bluuebunny/crossref_metadata_2025 huggingface dataset:
Imatinib Mesylate Inhibits Proliferation and Exerts an Antifibrotic Effect in Human Breast Stroma Fibroblasts
Authors: Gioni Vassiliki , Karampinas Theodoros , Voutsinas Gerassimos , Roussidis Andreas E. , Papadopoulos Savvas , Karamanos Nikos K. , Kletsas Dimitris
Published: May 2008

Tumor stroma plays an important role in cancer development. In a variety of tumors, such as breast carcinomas, a desmoplastic response, characterized by stromal fibroblast and collagen accumulation, is observed having synergistic effects on tumor progression. However, the effect of known anticancer drugs on stromal cells has not been thoroughly investigated. Imatinib mesylate is a selective inhibitor of several protein tyrosine kinases, including the receptor of platelet-derived growth factor, an important mediator of desmoplasia. Recently, we have shown that imatinib inhibits the growth and invasiveness of human epithelial breast cancer cells. Here, we studied the effect of imatinib on the proliferation and collagen accumulation in breast stromal fibroblasts. We have shown that it blocks the activation of the extracellular signal-regulated kinase and Akt signaling pathways and up-regulates cyclin-dependent kinase inhibitor p21WAF1, leading to the inhibition of fibroblast proliferation, by arresting them at the G0/G1 phase of the cell cycle. Imatinib inhibits more potently the platelet-derived growth factor–mediated stimulation of breast fibroblast proliferation. By using specific inhibitors, we have found that this is due to the inhibition of the Akt pathway. In addition, imatinib inhibits fibroblast-mediated collagen accumulation. Conventional and quantitative PCR analysis, as well as gelatin zymography, indicates that this is due to the down-regulation of mRNA synthesis of collagen I and collagen III—the main collagen types in breast stroma—and not to the up-regulation or activation of collagenases matrix metalloproteinase 2 and matrix metalloproteinase 9. These data indicate that imatinib has an antifibrotic effect on human breast stromal fibroblasts that may inhibit desmoplastic reaction and thus tumor progression. (Mol Cancer Res 2008;6(5):706–14)
A Suppressive Role of Mitogen Inducible Gene-2 in Mesenchymal Cancer Cell Invasion
Authors: Shi Xiaohua , Wu Chuanyue
Published: May 2008

Cancer cell invasion of extracellular matrix (ECM) is essential for dissemination of cancer cells and metastasis. In this study, we have investigated the role of mitogen inducible gene-2 (Mig-2, also known as kindlin-2), a focal adhesion protein whose expression is altered in several types of human cancers, in mesenchymal cancer cell invasion. Mig-2 is abundantly expressed in SK-LMS-1 leiomyosarcoma cells. The level of Mig-2, however, is considerably lower in more invasive HT-1080 fibrosarcoma cells. Overexpression of Mig-2 in HT-1080 and SK-LMS-1 cells substantially reduced their ability to invade ECM in an in vitro Matrigel invasion assay. Conversely, knockdown of Mig-2 markedly increased the invasiveness of these cells. Consistent with a suppressive role in mesenchymal cancer cell invasion, Mig-2 inhibits urokinase-type plasminogen activator (uPA) secretion and pericellular proteolysis. Overexpression of Mig-2 increased uPA accumulation at the intracellular face of cell-ECM adhesions and reduced the level of secreted uPA. Conversely, knockdown of Mig-2 reduced uPA accumulation at the intracellular face of cell-ECM adhesions and increased uPA secretion. Our results reveal an important role of Mig-2 in suppression of mesenchymal cancer cell invasion and shed new light on how altered Mig-2 expression could influence cancer cell invasion. (Mol Cancer Res 2008;6(5):715–24)
Mechanisms Altering Airway Smooth Muscle Cell Ca2+ Homeostasis in Two Asthma Models
Authors: Kellner Julia , Tantzscher Juliane , Oelmez Hamza , Edelmann Martin , Fischer Rainald , Huber Rudolf Maria , Bergner Albrecht
Published: May 2008

Background: Asthma is characterized by airway remodeling, altered mucus production and airway smooth muscle cell (ASMC) contraction causing extensive airway narrowing. In particular, alterations of ASMC contractility seem to be of crucial importance. The elevation of the cytoplasmic Ca2+ concentration is a key event leading to ASMC contraction and changes in the agonist-induced Ca2+ increase in ASMC have been reported in asthma. Objective: The aim of this study was to investigate mechanisms underlying these changes. Methods: Murine tracheal smooth muscle cells (MTSMC) from T-bet KO mice and human bronchial smooth muscle cells (HBSMC) incubated with IL-13 and IL-4 served as asthma models. Acetylcholine-induced changes in the cytoplasmic Ca2+ concentration were recorded using fluorescence microscopy and the expression of Ca2+ homeostasis regulating proteins was investigated with Western blot analysis. Results: Acetylcholine-induced Ca2+ transients were elevated in both asthma models. This correlated with an increased Ca2+ content of the sarcoplasmic reticulum (SR). In MTSMC from T-bet KO mice, the expression of the SR Ca2+ buffers calreticulin and calsequestrin was higher compared to wild-type mice. In HBSMC incubated with IL-13 or IL-4, the expression of ryanodine receptors, inositol-3-phosphate receptors and sarcoplasmic/endoplasmic reticulum Ca2+ ATPases 2 was increased compared to HBSMC without incubation with interleukins. The enlarged acetylcholine-induced Ca2+ transients could be reversed by blocking inositol-3-phosphate receptors. Conclusions: We conclude that in the murine asthma model the SR Ca2+ buffer capacity is increased, while in the human asthma model the expression of SR Ca2+ channels is altered. The investigation of the Ca2+ homeostasis of ASMC has the potential to provide new therapeutical options in asthma.
Precision of Biomarkers to Define Chronic Inflammation in CKD
Authors: LaClair Robert , O’Neal Kalisha , Ofner Susan , Sosa Marcelo J. , Labarrere Carlos A. , Moe Sharon M.
Published: May 2008

