bluuebunny/crossref_metadata_2025 huggingface dataset:
Protective Effect of Erythropoietin against Radiocontrast-Induced Renal Tubular Epithelial Cell Injury
Authors: Kolyada Alexey Y. , Liangos Orfeas , Madias Nicolaos E. , Jaber Bertrand L.
Published: October 2007

Background/Aims: Recombinant human erythropoietin (rhEpo) has been shown to reduce tissue injury following ischemia-reperfusion. We examined whether rhEpo protects in vitro renal tubular epithelial cells against radiocontrast media-induced injury. Methods: LLC-PK1 renal tubular epithelial cells were exposed to non-ionic radiocontrast agent iohexol (low-osmolar) or iodixanol (iso-osmolar), with or without rhEpo (200 U/ml). Following a 6-hour exposure, cells were incubated for 24 h in radiocontrast-free culture medium. Cell viability was then assessed by the MTT assay. We also assessed cell apoptosis by the TUNEL assay, and activities of caspase-3, caspase-8, and caspase-9 were determined by a luminescence assay. Results: rhEpo improved viability of iohexol-treated LLC-PK1 cells by 27 ± 6% (88.1 ± 1.5 vs. 70.8 ± 3.3%, p = 0.008). Similarly, rhEpo improved the viability of iodixanol-treated LLC-PK1 cells by 26 ± 4% (82.5 ± 2.1vs. 65.7 ± 1.7%, p = 0.028). rhEpo also decreased apoptosis rates of iohexol-treated LLC-PK1 cells (6.4 ± 0.9/1,000 cells vs. 14.8 ± 2.4/1,000 cells, p = 0.028), and iodixanol-treated LLC-PK1 cells (8.0 ± 1.2/1,000 cells vs. 13.5 ± 1.9/1,000 cells, p = 0.028). In iohexol-treated LLC-PK1 cells, rhEpo attenuated activation of caspase-3 (p = 0.003), caspase-8 (p = 0.033) and caspase-9 (p = 0.055). Conclusion: rhEpo attenuates in vitro renal tubular epithelial cell injury induced by low- and iso-osmolar radiocontrast media, possibly by reduction of caspases activation and apoptosis rates.
Olopatadine Ameliorates Rat Experimental Cutaneous Inflammation by Improving Skin Barrier Function
Authors: Tamura Tadafumi , Matsubara Masahiro , Amano Toru , Chida Michihiro
Published: October 2007

Olopatadine hydrochloride (olopatadine) is an antiallergic agent with histamine H1 receptor antagonistic action. We investigated the possible efficacies of olopatadine on the chronic inflammatory dermatitis and the impaired skin barrier functions induced by repeated application of oxazolone in rats. Oxazolone-sensitized rats were challenged with oxazolone applied to the ear every 3 days. Olopatadine was orally administered once daily (1 and 3 mg/kg/day). The effects of the drug were quantified by measurements of ear thickness, levels of cytokines in the lesioned ear and the number of scratching episodes. As parameters of skin barrier function, transepidermal water loss (TEWL) and hyaluronic acid (HA) levels in the lesioned ear were measured. The effect of olopatadine on the production of HA by cultured dermal fibroblasts was also measured. Repeated topical application of oxazolone to rat ears induced local inflammation that was exemplified by swelling. In inflamed ears, the amount of IFNγ increased at both the protein and mRNA level, but IL-4 levels changed minimally. Olopatadine significantly decreased ear swelling and the number of scratching episodes. The drug also significantly inhibited the increase of IFNγ and nerve growth factor production in inflamed ears. Olopatadine significantly inhibited the increase in TEWL and the decrease in HA in lesioned ears. Furthermore, the drug stimulated the production of HA by cultured dermal fibroblasts. These results suggest that olopatadine suppressed inflammation and scratching not only by inhibiting cytokine production, but also by repairing skin barrier function.
Cardiopulmonary Exercise Test Characteristics in Patients with Chronic Obstructive Pulmonary Disease and Associated Pulmonary Hypertension
Authors: Holverda Sebastiaan , Bogaard Harm J. , Groepenhoff H. , Postmus Pieter E. , Boonstra Anco , Vonk-Noordegraaf Anton
Published: October 2007

Background: Pulmonary hypertension (PH) is a well-known complication of chronic obstructive pulmonary disease (COPD). It remains unclear whether exercise parameters can be used to discriminate between COPD patients with associated PH (COPD-PH) and COPD patients without associated PH (COPD-nonPH). Objective: To study whether the existence of pulmonary hypertension in COPD is related to characteristic findings in gas exchange and circulatory parameters during cardiopulmonary exercise testing (CPET). Methods: We retrospectively analyzed CPET data in 25 COPD patients in whom right heart catheterization had been performed. Differences were assessed between COPD-PH and COPD-nonPH patients in peak oxygen uptake (VO2 peak), ventilatory efficiency (VE/VCO2), oxygen pulse, maximal ventilation and pulse oximetry (SpO2). Results: PH was found in 10 of 25 patients (mPpa = 33 ± 7 mm Hg), in 15 patients mean pulmonary artery pressure (mPpa) was below 25 mm Hg (18 ± 3 mm Hg). CPET in COPD-PH was characterized by a higher VE/VCO2 at nadir, a higher VE/VCO2 slope, and a lower SpO2 at rest and during exercise, but values in both groups were overlapping considerably. In the whole group mPpa was associated with resting PaO2 (r = –0.70, p < 0.001), VE/VCO2 nadir (r = 0.43, p < 0.05), and inversely related to SpO2 at rest and during exercise (r = –0.58 and r = –0.64, p < 0.01, respectively). Conclusion: Although CPET characteristics showed a large overlap in both groups, the existence of PH in COPD is associated with a significantly reduced ventilatory efficiency during CPET. However, a low SpO2 at rest and a further decrease during exercise similarly suggest the presence of PH in COPD.
Skin Blood Flow and Its Oscillatory Components in Patients with Acute Myocardial Infarction
Authors: Ažman-Juvan Katja , Bernjak Alan , Urbančič-Rovan Vilma , Stefanovska Aneta , Štajer Dušan
Published: October 2007

