bluuebunny/crossref_metadata_2025 huggingface dataset:
Lineage and Morphogenetic Analysis of the Cardiac Valves
Authors: de Lange Frederik J. , Moorman Antoon F.M. , Anderson Robert H. , Männer Jörg , Soufan Alexandre T. , Vries Corrie de Gier-de , Schneider Michael D. , Webb Sandra , van den Hoff Maurice J.B. , Christoffels Vincent M.
Published: August 2004

We used a genetic lineage-labeling system to establish the material contributions of the progeny of 3 specific cell types to the cardiac valves. Thus, we labeled irreversibly the myocardial (αMHC-Cre+), endocardial (Tie2-Cre+), and neural crest (Wnt1-Cre+) cells during development and assessed their eventual contribution to the definitive valvar complexes. The leaflets and tendinous cords of the mitral and tricuspid valves, the atrioventricular fibrous continuity, and the leaflets of the outflow tract valves were all found to be generated from mesenchyme derived from the endocardium, with no substantial contribution from cells of the myocardial and neural crest lineages. Analysis of chicken-quail chimeras revealed absence of any substantial contribution from proepicardially derived cells. Molecular and morphogenetic analysis revealed several new aspects of atrioventricular valvar formation. Marked similarities are seen during the formation of the mural leaflets of the mitral and tricuspid valves. These leaflets form by protrusion and growth of a sheet of atrioventricular myocardium into the ventricular lumen, with subsequent formation of valvar mesenchyme on its surface rather than by delamination of lateral cushions from the ventricular myocardial wall. The myocardial layer is subsequently removed by the process of apoptosis. In contrast, the aortic leaflet of the mitral valve, the septal leaflet of the tricuspid valve, and the atrioventricular fibrous continuity between these valves develop from the mesenchyme of the inferior and superior atrioventricular cushions. The tricuspid septal leaflet then delaminates from the muscular ventricular septum late in development.
Critical Roles for the Fas/Fas Ligand System in Postinfarction Ventricular Remodeling and Heart Failure
Authors: Li Yiwen , Takemura Genzou , Kosai Ken-ichiro , Takahashi Tomoyuki , Okada Hideshi , Miyata Shusaku , Yuge Kentaro , Nagano Satoshi , Esaki Masayasu , Khai Ngin Cin , Goto Kazuko , Mikami Atsushi , Maruyama Rumi , Minatoguchi Shinya , Fujiwara Takako , Fujiwara Hisayoshi
Published: August 2004

In myocardial infarction (MI), granulation tissue cells disappear via apoptosis to complete a final scarring with scanty cells. Blockade of this apoptosis was reported to improve post-MI ventricular remodeling and heart failure. However, the molecular biological mechanisms for the apoptosis are unknown. Fas and Fas ligand were overexpressed in the granulation tissue at the subacute stage of MI (1 week after MI) in mice, where apoptosis frequently occurred. In mice lacking functioning Fas (
lpr
strain) and in those lacking Fas ligand (
gld
strain), apoptotic rate of granulation tissue cells was significantly fewer compared with that of genetically controlled mice, and post-MI ventricular remodeling and dysfunction were greatly attenuated. Mice were transfected with adenovirus encoding soluble Fas (sFas), a competitive inhibitor of Fas ligand, on the third day of MI. The treatment resulted in suppression of granulation tissue cell apoptosis and produced a thick, cell-rich infarct scar containing rich vessels and bundles of smooth muscle cells with a contractile phenotype at the chronic stage (4 weeks after MI). This accompanied not only alleviation of heart failure but also survival improvement. However, the sFas gene delivery during scar tissue phase was ineffective, suggesting that beneficial effects of the sFas gene therapy owes to inhibition of granulation tissue cell apoptosis. The Fas/Fas ligand interaction plays a critical role for granulation tissue cell apoptosis after MI. Blockade of this apoptosis by interfering with the Fas/Fas ligand interaction may become one of the therapeutic strategies against chronic heart failure after large MI.
The
small heart
Mutation Reveals Novel Roles of Na
+
/K
+
-ATPase in Maintaining Ventricular Cardiomyocyte Morphology and Viability in Zebrafish
Authors: Yuan Shipeng , Joseph Elaine M.
Published: August 2004

Forward genetic screens in zebrafish have been used to identify mutations in genes with important roles in organogenesis. One of these mutants,
small heart
, develops a diminutive and severely malformed heart and multiple developmental defects of the brain, ears, eyes, and kidneys. Using a positional cloning approach, we identify that the mutant gene encodes the zebrafish Na
+
/K
+
-ATPase α1B1 protein. Disruption of Na
+
/K
+
-ATPase α1B1 function via morpholino “knockdown” or pharmacological inhibition with ouabain phenocopies the mutant phenotype, in a dose-dependent manner. Heterozygosity for the mutation sensitizes embryos to ouabain treatment. Our findings present novel genetic and morphological details on the function of the

Na
+
/K
+
-ATPase α1B1

in early cardiac morphogenesis and the pathogenesis of the small heart malformation. We demonstrate that the reduced size of the mutant heart is caused by dysmorphic ventricular cardiomyocytes and an increase in ventricular cardiomyocyte apoptosis. This study provides a new insight that Na
+
/K
+
-ATPase α1B1 is required for maintaining ventricular cardiomyocyte morphology and viability.
Functional MRI Follow-Up Study of Language Processes in Healthy Subjects and During Recovery in a Case of Aphasia
Authors: Fernandez Bruno , Cardebat Dominique , Demonet Jean-François , Joseph Pierre Alain , Mazaux Jean-Michel , Barat Michel , Allard Michèle
Published: August 2004

Background and Purpose—

The goal of this study was to develop a functional MRI (fMRI) paradigm robust and reproducible enough in healthy subjects to be adapted for a follow-up study aiming at evaluating the anatomical substratum of recovery in poststroke aphasia.

