bluuebunny/crossref_metadata_2025 huggingface dataset:
Tamm-Horsfall Protein Acts as a General Host-Defense Factor against Bacterial Cystitis
Authors: Raffi Hajamohideen S. , Bates, Jr. James M. , Laszik Zoltan , Kumar Satish
Published: October 2005

Aims: Tamm-Horsfall protein (THP) is urine’s most abundant protein, but its biological function has remained elusive. Recently, THP-deficient (THP–/–) mice were shown to have difficulty clearing Escherichia coli from the urinary bladder. It has remained unclear if interaction between THP and E. coli is specific for E. coli or if THP has a versatile ability to clear a variety of bacteria from the bladder, and act as a broad host-defense mechanism against urinary tract infection (UTI). In this study, we examined the role of THP as a protective factor against UTI caused by bacteria other than E. coli, namely Klebsiella pneumoniae and Staphylococcus saprophyticus by determining if the THP–/– mouse has difficulty clearing these bacteria from its bladder. Methods: THP gene knockout mice were generated by the technique of homologous recombination. K. pneumoniae and S. saprophyticus were introduced transurethrally, in separate experiments, into the bladders of the THP–/– and genetically similar wild-type (THP+/+) mice. Urine was collected at periodic intervals and cultured to quantitate the degree of bacteriuria. Bladders were surgically removed and examined histomorphometrically to determine the intensity of inflammation. Results: Results showed that both with K. pneumoniae and with S. saprophyticus, the THP–/– mice had more severe bacteriuria in comparison with THP+/+ mice. The inflammatory changes in the bladder were also markedly more intense in THP–/– mice with each of the bacterial species. Conclusions: These findings support the hypothesis that THP helps eliminate K. pneumoniae and S. saprophyticus from the urinary tract and acts as a general host-defense factor against UTI.
Cortisol and Heart Rate Measures during Casino Gambling in Relation to Impulsivity
Authors: Krueger Tillmann H.C. , Schedlowski Manfred , Meyer Gerhard
Published: October 2005

Problematic gambling behavior is thought to be influenced by neurobiological as well as environmental factors. In this study, we investigated the relationship among impulsivity, gambling behavior, the cardiovascular system and the hypothalamic-pituitary-adrenal axis activity in blackjack gamblers. Twenty-nine males were continuously monitored before, during and after a 90-min blackjack session in a casino wagering their own money and during a control condition where subjects played cards for accumulation of points. Heart rate and cortisol levels significantly increased with the onset of gambling and remained elevated throughout the test session compared to the control condition. After median split of impulsivity scores, high impulsivity subjects revealed significantly higher heart rate levels compared to the low impulsivity subgroup. Correlation analyses revealed a positive relationship between impulsivity scores and severity of pathological gambling. Impulsivity may be one important factor mediating gambling behavior and its accompanying autonomic response.
Leptin Directly Controls Secretory Activity of Human Ovarian Granulosa Cells: Possible Inter-Relationship with the IGF/IGFBP System
Authors: Sirotkin A.V. , Mlynček M. , Kotwica J. , Makarevich A.V. , Florkovičová I. , Hetényi L.
Published: October 2005

Aims: The aim of our in vitro studies was to understand the role of leptin and the insulin-like growth factor I/insulin-like growth factor protein (IGF/IGFBP) system in controlling human ovarian function. Methods: We studied the action of leptin (0, 1, 10, or 100 ng/ml) and immunoneutralization of IGF-I using specific antiserum (0.1%) on the release of progesterone (P), estradiol (E), oxytocin (OT), IGF-I, IGFBP-3, and prostaglandins F (PGF) by these cells using radioimmunoassay/immunoradiometric assay. Results: It was found that leptin stimulated the secretion of OT, IGFBP-3, and PGF. It suppressed the secretion of E and IGF-I, but not P, into the medium. The addition of antiserum against IGF-I decreased IGF-I output, increased P, OT, IGFBP-3, and PGF secretion, and had no effect on E release. Immunoneutralization of IGF-I also prevented or reversed the effects of leptin on P, E, IGF-I, IGFBP-3, PGF, but not on OT. Conclusions: These observations (1) demonstrate that leptin directly controls the secretory activity of human ovarian cells, (2) confirm the involvement of IGF-I in the regulation of ovarian cells, and (3) suggest an inter-relationship between leptin and the IGF/IGFBP system in the control of these functions and the involvement of IGF/IGFBP system in mediating leptin action on the ovary.
A Rapidly Growing Paraurethral Myoma with Profuse Bleeding from a Mucosal Vessel: Report of a Case
Authors: Chong Kian Mei , Chuang Jesse , Tsai Yieh-Loong , Hwang Jiann-Loung
Published: October 2005

Paraurethral leiomyoma is rare. This is the first reported case of a woman with a 6 × 7 × 5 cm urethral leiomyoma with profuse vaginal bleeding. The site of bleeding was identified as the anterior vaginal mucosal vein, which we subsequently electrocauterized. Enucleation of the mass was performed smoothly with a Foley catheter to avoid damage to the urethra. Profuse bleeding from a paraurethral myoma is possibly due to increased vascularity and the prolapsed nature of the tumor in this area.
Association of Endometriosis Risk and Genetic Polymorphisms Involving Sex Steroid Biosynthesis and Their Receptors: A Meta-Analysis
Authors: Guo Sun-Wei
Published: October 2005