Background/Aims: Laser Doppler flowmetry (LDF) was used to determine the influence of acute myocardial infarction (AMI) and of successful reperfusion treatment on basal skin blood flow and its oscillatory components. Methods: Skin LDF was performed on all extremities in 58 patients 4–9 days after AMI (Killip class I), and in 71 healthy age- and sex-matched controls. Wavelet analysis was applied to evaluate oscillatory components within the interval 0.005–2 Hz. Results: AMI patients had reduced mean flow (p < 0.01) and oscillatory components (p < 0.04) in all extremities. Reperfused (n = 40), compared to nonreperfused (n = 18), patients had higher mean flow and total spectral amplitude at all recording points. The difference was statistically significant only in legs (group median LDF in the left leg was 9.68 AU for reperfused and 5.71 AU for nonreperfused patients, p < 0.04, and 11.47 and 4.24 AU in the right leg, p < 0.01). Reperfused patients had significantly higher total spectral amplitude in both legs (p < 0.04). Conclusions: In AMI patients, reduced skin blood flow and its oscillatory components may reflect ongoing neurohumoral activation despite absence of clinically apparent heart failure. The reduction of blood flow and its oscillatory components was larger in nonreperfused AMI patients, although they had a comparable left ventricular function.
Recombinant Erythropoietin Is Neuroprotective in a Novel Mouse Oxidative Injury Model
Authors: Juul Sandra E. , McPherson Ronald J. , Bammler Theodor K. , Wilkerson Jasmine , Beyer Richard P. , Farin Federico M.
Published: October 2007

To identify neuroprotective changes in gene expression, we developed a neonatal mouse model of moderate to severe oxidative brain injury and hypothesized that recombinant erythropoietin (rEpo) would decrease the expression of proapoptotic and proinflammatory genes 24 and 48 h, respectively, after injury and increase the expression of neurogenic and angiogenic genes 168 h after injury. Postnatal day 10 BALB-c mice underwent sham surgery or right common carotid artery occlusion followed by alternating hypoxia and hyperoxia and were then treated with rEpo (5,000 U/kg s.c.) or saline (vehicle) daily for up to three doses. At death, gross brain injury was assessed, then hippocampus, cortex, and thalamus were isolated for RNA or protein extraction. Microarray analysis, real-time polymerase chain reaction, and Bio-Plex® suspension array system validation were performed. rEpo decreased both incidence and severity of brain injury (median injury score 3 vs. 0, p < 0.0001) and reduced the injury-induced increases in interleukin-1α and interleukin-6 gene expression (p < 0.001), with corresponding effects on protein translation. Similarly, the expression of caspase-1, caspase-4, and caspase-6 and of p53 was increased by brain injury at 24 h, but mitigated by rEpo (p < 0.01). The interleukin-10 expression was higher in the rEpo-treated animals. Apoptotic and proinflammatory gene expression persisted for 168 h. There was no increase in angiogenic gene expression at the time points studied.
Expression of Intercellular Adhesion Molecule-1 and Vascular Cell Adhesion Molecule-1 in Human Endometrial Stromal and Epithelial Cells Is Regulated by Interferon-Gamma but Not Iron
Authors: Defrère Sylvie , Donnez Jacques , Moulin Pierre , Befahy Philippe , Gonzalez-Ramos Reinaldo , Lousse Jean-Christophe , Van Langendonckt Anne
Published: October 2007

Background/Aims: Endometrial cells are chronically exposed to iron due to cyclic menstrual bleeding. Iron induces expression of adhesion molecules in endothelial cells. The purpose of this study was to investigate iron incorporation by human endometrial cells and to test whether iron may stimulate expression of intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1. Methods: Endometrial stromal and epithelial cells were cultured in medium alone or supplemented with INF-γ or transferrin (Tf). Iron incorporation by cells was quantified by densitometry of ferritin immunostaining. ICAM-1 and VCAM-1 expression were evaluated at the transcriptional level by real-time RT-PCR. Membrane-bound and soluble protein levels of ICAM-1 were measured by quantitative immunohistochemistry and ELISA, respectively. Results: Tf induced a significant increase in ferritin immunostaining in both endometrial cell types. Endometrial cells treated with INF-γ expressed more ICAM-1 and VCAM-1 than untreated cells. By contrast, Tf treatment did not alter ICAM-1 and VCAM-1 expression in cultured endometrial cells. Conclusions: Endometrial cells are able to incorporate iron from Tf and to metabolize it to ferritin. Iron, unlike interferon-γ, does not appear to be involved in the regulation of ICAM-1 and VCAM-1 expression in cultured endometrial cells.
Smoking during Pregnancy: A Risk Factor for Peripheral Neuropathy?
Authors: Carratù M.R. , Coluccia A. , Borracci P. , Fasano A. , Riccio P. , Cuomo V.
Published: October 2007