Background/Aims: Several inflammatory biomarkers have been found to be associated with cardiovascular disease or all-cause mortality in dialysis patients, but their usefulness in clinical practice or as surrogate endpoints is not certain. The purpose of the present study was to determine the intrapatient variation of C-reactive protein, IL-6, fetuin-A and albumin in a population of dialysis patients. Methods: Apparently healthy dialysis patients with either a tunneled dialysis catheter or fistula had monthly assessments of these biomarkers for a total of four determinations, and the intraclass correlation coefficients were calculated as measures of intersubject variance. Results: Our results showed large within-subject variation relative to the total variation in the measurements (31–46%). Having a tunneled catheter as opposed to a fistula was not significantly associated with mean levels, suggesting that chronic subclinical catheter infection does not explain the variation seen in the biomarkers. In contrast, there was a rapid change in these biomarkers with a clinically apparent acute infection. Conclusion: These results suggest that these biomarkers have limitations for use as surrogate endpoints in clinical trials due to wide fluctuations, even in apparently clinically healthy individuals.
Waltz Dancing in Patients With Chronic Heart Failure
Authors: Belardinelli Romualdo , Lacalaprice Francesca , Ventrella Chiara , Volpe Loretta , Faccenda Ernesto
Published: May 2008

Background—

There is evidence that aerobic exercise improves functional capacity in patients with New York Heart Association (NYHA) class II and III chronic heart failure. However, it is unknown whether dancing is safe and able to improve functional capacity in patients with chronic heart failure.

Methods and Results—

We prospectively studied 130 patients with stable chronic heart failure (107 men; mean age, 59�11 years) in New York Heart Association class II and III and left ventricle ejection fraction <40%. Patients were randomized to supervised aerobic exercise training at 70% of peak V̇
o
2
3 times a week for 8 weeks (group E, n=44) or to a dance protocol of alternate slow (5 minutes) and fast (3 minutes) waltz lasting 21 minutes (group D, n=44). A group that did not undergo exercise training served as control (group C, n=42). On study entry and at 8 weeks, all patients underwent cardiopulmonary exercise testing on a cycle ergometer until volitional fatigue, 2D-echo with Doppler, and endothelium-dependent dilation of the brachial artery. Heart rate was 111�15 bpm during exercise training and 113�19 bpm during dancing (
P
=0.59). Peak V̇
o
2
, anaerobic threshold, V̇
e
/V̇
co
2
slope, and V̇
o
2
/W slope were all similarly improved in both E and D groups (+16% and 18%, 20% and 21%, 14% and 15%, 18% and 19%, respectively;
P
not significant for all comparisons;
P
<0.001 versus controls). Endothelium-dependent relaxation was also similarly improved (group E, from 2.6�1.3% to 5.2�1.5%,
P
<0.001 versus control; group D, from 2.2�1.4% to 5.0�1.5%,
P
<0.001 versus control for both E and D). The change in peak V̇
o
2
in E and D groups was correlated with changes in peak velocity of early filling wave/peak velocity of late filling ratio (
r
=−0.58,
P
<0.001) and endothelium-dependent dilation (
r
=0.64,
P
<0.001). Untoward events were rare in both E and D groups.

Conclusions—

In patients with stable chronic heart failure, waltz dancing is safe and able to improve functional capacity and endothelium-dependent dilation similar to traditional aerobic exercise training. Waltz dancing may be considered in clinical practice in combination with aerobic exercise training or as an alternative to it.
Interaction Between Renal Function and Microalbuminuria for Cardiovascular Risk in Hypertension
Authors: Färbom Patrik , Wahlstrand Björn , Almgren Peter , Skrtic Stanko , Lanke Jan , Weiss Lars , Kjeldsen Sverre , Hedner Thomas , Melander Olle
Published: May 2008

We investigated whether renal function and microalbuminuria are independent predictors and whether any interaction exists between them, regarding future cardiovascular disease in hypertensive patients (n=10 881) followed for 4.5 years. The primary end points (PEs) were fatal and nonfatal myocardial infarction and stroke and other cardiovascular deaths. Creatinine and glomerular filtration rate (GFR), estimated using the formulas of the Modification of Diet in Renal Disease study group and Cockroft and Gault and in a subsample (n=4929) of microalbuminuria and interaction terms of microalbuminuria and renal function, were related to the risk of the PE using Cox proportional hazards model after full adjustment. Increased creatinine (
P
<0.001), decreased GFR from Cockroft and Gault (
P
=0.001), and decreased GFR from the Modification of Diet in Renal Disease study group (
P
=0.001) were all independent risk factors for the PE. Stepwise exclusion of patients with the poorest renal function excluded the possibility that the relationship between decreasing renal function and the PE was driven only by patients with severely impaired renal function. Microalbuminuria and all 3 of the indices of renal function predicted the PE independent of each other. There was a significant interaction between microalbuminuria and GFR from Cockroft and Gault (
P
=0.040) in prediction of the PE. Both renal function and microalbuminuria add independent prognostic information regarding cardiovascular risk in hypertensive patients. The cardiovascular risk associated with microalbuminuria increases with a decline in GFR, as demonstrated by a significant interaction between microalbuminuria and GFR from Cockroft and Gault. Because estimation of the total cardiovascular risk is essential for the aggressiveness of risk factor interventions, simultaneous inclusion of GFR and microalbuminuria in global cardiovascular risk assessment is essential.
Attentional Selection of Multiple Goal Positions Before Rapid Hand Movement Sequences: An Event-related Potential Study
Authors: Baldauf Daniel , Deubel Heiner
Published: May 2008