Endometriosis is a sex steroids-dependent disease. It has been postulated that certain genetic polymorphisms involved in sex steroids biosynthesis and metabolisms may be associated with increased risk of developing endometriosis. Despite a deluge of reports of positive associations of endometriosis with numerous polymorphisms involving sex steroids production and metabolism, the results are often conflicting. We performed a meta-analysis of 12 association studies on 5 genes (CYP17, CYP19, AR, PR and ER). We found that many reported positive findings were not supported by the data due to faulty analysis. There have been no functional data that support a putative relationship of these genetic polymorphisms with endometriosis. A handful of positive findings so far have not been independently replicated, and should be viewed as preliminary. In addition, these findings should be counterbalanced by legitimate concerns of multiple comparisons, small prior probability of association with a particular polymorphism, proper selection of controls, and lack of replication (at least until now). In future association studies, it may be productive to put more thought to study design, execution, and data analysis.
Théodore Herpin: Neglected Contributions in the Understanding of Epilepsy
Authors: Pearce J.M.S.
Published: October 2005

In contrast to earlier notions of epilepsy, Théodore Herpin considered that when symptoms started in the periphery or the viscera their origin was in the brain, a view later lauded by Hughlings Jackson. His respected book Des accès incomplets did not attempt to deal with disordered physiology but was designed to help physicians to recognise and treat epilepsy early in its course. He described juvenile myoclonic epilepsy and vividly and accurately portrayed many other clinical phenomena of epilepsy.
The Role of Menthol in Skin Penetration from Topical Formulations of Ibuprofen 5% in vivo
Authors: Brain K.R. , Green D.M. , Dykes P.J. , Marks R. , Bola T.S.
Published: October 2005

In vivo plasma profiles from formulations containing 5% ibuprofen were compared after a single topical application in a randomised, double-blind, cross-over trial. IbuleveTM gel (Dermal Laboratories, UK) contained only ibuprofen whilst Deep ReliefTM gel (Mentholatum, UK) also contained 3% menthol. In contrast to results obtained when these products were compared under in vitro conditions, there was no statistically significant difference in vivo between delivery of ibuprofen. Estimated relative bioavailability fraction (Deep Relief gel/Ibuleve gel) from log-transformed AUC(0–24h) was 0.99 (95% CI: 0.94–1.04), estimated Cmax ratio was 0.96 (95% CI: 0.91–1.00) and estimated tmax ratio was 1.01 (95% CI: 0.81–1.20). Menthol produces local vasodilation, which reduces skin barrier function, and these data demonstrate that it is inappropriate to extrapolate from in vitro data where formulation components produce biologically-mediated enhancement of permeation which cannot be modelled ex vivo. In clinical use, these products deliver comparable amounts of ibuprofen, but only Deep Reliefgel provides the secondary immediate benefit of the direct analgesic action of menthol.
Are Sweat Glands an Alternate Penetration Pathway? Understanding the Morphological Complexity of the Axillary Sweat Gland Apparatus
Authors: Wilke K. , Wepf R. , Keil F.J. , Wittern K.-P. , Wenck H. , Biel S.S.
Published: October 2005

To build an effective barrier against the penetration of extrinsic agents is one of the skin’s main functions. The barrier properties of the stratum corneum and the epidermis have been subject to extensive studies in the past while the role of skin appendages as possible pathways of penetration are only rarely described. In order to study the possible penetration barriers in these complex appendages, a careful investigation of their morphology and ultrastructure has to be done. Studying the morphology of axillary skin appendages requires clear-cut criteria for the differentiation between eccrine, apocrine and apoeccrine glands. Therefore we studied the distribution of proteins described to be specific for either eccrine or apocrine glands (CD15, CD44, S-100 and milk fat globulin) on axillary skin samples from healthy young adults by immunofluorescence. Additionally, we examined the distribution of cytoskeletal proteins such as cytokeratins (1/10/11, 14, 18) and F-actin. For a more detailed understanding of the possible versatile barrier elements of the axillary sweat glands, we studied the distribution of tight-junction-associated proteins (occludin, claudin 1, claudin 4). The coils and the dermal duct may provide an active barrier built of tight junctions as occludin and claudin 4 are co-localized. However, the intra-epidermal duct did not show any co-localization of the investigated proteins. By combining morphological features as revealed by F-actin staining and the distribution of the above-mentioned proteins, immunocytochemical typing of eccrine and apocrine glands becomes possible. With this tool, we could also confirm the existence of apoeccrine glands and locate them in their ‘natural environment’.
MRI Evaluation of the Uterine Structure after Myomectomy
Authors: Tsuji Shunichiro , Takahashi Kentaro , Imaoka Izumi , Sugimura Kazuro , Miyazaki Kohji , Noda Yoichi
Published: October 2005