Studies dealing with the outcomes of developmental carbon monoxide (CO) exposure on myelination in rat offspring are reviewed. Prenatal CO exposure from gestational day 0 to gestational day 20 impairs myelin deposition around peripheral axons resulting in a significant hypomyelination in juvenile and adult rats. Myelin protein patterns analyzed by SDS-polyacrylamide gel electrophoresis and lipid patterns analyzed by the HPTLC method are not altered in both peripheral and central nervous systems of CO-exposed offspring. Interestingly, when sphingomyelin is extracted and purified, the derivatization by OPA reagent and analysis by reversed-phase HPLC reveal a significant increase in sphingosine levels in peripheral nervous system but not in central nervous system of CO-exposed rats. The above morphological and biochemical alterations are not accompanied by motor disabilities.
Turning Stakeseekers Into Stakeholders
Authors: Holzer Boris
Published: October 2007

Many firms, especially transnational corporations, find it increasingly difficult to predict and handle conflicts with external interest groups. In addition to a set of established stakeholders, they face a complex arena of newly emerging “stakeseekers” who also claim to have a stake in the corporation's decision making. Corporations seek to establish relationships with such groups to anticipate and prevent conflicts that could otherwise wreak havoc on their reputation. Through stakeholder engagement, stakeseekers may be turned into stakeholders. It has been argued that in the process, critics are simply co-opted. However, corporate objectives are not necessarily monolithic. Conceived as a political coalition of various interests, the structure of the corporation often reflects societal divisions. Parts of the corporation can thus serve as sounding boards for external claims. Referring to the case of Royal Dutch/Shell in particular, this article shows how a political coalition perspective on the corporation may help to understand and analyze the politics of stakeholder influence.
Counseling Adopted Persons in Adulthood
Authors: Baden Amanda L. , O'Leary Wiley Mary
Published: October 2007

For the past 50 years, adults who were adopted during infancy have been research participants for empirical studies with goals ranging from twin studies for heritability, to adjustment following adoption, to attachment. While the research body is broad, it has given little attention to counseling practices with adopted adults. Because empirical research and clinical practice can inform each other, this article integrates literatures in both areas so that counseling practice with adopted adults can guide research, just as research guides practice. The authors grouped the clinically relevant literature into three main areas: identity (including genealogical and transracial adoption issues), search and reunion, and long-term outcomes. Within each section, the authors critiqued the literature as it informs counseling practice, used case studies to depict clinical implications, and suggested treatment strategies for use with adult adoptees. Epidemiological research found adequate adjustment for adopted adults. However, clinicians and researchers must address the consistent finding that a subset of adoptees struggles and copes with issues different than their nonadopted counterparts. The authors identify best clinical practices and a future research agenda related to adult adoptees and propose an adoption-sensitive paradigm for research and practice.
Culture, Prevention, and the Politics of Disparities
Authors: Vera Elizabeth M.
Published: October 2007

The following article is a response to Adams's (2007 [this issue]) and Griffin and Miller's (2007 [this issue]) reactions to the Major Contribution articles presented on Culturally Relevant Prevention. Each reaction article identifies important implications for engaging in culturally relevant prevention efforts that are relevant to developing this line of research in the future and integrating such work into our training programs. In particular, the importance of defining culture carefully and the politics of disparities are further discussed in this response article. In addition, the importance of learning from other disciplines how to integrate prevention practice and research into counseling psychology training is discussed.
Testing Murphy's Strategic Model
Authors: Arrington Theodore S. , Brenner Saul
Published: October 2007

Murphy contended that an astute chief justice could assign the majority opinion to the “most moderate member” of the conference coalition, hoping that such an assignment “might prevent defection or even gain adherents.” We discovered that Murphy's model was partially supported with data from the Vinson, Warren, and Burger Courts. When the conference coalition was larger than minimum winning (mw), assignment of the majority opinion to the marginal justice in the conference coalition was more likely to result in an opinion joined by the other justices in the conference coalition, making it an effective defensive strategy. It may not, however, be a very effective offensive strategy.
Physicochemical conditions and timing of rodingite formation: evidence from rodingite-hosted fluid inclusions in the JM Asbestos mine, Asbestos, Québec
Authors: Normand Charles , Williams-Jones Anthony E
Published: October 2007

Fluid inclusions and geological relationships indicate that rodingite formation in the Asbestos ophiolite, Québec, occurred in two, or possibly three, separate episodes during thrusting of the ophiolite onto the Laurentian margin, and that it involved three fluids. The first episode of rodingitization, which affected diorite, occurred at temperatures of between 290 and 360°C and pressures of 2.5 to 4.5 kbar, and the second episode, which affected granite and slate, occurred at temperatures of between 325 and 400°C and pressures less than 3 kbar. The fluids responsible for these episodes of alteration were moderately to strongly saline (~1.5 to 6.3 m eq. NaCl), rich in divalent cations and contained appreciable methane. A possible third episode of alteration is suggested by primary fluid inclusions in vesuvianite-rich bodies and secondary inclusions in other types of rodingite, with significantly lower trapping temperatures, salinity and methane content. The association of the aqueous fluids with hydrocarbon-rich fluids containing CH4 and higher order alkanes, but no CO2, suggests strongly that the former originated from the serpentinites. The similarities in the composition of the fluids in all rock types indicate that the ophiolite had already been thrust onto the slates when rodingitization occurred.
InPrePPI: an integrated evaluation method based on genomic context for predicting protein-protein interactions in prokaryotic genomes
Authors: Sun Jingchun , Sun Yan , Ding Guohui , Liu Qi , Wang Chuan , He Youyu , Shi Tieliu , Li Yixue , Zhao Zhongming
Published: October 2007

Background

Although many genomic features have been used in the prediction of protein-protein interactions (PPIs), frequently only one is used in a computational method. After realizing the limited power in the prediction using only one genomic feature, investigators are now moving toward integration. So far, there have been few integration studies for PPI prediction; one failed to yield appreciable improvement of prediction and the others did not conduct performance comparison. It remains unclear whether an integration of multiple genomic features can improve the PPI prediction and, if it can, how to integrate these features.