Methods—

Ten right-handed subjects were studied longitudinally using fMRI (7 of them being scanned twice) and compared with a patient with conduction aphasia during the first year of stroke recovery.

Results—

Controls exhibited reproducible activation patterns between subjects and between sessions during language tasks. In contrast, the patient exhibited dynamic changes in brain activation pattern, particularly in the phonological task, during the 2 fMRI sessions. At 1 month after stroke, language homotopic right areas were recruited, whereas large perilesional left involvement occurred later (12 months).

Conclusions—

We first demonstrate intersubject robustness and intrasubject reproducibility of our paradigm in 10 healthy subjects and thus its validity in a patient follow-up study over a stroke recovery time course. Indeed, results suggest a spatiotemporal poststroke brain reorganization involving both hemispheres during the recovery course, with an early implication of a new contralateral functional neural network and a later implication of an ipsilateral one.
Increased Expression of Transforming Growth Factor-β1 as a Stabilizing Factor in Human Atherosclerotic Plaques
Authors: Cipollone Francesco , Fazia Maria , Mincione Gabriella , Iezzi Annalisa , Pini Barbara , Cuccurullo Chiara , Ucchino Sante , Spigonardo Francesco , Di Nisio Marcello , Cuccurullo Franco , Mezzetti Andrea , Porreca Ettore
Published: August 2004

Background and Purpose—

Transforming growth factor-β (TGF-β) is a growth factor/cytokine involved in vascular remodeling and atherogenesis. Recent studies in apolipoprotein E-deficient mice have demonstrated a pivotal role of TGF-β in the maintenance of the balance between inflammation and fibrosis in atherosclerotic plaques. Furthermore, inhibition of TGF-β signaling has been shown to accelerate plaque formation and its progression toward an unstable phenotype in mice. However, if this mechanism is operative also in humans is still unknown. The aim of this study was to characterize the expression of TGF-β1 in human carotid plaque and to correlate it with the extent of inflammatory infiltration and collagen content with the clinical signs of plaque instability.

Methods—

Plaques were obtained from patients undergoing carotid endoarterectomy and divided into symptomatic and asymptomatic according to clinical evidence of recent transient ischemic attack or stroke. Plaques were analyzed for TGF-β1 expression by Immunocytochemistry, Western, and Northern blotting analysis. Immunocytochemistry was used to identify CD68
+
macrophages, CD3 T lymphocytes, HLA-DR
+
cells, and α-smooth muscle cells. Procollagen and interstitial collagen content were analyzed by immunohistochemistry and Sirius Red staining, respectively.

Results—

Plaque TGF-β1 mRNA was increased up to 3-fold in asymptomatic as compared with symptomatic plaques. Plaques from asymptomatic group had fewer (
P
<0.0001) macrophages and T lymphocytes compared with symptomatic plaques. TGF-β1 gene was transcriptionally active as demonstrated by increased (
P
<0.0001) TGF-β1 protein expression in asymptomatic plaques. Immunohistochemistry showed that TGF-β was mainly expressed in plaque shoulder and was associated with a comparable increase (
P
<0.0001) in plaque procollagen and collagen content.

Conclusions—

In conclusion, this study demonstrates the higher expression of TGF-β1 in human asymptomatic lesions and provides evidence that TGF-β1 may play an important role in the process of plaque stabilization.
Ko Hung's Discourse of Hsien-Immortality: A Taoist Configuration of an Alternate Ideal Self-Identity
Authors: Lai Chi-Tim
Published: August 2004

This study seeks to investigate the ideological base underlying Ko Hung's new Taoist discourse of Hsien-immortality in early Six Dynasties China. Our analysis will show the symbolic and discursive complex of Ko Hung's Hsien-immortality, his self-understanding, and his reappropriation of the ancient Chinese religious tradition of physical immortality, in particular in relation to his own emerging historical consciousness. In so doing, it can serve as one of the keys to the understanding of an early Taoist master's religious discourse of Hsien-immortality which functions as an organizing principle that orders the way he experiences the social and cultural world, as an ideological resolution for his feelings of incongruence between outer reality and inner world, and, finally, as a soteriological vision of an alternate ideal self-identity in contrast to the Han idealized Confucian sage. The construction of Taoist discourse of Hsien-identity, instanced in Ko Hung's Pao-p'u tzu nei-p'ien, should, therefore, not be seen as something abstract and arbitrary without the roots in socio-cultural reality, but a new formation and shape for the Six Dynasties literati of an alternate idealized self-identity.
Close Harmonies: The Science of Religion in Dutch Duplex Ordo Theology, 1860-1960
Authors: Platvoet Jan
Published: August 2004

In this article, the history of the Science of Religion in the Netherlands in the period 1860 to 1960 is surveyed at the time when it was an integral part of Dutch liberal academic theology as pursued in the faculties of theology at the universities of Leiden, Groningen, Utrecht and Amsterdam. In 1876, these faculties were given a special statute, the so-called Duplex Ordo, in a law that separated the ‘confessional’ theological disciplines from the ‘scientific’ ones. It also introduced the new disciplines of the Science of Religion and the Philosophy of Religion into these reconstituted faculties. I discuss Tiele's plan to make the Science of Religion their central discipline, and why it was ultimately given only a marginal place in them. My main concern, however, is to outline the theology which inspired the Science of Religion of Tiele, Chantepie, Van der Leeuw and Bleeker and to demonstrate its ‘close harmony’ with the liberal theology prevailing in these Duplex Ordo faculties, as also in at least some of the modalities of the Nederlandse Hervormde Kerk whose ministers were trained, again by law, in these faculties. It was that close harmony which allowed Van der Leeuw to disregard the Duplex Ordo and to establish a full harmony between the Science of Religion and confessional theology. I also discuss dissonant voices, Kraemer's especially, calling for the abrogation of the Duplex Ordo and the integration of the Science of Religion into a militantly confessional theology.
How Do You Say “God” in Dakota?
Authors: Siems Monica
Published: August 2004