A dot-probe paradigm was used to provide physiological evidence for the parallel selection of multiple movement goals before rapid hand movement sequences. Participants executed a sequence of manual pointing movements to two out of three possible goal positions. During movement preparation, a task-irrelevant visual transient (a dot probe) was flashed either at one of both movement goals, or at the third, movement-irrelevant location. The results revealed that the N1 component induced by the presentation of the dot was enhanced if the dot was flashed at one of the movement goals, indicating that both target positions were attended before the initialization of the movement sequence. A second experiment showed that movement-irrelevant locations between the movement goals were not attended, suggesting that attention splits into spatially distinct foci.
Difficulty of Discrimination Modulates Attentional Capture by Regulating Attentional Focus
Authors: Sawaki Risa , Katayama Jun'ichi
Published: May 2008

Attentional capture for distractors is enhanced by increasing the difficulty of discrimination between the standard and the target in the three-stimulus oddball paradigm. In this study, we investigated the cognitive mechanism of this modulation of attentional capture. Event-related brain potentials were recorded from participants while they performed a visual three-stimulus oddball paradigm (frequent standard, rare target, and rare distractor). The discrimination difficulty between standard and target was manipulated in the central location. Distractor stimuli were presented in the central or surrounding locations. The P3a component was elicited by distractor stimuli and was used as a measure of attentional capture. The results revealed that discrimination difficulty had opposite effects on the P3a response between central and surrounding locations. With an increase in the difficulty of discrimination, the P3a response was enhanced when distractor stimuli were presented in the central location. In contrast, the P3a response was reduced when distractor stimuli were presented in a surrounding location. This finding suggests that spatial attention was focused by the difficulty of discrimination, and deviant processing was increased within its focus but decreased outside its focus. Therefore, attentional capture for deviant distractors is modulated by top–down controlled attentional focus.
Distinctive Neural Mechanisms Supporting Visual Object Individuation and Identification
Authors: Xu Yaoda
Published: May 2008

Many everyday activities, such as driving on a busy street, require the encoding of distinctive visual objects from crowded scenes. Given resource limitations of our visual system, one solution to this difficult and challenging task is to first select individual objects from a crowded scene (object individuation) and then encode their details (object identification). Using functional magnetic resonance imaging, two distinctive brain mechanisms were recently identified that support these two stages of visual object processing. While the inferior intraparietal sulcus (IPS) selects a fixed number of about four objects via their spatial locations, the superior IPS and the lateral occipital complex (LOC) encode the features of a subset of the selected objects in great detail (object shapes in this case). Thus, the inferior IPS individuates visual objects from a crowded display and the superior IPS and higher visual areas participate in subsequent object identification. Consistent with the prediction of this theory, even when only object shape identity but not its location is task relevant, this study shows that object individuation in the inferior IPS treats four identical objects similarly as four objects that are all different, whereas object shape identification in the superior IPS and the LOC treat four identical objects as a single unique object. These results provide independent confirmation supporting the dissociation between visual object individuation and identification in the brain.
Saccades to a Remembered Location Elicit Spatially Specific Activation in Human Retinotopic Visual Cortex
Authors: Geng Joy J. , Ruff Christian C. , Driver Jon
Published: May 2008

The possible impact upon human visual cortex from saccades to remembered target locations was investigated using functional magnetic resonance imaging (fMRI). A specific location in the upper-right or upper-left visual quadrant served as the saccadic target. After a delay of 2400 msec, an auditory signal indicated whether to execute a saccade to that location (go trial) or to cancel the saccade and remain centrally fixated (no-go). Group fMRI analysis revealed activation specific to the remembered target location for executed saccades, in the contralateral lingual gyrus. No-go trials produced similar, albeit significantly reduced, effects. Individual retinotopic mapping confirmed that on go trials, quadrant-specific activations arose in those parts of ventral V1, V2, and V3 that coded the target location for the saccade, whereas on no-go trials, only the corresponding parts of V2 and V3 were significantly activated. These results indicate that a spatial–motor saccadic task (i.e., making an eye movement to a remembered location) is sufficient to activate retinotopic visual cortex spatially corresponding to the target location, and that this activation is also present (though reduced) when no saccade is executed. We discuss the implications of finding that saccades to remembered locations can affect early visual cortex, not just those structures conventionally associated with eye movements, in relation to recent ideas about attention, spatial working memory, and the notion that recently activated representations can be “refreshed” when needed.
Structural and Metabolic Correlates of Episodic Memory in Relation to the Depth of Encoding in Normal Aging
Authors: Kalpouzos Grégoria , Chételat Gaël , Landeau Brigitte , Clochon Patrice , Viader Fausto , Eustache Francis , Desgranges Béatrice
Published: May 2008