Results

In this study, we first performed a systematic evaluation on the PPI prediction in Escherichia coli (E. coli) by four genomic context based methods: the phylogenetic profile method, the gene cluster method, the gene fusion method, and the gene neighbor method. The number of predicted PPIs and the average degree in the predicted PPI networks varied greatly among the four methods. Further, no method outperformed the others when we tested using three well-defined positive datasets from the KEGG, EcoCyc, and DIP databases. Based on these comparisons, we developed a novel integrated method, named InPrePPI. InPrePPI first normalizes the AC value (an integrated value of the accuracy and coverage) of each method using three positive datasets, then calculates a weight for each method, and finally uses the weight to calculate an integrated score for each protein pair predicted by the four genomic context based methods. We demonstrate that InPrePPI outperforms each of the four individual methods and, in general, the other two existing integrated methods: the joint observation method and the integrated prediction method in STRING. These four methods and InPrePPI are implemented in a user-friendly web interface.

Conclusion

This study evaluated the PPI prediction by four genomic context based methods, and presents an integrated evaluation method that shows better performance in E. coli.
ProCKSI: a decision support system for Protein (Structure) Comparison, Knowledge, Similarity and Information
Authors: Barthel Daniel , Hirst Jonathan D , Błażewicz Jacek , Burke Edmund K , Krasnogor Natalio
Published: October 2007

Background

We introduce the decision support system for Protein (Structure) Comparison, Knowledge, Similarity and Information (ProCKSI). ProCKSI integrates various protein similarity measures through an easy to use interface that allows the comparison of multiple proteins simultaneously. It employs the Universal Similarity Metric (USM), the Maximum Contact Map Overlap (MaxCMO) of protein structures and other external methods such as the DaliLite and the TM-align methods, the Combinatorial Extension (CE) of the optimal path, and the FAST Align and Search Tool (FAST). Additionally, ProCKSI allows the user to upload a user-defined similarity matrix supplementing the methods mentioned, and computes a similarity consensus in order to provide a rich, integrated, multicriteria view of large datasets of protein structures.

Results

We present ProCKSI's architecture and workflow describing its intuitive user interface, and show its potential on three distinct test-cases. In the first case, ProCKSI is used to evaluate the results of a previous CASP competition, assessing the similarity of proposed models for given targets where the structures could have a large deviation from one another. To perform this type of comparison reliably, we introduce a new consensus method. The second study deals with the verification of a classification scheme for protein kinases, originally derived by sequence comparison by Hanks and Hunter, but here we use a consensus similarity measure based on structures. In the third experiment using the Rost and Sander dataset (RS126), we investigate how a combination of different sets of similarity measures influences the quality and performance of ProCKSI's new consensus measure. ProCKSI performs well with all three datasets, showing its potential for complex, simultaneous multi-method assessment of structural similarity in large protein datasets. Furthermore, combining different similarity measures is usually more robust than relying on one single, unique measure.

Conclusion

Based on a diverse set of similarity measures, ProCKSI computes a consensus similarity profile for the entire protein set. All results can be clustered, visualised, analysed and easily compared with each other through a simple and intuitive interface.

ProCKSI is publicly available at http://www.procksi.net for academic and non-commercial use.
Database of Trypanosoma cruzi repeated genes: 20 000 additional gene variants
Authors: Arner Erik , Kindlund Ellen , Nilsson Daniel , Farzana Fatima , Ferella Marcela , Tammi Martti T , Andersson Björn
Published: October 2007

Background

Repeats are present in all genomes, and often have important functions. However, in large genome sequencing projects, many repetitive regions remain uncharacterized. The genome of the protozoan parasite Trypanosoma cruzi consists of more than 50% repeats. These repeats include surface molecule genes, and several other gene families. In the T. cruzi genome sequencing project, it was clear that not all copies of repetitive genes were present in the assembly, due to collapse of nearly identical repeats. However, at the time of publication of the T. cruzi genome, it was not clear to what extent this had occurred.

Results

We have developed a pipeline to estimate the genomic repeat content, where shotgun reads are aligned to the genomic sequence and the gene copy number is estimated using the average shotgun coverage. This method was applied to the genome of T. cruzi and copy numbers of all protein coding sequences and pseudogenes were estimated. The 22 640 results were stored in a database available online. 18% of all protein coding sequences and pseudogenes were estimated to exist in 14 or more copies in the T. cruzi CL Brener genome. The average coverage of the annotated protein coding sequences and pseudogenes indicate a total gene copy number, including allelic gene variants, of over 40 000.

