bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of ‘tongue tie’ use on racing performance of Thoroughbreds in the United Kingdom
Authors: Barakzai S. Z. , Finnegan C. , Boden L. A.
Published: November 2009

Reasons for performing study: There is contradictory published evidence on the potential efficacy of ‘tongue ties’ (TTs) for treatment of intermittent dorsal displacement of the soft palate (DDSP) in racehorses.

Objectives: To evaluate the effect of TTs on racing performance in Thoroughbred racehorses in the UK using a retrospective cohort study.

Methods: Data on individual horses' lifetime racing performance and TT use were retrieved from the Racing Post Online Database. Exposed cases were horses that ran with a TT in randomly chosen race meetings on one of 60 randomly chosen dates from 2001–2003. Unexposed (control) horses were matched to each exposed horse. Various measures of racing performance were analysed both within and between exposed and unexposed groups. Subsets of exposed horses that ran for 3 or 5 consecutive starts wearing TTs and their matched controls were analysed separately to examine the effect of repeated TT use.

Results: The inclusion criteria were fulfilled by 108 horses. The odds ratio for ‘improvement’ in race earnings between exposed and unexposed horses was 1.85 for horses that ran at least once with a TT, and 3.60 and 4.24, respectively, for horses that ran in 3 or 5 consecutive races wearing a TT. After instigation of TT use, horses that ran in 3 or 5 consecutive races wearing a TT had a significant increase in earnings when they ran wearing a TT compared to their pre‐TT races.

Conclusions and potential relevance: The use of a TT appears to have a beneficial effect on racing performance in a selected population of Thoroughbred racehorses.
Radiographic and scintigraphic evaluation of spondylosis in the equine thoracolumbar spine: A retrospective study
Authors: Meehan L. , Dyson S. , Murray R.
Published: November 2009

Reasons for performing study: Clinical, radiographic and scintigraphic signs associated with spondylosis of the equine thoracolumbar spine have been poorly documented.

Objectives: To establish an objective radiographic and scintigraphic grading system for spondylosis lesions; to estimate the prevalence of spondylosis in a population of horses with back pain; and to compare the results of radiography and scintigraphy

Methods: Radiographic images of the thoracolumbar spine from 670 horses with clinical signs of back pain were graded. Scintigraphic images from horses with spondylosis lesions underwent subjective and objective analysis. Sensitivity and specificity of scintigraphy for detection of spondylosis relative to radiography for identification of spondylosis were calculated, and Chi‐squared analysis was performed to test for an association between location and severity of lesions.

Results: Twenty‐three of 670 horses (3.4%) with back pain had radiographic evidence of spondylosis. Of these horses, 14 (61%) had more than one lesion and 44% (n = 22) of lesions occurred between T11‐T13 vertebral bodies. Only 33% (n = 28) of locations with radiographic changes had increased radiopharmaceutical uptake.

Conclusion: Spondylosis occurs at a low prevalence in horses with back pain. It may be present alone or in association with other osseous abnormalities. The clinical significance of spondylosis needs further investigation.

Potential relevance: Spondylosis is uncommon but may be a contributor to back pain in the horse.
The Rate of Missed Test Results in an Emergency Department
Authors: Paoloni R. , Georgiou A. , Prgomet M. , Westbrook J. , Callen J.
Published: November 2009

Objectives: 1) To measure the incidence and impact of missed radiology and microbiology test results in an emergency department with an electronic test order and results viewing system, and 2) to assess the average times from test order to test availability.

Methods: The study was conducted in the emergency department (ED) of a 370-bed metropolitan teaching hospital in Sydney, Australia. A computerised provider order entry (CPOE) system was used to order all diagnostic tests and view all test results. For microbiology and radiology tests electronic results were then printed for ED patients not admitted to the hospital to allow ED physicians to document follow-up. All radiology (n = 197) and microbiology (n = 66) tests ordered and results received for discharged ED patients were collected for a seven-day period. We measured the: 1) proportion of radiology and microbiology test results without follow-up for discharged patients; 2) impact of non follow-up on patient outcomes; 3) average time from radiological examination and microbiology specimen collection to reporting of results; and 4) average time from reporting of results to follow-up.

Results: Two radiology (1.0%) and two microbiology reports (3.0%), all of which had negative findings, were never followed-up. Review of these patients’ medical records indicated there was no impact on patient outcomes or management. The average time from radiological examination to reporting of a result was 1.5 days, and from microbiology specimen collection to reporting was 2.5 days. Eighty-nine percent of radiology and 68% of microbiology results were followed-up on the same day that they were available to physicians.

Conclusions: Our rates of missed test results are lower than those reported from studies where paper ordering and reporting systems were used. This suggests that the availability of CPOE systems may reduce the risk of these events. Electronic result delivery, with electronic endorsement to allow documentation of follow-up of test results, may provide additional efficiency benefits and further reduce the risk of test results which are not followed up.
Development of a System for Measurement and Analysis of Tremor Using a Three-axis Accelerometer
Authors: Iizuka T. , Takeuchi A. , Shirataka M. , Ikeda N. , Mamorita N.
Published: November 2009

Objectives: The aim of the study was to develop a low-cost and compact system for analysis of tremor using a three-axis accelerometer (the Wii Remote (Nintendo)). To analyze tremor, we hypothesized that the influence of gravitational acceleration should be separated from that of movement. This hypothesis was tested experimentally and we also attempted to record and analyze tremor using our system in a clinical ward.