Myomectomy is a good indication for women with uterine leiomyoma who desire to preserve their child-bearing potential. However, there are still no reports about how long it takes the uterus to reach a stable state after myomectomy. We evaluated the changes in uterine structure during the recovery process after myomectomy by MR images. MR images were used to analyze the time-dependant changes in the length of the uterine cavity, the volume of the uterus, recovery of the junctional zone, prevalence of modification of the endometrium, and uterine structure in the region of the enucleated myoma. The cavity length and the volume of the uterus, and the myometrium were stabilized at six weeks after the myomectomy. With regard to the endometrium, 12 weeks were required for it to achieve a stable state after myomectomy. However, even at 12 weeks postoperatively, 14.2% of the cases showed an unusual view near the uterine incision on MR images. We concluded that the recovery process is complete at 12 weeks after the operation if there are no clear findings of hematoma or edema formation in the myometrium on MR images.
Two Successful Pregnancies following Eight Miscarriages in a Patient with Antithrombin Deficiency
Authors: Szilágyi A. , Nagy A´. , Tamás P. , Vizer M. , Szabó I. , Losonczy H.
Published: October 2005

Inherited thrombophilias are associated with an increased risk of maternal thromboembolism and certain adverse pregnancy outcomes, including second- and third-trimester fetal loss, placental abruption, severe intrauterine growth restriction, and early-onset, severe preeclampsia. Pregnant patients with severe thrombophilias, especially antithrombinopathies are at very high risk for both thromboembolism and adverse pregnancy outcomes. A case of a patient with antithrombin deficiency is reported, who had two successful pregnancies after eight miscarriages. Our case shows that a combined treatment with antithrombin substitution and a prophylactic, body-weight-adjusted dose of low-molecular-weight heparin may be successful in preventing pregnancy loss and thromboembolism in antithrombin deficiency during pregnancy, although other complications, such as preeclampsia and intrauterine growth restriction cannot always be prevented.
What Observation Scheme Should We Use for Profiling Floats to Achieve the Argo Goal for Salinity Measurement Accuracy? Suggestions from Software Calibration
Authors: Kobayashi Taiyo , Minato Shinya
Published: October 2005

To meet the Argo Project’s accuracy goal for salinity measurements (±0.01), several measurement schemes for profiling float observations were examined using the standard Argo software calibration for salinity. In this study, salinity calibration errors are evaluated in data series observed by employing several different measurement schemes. In the North Pacific the accuracy goal can be achieved by programming for constant measurements to a depth of 1500 db or more; however, measurements in the mixed water region require the adoption of a scheme that constantly measures salinity profiles to the marginal capability depth of floats (e.g., 2000 db), so that calibration errors can be minimized. In contrast, in the tropical regions of the Pacific and eastern Indian Ocean, 1000-db profiles provide salinity data of sufficient quality. When floats use a Park and Profile scheme, it is recommended that they be programmed to measure deeper profiles every observation cycle. Otherwise, it is impossible to detect and correct suspicious data, such as a salinity jump in which a salinity measurement suddenly differs from previous measurements by ±0.02–0.03 or more.
A Self-Contained Acoustic Scintillation Instrument for Path-Averaged Measurements of Flow and Turbulence with Application to Hydrothermal Vent and Bottom Boundary Layer Dynamics
Authors: Di Iorio D. , Lemon D. , Chave R.
Published: October 2005

A self-contained acoustical scintillation instrument is described that has been used to measure flow and turbulence characteristics in two diverse oceanographic settings. This instrument is a battery-operated and internally logging acoustic propagation system that is ideally suited to monitor long-term flow and small-scale effective refractive index fluctuations. When the temperature variability dominates the acoustic scattering, as is the case of a hydrothermal vent plume, then a measure of the vertical buoyancy-driven flow, together with the root-mean-square temperature fluctuations, can be obtained. Results for vent structure Hulk of the Main Endeavour vent field of the Juan de Fuca Ridge show that the long-term (71 days) temperature fluctuations, together with the vertical flow, can be used to estimate heat flux density. Measurements also show oscillations in the log-amplitude variance that result from plume advection by the ambient tidal currents and demonstrate the need for a long time series measurement. When the turbulent velocity dominates the acoustic scattering, as is the case in some energetic bottom boundary layer flows, then the turbulent kinetic energy dissipation rate is derived, assuming isotropic and homogeneous models. The methodology and results are summarized from an application to the Bosporus Canyon of the Black Sea, to monitor the flow and turbulence associated with Mediterranean seawater inflow.
Representing the Public in Campaign Media
Authors: Edwards Janis L. , Ware Laura
Published: October 2005

Although voters are central to the processes of democracy, scholarly attention to voters, with few exceptions, is limited to statistical analysis of decision-making and opinionformation processes, treating voters as a mass audience or audience segment. This article argues for more nuanced interpretations of voter representations in the media. Citing key studies of news media representation, the authors explore visual representation of voters in political cartoon commentary and argue that cartoons provide a viewof voters that supports and supplements views evident in the news media but question the role of voter depictions in cartoons as sources of understanding about political publics.
The Passion of the Christ Meets Fahrenheit 9/11
Authors: Goodnight G. Thomas
Published: October 2005

Why are The Passion of the Christand Fahrenheit 9/11mentioned in the same breath when referring to the political campaigns of 2004? What role did these films play in the election debate? How did television code reception, at once expanding and disciplining, tantalizing and moralizing the public? The article examines the entwinement of entertainment, media, and politics in national debate, as the most talked about films of 2004 transmute wartime suffering into political purpose, each serving a similar function for respective Republican and Democratic campaigns and each generating controversy stirred and strangled by televisual critics. The article concludes by advocating a role for critical inquiry into celebrity advocacy—as artistic enactment, stylistic vision, and mass media promotion. Studies of the multiple dimensions of celebrity advocacy should augment significantly our contemporary discussions of deliberative democracy.
A Typology for the Study of Entertainment Television and Politics
Authors: Holbert R. Lance
Published: October 2005