This study set out to establish the relationship between changes in episodic memory retrieval in normal aging on the one hand and gray matter volume and 18FDG uptake on the other. Structural MRI, resting-state 18FDG-PET, and an episodic memory task manipulating the depth of encoding and the retention interval were administered to 46 healthy subjects divided into three groups according to their age (young, middle-aged, and elderly adults). Memory decline was found not to be linear in the course of normal aging: Whatever the retention interval, the retrieval of shallowly encoded words was impaired in both the middle-aged and the elderly, whereas the retrieval of deeply encoded words only declined in the elderly. In middle-aged and elderly subjects, the reduced performance in the shallow encoding condition was mainly related to posterior mediotemporal volume and metabolism. By contrast, the impaired retrieval of deeply encoded words in the elderly group was mainly related to frontal and parietal regions, suggesting the adoption of inefficient strategic processes.
Development of Spatial and Verbal Working Memory Capacity in the Human Brain
Authors: Thomason Moriah E. , Race Elizabeth , Burrows Brittany , Whitfield-Gabrieli Susan , Glover Gary H. , Gabrieli John D. E.
Published: May 2008

A core aspect of working memory (WM) is the capacity to maintain goal-relevant information in mind, but little is known about how this capacity develops in the human brain. We compared brain activation, via fMRI, between children (ages 7–12 years) and adults (ages 20–29 years) performing tests of verbal and spatial WM with varying amounts (loads) of information to be maintained in WM. Children made disproportionately more errors than adults as WM load increased. Children and adults exhibited similar hemispheric asymmetry in activation, greater on the right for spatial WM and on the left for verbal WM. Children, however, failed to exhibit the same degree of increasing activation across WM loads as was exhibited by adults in multiple frontal and parietal cortical regions. Thus, children exhibited adult-like hemispheric specialization, but appeared immature in their ability to marshal the neural resources necessary to maintain large amounts of verbal or spatial information in WM.
The Common Neural Basis of Autobiographical Memory, Prospection, Navigation, Theory of Mind, and the Default Mode: A Quantitative Meta-analysis
Authors: Spreng R. Nathan , Mar Raymond A. , Kim Alice S. N.
Published: May 2008

A core brain network has been proposed to underlie a number of different processes, including remembering, prospection, navigation, and theory of mind [Buckner, R. L., & Carroll, D. C. Self-projection and the brain. Trends in Cognitive Sciences, 11, 49–57, 2007]. This purported network—medial prefrontal, medial-temporal, and medial and lateral parietal regions—is similar to that observed during default-mode processing and has been argued to represent self-projection [Buckner, R. L., & Carroll, D. C. Self-projection and the brain. Trends in Cognitive Sciences, 11, 49–57, 2007] or scene-construction [Hassabis, D., & Maguire, E. A. Deconstructing episodic memory with construction. Trends in Cognitive Sciences, 11, 299–306, 2007]. To date, no systematic and quantitative demonstration of evidence for this common network has been presented. Using the activation likelihood estimation (ALE) approach, we conducted four separate quantitative meta-analyses of neuroimaging studies on: (a) autobiographical memory, (b) navigation, (c) theory of mind, and (d) default mode. A conjunction analysis between these domains demonstrated a high degree of correspondence. We compared these findings to a separate ALE analysis of prospection studies and found additional correspondence. Across all domains, and consistent with the proposed network, correspondence was found within the medial-temporal lobe, precuneus, posterior cingulate, retrosplenial cortex, and the temporo-parietal junction. Additionally, this study revealed that the core network extends to lateral prefrontal and occipital cortices. Autobiographical memory, prospection, theory of mind, and default mode demonstrated further reliable involvement of the medial prefrontal cortex and lateral temporal cortices. Autobiographical memory and theory of mind, previously studied as distinct, exhibited extensive functional overlap. These findings represent quantitative evidence for a core network underlying a variety of cognitive domains.
Compressed Scaling of Abstract Numerosity Representations in Adult Humans and Monkeys
Authors: Merten Katharina , Nieder Andreas
Published: May 2008

There is general agreement that nonverbal animals and humans endowed with language possess an evolutionary precursor system for representing and comparing numerical values. However, whether nonverbal numerical representations in human and nonhuman primates are quantitatively similar and whether linear or logarithmic coding underlies such magnitude judgments in both species remain elusive. To resolve these issues, we tested the numerical discrimination performance of human subjects and two rhesus monkeys (Macaca mulatta) in an identical delayed match-to-numerosity task for a broad range of numerosities from 1 to 30. The results demonstrate a noisy nonverbal estimation system obeying Weber's Law in both species. With average Weber fractions in the range of 0.51 and 0.60, nonverbal numerosity discriminations in humans and monkeys showed similar precision. Moreover, the detailed analysis of the performance distributions exhibited nonlinearly compressed numerosity representations in both primate species. However, the difference between linear and logarithmic scaling was less pronounced in humans. This may indicate a gradual transformation of a logarithmic to linear magnitude scale in human adults as the result of a cultural transformation process during the course of mathematical education.
Behavioural responses of Phasmarhabditis hermaphrodita (Nematoda: Rhabditida) to mucus from potential hosts
Authors: Small Richard , Bradford Claire
Published: May 2008

