobutyric acid-A (GABAA) agonist, muscimol (MUS), and the GABAA antagonist, bicuculline (BIC). Rats (90-day-old male Sprague-Dawley) were implanted with electrodes for recording electroencephalographs (EEG) and electromyographs (EMG). Guide cannulae were aimed into RPO ( n = 9) and RPC ( n = 8) for microinjecting MUS (200, 1,000.0 μM) and BIC (0.056, 0.333, 1.0, 1,000.0, and 10,000.0 μM). Animals received bilateral microinjections of saline, MUS, and BIC (0.2 μl microinjected at 0.1 μl/min) into each region followed by 6-h sleep recordings. In RPO, MUS (1,000.0 μM) suppressed REM and BIC (1,000.0 μM) enhanced REM. In RPC, MUS (200, 1,000.0 μM) suppressed REM, but BIC (1,000.0 μM and less) did not significantly affect REM. Higher concentrations of BIC (10,000.0 μM) injected into RPO ( n = 9) and RPC ( n = 4) produced wakefulness and escape behavior. The results indicate that GABA in RPO/RPC is involved in the regulation of REM and suggest site-specific differences in this regulation.
NMDA Receptor Blockade in the Superior Colliculus Increases Receptive Field Size Without Altering Velocity and Size Tuning
Authors: Razak Khaleel A. , Huang Lihua , Pallas Sarah L.
Published: May 2006

Neonatal brain injury triggers compensatory processes that can be adaptive or detrimental, but little is known about the mechanisms of compensation or how they might affect the response properties of neurons within the injured region. We have studied this issue in a rodent model. Partial ablation of the hamster superior colliculus (SC) at birth results in a compressed but complete visual field map in the remaining SC and a compensatory conservation of receptive field (RF) size and stimulus velocity and size tuning. The circuit underlying stimulus tuning in this system or its preservation after brain lesions is not known. Our previous work has shown that N-methyl-d-aspartate (NMDA) receptors are necessary for the development and conservation of RF size after partial SC ablation. In this study, we examined whether NMDA receptor function is also necessary for the development and conservation of stimulus velocity and size tuning. We found that velocity and size tuning were unaffected by chronic postnatal blockade of NMDA receptors and the resulting increases in RF size. Thus NMDA receptors in the SC are not necessary for the development of stimulus velocity and size tuning or in the compensatory maintenance of these properties following brain damage. These results suggest that stimulus velocity and size tuning may arise in the retina or from NMDA receptor-independent circuitry intrinsic to SC. The lack of conflict between NMDA receptor activity-dependent and -independent processes may allow conservation of some RF properties while others change during injury-induced or evolutionary changes in afferent/target convergence.
Protein Phosphatases Mediate Depotentiation Induced by High-Intensity Theta-Burst Stimulation
Authors: Kang-Park Maeng-Hee , Sarda Meredith A. , Jones Katherine H. , Moore Scott D. , Shenolikar Shirish , Clark Suzanne , Wilson Wilkie A.
Published: May 2006

We have previously reported that varying stimulus intensity produces qualitatively different types of synaptic plasticity in area CA1 of hippocampal slices: brief low-intensity (LI) theta-burst (TB) stimuli induce long-term potentiation (LTP), but if the stimulus intensity is increased (to mimic conditions that may exist during seizures), LTP is not induced; instead, high-intensity (HI) TB stimuli erase previously induced LTP (“TB depotentiation”). We now have explored the mechanisms underlying TB depotentiation using extracellular field recordings with pharmacological manipulations. We found that TB depotentiation was blocked by okadaic acid and calyculin A (inhibitors of serine/threonine protein phosphatases PP1 and PP2A), FK506 (a specific blocker of calcineurin, a Ca2+/calmodulin (CaM) protein phosphatase), and 8-Br-cAMP (an activator of protein kinase A) with 3-isobutyl-1-methylxanthine (IBMX, a phosphodiesterase inhibitor). These results suggest that protein phosphatase pathways are involved in the TB depotentiation similar to other type of down-regulating synaptic plasticity such as low-frequency stimulation (LFS)-induced long-term depression (LTD) and depotentiation in the rat hippocampus. However, TB depotentiation and LFS depotentiation could have differential functional significance.
Mutation and Activation of Gαs Similarly Alters Pre- and Postsynaptic Mechanisms Modulating Neurotransmission
Authors: Renden Robert B. , Broadie Kendal
Published: May 2006