General questions concerning the role of entertainment media in politics permeated the 2004 election cycle. Political communication scholars are beginning to systematically analyze potential sociopolitical ramifications stemming from entertainment television viewing, but one difficulty with this emerging area of research is an inability to forge connections between seemingly disparate research agendas. Several distinct lines of research are summarized in this article and forged into a coherent whole through the lens of Zillmann's disposition theory and the broader study of media enjoyment. Extantworks devoted to the study of entertainment television and politics are summarized, and a ninepart typology is introduced. This article closes with a discussion of howthe typology can serve to advance the study of entertainment television and politics.
Beyond Pseudoevents
Authors: Bennett W. Lance
Published: October 2005

This analysis examines the rise of reality TV formats in news reporting. These formats put journalists in active roles in the dramatic framing of events, thus, moving news logic beyond the passive documentation of pseudoevents. Pseudoevents are made-for-media fabrications designed entirely by newsmakers to be reported as reality with little news value added by journalists. By contrast, journalistically driven reality TV formats are either coproduced actively with newsmakers or spun entirely by journalists. Coproduced reality news generally involves journalistic decisions to underplay or not report known facts or contradictory evidence that conflicts with the dramatic impact of staged events. Reality news that is initiated by journalists typically involves rumors or challenges thrown at political contestants to see who can survive the hostile media game, and who, in the case of elections, will be voted off the program.
Agenda Setting in a Culture of Fear
Authors: Matsaganis Matthew D. , Payne J. Gregory
Published: October 2005

Agenda setting has been developed, expanded, and employed in numerous studies as an analytical tool that affords an understanding of not only how our political reality is formulated but also how “realities” can be manufactured. However, as the authors argue, by grafting agenda setting and media systems dependency theory—two different traditions in mass communication theory—it is possible to better account for changes in the agenda-setting process because of shifts in the power relationships between all actors involved, especially under conditions of increased threat; conditions similar to those the American public has lived in since the September 11 terrorist attacks. Illustrations from Campaign 2004 complement this analysis. The authors suggest that it is critical to understand the dynamics of the making of “mediated realities” so as to alert readers of the importance in furthering critical media literacy skills necessary for the public to distinguish between facades and facts.
Reducing Violence and Aggression in Schools
Authors: Greene Michael B.
Published: October 2005

This article offers a framework for understanding and responding to school-based aggression and violence. The term school violence is defined, epidemiological data are summarized, a typology of violence reduction strategies is presented, and procedures to effectively implement evidence-based programs are discussed. Although many evidence-based violence prevention programs are now available to schools, much work remains in three critical areas. First, additional research is needed to evaluate the impact of security strategies, peer-led programs, and threat assessment and crisis response initiatives. More generally, multi-level evaluations of integrated arrays of school-based violence prevention strategies needtobeundertaken. Second, effective and realistic school-based assessment strategies need to be established to identify and address organizational barriers to the selection and adoption of an integrated and comprehensive array of targeted evidence-based violence prevention strategies in schools. And third, realistic guidelines through which schools can effectively monitor and implement evidence-based programs need to be developed.
Trauma, PTSD, and Resilience
Authors: Agaibi Christine E. , Wilson John P.
Published: October 2005

Based on the available literature, this review article investigates the issue of resilience in relation to trauma and posttraumatic stress disorder. Resilient coping to extreme stress and trauma is a multifaceted phenomena characterized as a complex repertoire of behavioral tendencies. An integrative Person × Situation model is developed based on the literature that specifies the nature of interactions among five classes of variables: (a) personality, (b) affect regulation, (c) coping, (d) ego defenses, and (e) the utilization and mobilization of protective factors and resources to aid coping.
Criminal Investigations of Child Abuse
Authors: Jones Lisa M. , Cross Theodore P. , Walsh Wendy A. , Simone Monique
Published: October 2005

This article reviews the research relevant to seven practices considered by many to be among the most progressive approaches to criminal child abuse investigations: multidisciplinary team investigations, trained child forensic interviewers, videotaped interviews, specialized forensic medical examiners, victim advocacy programs, improved access to mental health treatment for victims, and Children’s Advocacy Centers (CACs). The review finds that despite the popularity of these practices, little outcome research is currently available documenting their success. However, preliminary research supports many of these practices or has influenced their development. Knowledge of this research can assist investigators and policy makers who want to improve the response to victims, understand the effectiveness of particular programs, or identify where assumptions about effectiveness are not empirically supported.
Is Intimate Partner Violence Associated with Unintended Pregnancy? A Review of the Literature
Authors: Pallitto Christina C. , Campbell Jacquelyn C. , O’Campo Patricia
Published: October 2005

Although a substantial body of literature explores the adverse physical and mental health consequences associated with intimate partner violence, only a limited body of international research has explored the effect of intimate partner violence on women’s fertility control. Yet a compelling argument can be made of the indirect mechanism through which the climate of fear and control surrounding abusive relationships could limit women’s ability to control their fertility. Lack of fertility control can lead to unintended pregnancies, which are also associated with adverse outcomes for women’s and infant health, especially in developing countries. The association between intimate partner violence and unintended pregnancy also suggests serious social effects spawned by a cycle of unintended childbearing in abusive households. Therefore, further investigation is warranted to explore the nature of the association as well as the mechanisms through which these phenomena operate in the United States and in developing countries.
Hematopoietic stem cell exhaustion impacted by p18INK4C and p21Cip1/Waf1 in opposite manners
Authors: Yu Hui , Yuan Youzhong , Shen Hongmei , Cheng Tao
Published: October 2005