Host recognition behaviours were investigated in the Rhabditid nematode Phasmarhabditis hermaphrodita. Infective juveniles were exposed to mucus samples from mollusc species of varying susceptibility (Arion hortensis agg., Deroceras reticulatum, Tandonia budapestensis, Helix aspersa ,Limax flavus, Limax marginatus). Behavioural responses to the mucus were analysed using video-taped sequences. During contact with the mucus samples, infective juveniles decreased duration and frequency of forward crawling and increased frequency and duration of head thrusting and duration of head waving. Other host recognition behaviours identified included body waving and pushing. Most significant differences in behaviour recorded were between controls and during exposure to mollusc mucus; there were few significant differences between mollusc species. The results suggest that P. hermaphrodita forages by the 'cruise' method until mucus from potential hosts is encountered, but then concentrates its search behaviour, characterised by the decrease in forward crawling. However, there were no recorded behaviours that would differentiate between readily susceptible species and those with low or unknown susceptibility.
Description of Neodiplogaster crenatae sp. n. and N. acaloleptae sp. n. (Nematoda: Diplogastridae) from Japan
Authors: Kubono Takanori , Masuya Hayato , Kanzaki Natsumi
Published: May 2008

Neodiplogaster crenatae Kanzaki, Masuya & Kubono sp. n. and N. acaloleptae Kanzaki sp. n. are described and figured. Neodiplogaster crenatae sp. n. was isolated from the bark beetles Scolytoplatypus daimio and S. shogun (Scolytidae) emerging from dead logs of Fagus crenata, and was cultured on Botrytis cinerea growing on malt extract agar. Neodiplogaster crenatae sp. n. has male spicules which are relatively stout and arcuate in lateral view but curved to appear Y-shaped in ventral view; the male tail tip is bifurcate and the female tail is long and tapered. Neodiplogaster crenatae sp. n. appears closest to N. velata but is distinguished from it by spicular morphology and female tail length. Neodiplogaster acaloleptae sp. n. was isolated from the cerambycid beetle Acalolepta luxuriosa and reared on B. cinerea growing on potato dextrose agar. Neodiplogaster acaloleptae sp. n. has a conical male tail with narrow bursa covering the entire tail, smoothly tapered and strongly arcuate male spicules, leaf-shaped gubernaculum with a pointed anterior end, and long female tail with a filiform terminus. The new species is close to N. pinicola, N. piniphili and N. wacheki, but may be distinguished by its long female tail and male gubernaculum shape. The partial SSU and D2/D3 LSU of N. crenatae sp. n. were sequenced and compared with those of other sequenced nematodes, revealing that SSU was closest to those of other diplogastrid species, but that D2/D3 LSU was close to Anisakis spp. Neodiplogaster crenatae sp. n. propagated on a fungal lawn of B. cinerea and ambrosia fungi associating with its carrier beetles, but the nematode did not propagate on bacteria. The main food resource of Neodiplogaster spp. is hypothesised to be fungi.
Evidence for pathotype mixtures on Solanum bulbocastanum in Meloidogyne chitwoodi but not in M. fallax
Authors: Poleij Leo , van der Beek J.G. (Hans)
Published: May 2008

Variability for pathogenicity on seven Solanum bulbocastanum clones was studied in a collection of isolates of Meloidogyne chitwoodi and M. fallax from a wide range of geographical origins. Four different pathotypes could be distinguished within M. chitwoodi, while no specialisation was found in M. fallax. Meloidogyne chitwoodi isolates from the USA showed the largest variation; those from Europe belonged to one pathotype. In several M. chitwoodi isolates, the occurrence of pathotype mixtures was verified and for the first time reported in automictic Meloidogyne spp. The infrequently formed egg masses in incompatible isolate-plant genotype combinations were not able to reproduce further on various S. bulbocastanum genotypes, suggesting some stability aspects of the resistance. The occurrence of pathotypes and mixtures of pathotypes requires special attention in breeding programmes as well as in disease management.
Conventional and real-time PCR-based species identification and diversity of potato cyst nematodes (Globodera spp.) from Victoria, Australia
Authors: Nambiar Lila , Cunnington James , Quader Motiul
Published: May 2008

PCR (conventional and real-time) and DNA sequence analysis were used to identify species and genotypes of potato cyst nematodes from sites in Victoria, Australia. Only Globodera rostochiensis was detected. Sequence analyses of these isolates of PCN have confirmed the PCR-based results and have revealed the presence of genetically diverse populations in infested fields. However, the sequence variation was not in the diagnostic primer binding sites. The melting peaks, from multiplex real-time PCR analysis, for Globodera pallida and G. rostochiensis were 83.3 and 88.7, respectively. The importance of DNA extraction, PCR and sequence analysis for the molecular identification of PCN is discussed. This study has significant implications for detecting species of PCN in order to monitor/develop control strategies for the PCN of quarantine importance.
A new entomopathogenic nematode, Steinernema colombiense n. sp. (Nematoda: Steinernematidae), from Colombia
Authors: López-Núñez Juan Carlos , Plichta Kathryn , Góngora-Botero Carmenza E. , Stock S. Patricia
Published: May 2008