This essay explores some of the interpretive problems posed by the missionary work of the American Board of Commissioners for Foreign Missions among Minnesota's Dakota Indians in the mid-nineteenth century. Since “success” in conversion has often been defined largely in terms of Native American assimilation to white American societal norms, I argue that the Dakotas' acceptance of a Christianity that was preached to them in their own language poses a problem for understanding what “conversion” meant in this case. The difficulty of knowing what Christianity meant to the Dakotas, a difficulty I see as rooted in the problems attendant upon translation, can be illustrated by showing how distorted some of the missionaries' translations of Dakota religious ideas into English were. Their translation errors took two forms: the postulation of a similarity in reference between Dakota and English terms where it did not exist (as with the English words “god” and “spirit” and what the missionaries identified as their Dakota counterparts), or, conversely, the heightening of differences between Dakota and Protestant Christian religiosity (an effect achieved by associating Dakota terms with English pejoratives). I argue that what unites these two differing types of distortive translations is the missionaries' failure to understand the conceptual universe, or “deep structure”, underlying the Dakota language. And since they failed to perceive the holistic character of this conceptual system, the missionaries may have concluded that they had effected conversions among the Dakotas when they had only scratched the surface of Dakota religiosity.
Job: another new thesis
Authors: Caesar Lael
Published: August 2004

In 1996, Andrew E. Steinmann proposed that integrity and faith, not theodicy and suffering, were the heart of the book of Job ("The Structure & Message of the Book of Job", VT 46, pp. 85-100). I do not dismiss the element of theodicy, but view the issue of integrity as critical to appreciating the book's split level setting (heaven and earth), its dual focus (divine and human), and its prose-poetry, frame-and-center, integration. Exchanges between Job and "friends" connect to the book's prologue as the human contribution to discussion of its earliest stated theme, human integrity. The epilogue's divine verdict (xlii 7, 8) brings closure upon this question which opens the book and provokes its drama. This essay focuses on the book's juxtaposition of Job and Eliphaz as illustrative of contrasting postures with regard to human integrity, a subject too long overshadowed, in Job scholarship, by questions on theodicy and suffering.
The comparative optative: a Homeric reminiscence in the Greek Pentateuch?
Authors: Evans Trevor
Published: August 2004

The potential optative subordinated in a clause of comparison is extremely rare in extra-Biblical Greek, though found already in Homeric Epic. In the Septuagint it is relatively frequent. There are nine examples in the third century B.C. Greek Pentateuch and a further nine in later books. It will inevitably be suspected that some sort of Hebraistic influence on these translation Greek documents prompts the usage. Yet analysis of the comparative optative's relationship to text components in the underlying Hebrew reveals no specific motivation from that quarter. We are dealing with an independent Greek phenomenon. The argument of this paper, based on consideration of a large sample of Ancient Greek, is that Homeric reminiscence, far fetched as it must seem prima facie, offers the likeliest explanation of the Pentateuchal usage.
Were the Ephraimites to Blame?
Authors: Gosling
Published: August 2004

The article is concerned with a textual and historical examination of Ps. lxxviii 9. It begins by considering the history of exegesis of this Psalm, after which it considers in depth the only attested textual variant to the reading of the verb n⋅q, namely, nq⋅. Having concluded the investigation of the textual variant, the work then carries on to consider the reliability and integrity of the Hebrew text. It achieves this objective by a consideration of 11QPsd, which is followed by a consideration of the Septuagint's text. Next in turn for an evaluation of its contribution is the text of the Targum, which is considered at length. Since Rashi often bases his exegesis on the Targum it has been thought best to consider his exegesis of this difficult verse. The final part of the article is concerned with the cumulative assessment of all the pieces of textual evidence thus considered.
The stories of the family troubles of Judah and David: a study of their literary links
Authors: Ho Craig
Published: August 2004

This article adds some newly discovered literary links between Gen. xxxviii and the Succession Narrative (SN) to those already observed by scholars in the past thirty or so years. Gen. xxxviii is found to be dependent on and owes to the SN most of its literary components like names, plot, theme, motifs and special locutions. The literary conclusion arrived, contrary to the widely accepted traditio-historical view, is that Gen. xxxviii has no oral tradition stage before being written down. Further analysis of the story does not confirm its belonging to the J source but re-affirms its genealogical concern. The article concludes with a conjectured raison d'être for Gen. xxxviii, namely to provide a "narrative evidence" for David's genealogical link to Judah, i.e. to prove David's Jewishness. It is further conjectured that Gen. xxxviii was probably written about the time of the emergence of the book of Ruth, which shows similar concern.
The case of the blasphemer revisited (Lev. xxiv 10-23)
Authors: Hutton Rodney
Published: August 2004

The case of the "blasphemer" in Lev. xxiv 10-23 is complicated by the ambiguity of the offense. The expressions "blaspheme the name" (nāqab 'et-haššēm) and "curse" (qillēl) have been translated in a wide variety of ways. The arguments are reviewed and the proposal made that the offense was that the offender used the divine name in an illegitimate manner with the result that the name was degraded and dishonored.
The nucleotide transporter MRP4 (ABCC4) is highly expressed in human platelets and present in dense granules, indicating a role in mediator storage
Authors: Jedlitschky Gabriele , Tirschmann Konstanze , Lubenow Lena E. , Nieuwenhuis Hendrik K. , Akkerman Jan W. N. , Greinacher Andreas , Kroemer Heyo K.
Published: August 2004