Transplantation-associated stress can compromise the hematopoietic potential of hematopoietic stem cells (HSCs). As a consequence, HSCs may undergo “exhaustion” in serial transplant recipients, for which the cellular and molecular bases are not well understood. Hematopoietic exhaustion appears to be accelerated in the absence of p21Cip1/Waf1 (p21), a cyclin-dependent kinase inhibitor (CKI) in irradiated hosts. Our recent study demonstrated that unlike loss of p21, deletion of p18INK4C (p18), a distinct CKI, results in improved long-term engraftment, largely because of increased self-renewing divisions of HSCs in vivo. We show here that HSCs deficient in p18 sustained their competitiveness to wild-type HSCs from unmanipulated young mice, and retained multilineage differentiation potential after multiple rounds of serial bone marrow transfer over a period of more than 3 years. Further, p18 absence significantly decelerated hematopoietic exhaustion caused by p21 deficiency. Such an effect was shown to occur at the stem cell level, likely by a counteracting mechanism against the cellular senescence outcome. Our current study provides new insights into the distinct impacts of these cell-cycle regulators on HSC exhaustion and possibly HSC aging as well under proliferative stress, thereby offering potential pharmacologic targets for sustaining the durability of stressed HSCs in transplantation or elderly patients.
A role for cofilin in the activation of store-operated calcium entry by de novo conformational coupling in human platelets
Authors: Redondo Pedro C. , Harper Matthew T. , Rosado Juan A. , Sage Stewart O.
Published: October 2005

Store-operated Ca2+ entry (SOCE) is a major mechanism for Ca2+ influx in platelets and other cells. De novo conformational coupling between elements in the plasma membrane and Ca2+ stores, where the actin cytoskeleton plays an important regulatory role, has been proposed as the most likely mechanism to activate SOCE in platelets. Here we have examined for the first time changes in platelet F-actin levels on a subsecond time scale. Using stopped-flow fluorimetry and a quenched-flow approach, we provide evidence for the involvement of cofilin in actin filament reorganization and SOCE in platelets. Thrombin (0.1 U/mL) evoked an initial decrease in F-actin that commenced within 0.1 second and reached a minimum 0.9 second after stimulation, prior to the activation of SOCE. F-actin then increased, exceeding basal levels approximately 2.5 seconds after stimulation. Thrombin also induced cofilin dephosphorylation and activation, which paralleled the changes observed in F-actin, and rapid Btk activation. Inhibition of cofilin dephosphorylation by LFM-A13 resulted in the loss of net actin depolymerization and an increased delay in SOCE initiation. These results suggest that cofilin is important for the rapid actin remodeling necessary for the activation of SOCE in platelets through de novo conformational coupling.
EFA (9-β-D-erythrofuranosyladenine) is an effective salvage agent for methylthioadenosine phosphorylase-selective therapy of T-cell acute lymphoblastic leukemia with L-alanosine
Authors: Batova Ayse , Cottam Howard , Yu John , Diccianni Mitchell B. , Carrera Carlos J. , Yu Alice L.
Published: October 2005

The deficiency of methylthioadenosine phosphorylase (MTAP) in T-cell acute lymphoblastic leukemia (T-ALL) and other cancers, while constitutively expressed in normal cells, allows for selective therapy using L-alanosine, an inhibitor of de novo AMP synthesis. We demonstrate that MTAP- T-ALL cells obtained at relapse are as sensitive to L-alanosine toxicity as diagnosis samples. The therapeutic index of L-alanosine can be increased by the use of a MTAP substrate, which protects MTAP+ normal cells. Since MTAP substrates MTA and 5′deoxyadenosine are prone to toxicities associated with adenosine, we synthesized and evaluated a potentially nontoxic MTAP substrate, 9-β-D-erythrofuranosyladenine (EFA). The cytotoxicity of EFA to hematopoietic progenitors erythroid burst-forming units (BFU-Es) and granulocyte-macrophage colony-forming units (CFU-GMs) was at least 26- to 41-fold less than that of MTA. In addition, EFA selectively rescued MTAP+ MOLT-4 cells from L-alanosine toxicity at 25 μM with negligible toxicity even at 100 μM. As for MTA, significant, albeit incomplete, rescue was achieved at 12.5 μM, but higher concentrations were toxic. EFA at 20 μM or less rescued primary MTAP+ T-ALL cells and normal lymphocytes from L-alanosine toxicity. Collectively, these data indicate that EFA is an effective agent for salvaging MTAP+ cells from L-alanosine toxicity and is superior to MTA due to lower cytotoxicity.
Use of trifunctional bispecific antibodies to prevent graft versus host disease induced by allogeneic lymphocytes
Authors: Morecki Shoshana , Lindhofer Horst , Yacovlev Elena , Gelfand Yael , Slavin Shimon
Published: October 2005