A new entomopathogenic nematode, Steinernema colombiense n. sp., is described
from Colombia. Morphological, molecular (28S and ITS rDNA sequence data) and
cross-hybridisation studies were used for diagnostics and identification
purposes. In addition, 28S and ITS rDNA sequence data were used to assess
evolutionary relationships of the new species with other Steinernema spp.
Morphological diagnostic features for S. colombiense n. sp. include
morphometric features of the third-stage infective juvenile, including body
length of 636 (549-732) μm, narrow body diam. (31 (22-36) μm), position of
the excretory pore (35 (31-40) μm), tail length (41 (32-53) μm), D% = 29
(25-33) and E% = 205 (138-284). In addition, males of first and second
generations are characterised by the morphology of the spicules and
gubernaculum, the number and arrangement of the genital papillae and the
excretory pore position (at 67 (56-76) and 54 (46-63) μm, for first and
second generations, respectively). In addition to these traits, 28S and ITS
rDNA sequences analyses both showed this species to be a distinct and unique
entity.
A Novel Method of Obtaining Prostate Tissue for Gene Expression Profiling
Authors: Sooriakumaran Prasanna , Henderson Alastair , Denham Philippa , Langley Stephen EM.
Published: May 2008

Gene expression profiling by DNA microarray analysis is a technique with great promise in cancer biology. The multifocality and heterogeneity of many prostate cancers makes the collection of adequate biological samples for such profiling particularly challenging. Current methods, such as laser capture microdissection, are not widely available and can have significant limitations. In this article, a novel method of prostatic sampling, which does not affect the histopathological assessment of the surgical specimen and provides adequate RNA yield for microarray analysis is described. This method is simple, inexpensive, easily reproducible, and has been validated as having >95% sensitivity and 99% specificity for histological prediction of tissue obtained. This method can be adopted by other investigators to perform DNA microarray analysis on prostate tumors.
Stance-taking in Hebrew casual conversation via be'emet (`really, actually, indeed', lit. `in truth')
Authors: Maschler Yael , Estlein Roi
Published: May 2008

In this article, we investigate the functional itinerary followed by Hebrew be'emet (`really, actually, indeed', lit. `in truth'), through a close exploration of its synchronic uses in the contemporary spoken language. Since this utterance, derived from the noun 'emet (`truth'), is so profoundly tied in with the speaker's beliefs and attitudes towards his or her discourse, we consider issues of metalanguage, modality, evidentiality, and stance. Be'emet is traditionally classified as `adverb', but in our corpus of naturally occurring Hebrew conversation, only 22 percent of all tokens function in this role. Whereas these tokens function referentially, the great majority of tokens (70%) function in the interpersonal realm of discourse, manifesting properties of discourse markers: 44.5 percent of all tokens function as full-fledged discourse markers (both semantically and structurally, Maschler, 1998) serving mirative (DeLancey, 2001), reprimanding, or negating any doubt roles; 22.5 percent function to ratify a previous stance; and three percent function to latch onto a new topic, requesting its elaboration. An intermediate category (8%) is composed of tokens functioning both referentially and interpersonally, mid-way between an adverb and a discourse marker, in a way that is particularly illuminating for understanding the changes undergone by be'emet as a result of its employment in discourse. The study thus lends support to previous studies of subjectification and intersubjectification in the process of grammaticization of discourse markers (Traugott, 2003; Traugott and Dasher, 2002).
A proposed 24 h lighting scheme for older adults
Authors: Figueiro MG
Published: May 2008

Alzheimer's disease (AD) patients exhibit random patterns of rest and activity rather than the consolidated sleep/wake cycle found in normal, older people. Light treatment has been shown to improve rest and activity rhythms and sleep efficiency of AD patients, presumably through consolidation of their circadian rhythms. The circadian system is maximally sensitive to short-wavelength radiation. Two independent studies summarised here show that 30 lux at the cornea of blue light (λ max = 470 nm) from light emitting diodes (LEDs) for 2 h in the early evening improved sleep efficiency of older adults, including those with AD compared to exposure to the same dose of red light. Because compliance to blue light treatment may be difficult for adults with AD, we conceived of a lighting scheme that might be more practical and as effective. White light dosages of different spectra and amounts for night and for day, based on a computational model for human circadian phototransduction, might be more readily accepted by seniors and by their caregivers. Implications for an improved visual environment and for better sleep efficiency of older adults are discussed.
Angiopoietin-1 mediates the proangiogenic activity of the bone morphogenic protein antagonist Drm
Authors: Mitola Stefania , Moroni Emanuela , Ravelli Cosetta , Andres German , Belleri Mirella , Presta Marco
Published: May 2008

Recent observations have shown that Drm, a member the Dan family of bone morphogenic protein (BMP) antagonists, induces endothelial cell (EC) sprouting in vitro and angiogenesis in vivo by interacting with signaling EC receptors in a BMP-independent manner. Here, recombinant Drm (rDrm) up-regulates angiopoientin-1 (Ang-1) expression in EC without affecting Ang-2 and Tie-2 receptor expression. Ang-1 up-regulation is mediated by the activation of the transcription factor NF-κB. Specific inhibition of Ang-1 activity by anti–Ang-1 antibodies, soluble Tie-2 receptor, or Ang-1 siRNA transfection significantly reduced the rDrm-mediated sprouting of EC in three-dimensional fibrin and type I collagen gels. In addition, Ang-1 antagonists inhibited the angiogenic activity exerted by rDrm in the chick embryo chorioallantoic membrane. Taken together, the data indicate that the proangiogenic activity of Drm is mediated by the activation of an Ang-1–dependent autocrine loop of stimulation in EC.
Expression, activation, and function of integrin αMβ2 (Mac-1) on neutrophil-derived microparticles
Authors: Pluskota Elzbieta , Woody Neil M. , Szpak Dorota , Ballantyne Christie M. , Soloviev Dmitry A. , Simon Daniel I. , Plow Edward F.
Published: May 2008