Platelet aggregation is initiated by the release of mediators as adenosine diphosphate (ADP) stored in platelet granules. Possible candidates for transport proteins mediating accumulation of these mediators in granules include multidrug resistance protein 4 (MRP4, ABCC4), a transport pump for cyclic nucleotides and nucleotide analogs. We investigated the expression of MRP4 in human platelets by immunoblotting, detecting a strong signal at 170 kDa. Immunofluorescence microscopy using 2 MRP4-specific antibodies revealed staining mainly in intracellular structures, which largely colocalized with the accumulation of mepacrine as marker for delta-granules and to a lower extent at the plasma membrane. Furthermore, an altered distribution of MRP4 was observed in platelets from a patient with Hermansky-Pudlak syndrome with defective delta-granules. Adenosine triphosphate (ATP)–dependent cyclic guanosine monophosphate (cGMP) transport codistributed with MRP4 detection in subcellular fractions, with highest activities in the dense granule and plasma membrane fractions. This transport was inhibited by dipyramidole, indomethacin, and MK571 with median inhibitory concentration (IC50) values of 12, 22, and 43 μM, and by ibuprofen. Transport studies with [3H]ADP indicated the presence of an orthovanadate-sensitive ADP transporting system, inhibited by dipyramidole, MK571, and cyclic nucleotides. The results indicate a function of MRP4 in platelet mediator storage and inhibition of MRP4 may represent a novel mechanism for inhibition of platelet function by some anti-inflammatory drugs.
Haploinsufficiency of AML1 results in a decrease in the number of LTR-HSCs while simultaneously inducing an increase in more mature progenitors
Authors: Sun Weili , Downing James R.
Published: August 2004

The AML1/CBFβ transcriptional complex is essential for the formation of definitive hematopoietic stem cells (HSCs). Moreover, development of the hematopoietic system is exquisitely sensitive to the level of this complex. To investigate the effect of AML1 dosage on adult hematopoiesis, we compared the hematopoietic systems of AML1+/– and AML1+/+ mice. Surprisingly, loss of a single AML1 allele resulted in a 50% reduction in long-term repopulating hematopoietic stem cells (LTR-HSCs). This decrease did not, however, extend to the next level of hematopoietic differentiation. Instead, AML1+/– mice had an increase in multilineage progenitors, an expansion that resulted in enhanced engraftment following transplantation. The expanded pool of AML1+/– progenitors remained responsive to homeostatic mechanisms and thus the number of mature cells in most lineages remained within normal limits. Two notable exceptions were a decrease in CD4+ T cells, leading to an inversion of the CD4+ to CD8+ T-cell ratio and a decrease in circulating platelets. These data demonstrate a dosage-dependent role for AML1/CBFβ in regulating the quantity of HSCs and their downstream committed progenitors, as well as a more restricted role in T cells and platelets. The latter defect mimics one of the key abnormalities in human patients with the familial platelet disorder resulting from AML1 haploinsufficiency.
CLEVER-1 mediates lymphocyte transmigration through vascular and lymphatic endothelium
Authors: Salmi Marko , Koskinen Kaisa , Henttinen Tiina , Elima Kati , Jalkanen Sirpa
Published: August 2004

Common lymphatic endothelial and vascular endothelial receptor-1 (CLEVER-1; also known as stabilin-1 or FEEL-1) is a large multifunctional glycoprotein implicated in scavenging, angiogenesis, and cell adhesion. Here we studied the function of human CLEVER-1 in leukocyte trafficking. Lymphatic vessels expressed CLEVER-1 constitutively in skin in vivo, whereas on vascular endothelium it appeared only upon inflammation. On isolated vascular endothelial cells, CLEVER-1 supported rolling and transmigration of peripheral blood mononuclear cells (PBMCs) under physiologically relevant laminar shear stress. Intriguingly, CLEVER-1 also mediated transmigration of leukocytes through cultured lymphatic endothelium under static conditions. Thus, synthesis of CLEVER-1 is differentially regulated on the 2 anatomically distinct vascular beds, and CLEVER-1 mediates the transmigration step of the leukocyte traffic in both of them. Notably, CLEVER-1 is the first adhesion molecule shown to be involved in the PBMC transmigration through the lymphatic arm of the immune system. (Blood. 2004;104:3849-3857)
Anaplastic large cell lymphomas lack the expression of T-cell receptor molecules or molecules of proximal T-cell receptor signaling
Authors: Bonzheim Irina , Geissinger Eva , Roth Sabine , Zettl Andreas , Marx Alexander , Rosenwald Andreas , Müller-Hermelink Hans Konrad , Rüdiger Thomas
Published: August 2004