A trifunctional bispecific antibody (BiLu) directed against murine CD3 and human epithelial-cell adhesion molecule (EpCAM) was tested for its ability to improve cell-mediated adoptive immunotherapy in a murine model of B16 melanoma cells transfected with human EpCAM. Intraperitoneal inoculation of naive C57BL/6 (C57) splenocytes induced lethal graft versus host disease (GVHD) in 85% to 97% of sublethally irradiated (BALB/c × C57BL/6) F1 (F1) hosts inoculated intraperitoneally with a sublethal or lethal dose of melanoma cells. BiLu antibodies given intraperitoneally concomitantly with alloreactive C57 cells effectively prevented GVHD-related and tumor-related death in 16 of 25 F1 mice inoculated with a sublethal tumor-cell dose and in 10 of 20 mice inoculated with a lethal tumor-cell dose over a follow-up period of more than 200 days. BiLu treatment also efficiently prevented severe GVHD, which was induced by high doses of BALB/c-derived splenocytes. Trifunctional bispecific antibodies (TbsAbs) capable of cross-linking T lymphocytes, natural killer, and other FcγR-positive effector cells, via their Fc region, to the tumor cells may be applied together with adoptive allogeneic-cell therapy to maximize antitumor responses while acting on GVHD in patients with minimal residual disease.
Phagocytosis of codeveloping megakaryocytic progenitors by dendritic cells in culture with thrombopoietin and tumor necrosis factor-α and its possible role in hemophagocytic syndrome
Authors: Saito Kunie , Hirokawa Makoto , Inaba Kayo , Fukaya Hiroshi , Kawabata Yoshinari , Komatsuda Atsushi , Yamashita Junsuke , Sawada Kenichi
Published: October 2005

Tumor necrosis factor-α (TNF-α) and thrombopoietin (TPO) have been shown to induce the differentiation and proliferation of CD34+ cells toward dendritic cells (DCs) in the presence of multiacting cytokines. We hypothesized that the costimulation of TPO and TNF-α generates megakaryocytic progenitors and DCs together from human CD34+ cells and that the interaction of these cells may indicate a physiologic and/or a pathologic role of DCs in megakaryopoiesis. When highly purified human CD34+ cells were cultured for 7 days with TPO alone, the generated cells expressed megakaryocytic markers, such as CD41, CD42b, and CD61. The addition of TNF-α with TPO remarkably decreased the number of megakaryocytic progenitor cells without affecting the cell yield. Almost half of the cells thus generated expressed CD11c, and most of them were positive for CD4 and CD123. Furthermore, CD11c+ cells were found to capture damaged CD61+ cells and to induce autologous T-cell proliferation, although the cytokine productions were low. We also confirmed an engulfment of CD61+ cells and their fragment by CD11c+ cells in bone marrow cells from patients with hemophagocytic syndrome. These findings suggest that DCs generated under megakaryocytic and inflammatory stimuli are involved in megakaryopoiesis and the subsequent immune responses to self-antigens.
Sub-chronic inhalation of high concentrations of manganese sulfate induces lower airway pathology in rhesus monkeys
Authors: Dorman David C , Struve Melanie F , Gross Elizabeth A , Wong Brian A , Howroyd Paul C
Published: October 2005

Background

Neurotoxicity and pulmonary dysfunction are well-recognized problems associated with prolonged human exposure to high concentrations of airborne manganese. Surprisingly, histological characterization of pulmonary responses induced by manganese remains incomplete. The primary objective of this study was to characterize histologic changes in the monkey respiratory tract following manganese inhalation.

Methods

Subchronic (6 hr/day, 5 days/week) inhalation exposure of young male rhesus monkeys to manganese sulfate was performed. One cohort of monkeys (n = 4–6 animals/exposure concentration) was exposed to air or manganese sulfate at 0.06, 0.3, or 1.5 mg Mn/m3for 65 exposure days. Another eight monkeys were exposed to manganese sulfate at 1.5 mg Mn/m3for 65 exposure days and held for 45 or 90 days before evaluation. A second cohort (n = 4 monkeys per time point) was exposed to manganese sulfate at 1.5 mg Mn/m3and evaluated after 15 or 33 exposure days. Evaluations included measurement of lung manganese concentrations and evaluation of respiratory histologic changes. Tissue manganese concentrations were compared for the exposure and control groups by tests for homogeneity of variance, analysis of variance, followed by Dunnett's multiple comparison. Histopathological findings were evaluated using a Pearson's Chi-Square test.

Results

Animals exposed to manganese sulfate at ≥0.3 mg Mn/m3for 65 days had increased lung manganese concentrations. Exposure to manganese sulfate at 1.5 mg Mn/m3for ≥15 exposure days resulted in increased lung manganese concentrations, mild subacute bronchiolitis, alveolar duct inflammation, and proliferation of bronchus-associated lymphoid tissue. Bronchiolitis and alveolar duct inflammatory changes were absent 45 days post-exposure, suggesting that these lesions are reversible upon cessation of subchronic high-dose manganese exposure.