Methods: A system for tremor measurement and analysis was developed using the three-axis accelerometer built into the Wii Remote. The frequency and amplitude of mechanical oscillation were calculated using methods for frequency analysis of the axis of largest variance and an estimation of tremor amplitude.

Results: The system consists of a program for measurement and analysis of Wii Remote acceleration (Tremor Analyzer), a Wii Remote, a Bluetooth USB adapter and a Web camera. The Tremor Analyzer has a GUI (graphical user interface) that is divided into five segments. The sampling period of the analyzer is 30 msec. To confirm the hypothesis, mechanical oscillations were fed to the Wii Remote. The peak frequency of the power spectrum and the frequency of the oscillation generator were in good agreement, except at 1 Hz (0.01 G) and 2 Hz (0.02 G). With a change in the sum of squares of the three axes from 1.0 to 1.8 (G), the estimated and generated amplitude (0.3 cm) were in close agreement.

Conclusions: This system using a Wii Remote is capable of analyzing frequency and estimated amplitude of tremor between 3 Hz and 15 Hz.
Female and Male Sex Hormones Differentially Regulate Expression ofIfi202, an Interferon-Inducible Lupus Susceptibility Gene within theNba2Interval
Authors: Panchanathan Ravichandran , Shen Hui , Bupp Melanie Gubbels , Gould Karen A. , Choubey Divaker
Published: November 2009

Increased expression of IFN-inducible Ifi202 gene in certain strains of female mice is associated with susceptibility to systemic lupus erythematosus (SLE). Although, the development of SLE is known to have a strong sex bias, the molecular mechanisms remain unknown. Here we report that in vivo treatment of orchiectomized (NZB × NZW)F1 male mice with the female sex hormone 17β-estradiol significantly increased steady-state levels of Ifi202 mRNA in splenic cells, whereas treatment with the male hormone dihydrotestosterone decreased the levels. Moreover, increased expression of Ifi202 in B6.Nba2 B cells and reduced expression in T cells were associated with increased levels of estrogen receptor-α (ERα) and androgen receptor, respectively. Furthermore, the steady-state levels of Ifi202 mRNA were higher in splenic cells from C57BL/6, B6.Nba2, NZB, and (NZB × NZW)F1 female mice as compared with males. 17β-estradiol treatment of B cells and WT276 cells increased Ifi202 mRNA levels, whereas treatment with dihydrotestosterone decreased the levels. Interestingly, overexpression of ERα in WT276 cells increased the expression of Ifi202 and stimulated the activity of the 202-luc-reporter through the c-Jun/AP-1 DNA-binding site. Accordingly, ERα preferentially associated with the regulatory region of the Ifi202 gene in female B6.Nba2 B cells than in males. Furthermore, Ifi202 mRNA levels were detectable in splenic cells of wild-type (Esr1+/+), but not null (Esr1−/−), (NZB × NZW)F1 female mice. Collectively, our observations demonstrate that the female and male sex hormones differentially regulate the expression of Ifi202, thus providing support for the role of Ifi202 in sex bias in SLE.
Natural Occurring IL-17 Producing T Cells Regulate the Initial Phase of Neutrophil Mediated Airway Responses
Authors: Tanaka Shinya , Yoshimoto Takayuki , Naka Tetsuji , Nakae Susumu , Iwakura Yo-ichi , Cua Daniel , Kubo Masato
Published: November 2009

Effector Th17 cells are a major source of IL-17, a critical inflammatory cytokine in autoimmune diseases and in host defenses during bacterial infections. Recently, splenic lymphoid tissue inducer-like cells have been reported to be a source of T cell independent IL-17. In this study, we report that the immune system contains a unique set of natural occurring IL-17 producing cell, “natural” Th17 (nTh17), which are a memory-like T cell subset. The nTh17 cells can develop in the absence of the IL-6/STAT3 signaling axis required by inducible Th17 cells. The nTh17 cell population is distinct from conventional inducible Th17 cells, since nTh17 cells express substantial amounts of IL-17A (IL-17), but not IL-17F, under the control of the master regulator, RORγt. The nTh17 cells simultaneously produce IFN-γ. DO11.10 transgenic mice with a Rag−/− background (DO11.10 Rag−/−) lack nTh17 cells, and, following intranasal administration of OVA, IL-17-dependent neutrophil infiltration occurs in DO11.10 transgenic mice, but not in DO11.10 Rag−/− mice. The impaired neutrophil-dependent airway response is restored by adaptive transfer of nTh17 cells into DO11.10 Rag−/− mice. These results demonstrate that a novel T cell subset, nTh17, facilitates the early phase of Ag-induced airway responses and host defenses against pathogen invasion before the establishment of acquired immunity.
Increased Inflammation, Impaired Bacterial Clearance, and Metabolic Disruption after Gram-Negative Sepsis in Mkp-1-Deficient Mice
Authors: Frazier W. Joshua , Wang Xianxi , Wancket Lyn M. , Li Xiang-An , Meng Xiaomei , Nelin Leif D. , Cato Andrew C. B. , Liu Yusen
Published: November 2009