Anaplastic large cell lymphoma (ALCL) designates a heterogeneous group of CD30+ (systemic or primary cutaneous) peripheral T-cell lymphomas (PTCLs). A subgroup of systemic ALCL is transformed by anaplastic lymphoma kinase (ALK). We compared 24 ALK+, 15 ALK- systemic, and 7 cutaneous ALCLs with 29 nonanaplastic PTCLs in terms of T-cell receptor (TCR) rearrangements, expression of TCRs and TCR-associated molecules (CD3, ZAP-70 [zeta-associated protein 70]). Despite their frequent clonal rearrangement for TCRβ, only 2 (4%) of 47 ALCLs expressed TCRβ protein, whereas TCRs were detected on 27 of 29 nonanaplastic PTCLs. Moreover, both TCRβ+ ALCLs lacked CD3 and ZAP-70 (ie, molecules indispensable for the transduction of cognate TCR signals). Defective expression of TCRs is a common characteristic of all types of ALCL, which may contribute to the dysregulation of intracellular signaling pathways controlling T-cell activation and survival. This molecular hallmark of ALCL is analogous to defective immunoglobulin expression distinguishing Hodgkin lymphoma from other B-cell lymphomas. (Blood. 2004; 104:3358-3360)
A shortened activated partial thromboplastin time is associated with the risk of venous thromboembolism
Authors: Tripodi Armando , Chantarangkul Veena , Martinelli Ida , Bucciarelli Paolo , Mannucci Pier Mannuccio
Published: August 2004

Hypercoagulability due to high coagulation factors XI, VIII, IX, II, and fibrinogen is recognized as a risk factor of venous thromboembolism (VTE). These factors are cumulatively explored by the activated partial thromboplastin time (APTT). To test the hypothesis that a short APTT increases the risk of VTE, a case-control study was carried out in 605 patients referred for thrombophilia testing after documented VTE and in 1290 controls. Median APTT ratio (coagulation time of test-to-reference plasma) values were 0.97 (range: 0.75-1.41) for patients and 1.00 (range: 0.72-1.33) for controls (P < .001). In patients who had an APTT ratio smaller than the fifth percentile of the distribution in controls, the odds ratio (OR) for VTE was 2.4 (95% confidence interval [CI]: 1.7-3.6) and was independent of inherited thrombophilic abnormalities. Further statistical analyses in 193 patients and 259 controls for whom factor VIII (FVIII) levels were available showed a decrease of the OR from 2.7 (95% CI: 1.4-5.3) to 2.1 (95% CI: 1.0-4.2), indicating that the risk was only partially mediated by high FVIII levels. In conclusion, hypercoagulability detected by a shortened APTT is independently associated with VTE. This inexpensive and simple test should be considered in the evaluation of the risk of VTE.
Stimulation of endothelial cell proliferation by FGF-2 in the presence of fibrinogen requires αvβ3
Authors: Sahni Abha , Francis Charles W.
Published: August 2004

We have shown previously that fibrin(ogen) binding potentiates the capacity of fibroblast growth factor 2 (FGF-2) to stimulate endothelial cell (EC) proliferation. We have now investigated the receptor requirement for EC proliferation by fibrinogen-bound FGF-2. ECs were cultured with 25 ng/mL FGF-2 with or without 10 μg/mL fibrinogen, and proliferation was measured as 3H-thymidine incorporation. Proliferation was increased 2.4 ± 0.5-fold over medium alone with FGF-2 and increased significantly more to 4.0 ± 0.7-fold with fibrinogen and FGF-2 (P < .005). Addition of 7E3 or LM609, antibodies to αvβ3, inhibited EC proliferation with fibrinogen-bound FGF-2 by 80% ± 8% (P < .001) or 67% ± 14% (P < .002), respectively, to levels significantly less than that observed with FGF-2 alone (P < .001). Neither LM609 nor 7E3 exhibited any inhibition of activity with FGF-2 alone. Peptide GRGDS caused dose-dependent inhibition of proliferation by fibrinogen-bound FGF-2 of 31% ± 8%, 45% ± 9%, and 68% ± 11% at 0.25, 0.5, and 1 mM, respectively. Coimmunoprecipitation and immunofluorescence studies demonstrated a direct specific association between αvβ3 and FGF receptor 1 (FGFR1) in ECs and fibroblasts when exposed to both FGF-2 and fibrinogen but not with vitronectin. We conclude that fibrinogen binding of FGF-2 enhances EC proliferation through the coordinated effects of colocalized αvβ3 and FGFR1.
Nonhematopoietic mesenchymal stem cells can be mobilized and differentiate into cardiomyocytes after myocardial infarction
Authors: Kawada Hiroshi , Fujita Jun , Kinjo Kentaro , Matsuzaki Yumi , Tsuma Mitsuyo , Miyatake Hiroko , Muguruma Yukari , Tsuboi Kosuke , Itabashi Yuji , Ikeda Yasuo , Ogawa Satoshi , Okano Hideyuki , Hotta Tomomitsu , Ando Kiyoshi , Fukuda Keiichi
Published: August 2004

Bone marrow (BM) cells are reported to contribute to the process of regeneration following myocardial infarction. However, the responsible BM cells have not been fully identified. Here, we used 2 independent clonal studies to determine the origin of bone marrow (BM)–derived cardiomyocytes. First, we transplanted single CD34– c-kit+Sca-1+ lineage– side population (CD34–KSL-SP) cells or whole BM cells from mice ubiquitously expressing enhanced green fluorescent protein (EGFP) into lethally irradiated mice, induced myocardial infarction (MI), and treated the animals with granulocyte colony-stimulating factor (G-CSF) to mobilize stem cells to the damaged myocardium. At 8 weeks after MI, from 100 specimens we counted only 3 EGFP+ actinin+ cells in myocardium of CD34– KSL-SP cells in mice that received transplants, but more than 5000 EGFP+ actinin+ cells in whole BM cell in mice that received transplants, suggesting that most of EGFP+ actinin+ cells were derived from nonhematopoietic BM cells. Next, clonally purified nonhematopoietic mesenchymal stem cells (MSCs), cardiomyogenic (CMG) cells, that expressed EGFP in the cardiomyocyte-specific manner were transplanted directly into BM of lethally irradiated mice, MI was induced, and they were treated with G-CSF. EGFP+ actinin+ cells were observed in the ischemic myocardium, indicating that CMG cells had been mobilized and differentiated into cardiomyocytes. Together, these results suggest that the origin of the vast majority of BM-derived cardiomyocytes is MSCs.
Combined disruption of both the MEK/ERK and the IL-6R/STAT3 pathways is required to induce apoptosis of multiple myeloma cells in the presence of bone marrow stromal cells
Authors: Chatterjee Manik , Stühmer Thorsten , Herrmann Pia , Bommert Kurt , Dörken Bernd , Bargou Ralf C.
Published: August 2004