Conclusion

High-dose subchronic manganese sulfate inhalation is associated with increased lung manganese concentrations and small airway inflammatory changes in the absence of observable clinical signs. Subchronic exposure to manganese sulfate at exposure concentrations (≤0.3 mg Mn/m3) similar to the current 8-hr occupational threshold limit value established for inhaled manganese was not associated with pulmonary pathology.
Dynamic modeling of cis-regulatory circuits and gene expression prediction via cross-gene identification
Authors: Lin Li-Hsieh , Lee Hsiao-Ching , Li Wen-Hsiung , Chen Bor-Sen
Published: October 2005

Background

Gene expression programs depend on recognition of cis elements in promoter region of target genes by transcription factors (TFs), but how TFs regulate gene expression via recognition of cis elements is still not clear. To study this issue, we define the cis-regulatory circuit of a gene as a system that consists of its cis elements and the interactions among their recognizing TFs and develop a dynamic model to study the functional architecture and dynamics of the circuit. This is in contrast to traditional approaches where a cis-regulatory circuit is constructed by a mutagenesis or motif-deletion scheme. We estimate the regulatory functions of cis-regulatory circuits using microarray data.

Results

A novel cross-gene identification scheme is proposed to infer how multiple TFs coordinate to regulate gene transcription in the yeast cell cycle and to uncover hidden regulatory functions of a cis-regulatory circuit. Some advantages of this approach over most current methods are that it is based on data obtained from intact cis-regulatory circuits and that a dynamic model can quantitatively characterize the regulatory function of each TF and the interactions among the TFs. Our method may also be applicable to other genes if their expression profiles have been examined for a sufficiently long time.

Conclusion

In this study, we have developed a dynamic model to reconstruct cis-regulatory circuits and a cross-gene identification scheme to estimate the regulatory functions of the TFs that control the regulation of the genes under study. We have applied this method to cell cycle genes because the available expression profiles for these genes are long enough. Our method not only can quantify the regulatory strengths and synergy of the TFs but also can predict the expression profile of any gene having a subset of the cis elements studied.
Human promoter genomic composition demonstrates non-random groupings that reflect general cellular function
Authors: McNutt Markey C , Tongbai Ron , Cui Wenwu , Collins Irene , Freebern Wendy J , Montano Idalia , Haggerty Cynthia M , Chandramouli GVR , Gardner Kevin
Published: October 2005

Background

The purpose of this study is to determine whether or not there exists nonrandom grouping of cis-regulatory elements within gene promoters that can be perceived independent of gene expression data and whether or not there is any correlation between this grouping and the biological function of the gene.

Results

Using ProSpector, a web-based promoter search and annotation tool, we have applied an unbiased approach to analyze the transcription factor binding site frequencies of 1400 base pair genomic segments positioned at 1200 base pairs upstream and 200 base pairs downstream of the transcriptional start site of 7298 commonly studied human genes. Partitional clustering of the transcription factor binding site composition within these promoter segments reveals a small number of gene groups that are selectively enriched for gene ontology terms consistent with distinct aspects of cellular function. Significance ranking of the class-determining transcription factor binding sites within these clusters show substantial overlap between the gene ontology terms of the transcriptions factors associated with the binding sites and the gene ontology terms of the regulated genes within each group.

Conclusion

Thus, gene sorting by promoter composition alone produces partitions in which the "regulated" and the "regulators" cosegregate into similar functional classes. These findings demonstrate that the transcription factor binding site composition is non-randomly distributed between gene promoters in a manner that reflects and partially defines general gene class function.
Novel splice variants associated with one of the zebrafish dnmt3genes
Authors: Smith Tamara HL , Dueck Christine C , Mhanni Aizeddin A , McGowan Ross A
Published: October 2005

Background

DNA methylation and the methyltransferases are known to be important in vertebrate development and this may be particularly true for the Dnmt3 family of enzymes because they are thought to be the de novo methyltransferases. Mammals have three Dnmt3 genes; Dnmt3a, Dnmt3b, and Dnmt3L, two of which encode active enzymes and one of which produces an inactive but necessary cofactor. However, due to multiple promoter use and alternative splicing there are actually a number of dnmt3 isoforms present. Six different dnmt3 genes have recently been identified in zebrafish.

Results

We have examined two of the dnmt3 genes in zebrafish that are located in close proximity in the same linkage group and we find that the two genes are more similar to each other than they are to the other zebrafish dnmt3 genes. We have found evidence for the existence of several different splice variants and alternative splice sites associated with one of the two genes and have examined the relative expression of these genes/variants in a number of zebrafish developmental stages and tissues.

Conclusion

The similarity of the dnmt3-1 and dnmt3-2 genes suggests that they arose due to a relatively recent gene duplication event. The presence of alternative splice and start sites, reminiscent of what is seen with the human DNMT3s, demonstrates strong parallels between the control/function of these genes across vertebrate species. The dynamic expression levels of these genes/variants suggest that they may well play a role in early development and this is particularly true for dnmt3-2-1 and dnmt3-1. dnmt3-2-1 is the predominantly expressed form prior to zygotic gene activation whereas dnmt3-1 predominates post zygotic gene activation suggesting a distinct developmental role for each.
Generation, annotation, analysis and database integration of 16,500 white spruce EST clusters
Authors: Pavy Nathalie , Paule Charles , Parsons Lee , Crow John A , Morency Marie-Josee , Cooke Janice , Johnson James E , Noumen Etienne , Guillet-Claude Carine , Butterfield Yaron , Barber Sarah , Yang George , Liu Jerry , Stott Jeff , Kirkpatrick Robert , Siddiqui Asim , Holt Robert , Marra Marco , Seguin Armand , Retzel Ernest , Bousquet Jean , MacKay John
Published: October 2005

Background

The sequencing and analysis of ESTs is for now the only practical approach for large-scale gene discovery and annotation in conifers because their very large genomes are unlikely to be sequenced in the near future. Our objective was to produce extensive collections of ESTs and cDNA clones to support manufacture of cDNA microarrays and gene discovery in white spruce (Picea glauca [Moench] Voss).