MAPKs are crucial for TNF-α and IL-6 production by innate immune cells in response to TLR ligands. MAPK phosphatase 1 (Mkp-1) deactivates p38 and JNK, abrogating the inflammatory response. We have previously demonstrated that Mkp-1−/− mice exhibit exacerbated inflammatory cytokine production and increased mortality in response to challenge with LPS and heat-killed Staphylococcus aureus. However, the function of Mkp-1 in host defense during live Gram-negative bacterial infection remains unclear. We challenged Mkp-1+/+ and Mkp-1−/− mice with live Escherichia coli i.v. to examine the effects of Mkp-1 deficiency on animal survival, bacterial clearance, metabolic activity, and cytokine production. We found that Mkp-1 deficiency predisposed animals to accelerated mortality and was associated with more robust production of TNF-α, IL-6 and IL-10, greater bacterial burden, altered cyclooxygenase-2 and iNOS expression, and substantial changes in the mobilization of energy stores. Likewise, knockout of Mkp-1 also sensitized mice to sepsis caused by cecal ligation and puncture. IL-10 inhibition by neutralizing Ab or genetic deletion alleviated increased bacterial burden. Treatment with the bactericidal antibiotic gentamicin, given 3 h after Escherichia coli infection, protected Mkp-1+/+ mice from septic shock but had no effect on Mkp-1−/− mice. Thus, during Gram-negative bacterial sepsis Mkp-1 not only plays a critical role in the regulation of cytokine production but also orchestrates the bactericidal activities of the innate immune system and controls the metabolic response to stress.
Inhibition of RANK Expression and Osteoclastogenesis by TLRs and IFN-γ in Human Osteoclast Precursors
Authors: Ji Jong-Dae , Park-Min Kyung-Hyun , Shen Zenxin , Fajardo Roberto J. , Goldring Steven R. , McHugh Kevin P. , Ivashkiv Lionel B.
Published: November 2009

TLRs have been implicated in promoting osteoclast-mediated bone resorption associated with inflammatory conditions. TLRs also activate homeostatic mechanisms that suppress osteoclastogenesis and can limit the extent of pathologic bone erosion associated with infection and inflammation. We investigated mechanisms by which TLRs suppress osteoclastogenesis. In human cell culture models, TLR ligands suppressed osteoclastogenesis by inhibiting expression of receptor activator of NF-κB (RANK), thereby making precursor cells refractory to the effects of RANKL. Similar but less robust inhibition of RANK expression was observed in murine cells. LPS suppressed generation of osteoclast precursors in mice in vivo, and adsorption of LPS onto bone surfaces resulted in diminished bone resorption. Mechanisms that inhibited RANK expression were down-regulation of RANK transcription, and inhibition of M-CSF signaling that is required for RANK expression. TLRs inhibited M-CSF signaling by rapidly down-regulating cell surface expression of the M-CSF receptor c-Fms by a matrix metalloprotease- and MAPK-dependent mechanism. Additionally, TLRs cooperated with IFN-γ to inhibit expression of RANK and of the CSF1R gene that encodes c-Fms, and to synergistically inhibit osteoclastogenesis. Our findings identify a new mechanism of homeostatic regulation of osteoclastogenesis that targets RANK expression and limits bone resorption during infection and inflammation.
Ineffective Vaccination against Solid Tumors Can Be Enhanced by Hematopoietic Cell Transplantation
Authors: Filatenkov Alexander , Müller Antonia M. S. , Tseng William Wei-Lin , Dejbakhsh-Jones Sussan , Winer Daniel , Luong Richard , Shizuru Judith A. , Engleman Edgar G. , Strober Samuel
Published: November 2009

Vaccination with tumor Ags has not been an effective treatment for solid tumors. The goal of the current study was to determine whether a combination of vaccination and hematopoietic cell transplantation (HCT) can effectively treat primary, disseminated, or metastatic CT26 and MC38 murine colon tumors. Vaccination of tumor-bearing mice with irradiated tumor cells and CpG adjuvant failed to alter progressive tumor growth. However, mice bearing primary, disseminated lung, or metastatic liver tumors were uniformly cured after administration of total body irradiation, followed by the transplantation of hematopoietic progenitor cells and T cells from syngeneic, but not allogeneic vaccinated donors. Requirements for effective treatment of tumors included irradiation of hosts, vaccination of donors with both tumor cells and CpG, transfer of both CD4+ and CD8+ T cells along with progenitor cells, and ability of donor cells to produce IFN-γ. Irradiation markedly increased the infiltration of donor T cells into the tumors, and the combined irradiation and HCT altered the balance of tumor-infiltrating cells to favor CD8+ effector memory T cells as compared with CD4+CD25+FoxP3+ T regulatory cells. The combination of vaccination and autologous hematopoietic cell transplantation was also effective in treating tumors. In conclusion, these findings show that otherwise ineffective vaccination to solid nonhematologic tumors can be dramatically enhanced by HCT.
Sex Steroid Ablation Enhances Hematopoietic Recovery following Cytotoxic Antineoplastic Therapy in Aged Mice
Authors: Dudakov Jarrod A. , Goldberg Gabrielle L. , Reiseger Jessica J. , Vlahos Katerina , Chidgey Ann P. , Boyd Richard L.
Published: November 2009