The interleukin-6 receptor (IL-6R)/signal transducer and activator of transcription 3 (STAT3) pathway contributes to the pathogenesis of multiple myeloma (MM) and protects MM cells from apoptosis. However, MM cells survive the IL-6R blockade if they are cocultured with bone marrow stromal cells (BMSCs), suggesting that the BM microenvironment stimulates IL-6–independent pathways that exert a pro-survival effect. The goal of this study was to investigate the underlying mechanism. Detailed pathway analysis revealed that BMSCs stimulate STAT3 via the IL-6R, and mitogen-activated protein (MAP) kinases via IL-6R–independent mechanisms. Abolition of MEK1,2 activity with PD98059, or ERK1,2 small interfering RNA knockdown, was insufficient to induce apoptosis. However, the combined disruption of the IL-6R/STAT3 and MEK1,2/ERK1,2 pathways led to strong induction of apoptosis even in the presence of BMSCs. This effect was observed with MM cell lines and with primary MM cells, suggesting that the BMSC-induced activation of MEK1,2/ERK1,2 renders MM cells IL-6R/STAT3 independent. Therefore, in the presence of cells from the BM micro-environment, combined targeting of different (and independently activated) pathways is required to efficiently induce apoptosis of MM cells. This might have direct implications for the development of future therapeutic strategies for MM.
Organization of the mitochondrial genomes of whiteflies, aphids, and psyllids (Hemiptera, Sternorrhyncha)
Authors: Thao MyLo L , Baumann Linda , Baumann Paul
Published: August 2004

Background

With some exceptions, mitochondria within the class Insecta have the same gene content, and generally, a similar gene order allowing the proposal of an ancestral gene order. The principal exceptions are several orders within the Hemipteroid assemblage including the order Thysanoptera, a sister group of the order Hemiptera. Within the Hemiptera, there are available a number of completely sequenced mitochondrial genomes that have a gene order similar to that of the proposed ancestor. None, however, are available from the suborder Sternorryncha that includes whiteflies, psyllids and aphids.

Results

We have determined the complete nucleotide sequence of the mitochondrial genomes of six species of whiteflies, one psyllid and one aphid. Two species of whiteflies, one psyllid and one aphid have mitochondrial genomes with a gene order very similar to that of the proposed insect ancestor. The remaining four species of whiteflies had variations in the gene order. In all cases, there was the excision of a DNA fragment encoding for cytochrome oxidase subunit III(COIII)-tRNAgly-NADH dehydrogenase subunit 3(ND3)-tRNAala-tRNAarg-tRNAasn from the ancestral position between genes for ATP synthase subunit 6 and NADH dehydrogenase subunit 5. Based on the position in which all or part of this fragment was inserted, the mitochondria could be subdivided into four different gene arrangement types. PCR amplification spanning from COIII to genes outside the inserted region and sequence determination of the resulting fragments, indicated that different whitefly species could be placed into one of these arrangement types. A phylogenetic analysis of 19 whitefly species based on genes for mitochondrial cytochrome b, NADH dehydrogenase subunit 1, and 16S ribosomal DNA as well as cospeciating endosymbiont 16S and 23S ribosomal DNA indicated a clustering of species that corresponded to the gene arrangement types.

Conclusions

In whiteflies, the region of the mitochondrial genome consisting of genes encoding for COIII-tRNA
gly
-ND3-tRNA
ala
-tRNA
arg
-tRNA
asn
can be transposed from its ancestral position to four different locations on the mitochondrial genome. Related species within clusters established by phylogenetic analysis of host and endosymbiont genes have the same mitochondrial gene arrangement indicating a transposition in the ancestor of these clusters.
Comparison of the accuracy of methods of computational haplotype inference using a large empirical dataset
Authors: Adkins Ronald M
Published: August 2004

Background

Analyses of genetic data at the level of haplotypes provide increased accuracy and power to infer genotype-phenotype correlations and evolutionary history of a locus. However, empirical determination of haplotypes is expensive and laborious. Therefore, several methods of inferring haplotypes from unphased genotypic data have been proposed, but it is unclear how accurate each of the methods is or which methods are superior. The accuracy of some of the leading methods of computational haplotype inference (PL-EM, Phase, SNPHAP, Haplotyper) are compared using a large set of 308 empirically determined haplotypes based on 15 SNPs, among which 36 haplotypes were observed to occur. This study presents several advantages over many previous comparisons of haplotype inference methods: a large number of subjects are included, the number of known haplotypes is much smaller than the number of chromosomes surveyed, a range in values of linkage disequilibrium, presence of rare SNP alleles, and considerable dispersion in the frequencies of haplotypes.

Results

In contrast to some previous comparisons of haplotype inference methods, there was very little difference in the accuracy of the various methods in terms of either assignment of haplotypes to individuals or estimation of haplotype frequencies. Although none of the methods inferred all of the known haplotypes, the assignment of haplotypes to subjects was about 90% correct for individuals heterozygous for up to three SNPs and was about 80% correct for up to five heterozygous sites. All of the methods identified every haplotype with a frequency above 1%, and none assigned a frequency above 1% to an incorrect haplotype.