Results

We produced 16 cDNA libraries from different tissues and a variety of treatments, and partially sequenced 50,000 cDNA clones. High quality 3' and 5' reads were assembled into 16,578 consensus sequences, 45% of which represented full length inserts. Consensus sequences derived from 5' and 3' reads of the same cDNA clone were linked to define 14,471 transcripts. A large proportion (84%) of the spruce sequences matched a pine sequence, but only 68% of the spruce transcripts had homologs in Arabidopsis or rice. Nearly all the sequences that matched the Populus trichocarpa genome (the only sequenced tree genome) also matched rice or Arabidopsis genomes. We used several sequence similarity search approaches for assignment of putative functions, including blast searches against general and specialized databases (transcription factors, cell wall related proteins), Gene Ontology term assignation and Hidden Markov Model searches against PFAM protein families and domains. In total, 70% of the spruce transcripts displayed matches to proteins of known or unknown function in the Uniref100 database (blastx e-value < 1e-10). We identified multigenic families that appeared larger in spruce than in the Arabidopsis or rice genomes. Detailed analysis of translationally controlled tumour proteins and S-adenosylmethionine synthetase families confirmed a twofold size difference. Sequences and annotations were organized in a dedicated database, SpruceDB. Several search tools were developed to mine the data either based on their occurrence in the cDNA libraries or on functional annotations.

Conclusion

This report illustrates specific approaches for large-scale gene discovery and annotation in an organism that is very distantly related to any of the fully sequenced genomes. The ArboreaSet sequences and cDNA clones represent a valuable resource for investigations ranging from plant comparative genomics to applied conifer genetics.
Pseudouridine modification in Caenorhabditis elegans spliceosomal snRNAs: unique modifications are found in regions involved in snRNA-snRNA interactions
Authors: Patton Jeffrey R , Padgett Richard W
Published: October 2005

Background

Pseudouridine (Ψ) is an abundant modified nucleoside in RNA and a number of studies have shown that the presence of Ψ affects RNA structure and function. The positions of Ψ in spliceosomal small nuclear RNAs (snRNAs) have been determined for a number of species but not for the snRNAs fromCaenorhabditis elegans(C. elegans), a popular experimental model system of development.

Results

As a prelude to determining the function of or requirement for this modification in snRNAs, we have mapped the positions of Ψ in U1, U2, U4, U5, and U6 snRNAs from worms using a specific primer extension method. As with other species,C. elegansU2 snRNA has the greatest number of Ψ residues, with nine, located in the 5' half of the U2 snRNA. U5 snRNA has three Ψs, in or near the loop of the large stem-loop that dominates the structure of this RNA. U6 and U1 snRNAs each have one Ψ, and two Ψ residues were found in U4 snRNA.

Conclusion

The total number of Ψs found in the snRNAs ofC. elegansis significantly higher than the minimal amount found in yeasts but it is lower than that seen in sequenced vertebrate snRNAs. When the actual sites of modification onC. eleganssnRNAs are compared with other sequenced snRNAs most of the positions correspond to modifications found in other species. However, two of the positions modified onC. eleganssnRNAs are unique, one at position 28 on U2 snRNA and one at position 62 on U4 snRNA. Both of these modifications are in regions of these snRNAs that interact with U6 snRNA either in the spliceosome or in the U4/U6 small nuclear ribonucleoprotein particle (snRNP) and the presence of Ψ may be involved in strengthening the intermolecular association of the snRNAs.
A simple and rapid approach for screening of SARS-coronavirus genotypes: an evaluation study
Authors: Chung Grace TY , Chiu Rossa WK , Cheung Jo LK , Jin Yongjie , Chim Stephen SC , Chan Paul KS , Lo YM Dennis
Published: October 2005

Background

The Severe Acute Respiratory Syndrome (SARS) was a newly emerged infectious disease which caused a global epidemic in 2002–2003. Sequence analysis of SARS-coronavirus isolates revealed that specific genotypes predominated at different periods of the epidemic. This information can be used as a footprint for tracing the epidemiology of infections and monitor viral evolution. However, direct sequencing analysis of a large number of clinical samples is cumbersome and time consuming. We present here a simple and rapid assay for the screening of SARS-coronavirus genotypes based on the use of fluorogenic oligonucleotide probes for allelic discrimination.

Methods

Thirty SARS patients were recruited. Allelic discrimination assays were developed based on the use of fluorogenic oligonucleotide probes (TaqMan). Genotyping of the SARS-coronavirus isolates obtained from these patients were carried out by the allelic discrimination assays and confirmed by direct sequencing.

Results

Genotyping based on the allelic discrimination assays were fully concordant with direct sequencing. All of the 30 SARS-coronavirus genotypes studied were characteristic of genotypes previously documented to be associated with the latter part of the epidemic. Seven of the isolates contained a previously reported major deletion but in patients not epidemiologically related to the previously studied cohort.

Conclusion

We have developed a simple and accurate method for the characterization and screening of SARS-coronavirus genotypes. It is a promising tool for the study of epidemiological relationships between documented cases during an outbreak.