Leukocyte-derived microparticles (MPs) are markers of cardiovascular diseases and contribute to pathogenesis by their interaction with various cell types. The presence and activation state of a multifunctional leukocyte receptor, integrin αMβ2 (CD11b/18), on MPs derived from human neutrophils (PMNs) were examined. αMβ2 expression was significantly enhanced on MPs derived from stimulated compared with resting PMNs. Furthermore, αMβ2 on MPs from stimulated but not resting PMNs was in an activated conformation because it was capable of binding activation-specific monoclonal antibodies (CBRM1/5 and mAb24) and soluble fibrinogen. MPs expressing active αMβ2 interacted with and were potent activators of resting platelets as assessed by induction of P-selectin expression and activation of αIIbβ3. With the use of function-blocking antibodies and MPs obtained from αM−/−-deficient mice, we found that engagement of GPIbα on platelets by αMβ2 on MPs plays a pivotal role in MP binding. Platelet activation by MPs occurs by a pathway dependent on Akt phosphorylation. PSGL-1/P-selectin interaction also is involved in the conjugation of MPs to platelets, and the combination of blocking reagents to both αMβ2/GPIbα and to PSGL-1/P-selectin completely abrogates MP-induced platelet activation. Thus, cooperation of these 2 receptor/counterreceptor systems regulates the prothrombotic properties of PMN-derived MPs.
c-Cbl expression levels regulate the functional responses of human central and effector memory CD4 T cells
Authors: Brembilla Nicolò C. , Weber Johann , Rimoldi Donata , Pradervand Sylvain , Schütz Frédéric , Pantaleo Giuseppe , Rüegg Curzio , Quadroni Manfredo , Harshman Keith , Doucey Marie-Agnès
Published: May 2008

The biochemical mechanisms controlling the diverse functional outcomes of human central memory (CM) and effector memory (EM) T-cell responses triggered through the T-cell receptor (TCR) remain poorly understood. We implemented reverse phase protein arrays to profile TCR signaling components in human CD8 and CD4 memory T-cell subsets isolated ex vivo. As compared with CD4 CM cells, EM cells express statistically significant increased amounts of SLP-76 and reduced levels of c-Cbl, Syk, Fyn, and LAT. Moreover, in EM cells reduced expression of negative regulator c-Cbl correlates with expression of c-Cbl kinases (Syk and Fyn), PI3K, and LAT. Importantly, consistent with reduced expression of c-Cbl, EM cells display a lower functional threshold than CM cells. Increasing c-Cbl content of EM cells to the same level as that of CM cells using cytosolic transduction, we impaired their proliferation and cytokine production. This regulatory mechanism depends primarily on c-Cbl E3 ubiquitin ligase activity as evidenced by the weaker impact of enzymatically deficient c-Cbl C381A mutant on EM cell functions. Our study reports c-Cbl as a critical regulator of the functional responses of memory T cell subsets and identifies for the first time in humans a mechanism controlling the functional heterogeneity of memory CD4 cells.
Retrovirally induced CTL degranulation mediated by IL-15 expression and infection of mononuclear phagocytes in patients with HTLV-I–associated neurologic disease
Authors: Enose-Akahata Yoshimi , Oh Unsong , Grant Christian , Jacobson Steven
Published: May 2008

CD8+ T cells contribute to central nervous system inflammation in human T-cell lymphotropic virus type I (HTLV-I)–associated myelopathy/tropical spastic paraparesis (HAM/TSP). We analyzed CD8+ T-cell dysfunction (degranulation and IFN-γ production) and have demonstrated that CD8+ T cells of patients with HAM/TSP (HAM/TSP patients) spontaneously degranulate and express IFN-γ in ex vivo unstimulated culture. CD8+ T cells of HTLV-I asymptomatic carriers and healthy donors did not. Spontaneous degranulation was detected in Tax11-19/HLA-A*201 tetramer+ cells, but not in CMV pp65 tetramer+ cells. Interestingly, degranulation and IFN-γ production in CD8+ T cells was induced by coculture with autologous CD14+ cells, but not CD4+ T cells, of HAM/TSP patients, which correlated with proviral DNA load in CD14+ cells of infected patients. Moreover, the expression of IL-15, which induced degranulation and IFN-γ production in infected patients, was enhanced on surface of CD14+ cells in HAM/TSP patients. Blockade of MHC class I and IL-15 confirmed these results. Thus, CD8+ T-cell dysregulation was mediated by both virus infection and enhanced IL-15 on CD14+ cells in HAM/TSP patients. Despite lower viral expression than in CD4+ T cells, HTLV-I–infected or –activated CD14+ cells may be a heretofore important but under recognized reservoir particularly in HAM/TSP patients.
Genome-wide gene expression profiling analysis of Leishmania major and Leishmania infantum developmental stages reveals substantial differences between the two species
Authors: Rochette Annie , Raymond Frédéric , Ubeda Jean-Michel , Smith Martin , Messier Nadine , Boisvert Sébastien , Rigault Philippe , Corbeil Jacques , Ouellette Marc , Papadopoulou Barbara
Published: May 2008

Background

Leishmaniaparasites cause a diverse spectrum of diseases in humans ranging from spontaneously healing skin lesions (e.g.,L. major) to life-threatening visceral diseases (e.g.,L. infantum). The high conservation in gene content and genome organization betweenLeishmania majorandLeishmania infantumcontrasts their distinct pathophysiologies, suggesting that highly regulated hierarchical and temporal changes in gene expression may be involved.