Conclusion

Our results indicate that the number of protein coding sequences and pseudogenes in the T. cruzi genome may be twice the previous estimate. We have constructed a database of the T. cruzi gene repeat data that is available as a resource to the community. The main purpose of the database is to enable biologists interested in repeated, unfinished regions to closely examine and resolve these regions themselves using all available shotgun data, instead of having to rely on annotated consensus sequences that often are erroneous and possibly misleading. Five repetitive genes were studied in more detail, in order to illustrate how the database can be used to analyze and extract information about gene repeats with different characteristics in Trypanosoma cruzi.
Depression in the elderly: Does family system play a role? A cross-sectional study
Authors: Taqui Ather M , Itrat Ahmed , Qidwai Waris , Qadri Zeeshan
Published: October 2007

Background

The most common geriatric psychiatric disorder is depression. The role of family systems in depression among the elderly has not been studied extensively. It has been suggested that urbanization promotes nucleation of family systems and a decrease in care and support for the elderly. We conducted this study in Karachi, a large urban city of Pakistan, to determine the relationship between the type of family system and depression. We also determined the prevalence of depression in the elderly, as well as correlation of depression with other important socio-demographic variables.

Methods

A cross-sectional study was carried out in the premises of a tertiary care hospital in Karachi, Pakistan. Questionnaire based interviews were conducted among the elderly people visiting the hospital. Depression was assessed using the 15-item Geriatric Depression Scale.

Results

Four hundred subjects aged 65 and above were interviewed. The age of majority of the subjects ranged from 65 to 74 years. Seventy eight percent of the subjects were male. The prevalence of depression was found to be 19.8%. Multiple logistic regression analysis revealed that the following were significant (p < 0.05) independent predictors of depression: nuclear family system, female sex, being single or divorced/widowed, unemployment and having a low level of education. The elderly living in a nuclear family system were 4.3 times more likely to suffer from depression than those living in a joint family system (AOR = 4.3 [95% CI = 2.4–7.6]).

Conclusion

The present study found that residing in a nuclear family system is a strong independent predictor of depression in the elderly. The prevalence of depression in the elderly population in our study was moderately high and a cause of concern. The transition in family systems towards nucleation may have a major deleterious effect on the physical and mental health of the elderly.
Hypertabastic survival model
Authors: Tabatabai Mohammad A , Bursac Zoran , Williams David K , Singh Karan P
Published: October 2007

A new two-parameter probability distribution called hypertabastic is introduced to model the survival or time-to-event data. A simulation study was carried out to evaluate the performance of the hypertabastic distribution in comparison with popular distributions. We then demonstrate the application of the hypertabastic survival model by applying it to data from two motivating studies. The first one demonstrates the proportional hazards version of the model by applying it to a data set from multiple myeloma study. The second one demonstrates an accelerated failure time version of the model by applying it to data from a randomized study of glioma patients who underwent radiotherapy treatment with and without radiosensitizer misonidazole. Based on the results from the simulation study and two applications, the proposed model shows to be a flexible and promising alternative to practitioners in this field.
Motavizumab, A Neutralizing Anti-Respiratory Syncytial Virus (Rsv) Monoclonal Antibody Significantly Modifies The Local And Systemic Cytokine Responses Induced By Rsv In The Mouse Model
Authors: Mejías Asunción , Chávez-Bueno Susana , Raynor Martin B , Connolly John , Kiener Peter A , Jafri Hasan S , Ramilo Octavio
Published: October 2007

Motavizumab (MEDI-524) is a monoclonal antibody with enhanced neutralizing activity against RSV. In mice, motavizumab suppressed RSV replication which resulted in significant reduction of clinical parameters of disease severity. We evaluated the effect of motavizumab on the local and systemic immune response induced by RSV in the mouse model. Balb/c mice were intranasally inoculated with 106.5 PFU RSV A2 or medium. Motavizumab was given once intraperitoneally (1.25 mg/mouse) as prophylaxis, 24 h before virus inoculation. Bronchoalveolar lavage (BAL) and serum samples were obtained at days 1, 5 (acute) and 28 (long-term) post inoculation and analyzed with a multiplex assay (Beadlyte Upstate, NY) for simultaneous quantitation of 18 cytokines: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, KC (similar to human IL-8), IL-10, IL-12p40, IL-12p70, IL-13, IL-17, TNF-α, MCP-1, RANTES, IFN-γ and GM-CSF. Overall, cytokine concentrations were lower in serum than in BAL samples. By day 28, only KC was detected in BAL specimens at low concentrations in all groups. Administration of motavizumab significantly reduced (p < 0.05) BAL concentrations of IL-1α, IL-12p70 and TNF-α on day 1, and concentrations of IFN-γ on days 1 and 5 compared with RSV-infected untreated controls. In the systemic compartment, the concentrations of IL-10, IFN-γ and KC were significantly reduced in the motavizumab-treated mice compared with the untreated controls. In summary, prophylactic administration of motavizumab was associated with significant reductions on RSV replication and concentrations of cytokine and chemokines, which are likely related to the improvement observed in clinical markers of disease severity.
Thiol-reactive reagents inhibits intracellular trafficking of human papillomavirus type 16 pseudovirions by binding to cysteine residues of major capsid protein L1
Authors: Ishii Yoshiyuki , Kondo Kazunari , Matsumoto Tamae , Tanaka Keiko , Shinkai-Ouchi Fumiko , Hagiwara Ken'ichi , Kanda Tadahito
Published: October 2007

Background

A human papillomavirus (HPV) virion is composed of capsid proteins L1 and L2. Several cysteine residues are located on L1 of various HPVs at markedly similar relative positions, suggesting their important functions. Although the authentic virions cannot be studied with cultured cells, surrogate pseudovirions consisting of capsid and reporter plasmid are available for studies dealing with infectivity.