Cytotoxic antineoplastic therapy is widely used in the clinic as a treatment for malignant diseases. The treatment itself, however, leads to long-term depletion of the adaptive immune system, which is more pronounced in older patients, predominantly due to thymic atrophy. We and others have previously shown that withdrawal of sex steroids is able to regenerate the aged thymus and enhance recovery from autologous and allogeneic hematopoietic stem cell transplant. In this study we have examined the effects of sex steroid ablation (SSA) on the recovery of lymphopoiesis in the bone marrow (BM) and thymus following treatment with the chemotherapeutic agent cyclophosphamide (Cy) in middle-aged and old mice. Furthermore, we have also examined the impact of this regeneration on peripheral immunity. SSA enhanced the recovery of BM resident hematopoietic stem cells and lymphoid progenitors and promoted lymphopoiesis. Interestingly, Cy alone caused a profound increase in the recently described common lymphoid progenitor 2 (CLP-2) population in the BM. In the thymus, SSA caused a profound increase in cellularity as well as all intrathymic T-lineage progenitors including early T-lineage progenitors (ETPs) and non-canonical T cell progenitors such as the CLP-2. We also found that these transferred into numerical increases in the periphery with enhanced B and T cell numbers. Furthermore, these lymphocytes were found to have an enhanced functional capacity with no perturbation of the TCR repertoire. Taken together, these results provide the basis for the use of SSA in the clinic to enhance treatment outcomes from cytotoxic antineoplastic therapy.
The Cutaneous Biochemical Redox Barrier: A Component of the Innate Immune Defenses against Sensitization by Highly Reactive Environmental Xenobiotics
Authors: Pickard Chris , Louafi Fethi , McGuire Carolann , Lowings Kelly , Kumar Pawan , Cooper Hywel , Dearman Rebecca J. , Cumberbatch Marie , Kimber Ian , Healy Eugene , Friedmann Peter S.
Published: November 2009

Contact allergy to environmental xenobiotics is a common and important problem, but it is unclear why some chemicals are potent sensitizers and others weak/nonsensitizers. We explored this by investigating why similar chemicals, 2,4-dinitrochlorobenzene (DNCB) and 2,4-dinitrothiocyanobenzene (DNTB), differ in their ability to induce contact hypersensitivity (CHS). DNCB induced CHS in humans, whereas at similar doses DNTB did not. However, following DNCB sensitization, DNTB elicited CHS in vivo and stimulated DNCB-responsive T cells in vitro, suggesting that differences in response to these compounds lie in the sensitization phase. In contrast to DNCB, DNTB failed to induce emigration of epidermal Langerhans cells in naive individuals. Examination for protein dinitrophenylation in skin revealed that DNCB penetrated into the epidermis, whereas DNTB remained bound to a thiol-rich band within the stratum corneum. DNTB reacted rapidly with reduced glutathione in vitro and was associated with a decrease in the free thiol layer in the stratum corneum, but not in the nucleated epidermis. By contrast, DNCB required GST facilitation to react with gluthathione and, following penetration through the stratum corneum, depleted thiols in the viable epidermis. Chemical depletion of the thiol-rich band or removing it by tape stripping allowed increased penetration of DNTB into the epidermis. Our results suggest that the dissimilar sensitizing potencies of DNCB and DNTB in humans are determined by a previously undescribed outer epidermal biochemical redox barrier, a chemical component of the innate immune defense mechanisms that defend against sensitization by highly reactive environmental chemicals.
Intranasal Immunization Promotes Th17 Immune Responses
Authors: Zygmunt Beata M. , Rharbaoui Faiza , Groebe Lothar , Guzman Carlos A.
Published: November 2009

Th17 cells are a lineage of CD4+ T cells characterized by IL-17 secretion, which plays a crucial role in immune responses against important respiratory pathogens, such as Mycobacterium tuberculosis. In this study, we demonstrated that intranasal (i.n.) immunization leads per se to Th17-biased immune responses, regardless of the adjuvant used. The activated CD4+ T cells also showed an up-regulated expression of the chemokine receptor CCR6, which is a marker for murine Th17 cells. These results have important implications in the context of optimizing rational vaccine design, since i.n. immunization appears to be the strategy of choice for situations where the induction of a Th17 phenotype would be beneficial.
Plasmacytoid Dendritic Cells Regulate Autoreactive B Cell Activation via Soluble Factors and in a Cell-to-Cell Contact Manner
Authors: Ding Chuanlin , Cai Yihua , Marroquin Jose , Ildstad Suzanne T. , Yan Jun
Published: November 2009

Plasmacytoid dendritic cells (pDCs) are specialized type I IFN producers, which play an important role in pathogenesis of autoimmune disorders. Dysregulated autoreactive B cell activation is a hallmark in most autoimmune diseases. This study was undertaken to investigate interactions between pDCs and autoreactive B cells. After coculture of autoreactive B cells that recognize self-Ag small nuclear ribonucleoprotein particles with activated pDCs, we found that pDCs significantly enhance autoreactive B cell proliferation, autoantibody production, and survival in response to TLR and BCR stimulation. Neutralization of IFN-α/β and IL-6 abrogated partially pDC-mediated enhancement of autoreactive B cell activation. Transwell studies demonstrated that pDCs could provide activation signals to autoreactive B cells via a cell-to-cell contact manner. The involvement of the ICAM-1-LFA-1 pathway was revealed as contributing to this effect. This in vitro enhancement effect was further demonstrated by an in vivo B cell adoptive transfer experiment, which showed that autoreactive B cell proliferation and activation were significantly decreased in MyD88-deficient mice compared with wild-type mice. These data suggest the dynamic interplay between pDCs and B cells is required for full activation of autoreactive B cells upon TLR or BCR stimulation.
Limited Role of CD4+Foxp3+ Regulatory T Cells in the Control of Experimental Cerebral Malaria
Authors: Steeg Christiane , Adler Guido , Sparwasser Tim , Fleischer Bernhard , Jacobs Thomas
Published: November 2009