Results

We used a multispecies DNA oligonucleotide microarray to compare whole-genome expression patterns of promastigote (sandfly vector) and amastigote (mammalian macrophages) developmental stages betweenL. majorandL. infantum. Seven per cent of the totalL. infantumgenome and 9.3% of theL. majorgenome were differentially expressed at the RNA level throughout development. The main variations were found in genes involved in metabolism, cellular organization and biogenesis, transport and genes encoding unknown function. Remarkably, this comparative global interspecies analysis demonstrated that only 10–12% of the differentially expressed genes were common toL. majorandL. infantum. Differentially expressed genes are randomly distributed across chromosomes further supporting a posttranscriptional control, which is likely to involve a variety of 3'UTR elements.

Conclusion

This study highlighted substantial differences in gene expression patterns betweenL. majorandL. infantum. These important species-specific differences in stage-regulated gene expression may contribute to the disease tropism that distinguishesL. majorfromL. infantum.
His-tag ELISA for the detection of humoral tumor-specific immunity
Authors: Goodell Vivian , McNeel Douglas , Disis Mary L
Published: May 2008

Background

The application of high throughput molecular techniques such as SEREX are resulting in the identification of a multitude of tumor associated antigens. As newly identified antigens are incorporated into a variety of clinical trials, standardization of immunologic monitoring methods becomes increasingly important. We questioned whether mammalian cell expression of a histadine-linked human protein could be used to produce antigen suitable for detecting tumor-specific humoral immunity and whether such an assay could be amenable to standardization for clinical use.

Methods

We designed a his-tagged capture ELISA based on lysate from genetically engineered CHO cells for detection of antibodies to insulin-like growth factor binding protein 2, a novel tumor antigen. We performed technical and preliminary clinical validation studies, including comparison to a standard indirect ELISA based on commercially prepared recombinant antigen.

Results

The his-tagged capture ELISA could be standardized. Precision experiments resulted in CVs < 15%. Linearity and calibration experiments demonstrated r2 values of 0.99. In comparison to Western blot analysis, his-tag and indirect ELISA accurately identified 88% and 93% of samples, respectively. Sample concordance between capture and indirect assays was highly significant (p = 0.003). Furthermore, significantly greater levels of IGFBP-2 antibody immunity were found in cancer patients compared to normal controls (p = 0.008).

Conclusion

A genetically engineered cell lysate based ELISA can be amenable to standardization and can detect increased levels of antibody immunity to tumor-associated antigen in cancer patients compared to non tumor-bearing healthy controls.
Oil palm (Elaeis guineensisJacq.) tissue culture ESTs: Identifying genes associated with callogenesis and embryogenesis
Authors: Low Eng-Ti L , Alias Halimah , Boon Soo-Heong , Shariff Elyana M , Tan Chi-Yee A , Ooi Leslie CL , Cheah Suan-Choo , Raha Abdul-Rahim , Wan Kiew-Lian , Singh Rajinder
Published: May 2008

Background

Oil palm (Elaeis guineensis Jacq.) is one of the most important oil bearing crops in the world. However, genetic improvement of oil palm through conventional breeding is extremely slow and costly, as the breeding cycle can take up to 10 years. This has brought about interest in vegetative propagation of oil palm. Since the introduction of oil palm tissue culture in the 1970s, clonal propagation has proven to be useful, not only in producing uniform planting materials, but also in the development of the genetic engineering programme. Despite considerable progress in improving the tissue culture techniques, the callusing and embryogenesis rates from proliferating callus cultures remain very low. Thus, understanding the gene diversity and expression profiles in oil palm tissue culture is critical in increasing the efficiency of these processes.

Results

A total of 12 standard cDNA libraries, representing three main developmental stages in oil palm tissue culture, were generated in this study. Random sequencing of clones from these cDNA libraries generated 17,599 expressed sequence tags (ESTs). The ESTs were analysed, annotated and assembled to generate 9,584 putative unigenes distributed in 3,268 consensi and 6,316 singletons. These unigenes were assigned putative functions based on similarity and gene ontology annotations. Cluster analysis, which surveyed the relatedness of each library based on the abundance of ESTs in each consensus, revealed that lipid transfer proteins were highly expressed in embryogenic tissues. A glutathione S-transferase was found to be highly expressed in non-embryogenic callus. Further analysis of the unigenes identified 648 non-redundant simple sequence repeats and 211 putative full-length open reading frames.

Conclusion

This study has provided an overview of genes expressed during oil palm tissue culture. Candidate genes with expression that are modulated during tissue culture were identified. However, in order to confirm whether these genes are suitable as early markers for embryogenesis, the genes need to be tested on earlier stages of tissue culture and a wider range of genotypes. This collection of ESTs is an important resource for genetic and genome analyses of the oil palm, particularly during tissue culture development.