Constitutive activation of Gαs in the Drosophilabrain abolishes associative learning, a behavioral disruption far worse than that observed in any single cAMP metabolic mutant, suggesting that Gαs is essential for synaptic plasticity. The intent of this study was to examine the role of Gαs in regulating synaptic function by targeting constitutively active Gαs to either pre- or postsynaptic cells and by examining loss-of-function Gαs mutants ( dgs) at the glutamatergic neuromuscular junction (NMJ) model synapse. Surprisingly, both loss of Gαs and activation of Gαs in either pre- or postsynaptic compartment similarly increased basal neurotransmission, decreased short-term plasticity (facilitation and augmentation), and abolished posttetanic potentiation. Elevated synaptic function was specific to an evoked neurotransmission pathway because both spontaneous synaptic vesicle fusion frequency and amplitude were unaltered in all mutants. In the postsynaptic cell, the glutamate receptor field was regulated by Gαs activity; based on immunocytochemical studies, GluRIIA receptor subunits were dramatically downregulated (>75% decrease) in both loss and constitutive active Gαs genotypes. In the presynaptic cell, the synaptic vesicle cycle was regulated by Gαs activity; based on FM1-43 dye imaging studies, evoked vesicle fusion rate was increased in both loss and constitutively active Gαs genotypes. An important conclusion of this study is that both increased and decreased Gαs activity very similarly alters pre- and postsynaptic mechanisms. A second important conclusion is that Gαs activity induces transynaptic signaling; targeted Gαs activation in the presynapse downregulates postsynaptic GluRIIA receptors, whereas targeted Gαs activation in the postsynapse enhances presynaptic vesicle cycling.
Effects of Ventrobasal Lesion and Cortical Cooling on Fast Oscillations (>200 Hz) in Rat Somatosensory Cortex
Authors: Staba Richard J. , Brett-Green Barbara , Paulsen Marcy , Barth Daniel S.
Published: May 2006

High-frequency oscillatory activity (>200 Hz) termed “fast oscillations” (FO) have been recorded in the rodent somatosensory cortex and may reflect very rapid integration of vibrissal information in sensory cortex. Yet, while electrophysiological correlates suggest that FO is generated within intracortical networks, contributions of subcortical structures along the trigeminal pathway remain uncertain. Using surface and laminar electrode arrays, in vivo recordings of vibrissal and electrically evoked FO were made within somatosensory cortex of anesthetized rodents before and after ablation of the ventrobasal thalamus (VB) or during reversible cortical cooling. In VB-lesioned animals, vibrissal stimulation failed to evoke FO, while epicortical stimulation in lesioned animals remained effective in generating FO. In nonlesioned animals, cortical cooling eliminated vibrissal-evoked FO despite the persistence of thalamocortical input. Vibrissal-evoked FO returned with the return to physiological temperatures. Results from this study indicate that somatosensory cortex alone is able to initiate and sustain FO. Moreover, these data suggest that cortical network interactions are solely responsible for the generation of FO, while synchronized thalamocortical input serves as the afferent trigger.
On the maximal cliques in c-max-tolerance graphs and their application in clustering molecular sequences
Authors: Lehmann Katharina A , Kaufmann Michael , Steigele Stephan , Nieselt Kay
Published: May 2006

Given a set S of n locally aligned sequences, it is a needed prerequisite to partition it into groups of very similar sequences to facilitate subsequent computations, such as the generation of a phylogenetic tree. This article introduces a new method of clustering which partitions S into subsets such that the overlap of each pair of sequences within a subset is at least a given percentage c of the lengths of the two sequences. We show that this problem can be reduced to finding all maximal cliques in a special kind of max-tolerance graph which we call a c-max-tolerance graph. Previously we have shown that finding all maximal cliques in general max-tolerance graphs can be done efficiently in O(n
3 + out). Here, using a new kind of sweep-line algorithm, we show that the restriction to c-max-tolerance graphs yields a better runtime of O(n
2 log n + out). Furthermore, we present another algorithm which is much easier to implement, and though theoretically slower than the first one, is still running in polynomial time. We then experimentally analyze the number and structure of all maximal cliques in a c-max-tolerance graph, depending on the chosen c-value. We apply our simple algorithm to artificial and biological data and we show that this implementation is much faster than the well-known application Cliquer. By introducing a new heuristic that uses the set of all maximal cliques to partition S, we finally show that the computed partition gives a reasonable clustering for biological data sets.
Regulation of cell proliferation by the antizyme inhibitor: evidence for an antizyme-independent mechanism
Authors: Kim S. W. , Mangold U. , Waghorne C. , Mobascher A. , Shantz L. , Banyard J. , Zetter B. R.
Published: May 2006

The antizyme inhibitor was discovered as a protein that binds to the regulatory protein antizyme and inhibits the ability of antizyme to interact with the enzyme ornithine decarboxylase (ODC). Blocking antizyme activity subsequently leads to increased intracellular levels of ODC and increased ODC enzymatic activity. We now report that antizyme inhibitor is a positive modulator of cell growth. Overexpression of antizyme inhibitor in NIH-3T3 mouse fibroblasts or in AT2.1 Dunning rat prostate carcinoma cells resulted in an increased rate of cell proliferation and an increase in saturation density of the cultured cells. This was accompanied by an increase in intracellular levels of the polyamine putrescine. In AT2.1 cells, antizyme inhibitor overexpression also increased the ability of the cells to form foci when grown under anchorage-independent conditions. In order to determine the role of antizyme on antizyme inhibitor activity we created an antizyme inhibitor mutant, AZIΔ117-140, which lacks the putative antizyme-binding domain. We show that this mutant fails to bind to antizyme, but remains capable of inducing increased rates of cell proliferation, suggesting that antizyme inhibitor has antizyme-independent functions. Silencing antizyme inhibitor expression leads to diminished levels of cyclin D1 and to reduced cell proliferation. Antizyme inhibitor is capable of preventing cyclin D1 degradation, and this effect is at least partially independent of antizyme. We show that wild-type antizyme inhibitor and the AZIΔY mutant are capable of direct interaction with cyclin D1 suggesting a potential mechanism for the antizyme-independent effects. Together, our data suggest a novel function for antizyme inhibitor in cellular growth control.
The Arf1p GTPase-activating protein Glo3p executes its regulatory function through a conserved repeat motif at its C-terminus
Authors: Yahara N. , Sato K. , Nakano A.
Published: May 2006