Cerebral malaria (CM) associated with Plasmodium berghei ANKA (PbA) infection is an accepted model of human CM. CM during PbA infection critically depends on sequestration of T cells into the brain. Several studies aimed to address the role of regulatory T cells (Treg) in modulating this pathogenic T cell response. However, these studies are principally hampered due to the fact that until recently no reagents were available to deplete Foxp3+ Treg specifically. To study the function of Treg in the genesis of CM, we used depletion of Treg mice that are transgenic for a bacterial artificial chromosome expressing a diphtheria toxin receptor-enhanced GFP fusion protein under the control of the foxp3 gene locus. These mice allow for a selective depletion of Foxp3+ Treg by diphtheria toxin injection, and also their specific detection and purification during an ongoing infection. Using depletion of Treg mice, we found only a small increase in the absolute numbers of Foxp3+ Treg during PbA infection and, consequently, the ratio of Treg to T effector cells (Teff) decreased due to the rapid expansion of Teff. Although the latter sequester in the brains of infected mice, almost no Treg were found in the brains of infected mice. Furthermore, we demonstrate that depletion of Treg has no influence on sequestration of Teff and on the clinical outcome, and only minor influence on T cell activation. Using ex vivo analysis of purified Treg from either naive mice or PbA-infected mice, we found that both exhibit similar inhibitory capacity on Teff.
Estrogen Regulates Transcription Factors STAT-1 and NF-κB to Promote Inducible Nitric Oxide Synthase and Inflammatory Responses
Authors: Dai Rujuan , Phillips Rebecca A. , Karpuzoglu Ebru , Khan Deena , Ahmed S. Ansar
Published: November 2009

Estrogen regulation of inflammatory responses has broad physiological and pathological consequences. However, the molecular mechanism of estrogen regulation of inflammation is still poorly understood. In this study, we report that activation of both STAT-1 and NF-κB signaling is essential for Con A-induced inducible NO synthase (iNOS) and NO in murine splenocytes. Estrogen enhances STAT-1 DNA-binding activity without increasing the expression of phosphorylated and total STAT-1 protein. We have recently reported that estrogen blocks the nuclear expression of NF-κB p65 and modifies nuclear NF-κBp50. Here, we demonstrated that both nuclear STAT-1 and NF-κB are modified by serine protease-mediated proteolysis, which resulted in altered STAT-1 and NF-κB activity/signaling in splenocytes from estrogen-treated mice. Inhibition of serine protease activity with 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) restores the nuclear expression of full-length STAT-1 and NF-κB proteins, and resulted in decreased STAT-1 DNA-binding activity and formation of NF-κB p65/p50 binding complexes in nuclei of splenocytes from estrogen-treated mice. Consequently, there is significantly decreased iNOS and IFN-γ production in AEBSF-treated splenocytes from estrogen-treated mice, which suggests a positive regulatory role of truncated STAT-1 and/or NF-κB. Interestingly, there is increased production of MCP-1 in STAT-1 or NF-κB small interfering RNA-transfected cells, as well as in AEBSF-treated splenocytes from estrogen-treated mice. These data suggest a differential role of truncated STAT-1 and NF-κB in regulation of various inflammatory molecules in splenocytes from estrogen-treated mice. Together, our data reveal a novel molecular mechanism of estrogen-mediated promotion of inflammatory responses, which involves posttranslational modification of STAT-1 and NF-κB proteins.
Dynamic Regulation of CXCR1 and CXCR2 Homo- and Heterodimers
Authors: Martínez Muñoz Laura , Lucas Pilar , Navarro Gemma , Checa Ana I. , Franco Rafael , Martínez-A. Carlos , Rodríguez-Frade José Miguel , Mellado Mario
Published: November 2009

Although homo- and heterodimerization are reported for some chemokine receptors, it remains unclear whether these functional states are in dynamic equilibrium and how receptor/ligand levels influence oligomerization. In human neutrophils and in cell lines that coexpress the chemokine receptors CXCR1 and CXCR2, we used fluorescence resonance energy transfer techniques to show that these two receptors form homo- and heterodimers. Receptor expression and ligand activation were found to regulate the balance between these complexes, adapting the response to changes in the milieu. CXCL8, a ligand for both receptors, alters heterodimeric complexes, whereas it stabilizes homodimers and promotes receptor internalization. Oligomerization of receptors, together with the regulation of their expression and desensitization, could thus contribute to the fine control of chemokine functions.
Th1, Th17, and Th9 Effector Cells Induce Experimental Autoimmune Encephalomyelitis with Different Pathological Phenotypes
Authors: Jäger Anneli , Dardalhon Valérie , Sobel Raymond A. , Bettelli Estelle , Kuchroo Vijay K.
Published: November 2009