Results

HPV type16-pseudovirions (16PVs) were found to lose their infectivity after incubation with thiol-reactive reagents [biotin polyethyleneoxide iodoacetamide (BPEOIA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), N-ethylmaleimide (NEM), 4-(N-maleimido)benzyl-trimethylammonium iodide (MBTA), and [2-(trimethylammonium)ethyl] methanethiosulfonate bromide (MTSET)]. A labelled streptavidin was detected to bind to the complex of BPEOIA and L1 of the 16PVs incubated with BPEOIA. The analysis of molecular mass of trypsin-fragments derived from the complex of the BPEOIA and L1 indicated that BPEOIA bound to at least C146, C225, and C229. No appreciable change of the 16PVs carrying DTNB or NEM was detected by sedimentation analysis or electron microscopy. The 16PVs carrying DTNB or NEM were able to bind to and enter HeLa cells but degraded before they reached the perinuclear region.

Conclusion

HPV16 L1 C146, C225, and C229 have free thiol, which are accessible to BPEOIA, DTNB, NEM, MBTA, and MTSET. Binding of DTNB or NEM to the thiols may cause conformational changes that result in the inhibition of the entry and trafficking of the 16PVs.
RAId_DbS: Peptide Identification using Database Searches with Realistic Statistics
Authors: Alves Gelio , Ogurtsov Aleksey Y , Yu Yi-Kuo
Published: October 2007

Background

The key to mass-spectrometry-based proteomics is peptide identification. A major challenge in peptide identification is to obtain realistic E-values when assigning statistical significance to candidate peptides.

Results

Using a simple scoring scheme, we propose a database search method with theoretically characterized statistics. Taking into account possible skewness in the random variable distribution and the effect of finite sampling, we provide a theoretical derivation for the tail of the score distribution. For every experimental spectrum examined, we collect the scores of peptides in the database, and find good agreement between the collected score statistics and our theoretical distribution. Using Student's t-tests, we quantify the degree of agreement between the theoretical distribution and the score statistics collected. The T-tests may be used to measure the reliability of reported statistics. When combined with reported P-value for a peptide hit using a score distribution model, this new measure prevents exaggerated statistics. Another feature of RAId_DbS is its capability of detecting multiple co-eluted peptides. The peptide identification performance and statistical accuracy of RAId_DbS are assessed and compared with several other search tools. The executables and data related to RAId_DbS are freely available upon request.
Cytofluorographic and molecular identification of a CD8-positive, TCR-α/β-negative intraocular T cell lymphoma: a case report and review of the literature
Authors: Saenz Alvaro D , Amador Alexandra , Ruiz Brianna M , Davis Janet , Ruiz Phillip
Published: October 2007

Introduction

Cytofluorographic and molecular techniques are effective adjuncts in diagnosing intraocular lymphoma. Primary intraocular lymphoma is an uncommon entity predominantly of B cell origin and rarely with a T cell phenotype. The aim of the present paper is to report a case of a CD8-positive, TCR-α/β-negative intraocular T cell lymphoma and review the literature.

Case presentation

T cell neoplasia was detected based on flow cytometric demonstration of an abnormal T cell population and polymerase chain reactions for immunoglobulin and T-cell receptor rearrangements demonstrating evidence of monoclonality. Flow cytometry revealed a T cell population aberrantly expressing T-cell lineage markers. This T cell population expressed CD2, bright CD3, CD8, bright CD7, CD38, CD69, and variable CD25. T-cell receptor γ gene rearrangement studies demonstrated evidence of T-cell gene rearrangement confirming that the T cells were monoclonal.

Conclusion

We herein report the rare case of a TCR α/β-negative CD8+ intraocular T-cell lymphoma suggestive of gamma/delta origin diagnosed by flow cytometry and polymerase chain reaction.
Importance of spliceosomal RNP1 motif for intermolecular T-B cell spreading and tolerance restoration in lupus
Authors: Monneaux Fanny , Parietti Véronique , Briand Jean-Paul , Muller Sylviane
Published: October 2007

We previously demonstrated the importance of the RNP1 motif-bearing region 131–151 of the U1-70K spliceosomal protein in the intramolecular T-B spreading that occurs in MRL/lpr lupus mice. Here, we analyze the involvement of RNP1 motif in the development and prevention of naturally-occurring intermolecular T-B cell diversification. We found that MRL/lpr peripheral blood lymphocytes proliferated in response to peptides containing or corresponding exactly to the RNP1 motif of spliceosomal U1-70K, U1-A and hnRNP-A2 proteins. We also demonstrated that rabbit antibodies to peptide 131–151 cross-reacted with U1-70K, U1-A and hnRNP-A2 RNP1-peptides. These antibodies recognized the U1-70K and U1-A proteins, and also U1-C and SmD1 proteins, which are devoid of RNP1 motif. Repeated administration of phosphorylated peptide P140 into MRL/lpr mice abolished T-cell response to several peptides from the U1-70K, U1-A and SmD1 proteins without affecting antibody and T-cell responses to foreign (viral) antigen in treated mice challenged with infectious virus. These results emphasized the importance of the dominant RNP1 region, which seems to be central in the activation cascade of B and T cells reacting with spliceosomal RNP1+ and RNP1- spliceosomal proteins. The tolerogenic peptide P140, which is recognized by lupus patients' CD4+ T cells and known to protect MRL/lpr mice, is able to thwart emergence of intermolecular T-cell spreading in treated animals.
Different measures of smoking exposure and mammographic density in postmenopausal Norwegian women: a cross-sectional study
Authors: Bremnes Yngve , Ursin Giske , Bjurstam Nils , Gram Inger T
Published: October 2007

Background

Recent cohort studies have suggested an increased risk of breast cancer with long duration of smoking, and with smoking initiation before first birth. Cigarette smoking may have both carcinogenic effects and antiestrogenic effects on the breast tissue. We decided to examine the relationship between different measures of smoking exposure and mammographic density.