ADP-ribosylation factors (Arfs), key regulators of intracellular membrane traffic, are known to exert multiple roles in vesicular transport. We previously isolated eight temperature-sensitive (ts) mutants of the yeast ARF1 gene, which showed allele-specific defects in protein transport, and classified them into three groups of intragenic complementation. In this study, we show that the overexpression of Glo3p, one of the GTPase-activating proteins of Arf1p (ArfGAP), suppresses the ts growth of a particular group of the arf1 mutants (arf1-16 and arf1-17). Other ArfGAPs do not show such a suppression activity. All these ArfGAPs show sequence similarity in the ArfGAP catalytic domain, but are divergent in the rest of molecules. By domain swapping analysis of Glo3p and another ArfGAP, Gcs1p, we have shown that the non-catalytic C-terminal region of Glo3p is required for the suppression of the growth defect in the arf1 ts mutants. Interestingly, Glo3p and its homologues from other eukaryotes harbor a well-conserved repeated ISSxxxFG sequence near the C-terminus, which is not found in Gcs1p and its homologues. We name this region the Glo3 motif and present evidence that the motif is required for the function of Glo3p in vivo.
Relativistic origins of emotional reactions to events happening to others and to ourselves
Authors: Smith Richard H. , Eyre Heidi L. , Powell Caitlin A. J. , Kim Sung Hee
Published: May 2006

The purpose of the present study was to show that people's emotional reactions to both good and bad events happening to others can be influenced by how their own experiences compare with these events. Female undergraduate students took a test of intellectual ability and received false feedback suggesting that they had done well or poorly. Later, they viewed written feedback apparently given to another participant suggesting that she had performed well or poorly. Finally, participants gave their emotional reactions to their own performance as well as the performance of the other participant. Results showed that participants' relative performance influenced how happy they felt for the high performing other participant and how sad they felt for the low performing other participant. Participants' self‐focused emotions of pride and shame were also affected by the relative performance of the other participant.
Exocytosis of Vesicular Zinc Reveals Persistent Depression of Neurotransmitter Release during Metabotropic Glutamate Receptor Long-Term Depression at the Hippocampal CA3–CA1 Synapse
Authors: Qian Jing , Noebels Jeffrey L.
Published: May 2006

Exocytosis can be directly measured in mammalian brain slices by fluorescence detection of vesicular zinc release. Detection of the low-level evoked zinc signal [Zn]twas first demonstrated at the zinc-rich hippocampal mossy fiber pathway and required the use of high-frequency presynaptic stimulation. Here, we show that release after individual action potentials can be reliably detected even at non-mossy fiber, zinc-poor synapses in the hippocampus, a major enhancement in the temporal resolution of the technique. Short-term facilitation of release properties of zinc-positive CA3–CA1 Schaffer collateral/commissural synapses in the stratum radiatum differ from those at mossy fibers but are similar to those measured for the EPSP [field EPSP (fEPSP)]. The N-type Ca2+channel toxin ω-conotoxin GVIA inhibited both the [Zn]tand fEPSP equally, and the modulation of neurotransmitter release by neuropeptide Y, baclofen, and adenosine as revealed by [Zn]tclosely resembles that measured for the fEPSP. A long-standing controversy in hippocampal synaptic plasticity involves the site of long-term depression (LTD) at these synapses. Using zinc release as a direct marker for exocytotic events and a surrogate marker for glutamate release, we demonstrate that persistent depression of presynaptic release occurs in the late expression of DHPG [(S)-3,5-dihydroxyphenylglycine]-induced LTD at this synapse. The ability to examine release dynamics with zinc fluorescence detection will facilitate exploration of the molecular pharmacology and plasticity of exocytosis at many CNS synapses.
Lithium Administration to Preadolescent Rats Causes Long-Lasting Increases in Anxiety-Like Behavior and Has Molecular Consequences
Authors: Youngs Rachael M. , Chu Melissa S. , Meloni Edward G. , Naydenov Alipi , Carlezon William A. , Konradi Christine
Published: May 2006