Experimental autoimmune encephalomyelitis (EAE) is a model of human multiple sclerosis induced by autoreactive Th cells that mediate tissue inflammation and demyelination in the CNS. Initially, IFN-γ-producing Th1 cells and, more recently, IL-17-producing Th17 cells with specificity for myelin Ags have been implicated in EAE induction, but whether Th17 cells are encephalitogenic has been controversial. Moreover, a new effector T cell subset, Th9 cells, has been identified; however, the ability of this T cell subset to induce EAE has not been investigated. Here, we have developed protocols to generate myelin oligodendrocyte glycoprotein-specific Th17, Th1, Th2, and Th9 cells in vitro, so that we could directly compare and characterize the encephalitogenic activity of each of these subsets upon adoptive transfer. We show that myelin oligodendrocyte glycoprotein-specific Th1, Th17, and Th9 cells but not Th2 cells induce EAE upon adoptive transfer. Importantly, each T cell subset induced disease with a different pathological phenotype. These data demonstrate that different effector T cell subsets with specificity for myelin Ags can induce CNS autoimmunity and that the pathological heterogeneity in multiple sclerosis lesions might in part be due to multiple distinct myelin-reactive effector T cells.
RREB-1 Is a Transcriptional Repressor of HLA-G
Authors: Flajollet Sébastien , Poras Isabelle , Carosella Edgardo D. , Moreau Philippe
Published: November 2009

The nonclassical HLA-G is a molecule specifically involved in immune tolerance with highly restricted tissue distribution in healthy conditions. Yet it is overexpressed in numerous tumors and in allografts with better acceptance. Major mechanisms involved in regulation of HLA-G transcription are still poorly described. Thus, to characterize these mechanisms we have developed a specific proteomic approach to identify proteins that bind differentially to the HLA-G gene promoter by promoter pull-down assay followed by spectrometry mass analysis. Among specific binding factors, we focused on RREB-1, a ras-responsive element binding protein 1. We demonstrated that RREB-1 represses HLA-G transcriptional activity and binds three ras response elements within the HLA-G promoter. RREB-1 protein, specifically in HLA-G-negative cells, interacts with subunits of CtBP complex implicated in chromatin remodeling. This demonstration is the first of a repressor factor of HLA-G transcriptional activity taking part in HLA-G repression by epigenetic mechanisms.
Innate Immune CD11b+Gr-1+ Cells, Suppressor Cells, Affect the Immune Response during Theiler’s Virus-Induced Demyelinating Disease
Authors: Bowen Jenna L. , Olson Julie K.
Published: November 2009

Multiple sclerosis is a demyelinating disease associated with an inflammatory immune response in the CNS. Theiler’s murine encephalomyelitis virus (TMEV)-induced demyelinating disease is a relevant mouse model for the study of multiple sclerosis. TMEV infection of susceptible mice leads to a persistent virus infection of the CNS which contributes to development of demyelinating disease. We have previously shown that the innate immune response can affect the development and progression of demyelinating disease. In the current studies, we determined that the predominant infiltrating cells during the innate immune response are CD11b+Ly6C+ cells. CD11b+Ly6C+ cells are immature myeloid cells that have exited the bone marrow without maturing and have been shown to suppress CD4+ and CD8+ T cell responses. Therefore, we wanted to determine what role these cells play in development and progression of demyelinating disease. TMEV-infected mice depleted of CD11b+Ly6C+ cells during the innate immune response developed a reduced demyelinating disease which was associated with a decreased myelin-specific CD4+ T cell response and a decreased inflammatory immune response in the CNS. TMEV-infected mice depleted of CD11b+Ly6C+ cells had increased virus-specific CD4+ and CD8+ T cell responses during early virus infection associated with increased expression of IFN-γ and IL-17 and decreased expression of IL-10 in the CNS. These results suggest that CD11b+Ly6C+ cells which infiltrate into the CNS during the innate immune response are myeloid-derived suppressor cells that suppress virus-specific T cell responses and contribute to the development of demyelinating disease.
Medication Timing Errors for Parkinson's Disease: Perspectives Held by Caregivers and People with Parkinson's in New Zealand
Authors: Buetow Stephen , Henshaw Jenny , Bryant Linda , O'Sullivan Deirdre
Published: November 2009

Background. Common but seldom published are Parkinson's disease (PD) medication errors involving late, extra, or missed doses. These errors can reduce medication effectiveness and the quality of life of people with PD and their caregivers.Objective. To explore lay perspectives of factors contributing to medication timing errors for PD in hospital and community settings.Design and Methods. This qualitative research purposively sampled individuals with PD, or a proxy of their choice, throughout New Zealand during 2008-2009. Data collection involved 20 semistructured, personal interviews by telephone. A general inductive analysis of the data identified core insights consistent with the study objective.Results. Five themes help to account for possible timing adherence errors by people with PD, their caregivers or professionals. The themes are the abrupt withdrawal of PD medication; wrong, vague or misread instructions; devaluation of the lay role in managing PD medications; deficits in professional knowledge and in caring behavior around PD in formal health care settings; and lay forgetfulness.Conclusions. The results add to the limited published research on medication errors in PD and help to confirm anecdotal experience internationally. They indicate opportunities for professionals and lay people to work together to reduce errors in the timing of medication for PD in hospital and community settings.
Congregational analysis: A theological and ministerial approach
Authors: Nel Malan
Published: November 2009