Methods

Lifetime smoking history was collected through interview and questionnaires among 907 postmenopausal participants in the Tromsø Mammography and Breast Cancer study. The mammograms were obtained from the governmental Norwegian Breast Cancer Screening Program. Mammograms were classified according to the percentage and absolute mammographic densities using a previously validated computer-assisted method.

Results

Sixty-five percent of the women reported having ever smoked cigarettes, while 34% were current smokers. After adjustment for age, age at first birth, parity, age at menopause, postmenopausal hormone therapy use, and body mass index, smoking was inversely associated with both measures of mammographic density (both trends P < 0.01). Both current smokers and former smokers had significantly lower adjusted mean percentage mammographic density compared with never smokers (P = 0.003 and P = 0.006, respectively). An inverse dose–response relationship with mammographic density was found between both the number of cigarettes and the number of pack-years smoked among current smokers. Current smokers who smoked 11 cigarettes or more daily had a 3.7% absolute (36% relative difference) lower percentage mammographic density compared with current smokers who smoked seven cigarettes or less daily (P = 0.008). When former smokers were stratified according to time since smoking cessation, we found that women who had stopped smoking less than 24 years ago had a significantly lower mean percentage mammographic density compared with never smokers (P < 0.001).

Conclusion

We found modest inverse dose–response associations between numbers of cigarettes and of pack-years smoked and both measures of mammographic density among current smokers. Former smokers who had stopped smoking less than 24 years ago also had a statistically significantly lower mean percentage mammographic density when compared with never smokers. These findings are consistent with an antiestrogenic effect of cigarette smoking on the breast tissue.
Impaired thymic selection in mice expressing altered levels of the SLP-76 adaptor protein
Authors: Ramsey Kimberley , Luckashenak Nancy , Koretzky Gary A , Clements James L
Published: October 2007

Intracellular signaling initiated by ligation of the TCR influences cell fate at multiple points during the lifespan of a T cell. This is especially evident during thymic selection, where the nature of TCR-dependent signaling helps to establish a MHC-restricted, self-tolerant T cell repertoire. The Src homology 2 domain-containing leukocyte-specific phosphoprotein of 76 kDa (SLP-76) adaptor protein is a required intermediate in multiple signaling pathways triggered by TCR engagement, several of which have been implicated in dictating the outcome of thymic selection (e.g., intracellular calcium flux and activation of ERK family MAPKs). To determine if thymocyte maturation and selection at later stages of development are sensitive to perturbations in SLP-76 levels, we analyzed these crucial events using several transgenic (Tg) lines of mice expressing altered levels of SLP-76 in the thymus. In Tg mice expressing low levels of SLP-76 in preselection thymocytes, the CD4:CD8 ratio in the thymus and spleen was skewed in a manner consistent with impaired selection and/or maturation of CD4+ thymocytes. Low SLP-76 expression also correlated with reduced CD5 expression on immature thymocytes, consistent with reduced TCR signaling potential. In contrast, reconstitution of SLP-76 at higher levels resulted in normal thymic CD5 expression and CD4:CD8 ratios in the thymus and periphery. It is curious that thymic deletion of TCR-Tg (HY) thymocytes was markedly impaired in both lines of Tg-reconstituted SLP-76−/− mice. Studies using chimeric mice indicate that the defect in deletion of HY+ thymocytes is intrinsic to the developing thymocyte, suggesting that maintenance of sufficient SLP-76 expression from the endogenous locus is a key element in the selection process.
Satellite and stem cells in muscle growth and repair
Authors: Le Grand Fabien , Rudnicki Michael
Published: October 2007

The FASEB summer research conference on Skeletal Muscle Satellite and Stem Cells, organized by Thomas Rando, Giulio Cossu and Jeffrey Chamberlain, was held in Indian Wells, California, in July. An international array of researchers gathered to share numerous new insights into the cellular and molecular regulation of stem cells and satellite cells in skeletal muscle biology. The conference is unique in that it brings together investigators from diverse backgrounds, who work on the growth and repair of skeletal muscle in humans and model systems, in health and disease.
Snail1a and Snail1b cooperate in the anterior migration of the axial mesendoderm in the zebrafish embryo
Authors: Blanco María J. , Barrallo-Gimeno Alejandro , Acloque Hervé , Reyes Ariel E. , Tada Masazumi , Allende Miguel L. , Mayor Roberto , Nieto M. Angela
Published: October 2007