Lithium (Li) is frequently used in the treatment of bipolar disorder (BPD), a debilitating condition that is increasingly diagnosed in children and adolescents. Because the symptoms of BPD in children are different from the typical symptoms in adulthood and have significant overlap with other childhood psychiatric disorders, this disorder is notoriously difficult to diagnose. This raises the possibility that some children not affected by BPD are treated with Li during key periods of brain development. The objective of this investigation was to examine the long-term effects of Li on the developing brain via a series of behavioral and molecular studies in rats. Rat pups were reared on Li chow for 3 weeks. Parallel groups were tested while on Li chow or 2 and 6 weeks after discontinuation of treatment. We found increased measures of anxiety-like behavior at all times tested. Gene microarray studies of the amygdala revealed that Li affected the expression of gene transcripts of the synapse and the cytoskeleton, suggesting that the treatment induced synaptic adjustments. Our study indicates that Li can alter the trajectory of brain development. Although the effects of Li on the normal brain seems unfavorable, effects on the abnormal brain cannot be determined from these studies alone and may well be therapeutic. Our results indicate that Li administration to the normal brain has the potential for lasting adverse effects.
Role of Substantia Nigra–Amygdala Connections in Surprise-Induced Enhancement of Attention
Authors: Lee Hongjoo J. , Youn Jina M. , O Mary J. , Gallagher Michela , Holland Peter C.
Published: May 2006

Coding of prediction error by midbrain dopamine neurons has been examined extensively in the framework of associative learning theory. Most of this research has focused on the role of prediction error in determining the reinforcement value of unconditioned stimuli: poorly predicted (“surprising”) outcomes are more effective reinforcers and produce a greater dopamine response than well predicted outcomes. However, surprise also enhances attention to cues that signal poorly predicted outcomes. Previous reports from our laboratories demonstrated that circuitry, including the amygdala central nucleus (CeA), the cholinergic neurons of the substantia innominata/nucleus basalis region, and their innervation of the posterior parietal cortex, is critical to these surprise-induced enhancements of attention in associative learning. The present study considered the origin of prediction error information important for the operation of this system by examining the effects of disrupting communication between the midbrain substantia nigra pars compacta (SNc) and the CeA. Rats received unilateral lesions of the SNc and lesions of the CeA in either the contralateral or ipsilateral hemisphere. Contralateral lesions eliminated the surprise-induced enhancement of attention and learning that was displayed by rats with ipsilateral control lesions. These results show that SNc–CeA communication is critical to mechanisms by which the coding of prediction error by midbrain dopamine neurons is translated into enhancement of attention and learning modulated by the cholinergic system.
The gum syndrome: predicaments in court interpreting
Authors: Morris Ruth
Published: May 2006

This article examines aspects of communication relevant to the process of interlingual interpretation, and considers the law's equivocal attitudes to interpreters. It identifies a tendency by many of the professional figures in the courtroom to consider the interpreter a mere instrument, and by many language-handicapped defendants to view the interpreter as a linguistic and psychological haven. The effects of variations in the physical courtroom setting are considered, and some aspects of interpreter role delineation are reviewed. A number of predicaments typically experienced by court interpreters are described, and their effects on interpreters as individuals analysed. Examples are drawn from a variety of contemporary and historical sources, including case reports and two email discussion forums. Although most of the material comes from the Anglo-Saxon legal tradition, it is argued that the interpreter's position on the one hand as a humble technical instrument and on the other as the potential saviour in a situation of language handicap is a constant in the legal arena irrespective of time frame or jurisdiction.
Chronic Stress, Depression, and Cortisol Levels as Risk Indicators of Elevated Plaque and Gingivitis Levels in Individuals Aged 50 Years and Older
Authors: Hugo Fernando N. , Hilgert Juliana B. , Bozzetti Mary C. , Bandeira Denise R. , Gonçalves Tonantzin R. , Pawlowski Josiane , de Sousa Maria da Luz R.
Published: May 2006

Background: The role of psychoneuroimmunological factors in oral hygiene has been overlooked in the dental literature. Our objective was to evaluate the effects of stress, depression, and cortisol levels in dental plaque accumulation and gingivitis in a population of individuals aged ≥50 years.

Methods: In this cross‐sectional study, 230 subjects, selected from caregivers of demented patient groups and from social activities groups of Porto Alegre, Brazil, were evaluated. Stress was evaluated with the Lipp stress inventory, whereas depressive symptoms were assessed using the Beck depression inventory. Three saliva samples (at 8, 12, and 20 hours) were collected for cortisol analysis by means of radioimmunoassay. Cortisol levels were expressed as the area under the curve of the three samples for each patient. Multivariate logistic regression was performed with the visible plaque index and gingival bleeding index as outcomes.

Results: The mean age of subjects was 61.6 ± 8.2 years, mean monthly income in United States dollars (US$) was US$668 ± 590, and mean cortisol level was 22.1 ± 33.7 nmol/l. A total of 51.5% of the individuals were caregivers; 9.4% were smokers, and 25.5% used interdental floss/brush. Being a caregiver (odds ratio [OR] = 3.97; 95% confidence interval [95% CI] = 2.08 to 7.54), using an interdental brush (OR = 0.23; 95% CI = 0.11 to 0.47), cortisol (OR = 2.03; 95% CI = 1.09 to 3.81), and stress (OR = 1.45; 95% CI = 1.03 to 2.02) were all significantly associated with visible plaque index >mean (43%). Being a caregiver (OR = 2.35; 95% CI = 1.13 to 4.86), using an interdental brush (OR = 0.32; 95% CI = 0.13 to 0.77), smoking (OR = 0.26; 95% CI = 0.08 to 0.89), stress (OR = 1.78; 95% CI =1.31 to 2.38), and visible plaque index (mean) (OR = 12.87; 95% CI = 6.29 to 26.31) were significantly associated with a gingival bleeding index > mean (37%).