The research problem concerned is: What criteria should be used when congregations are analysed? Congregations as faith communities are defined differently. Identifying the local congregation as a defined and as an empirical subject plays a major role in answering the research question. The theological points of departure taken are that any measure of a local congregation has to deal with issues such as faithfulness to the gospel and the missional identity and integrity of the congregation as a contextual faith community. The hypothesis is that theologically informed and motivated congregations can and should be analysed in the process of continuing reformation. While the first article described the approach and outcome of empirical research, this article focuses mainly on understanding the church as intermediary in its ministerial role-fulfilment. Ministries are modes by which God is coming to his own and through them to his world. Eventually a case is made for congregational analysis as a test of faithfulness to this understanding of ministry.
In Vitro Assays of Inorganic Arsenic Methylation
Authors: Drobna Zuzana , Styblo Miroslav , Thomas David J.
Published: November 2009

Inorganic arsenic is extensively metabolized to produce mono‐, di‐, and trimethylated products. The formation of these metabolites produces a variety of intermediates that differ from inorganic arsenic in terms of patterns of distribution and retention and in toxic effects. In order to elucidate the pathway for arsenic methylation, it was necessary to develop a reliable in vitro assay system in which the formation of methylated metabolites could be monitored. Here, we describe an in vitro assay system that uses the postmicrosomal supernatant from rat liver as the source of the enzymatic activity that catalyzes methylation reactions. This system can be used to study the requirements for methylation reactions (e.g., identifying the donor of methyl groups) and for screening of compounds as potential activators or inhibitors of arsenic methylation. Curr. Protoc. Toxicol. 42:4.32.1‐4.32.10. © 2009 by John Wiley & Sons, Inc.
Analysis of Arsenical Metabolites in Biological Samples
Authors: Hernandez‐Zavala Araceli , Drobna Zuzana , Styblo Miroslav , Thomas David J.
Published: November 2009

Quantitation of iAs and its methylated metabolites in biological samples provides dosimetric information needed to understand dose‐response relations. Here, methods are described for separation of inorganic and mono‐, di‐, and trimethylated arsenicals by thin‐layer chromatography. This method has been extensively used to track the metabolism of the radionuclide [73As] in a variety of in vitro assay systems. In addition, a hydride generation‐cryotrapping‐gas chromatography‐atomic absorption spectrometric method is described for the quantitation of arsenicals in biological samples. This method uses pH‐selective hydride generation to differentiate among arsenicals containing trivalent or pentavalent arsenic. Curr. Protoc. Toxicol. 42:4.33.1‐4.33.17. © 2009 by John Wiley & Sons, Inc.
Purification of Arsenic (+3 Oxidation State) Methyltransferase from Rat Liver Cytosol
Authors: Drobna Zuzana , Styblo Miroslav , Thomas David J.
Published: November 2009

Demonstrating the enzymatic basis of arsenic methylation is critical to further studies of the pathway for the conversion of inorganic arsenic into a variety of methylated metabolites. This protocol describes a procedure for the purification of an arsenic methyltransferase from rat liver cytosol. Purification of this enzyme and subsequent cloning of its gene has permitted studies of enzyme structure and function, and has lead to the identification of orthologous genes in genomes of organisms ranging in complexity from sea urchins to humans. These proteins are referred to as arsenic (+3 oxidation state) methyltransferases. Curr. Protoc. Toxicol. 42:4.34.1‐4.34.13. © 2009 by John Wiley & Sons, Inc.
Olefin Polymers
Authors: Kissin Yury V. , Pebsworth Lloyd W. , Kissin Yury V. , Kissin Yury V. , Lieberman Richard B. , Kissin Yury V. , Lappin G. R. , Nemec L. H. , Sauer J. D. , Wagner J. D.
Published: November 2009

POLYETHYLENE

LOW DENSITY POLYETHYLENE

HIGH DENSITY POLYETHYLENE

LINEAR LOW DENSITY POLYETHYLENE

POLYPROPYLENE

POLYMERS OF HIGHER OLEFINS

OLEFINS, HIGHER
Differential Staining of Bacteria: Acid Fast Stain
Authors: Reynolds Jackie , Moyes Rita B. , Breakwell Donald P.
Published: November 2009

Acid‐fastness is an uncommon characteristic shared by the genera Mycobacterium (Section 10A) and Nocardia. Because of this feature, this stain is extremely helpful in identification of these bacteria. Although Gram positive, acid‐fast bacteria do not take the crystal violet into the wall well, appearing very light purple rather than the deep purple of normal Gram‐positive bacteria. Curr. Protoc. Microbiol. 15:A.3H.1‐A.3H.5. © 2009 by John Wiley & Sons, Inc.
Absorber intercooling in CO2 absorption by piperazine‐promoted potassium carbonate
Authors: Plaza Jorge M. , Chen Eric , Rochelle Gary T.
Published: November 2009

Intercooling was evaluated as a process option in CO2 absorption by piperazine (PZ) promoted potassium carbonate. The system performance with 4.5 m K+/4.5 m PZ was simulated by a model in Aspen Plus® RateSep™. The absorber was evaluated for use with a double matrix stripper by optimizing the position of the semilean feed and intercooling stages to maximize CO2 removal. Additionally, a simple absorber system was modeled to observe the effect of intercooling on systems with variable CO2 lean loading. Intercooling increases CO2 removal by as much as 10% with the double matrix configuration. With a simple absorber, the effectiveness of intercooling depends on solvent rate. Near a critical liquid/gas ratio (L/G) there is a large improvement with intercooling. This is related to the position of the temperature bulge. An approximation is proposed to estimate the critical L/G where intercooling may maximize removal. © 2009 American Institute of Chemical Engineers AIChE J, 2010
Prediction of the enantioselectivity of lipases and esterases by molecular docking method with modified force field parameters
Authors: Ji Liu , Xiaoling Tang , Hongwei Yu
Published: November 2009