Conclusions

All of the methods of haplotype inference have high accuracy and one can have confidence in inferences made by any one of the methods. The ability to identify even rare (≥ 1%) haplotypes is reassuring for efforts to identify haplotypes that contribute to disease in a significant proportion of a population. Assignment of haplotypes is relatively accurate among subjects heterozygous for up to 5 sites, and this might be the largest number of SNPs for which one should define haplotype blocks or have confidence in haplotype assignments.
Immobilized probe and glass surface chemistry as variables in microarray fabrication
Authors: Hessner Martin J , Meyer Lisa , Tackes Jennifer , Muheisen Sanaa , Wang Xujing
Published: August 2004

Background

Global gene expression studies with microarrays can offer biological insights never before possible. However, the technology possesses many sources of technical variability that are an obstacle to obtaining high quality data sets. Since spotted microarrays offer design/content flexibility and potential cost savings over commercial systems, we have developed prehybridization quality control strategies for spotted cDNA and oligonucleotide arrays. These approaches utilize a third fluorescent dye (fluorescein) to monitor key fabrication variables, such as print/spot morphology, DNA retention, and background arising from probe redistributed during blocking. Here, our labeled cDNA array platform is used to study, 1) compression of array data using known input ratios of Arabidopsis in vitro transcripts and arrayed serial dilutions of homologous probes; 2) how curing time of in-house poly-L-lysine coated slides impacts probe retention capacity; and 3) the retention characteristics of 13 commercially available surfaces.

Results

When array element fluorescein intensity drops below 5,000 RFU/pixel, gene expression measurements become increasingly compressed, thereby validating this value as a prehybridization quality control threshold. We observe that the DNA retention capacity of in-house poly-L-lysine slides decreases rapidly over time (~50% reduction between 3 and 12 weeks post-coating; p < 0.0002) and that there are considerable differences in retention characteristics among commercially available poly-L-lysine and amino silane-coated slides.

Conclusions

High DNA retention rates are necessary for accurate gene expression measurements. Therefore, an understanding of the characteristics and optimization of protocols to an array surface are prerequisites to fabrication of high quality arrays.
Short-term cytotoxic effects and long-term instability of RNAi delivered using lentiviral vectors
Authors: Fish Richard J , Kruithof Egbert KO
Published: August 2004

Background

RNA interference (RNAi) can potently reduce target gene expression in mammalian cells and is in wide use for loss-of-function studies. Several recent reports have demonstrated that short double-stranded RNAs (dsRNAs), used to mediate RNAi, can also induce an interferon-based response resulting in changes in the expression of many interferon-responsive genes. Off-target gene silencing has also been described, bringing into question the validity of certain RNAi-based approaches for studying gene function. We have targeted the plasminogen activator inhibitor-2 (PAI-2 or SERPINB2) mRNA using lentiviral vectors for delivery of U6 promoter-driven PAI-2-targeted short hairpin RNA (shRNA) expression. PAI-2 is reported to have anti-apoptotic activity, thus reduction of endogenous expression may be expected to make cells more sensitive to programmed cell death.

Results

As expected, we encountered a cytotoxic phenotype when targeting the PAI-2 mRNA with vector-derived shRNA. However, this predicted phenotype was a potent non-specific effect of shRNA expression, as functional overexpression of the target protein failed to rescue the phenotype. By decreasing the shRNA length or modifying its sequence we maintained PAI-2 silencing and reduced, but did not eliminate, cytotoxicity. ShRNA of 21 complementary nucleotides (21 mers) or more increased expression of the oligoadenylate synthase-1 (OAS1) interferon-responsive gene. 19 mer shRNA had no effect on OAS1 expression but long-term selective pressure on cell growth was observed. By lowering lentiviral vector titre we were able to reduce both expression of shRNA and induction of OAS1, without a major impact on the efficacy of gene silencing.

Conclusions

Our data demonstrate a rapid cytotoxic effect of shRNAs expressed in human tumor cell lines. There appears to be a cut-off of 21 complementary nucleotides below which there is no interferon response while target gene silencing is maintained. Cytotoxicity or OAS1 induction could be reduced by changing shRNA sequence or vector titre, but stable gene silencing could not be maintained in extended cell culture despite persistent marker gene expression from the RNAi-inducing transgene cassette. These results underscore the necessity of careful controls for immediate and long-term RNAi use in mammalian cell systems.
Arrhythmia-provoking factors and symptoms at the onset of paroxysmal atrial fibrillation: A study based on interviews with 100 patients seeking hospital assistance
Authors: Hansson Anders , Madsen-Härdig Bjarne , Bertil Olsson S
Published: August 2004

Background

Surprisingly little information on symptoms of paroxysmal atrial fibrillation is available in scientific literature. Using questionnaires, we have analyzed the symptoms associated with arrhythmia attacks.

Methods

One hundred randomly-selected patients with idiopathic paroxysmal atrial fibrillation filled in a structured questionnaire.

Results

Psychic stress was the most common factor triggering arrhythmia (54%), followed by physical exertion (42%), tiredness (41%) coffee (25%) and infections (22%). Thirty-four patients cited alcohol, 26 in the form of red wine, 16 as white wine and 26 as spirits. Among these 34, red wine and spirits produced significantly more episodes of arrhythmia than white wine (p = 0.01 and 0.005 respectively).

Symptoms during arrhythmia were palpitations while exerting (88%), reduced physical ability (87%), palpitations at rest (86%), shortage of breath during exertion (70%) and anxiety (59%). Significant differences between sexes were noted regarding swollen legs (women 21%, men 6%, p = 0.027), nausea (women 36%, men 13%, p = 0.012) and anxiety (females 79%, males 51%, p = 0.014).