Conclusion: Stress was a significant risk indicator of elevated levels of plaque and gingivitis, whereas cortisol was a risk indicator of plaque in the sample after controlling for confounders.
Cessation of Hormone Replacement Therapy After Reports of Adverse Findings From Randomized Controlled Trials: Evidence From a British Birth Cohort
Authors: Mishra Gita , Kok Helen , Ecob Russell , Cooper Rachel , Hardy Rebecca , Kuh Diana
Published: May 2006

Objectives. We examined the cessation of hormone replacement therapy (HRT) among British women, by educational level, social class, and cardiovascular risk factors, at the time of publicity about 2 clinical trials of HRT that were halted after adverse findings.

Methods. A total of 1387 women aged 57 years reported their monthly HRT use between January 2002 and February 2003. A succession of regression-based time-series models were fitted to detect changes in the proportion of HRT users stratified by education level, social class, hypertension, and obesity.

Results. The overall percentage of HRT users declined from 31% in January 2002 to less than 26% by February 2003. Changes in trends of HRT use were first detected in June 2002 (for women with advanced secondary educational qualification or higher) and in July 2002 (for all other groups). The rate of decline was greatest for women with no formal educational qualifications, from the manual social class, or who were hypertensive or obese.

Conclusions. These decreases coincided with the announced cessation of a large US clinical trial of HRT. This publicity may have had a differential influence on the immediate decline in HRT use by various groups of British women.
Formatted anti–tumor necrosis factor α VHH proteins derived from camelids show superior potency and targeting to inflamed joints in a murine model of collagen‐induced arthritis
Authors: Coppieters Ken , Dreier Torsten , Silence Karen , Haard Hans De , Lauwereys Marc , Casteels Peter , Beirnaert Els , Jonckheere Heidi , Wiele Christophe Van De , Staelens Ludovicus , Hostens Jeroen , Revets Hilde , Remaut Erik , Elewaut Dirk , Rottiers Pieter
Published: May 2006

Objective

The advent of tumor necrosis factor (TNF)–blocking drugs has provided rheumatologists with an effective, but highly expensive, treatment for the management of established rheumatoid arthritis (RA). Our aim was to explore preclinically the application of camelid anti‐TNF VHH proteins, which are single‐domain antigen binding (VHH) proteins homologous to human immunoglobulin VHdomains, as TNF antagonists in a mouse model of RA.

Methods

Llamas were immunized with human and mouse TNF, and antagonistic anti‐TNF VHH proteins were isolated and cloned for bacterial production. The resulting anti‐TNF VHH proteins were recombinantly linked to yield bivalent mouse and human TNF‐specific molecules. To increase the serum half‐life and targeting properties, an anti–serum albumin anti‐TNF VHH domain was incorporated into the bivalent molecules. The TNF‐neutralizing potential was analyzed in vitro. Mouse TNF‐specific molecules were tested in a therapeutic protocol in murine collagen‐induced arthritis (CIA). Disease progression was evaluated by clinical scoring and histologic evaluation. Targeting properties were evaluated by99mTc labeling and gamma camera imaging.

Results

The bivalent molecules were up to 500 times more potent than the monovalent molecules. The antagonistic potency of the anti‐human TNF VHH proteins exceeded even that of the anti‐TNF antibodies infliximab and adalimumab that are used clinically in RA. Incorporation of binding affinity for albumin into the anti‐TNF VHH protein significantly prolonged its serum half‐life and promoted its targeting to inflamed joints in the murine CIA model of RA. This might explain the excellent therapeutic efficacy observed in vivo.

Conclusion

These data suggest that because of the flexibility of their format, camelid anti‐TNF VHH proteins can be converted into potent therapeutic agents that can be produced and purified cost‐effectively.
Functional assay of type I interferon in systemic lupus erythematosus plasma and association with anti–RNA binding protein autoantibodies
Authors: Hua Jing , Kirou Kyriakos , Lee Christina , Crow Mary K.
Published: May 2006

Objective

Peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) have increased expression of genes typically induced by type I interferon (IFN). However, it has been difficult to identify and quantify the factors responsible for activation of the IFN pathway in SLE. To characterize these mediators, we developed an assay that measures the functional effects of plasma or serum components on the gene expression of cultured target cells.

Methods

WISH epithelial cell line cells were cultured with medium, with recombinant IFNα, IFNβ, or IFNγ, or with 50% plasma from SLE patients (n = 73), rheumatoid arthritis (RA) patients (n = 19), or healthy donors (n = 30). Real‐time quantitative polymerase chain reaction was used to determine WISH cell expression of IFN target genes, including PRKR, IFIT1, IFI44, MX1, and C1orf29 (preferentially induced by IFNα) and CXCL9 (Mig) (preferentially induced by IFNγ).