The Snail genes are implicated in processes that involve cell movement,both during embryonic development and tumour progression. In teleosts, the vertebrate Snail1 gene is represented by two distinct genes, snail1a and snail1b (previously snail1 and snail2). These genes are expressed in complementary mesodermal domains and their combined expression matches that of their mammalian counterpart. By analysing their loss and gain of function, we found that the most-anterior axial mesendodermal cells, the precursors of the polster, move in a cohesive manner directed by the activity of snail1a- and snail1b-expressing cells surrounding these precursors. The cell-autonomous function of Snail1 proteins regulates cell motility and influences the behaviour of Snail-negative neighbouring cells. Snail1a is required by the prechordal plate for it to reach its normal position, whereas Snail1b controls the acquisition of its normal shape. These non-redundant functions of Snail1a and Snail1b in controlling axial mesendoderm migration comply with the duplication-degeneration-complementation model, and indicate that Snail genes not only act as inducers of epithelial-to-mesenchymal transition, but also as more general regulators of cell adhesion and movement.
Arabidopsis GLAUCEpromotes fertilization-independent endosperm development and expression of paternally inherited alleles
Authors: Ngo Quy A. , Moore James M. , Baskar Ramamurthy , Grossniklaus Ueli , Sundaresan Venkatesan
Published: October 2007

Early seed development of sexually reproducing plants requires both maternal and paternal genomes but is prominently maternally influenced. A novel gametophytic maternal-effect mutant defective in early embryo and endosperm development, glauce (glc), has been isolated from a population of Arabidopsis Ds transposon insertion lines. The glc mutation results from a deletion at the Ds insertion site, and the molecular identity of GLC is not known. glcembryos can develop up to the globular stage in the absence of endosperm and glc central cells appear to be unfertilized. glc suppresses autonomous endosperm development observed in the fertilization-independent seed (fis) class mutants. glc is also epistatic to mea, one of the fis class mutants, in fertilized seeds, and is essential for the biparental embryonic expression of PHE1, a repressed downstream target of MEA. In addition, maternal GLC function is required for the paternal embryonic expression of the ribosome protein gene RPS5a and the AMP deaminase gene FAC1,both of which are essential for early embryo and endosperm development. These results indicate that factors derived from the female gametophyte activate a subset of the paternal genome of fertilized seeds.
Management of L-Dopa related hyperhomocysteinemia: catechol-O-methyltransferase (COMT) inhibitors or B vitamins? Results from a review
Authors: Zoccolella Stefano , Iliceto Giovanni , deMari Michele , Livrea Paolo , Lamberti Paolo
Published: October 2007

In recent years, L-Dopa treatment has been indicated as an acquired cause of hyperhomocysteinemia (HHcy). The mechanism underlying L-Dopa-related HHcy is the O-methylation of the drug catalyzed by the enzyme catechol-O-methyltransferase (COMT). Folate and cobalamin status also influences the effects of L-Dopa on plasma homocysteine (Hcy) levels. Although clinical correlations of HHcy in Parkinson's disease still remain uncertain, management of elevated plasma Hcy levels has been advocated, due to multiple cytotoxic effects of Hcy on neurons. This review summarizes data available in the literature concerning the two main therapeutic approaches to L-Dopa-related HHcy (use of COMT inhibitors or B vitamins diet supplementation).

Clin Chem Lab Med 2007;45:1607–13.
Quantitative profiling of folate and one-carbon metabolism in large-scale epidemiological studies by mass spectrometry
Authors: Ueland Per Magne , Midttun Øivind , Windelberg Amrei , Svardal Asbjørn , Skålevik Rita , Hustad Steinar
Published: October 2007

Clin Chem Lab Med 2007;45:1737–45.
Metabolic regulatory properties of S-adenosylmethionine and S-adenosylhomocysteine
Authors: Finkelstein James D.
Published: October 2007

In mammalian liver, two intersecting pathways, remethylation and transsulfuration, compete for homocysteine that has been formed from methionine. Remethylation of homocysteine, employing either methyltetrahydrofolate or betaine as the methyl donor, forms a methionine cycle that functions to conserve methionine. In contrast, the transsulfuration sequence – cystathionine synthase and cystathionase – serves to irreversibly catabolize the homocysteine while synthesizing cysteine. The rate of homocysteine formation and its distribution between these two pathways are the sites for metabolic regulation and coordination. The mechanisms for regulation include both the tissue content and the kinetic properties of the component enzymes as well as the concentrations of their substrates and other metabolic effectors. Adenosylmethionine and adenosylhomocysteine are important regulatory metabolites and may use one or more mechanisms to affect the enzymes. Adenosylmethionine is a positive effector of its own synthesis, cystathionine synthase and glycine methyltransferase but impairs both homocysteine methylases. Thus, the concentration of adenosylmethionine may be self-regulatory in mammalian liver. By means of other enzymatic mechanisms, the hepatic concentration of adenosylhomocysteine, an index of homocysteine accumulation, is also self-regulated. These considerations pertain primarily to liver, which has the unique capacity to synthesize more adenosylmethionine in the presence of excess methionine. However, there are organ-specific patterns of methionine metabolism and its regulation. All tissues possess the methionine cycle with methyltetrahydrofolate as the methyl donor but only liver, kidney, pancreas, intestine and brain also contain the transsulfuration pathway. The limitation of adenosylmethionine concentrations may make adenosylhomocysteine a more significant metabolic regulator in extrahepatic tissues. However, estimates of regulatory changes based on determinations of the plasma concentrations of the two metabolites are of limited value and must be used with caution. In addition, the recent description of “cystathionine (CBS) domains” in proteins not involved with methionine metabolism raises the possibility that abnormal concentrations of the adenosyl metabolites may impact on other metabolic pathways.

Clin Chem Lab Med 2007;45:1694–9.