Conclusion

Psychic stress was the commonest triggering factor in hospitalized patients with paroxysmal atrial fibrillation. Red wine and spirits were more proarrhythmic than white wine. Symptoms in women in connection with attacks of arrhythmia vary somewhat from those in men.
Liver, spleen, pancreas and kidney involvement by human fascioliasis: imaging findings
Authors: Zali Mohammad Reza , Ghaziani Tahereh , Shahraz Saeed , Hekmatdoost Azita , Radmehr Ali
Published: August 2004

Background

Fasciola hepatica primarily involves the liver, however in some exceptional situations other organs have been reported to be involved. The ectopic involvement is either a result of Parasite migration or perhaps eosinophilic reaction.

Case presentation

Here we report a known case of multiple myeloma who was under treatment with prednisolone and melphalan. He was infected by Fasciola hepatica, which involved many organs and the lesions were mistaken with metastatic ones.

Discussion

Presented here is a very unusual case of the disease, likely the first case involving the pancreas, spleen, and kidney, as well as the liver.
'Perinatal outcome in preterm premature rupture of membranes with Amniotic fluid index < 5 (AFI < 5)
Authors: Borna Sedigheh , Borna Hajieh , khazardoost Soghra , Hantoushzadeh Sedigheh
Published: August 2004

Background

Our purpose was to determine whether AFI<5 cm after preterm premature rupture of the membranes (PPROM) is associated with an increased risk of perinatal morbidity.

Methods

We performed a prospective cohort study of 95 singleton pregnancies complicated by preterm premature rupture of the membranes (PPROM) with delivery between 26 and 34 weeks gestation.

Patients were categorized in two groups on the basis of amniotic fluid index<5, (AFI<5 cm)(n = 26) or AFI ≥ 5 cm (n = 69). Categorical data were tested for significance with the χ2 and Fisher exact tests. Continuous data were evaluated for normal distribution and tested for significance with the student t test.

All 2-sided p values < 0.05 were considered significant.

Results

Both groups were similar with respect to selected demographics, gestational age at rupture of the membranes, gestational age at the delivery, birth weight. Both groups were similar with respect to selected variable, latency until delivery, early onset neonatal sepsis, RDS and neonatal death. Patients with AFI<5 cm demonstrated greater frequency of C/S delivery for non reassuring fetal tests (23%vs 2.8%) (p = 0.001). Our study demonstrated that patients in group I had a significant increase in the frequency of clinical chorioamnionitis (P < 0/001). Post partum infections were not seen in 2 groups.

Conclusions

An AFI<5 cm after PPROM between 26 and 34 weeks gestation is associated with an increased risk of maternal infections and frequency of C/S.
Estimating the beginning of the waterpipe epidemic in Syria
Authors: Rastam Samer , Ward Kenneth D , Eissenberg Thomas , Maziak Wasim
Published: August 2004

Background

Waterpipe smoking is becoming a global public health problem, especially in the Eastern Mediterranean region (EMR).

Methods

We try in this study, which is a cross sectional survey among a representative sample of waterpipe smokers in cafes/restaurants in Aleppo-Syria, to assess the time period for the beginning of this new smoking hype. We recruited 268 waterpipe smokers (161 men, 107 women; mean age ± standard deviation (SD) 30.1 ± 10.2, response rate 95.3%). Participants were divided into 4 birth cohorts (≤ 1960, 1961–1970, 1971–1980, >1980) and year of initiation of waterpipe smoking and daily cigarette smoking were plotted according to these birth cohorts.

Results

Data indicate that unlike initiation of cigarette smoking, which shows a clear age-related pattern, the nineties was the starting point for most of waterpipe smoking implicating this time period for the beginning of the waterpipe epidemic in Syria.

Conclusion

The introduction of new flavored and aromatic waterpipe tobacco (Maassel), and the proliferation of satellite and electronic media during the nineties may have helped spread the new hype all over the Arab World.
Altered mRNA expression of genes related to nerve cell activity in the fracture callus of older rats: A randomized, controlled, microarray study
Authors: Meyer Martha H , Etienne Wiguins , Meyer Ralph A
Published: August 2004

Background

The time required for radiographic union following femoral fracture increases with age in both humans and rats for unknown reasons. Since abnormalities in fracture innervation will slow skeletal healing, we explored whether abnormal mRNA expression of genes related to nerve cell activity in the older rats was associated with the slowing of skeletal repair.

Methods

Simple, transverse, mid-shaft, femoral fractures with intramedullary rod fixation were induced in anaesthetized female Sprague-Dawley rats at 6, 26, and 52 weeks of age. At 0, 0.4, 1, 2, 4, and 6 weeks after fracture, a bony segment, one-third the length of the femur, centered on the fracture site, including the external callus, cortical bone, and marrow elements, was harvested. cRNA was prepared and hybridized to 54 Affymetrix U34A microarrays (3/age/time point).

Results

The mRNA levels of 62 genes related to neural function were affected by fracture. Of the total, 38 genes were altered by fracture to a similar extent at the three ages. In contrast, eight neural genes showed prolonged down-regulation in the older rats compared to the more rapid return to pre-fracture levels in younger rats. Seven genes were up-regulated by fracture more in the younger rats than in the older rats, while nine genes were up-regulated more in the older rats than in the younger.

Conclusions

mRNA of 24 nerve-related genes responded differently to fracture in older rats compared to young rats. This differential expression may reflect altered cell function at the fracture site that may be causally related to the slowing of fracture healing with age or may be an effect of the delayed healing.