Results

IFNα‐regulated genes were induced by SLE plasma samples, but not by most of the RA or healthy control plasma samples. The activity in SLE plasma was inhibited >90% by anti‐IFNα antibody, but not by anti‐IFNβ or anti‐IFNγ antibodies. The expression of each IFNα target gene induced by SLE plasma correlated with the expression of that gene studied ex vivo in PBMCs from the same patients and with the titer of anti–RNA binding protein (anti‐RBP)–specific autoantibodies. Plasma activity paralleled PBMC expression of IFNα‐inducible genes over time.

Conclusion

IFNα in SLE plasma is a major stimulus of IFN target gene expression and is related to expression of those genes in PBMCs from SLE patients and to the titer of anti‐RBP autoantibodies. These data provide additional support for the view that IFNα mediates immune system activation and dysregulation in SLE.
Quality of life after surgical treatment of colorectal liver metastases
Authors: Langenhoff B S , Krabbe P F M , Peerenboom L , Wobbes T , Ruers T J M
Published: May 2006

Background

The surgical approach to colorectal liver metastases is becoming increasingly aggressive. The aim of this prospective study was to evaluate the impact of surgery on health-related quality of life (HRQoL) of patients with colorectal liver metastases.

Methods

HRQoL data from 97 patients with colorectal liver metastases were analysed. Sixty patients (group 1) had surgical treatment of the liver metastases. Seventeen patients (group 2) were shown to have inoperable disease at laparotomy. Twenty outpatients with inoperable disease were included as a control group (group 3). Two validated HRQoL instruments, the European Organization for Research and Treatment of Cancer Core questionnaire (QLQ C-30) and the EuroQol-5D, were applied.

Results

By 2 weeks after operation patients in group 1 showed a clear overall deterioration in HRQoL, but after 3 months most HRQoL scores had returned to baseline levels. At 2 weeks after surgery there was clear deterioration in almost all HRQoL domains in group 2, and several symptoms were still being reported at 3 months. Patients in group 3 showed hardly any deterioration in HRQoL over the 3 months.

Conclusion

The fast recovery of HRQoL, generally within 3 months, justifies an aggressive surgical approach to colorectal liver metastases. However, careful preoperative evaluation is crucial to avoid needless laparotomy, considering the ongoing deteriorated HRQoL of group 2.
Bevacizumab combination therapy in recurrent, platinum‐refractory, epithelial ovarian carcinoma
Authors: Wright Jason D. , Hagemann Andrea , Rader Janet S. , Viviano Dana , Gibb Randall K. , Norris Lori , Mutch David G. , Powell Matthew A.
Published: May 2006

BACKGROUND

The study was undertaken to determine the safety and efficacy of the monoclonal, antivascular endothelial growth factor antibody bevacizumab in combination with cytotoxic chemotherapy for women with platinum‐refractory ovarian cancer.

METHODS

A retrospective analysis of women who received bevacizumab in combination with a cytotoxic agent was performed. Response was determined by measurable disease or assessment of serial cancer antigen (CA) 125 measurements.

RESULTS

Twenty‐three patients were identified. The patients were heavily pretreated with a median of 7 prior regimens including a median of 3 prior platinum regimens. The combination regimen included cyclophosphamide in 15 (65%), 5‐fluorouracil (5‐FU) in 6 (26%), docetaxel in 1 (4%), and gemcitibine/liposomal doxorubicin in 1 (4%). Two (9%) women developed chylous ascites during treatment. CTC Grade 4–5 toxicities occurred in 4 (17%) subjects. Gastrointestinal perforation occurred in 2 (9%) patients. Measurable disease was present in 22. The overall best response rate was 35% and all 8 were partial responses (PRs). Stable disease was found in a further 10 (44%) women, whereas progressive disease was observed in 5 (22%). The median time to progression was 5.6 months in patients with a PR and 2.3 months in subjects with stable disease. Three (13%) women experienced a progression‐free interval (PFI) of >6 months. At last follow‐up, 8 (35%) subjects had died of disease, whereas 15 (65%) women were alive with disease.

CONCLUSIONS

Combination bevacizumab therapy demonstrated activity in heavily pretreated women with ovarian cancer. Gastrointestinal perforations were identified in 9%. Despite the toxicity of the regimen, prospective studies, particularly in less heavily pretreated patients, are warranted. Cancer 2006. © 2006 American Cancer Society.
Cover Picture: High Density Amorphous Ice from Cubic Ice (ChemPhysChem 6/2006)
Authors: Loerting Thomas , Kohl Ingrid , Schustereder Werner , Winkel Katrin , Mayer Erwin
Published: May 2006

The cover picture shows two powder X‐ray diffractograms recorded from two different "ice" samples obtained by pressurizing cubic (red) and hexagonal (black) ice crystals at ≈80 K. Sharp peaks marked with an asterisk are from traces of hexagonal ice. In their Communication on page 1203 Loerting et al. demonstrate that the two types of ice I obtainable at ambient pressure result in two different structural states of high density amorphous ice (HDA) when pressurized to 1.5 GPa at 77 K. The background image is a high‐resolution photograph of a hexagonal snowflake provided by Professor Kenneth G. Libbrecht (California Institute of Technology).
Optimization of a fibrin scaffold for sustained release of an adenoviral gene vector
Authors: Breen Ailish , Strappe Padraig , Kumar Anil , O'Brien Timothy , Pandit Abhay
Published: May 2006

One of the major limitations of adenovirus as an efficient gene delivery vehicle is its transient nature of transgene expression. At present, no biodegradable release system exists that permits the sustained release of gene delivery. This study aimed to optimize a biodegradable fibrin scaffold for sustained adenoviral mediated gene delivery. Scaffold formulations were fabricated incorporating an adenoviral vector encoding β‐galactosidase, and each scaffold was incubated in elution fluid. The level of transfection of cells was measured to assess virus diffusion from each scaffold at different time periods. Results showed that viral particles were present in the elution fluid and had the ability to transfect cells at different time points. Depending on the scaffold formulation, variable rates of diffusion were seen. Analysis of scaffold microarchitecture using stereo pairs showed that variable pore depth is seen in different fibrin scaffold concentration, which may be a determining factor in the diffusion rate. It was concluded that sustained release of the adenovirus could be attained up to 192 h at the optimal concentration of 60 mg/mL fibrinogen and 4 IU thrombin. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2006
Applying Darwinian principles in designing effective intervention strategies: The case of sun tanning
Authors: Saad Gad , Peng Albert
Published: May 2006

Public‐service announcements typically seek to educate consumers regarding a given unhealthy practice, the assumption being that individuals will cease the harmful behavior once they are fully informed. Many intervention strategies have failed in curbing the targeted behaviors because these are not due to incomplete information but instead may also have a Darwinian‐based etiology. Using sunbathing as a case analysis, it is shown how Darwinian theorizing (evolutionary psychology, life‐history theory, gene‐culture co‐evolution, and memetic theory) can augment social marketers' ability to develop efficacious intervention strategies. © 2006 Wiley Periodicals, Inc.
Jaw1/LRMP, a germinal centre‐associated marker for the immunohistological study of B‐cell lymphomas
Authors: Tedoldi S , Paterson JC , Cordell J , Tan S‐Y , Jones M , Manek S , Dei Tos AP , Roberton H , Masir N , Natkunam Y , Pileri SA , Facchetti F , Hansmann M‐L , Mason DY , Marafioti T
Published: May 2006

Jaw1, also known as lymphoid‐restricted membrane protein (LRMP), is an endoplasmic reticulum‐associated protein. High levels of Jaw1/LRMP mRNA have been found in germinal centre B‐cells and in diffuse large B‐cell lymphomas of ‘germinal centre’ subtype. This paper documents Jaw1/LRMP expression at the protein level in human tissues by immunohistochemical and western blotting analysis using an antibody reactive with paraffin‐embedded tissues. Jaw1/LRMP was highly expressed in germinal centre B‐cells (in keeping with gene expression data), in ‘monocytoid B‐cells’, and in splenic marginal zone B‐cells. It was absent, or present at only low levels, in mature T‐cells, although cortical thymocytes were weakly positive. Among lymphoid neoplasms, Jaw1/LRMP was found in germinal centre‐derived lymphomas (follicle centre lymphoma, Burkitt's lymphoma, lymphocyte‐predominant Hodgkin's disease) but not in T‐cell neoplasms (with the exception of a single T lymphoblastic lymphoma). Classical Hodgkin's disease and myeloma lacked Jaw1/LRMP but many cases of chronic lymphocytic leukaemia (but not mantle zone lymphoma) were Jaw1/LRMP‐positive. Approximately half of the marginal zone lymphomas were Jaw1/LRMP‐positive. In diffuse large B‐cell lymphomas, Jaw1/LRMP was found in three‐quarters (24/32) of the cases classified phenotypically as being of ‘germinal centre’ type, but it was also expressed in almost half (13/28) of the ‘non‐germinal centre’ cases. A similar proportion of ‘non‐germinal centre’ cases were positive for the protein products of two other genes expressed highly in germinal centre cells (HGAL/GCET2 and PAG). The fact that all three of these proteins are expressed in a significant proportion of diffuse large B‐cell lymphomas assigned to the ‘non‐germinal centre’ category indicates that the immunophenotypic categorization of diffuse large B‐cell lymphoma according to cellular origin may be more complicated than currently understood. Finally, the expression of Jaw1/LRMP in other types of lymphoma and in non‐lymphoid tissues/tumours may be of interest in differential diagnosis and research. Copyright © 2006 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Oligonucleotide microarrays for studying the effects of Ras signal transduction on the genetic program
Authors: Tchernitsa Oleg I. , Sers Christine , Geflitter Anita , Schäfer Reinhold
Published: May 2006

Cytoplasmic signal transduction processes are tightly coupled with the transcriptional regulation of genes executing cellular phenotypes in normal conditions and in disease. A major challenge in signal transduction research is to understand how the specificity of the transcriptional response is determined by complex networks of signaling effectors and transcription factors. The Ras signal transduction pathway belongs to the few characterized pathways triggering malignant phenotypes both in experimental and human cancer. We have developed an array‐based tool which permits analysis of Ras signaling on transcription under various experimental conditions as well as in tumor specimens. The composition of targets represented on the Ras signaling target array (RASTA) is based on previously validated gene expression profiles and published functional studies. The Ras signaling target array was used to compare the transcriptional response toward oncogenic Ras in fibroblasts and epithelial cells and to assess the time‐dependent deregulation of target genes.
boc-Aspartyl(OMe)-Fluoromethylketone Attenuates Mitochondrial Release of Cytochrome c and Delays Brain Tissue Loss after Traumatic Brain Injury in Rats
Authors: Clark Robert SB , Nathaniel Paula D , Zhang Xiaopeng , Dixon C Edward , Alber Sean M , Watkins Simon C , Melick John A , Kochanek Patrick M , Graham Steven H
Published: May 2006

The pathobiology of traumatic brain injury (TBI) includes activation of multiple caspases followed by cell death with a spectrum of apoptotic phenotypes. There are initiator (e.g. caspase-2, −8, and −9) and effector (e.g. caspase-3 and −7) caspases. Recently, caspase-2 and −8 have been shown to regulate cell death via provoking cytochrome c release from the mitochondria upstream of caspase-9. Here, we show that an intracerebral injection of the pan-caspase inhibitor boc-Aspartyl(OMe)-fluoromethylketone (BAF; 1 μmol) 1 min after TBI in rats reduces caspase-3-like activity, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and tissue damage, and cytochrome c release in ipsilateral cortex at 24 h versus vehicle. To investigate whether either caspase-2 and/or caspase-8 activation may contribute to cytochrome release, the effect of BAF treatment on caspase-2 and caspase-8 proteolysis was also examined. boc-aspartyl(OMe)-fluoromethylketone treatment inhibited proteolysis of caspase-2 but not caspase-8 24 h after TBI in rats versus vehicle. However, BAF with or without nerve growth factor (12.5 ng/hx14 days intracerebrally via osmotic pump) did not result in differences in motor function, Morris water maze performance, hippocampal neuron survival, nor contusion volume at 14 days. These data suggest that BAF treatment reduces acute cell death after TBI by inhibiting mitochondrial release of cytochrome c, possibly via a mechanism involving initiator caspases; however, BAF appears to delay cell death, rather than result in permanent protection.
Single intrinsic Josephson junction with double-sided fabrication technique
Authors: You L. X. , Torstensson M. , Yurgens A. , Winkler D. , Lin C. T. , Liang B.
Published: May 2006

We make stacks of intrinsic Josephson junctions (IJJs) embedded in the bulk of very thin (d⩽100nm) Bi2Sr2CaCu2O8+x single crystals. By precisely controlling the etching depth during the double-sided fabrication process, the stacks can be reproducibly tailor-made to be of any microscopic height (0−9nm<d), i.e., enclosing a specified number of IJJ (0–6), including the important case of a single junction. We discuss reproducible gaplike features in the current-voltage characteristics of the samples at high bias.
X-ray photoconductivity due to trap-sensitive relaxation of hot carriers
Authors: Konovalov I. , Makhova L. , Wett D. , Hallmeier K.-H. , Szargan R. , Mitdank R.
Published: May 2006

We observed the temperature-dependent modulation of the electrical conductivity in ZnO thin films under periodic illumination by soft x rays. At specific temperatures, small variations of the excitation energy near the x-ray absorption edges resulted in large element-specific variations of the conductivity modulation. The emission rate of electrons at traps E1/E2 and E3/E4 in ZnO at these specific temperatures roughly equals the excitation frequency. We conclude that relaxation of electrons, excited from localized core levels into the conduction band, predominantly happens into trap states with the same localization. The experimental results were explained using symmetry selection rules and local transition probabilities.
Observation of magnetism in Au thin films
Authors: Reich S. , Leitus G. , Feldman Y.
Published: May 2006

Direct magnetization measurements of thin gold films are presented. These measurements integrate the signal from the thin film under study and the magnetic contribution of the film’s interface with the substrate. The diamagnetic contribution to the signal from the bulk substrate is of the same order as the noise level. We find that thin gold films can exhibit positive magnetization. The character of their magnetic behavior is strongly substrate dependent.
Establishment of non-native plant species after wildfires: effects of fuel treatments, abiotic and biotic factors, and post-fire grass seeding treatments
Authors: Hunter Molly E. , Omi Philip N. , Martinson Erik J. , Chong Geneva W.
Published: May 2006

Establishment and spread of non-native species following wildfires can pose threats to long-term native plant recovery. Factors such as disturbance severity, resource availability, and propagule pressure may influence where non-native species establish in burned areas. In addition, pre- and post-fire management activities may influence the likelihood of non-native species establishment. In the present study we examine the establishment of non-native species after wildfires in relation to native species richness, fire severity, dominant native plant cover, resource availability, and pre- and post-fire management actions (fuel treatments and post-fire rehabilitation treatments). We used an information-theoretic approach to compare alternative hypotheses. We analysed post-fire effects at multiple scales at three wildfires in Colorado and New Mexico. For large and small spatial scales at all fires, fire severity was the most consistent predictor of non-native species cover. Non-native species cover was also correlated with high native species richness, low native dominant species cover, and high seeded grass cover. There was a positive, but non-significant, association of non-native species with fuel-treated areas at one wildfire. While there may be some potential for fuels treatments to promote non-native species establishment, wildfire and post-fire seeding treatments seem to have a larger impact on non-native species.