In the work, molecular docking method was applied to extensively predict the enantioselectivity of lipases and esterases. A ligand library consisted of 69 chiral substrates was docked to four lipases and two esterases to set up the prediction model. During the docking process, necessary modification was carried out on van de Waals and hydrogen bond parameters of enzyme/substrate pair so that the ligands were able to adopt productive geometry in the enzymes. The docking results correctly indicated the enantiopreference for 91% (63/69) of docking pairs and the docking energy difference between substrate enantiomers (ΔΔGdocking) was significantly (correlation coefficient = 0.72, P < 0.05) correlated with the activation free energy difference (ΔΔG≠) that was quantitatively correlated with enantioselectivity of the enzymes. The prediction method was further validated by docking with another 12 enzyme/substrate pairs. Moreover, the prediction error was susceptible to the size of groups bonded to substrate's chiral center and expected ΔΔG≠ values but was not related to the substrate type and reaction medium. The possible reasons of observed error were discussed. It is demonstrated that the docking method has great application potential in high performance prediction of enzyme enantioselectivity. Biotechnol. Bioeng. 2010. 105: 687–696. © 2009 Wiley Periodicals, Inc.
Solvent‐Dependent Dihydrogen/Dihydride Stability for [Mo(CO)(Cp*)H2(PMe3)2]+[BF4]− Determined by Multiple Solvent⋅⋅⋅Anion⋅⋅⋅Cation Non‐Covalent Interactions
Authors: Dub Pavel A. , Belkova Natalia V. , Filippov Oleg A. , Daran Jean‐Claude , Epstein Lina M. , Lledós Agustí , Shubina Elena S. , Poli Rinaldo
Published: November 2009

Low‐temperature (200 K) protonation of [Mo(CO)(Cp*)H(PMe3)2] (1) by Et2O⋅HBF4 gives a different result depending on a subtle solvent change: The dihydrogen complex [Mo(CO)(Cp*)(η2‐H2)(PMe3)2]+ (2) is obtained in THF, whereas the tautomeric classical dihydride [Mo(CO)(Cp*)(H)2(PMe3)2]+ (3) is the only observable product in dichloromethane. Both products were fully characterised (νCO IR; 1H, 31P, 13C NMR spectroscopies) at low temperature; they lose H2 upon warming to 230 K at approximately the same rate (ca. 10−3 s−1), with no detection of the non‐classical form in CD2Cl2, to generate [Mo(CO)(Cp*)(FBF3)(PMe3)2] (4). The latter also slowly decomposes at ambient temperature. One of the decomposition products was crystallised and identified by X‐ray crystallography as [Mo(CO)(Cp*)(FH⋅⋅⋅FBF3)(PMe3)2] (5), which features a neutral HF ligand coordinated to the transition metal through the F atom and to the BF4− anion through a hydrogen bond. The reason for the switch in relative stability between 2 and 3 was probed by DFT calculations based on the B3LYP and M05‐2X functionals, with inclusion of anion and solvent effects by the conductor‐like polarisable continuum model and by explicit consideration of the solvent molecules. Calculations at the MP4(SDQ) and CCSD(T) levels were also carried out for calibration. The calculations reveal the key role of non‐covalent anion–solvent interactions, which modulate the anion–cation interaction ultimately altering the energetic balance between the two isomeric forms.
Hydrophilic Interaction Chromatography Based Enrichment of Glycopeptides by Using Click Maltose: A Matrix with High Selectivity and Glycosylation Heterogeneity Coverage
Authors: Yu Long , Li Xiuling , Guo Zhimou , Zhang Xiuli , Liang Xinmiao
Published: November 2009

Glycosylation analysis based on mass spectrometry (MS) of glycopeptides requires the isolation of glycopeptides from complex glycoprotein digests to facilitate structural determination of the glycopeptides. To this end, hydrophilic interaction chromatography (HILIC)‐based methods have been developed to selectively enrich glycopeptides by utilizing the hydrophilicity of the glycans. However, the application of these methods is limited by the medium selectivity of HILIC matrices. To improve the effectiveness of HILIC‐based methods, we introduced a customized hydrophilic matrix named “click maltose” and characterized its selectivity and glycosylation heterogeneity coverage. In the selectivity assessment, the non‐glycopeptides causing ion suppression to the glycopeptides were effectively removed by click maltose, leading to the identification of 27 glycopeptides in the fractions enriched from human serum immunoglobulin G digest, compared to 13 glycopeptides enriched using Sepharose CL‐6B, a commercially available matrix. For the assessment of glycosylation heterogeneity coverage, more than 140 glycopeptides covering all the five glycosites of human serum α1‐acid glycoprotein were captured using click maltose. Click maltose was synthesized by linking alkynyl‐derivatized maltose to azide‐derivatized silica through click chemistry. The resulting flexible saccharide chain structure remarkably enhances the hydrogen‐bonding interactions between the glycans of the glycopeptides and the matrix, which are responsible for the increased selectivity and glycosylation heterogeneity coverage of click maltose.
A Nucleic Acid Dependent Chemical Photocatalysis in Live Human Cells
Authors: Arian Dumitru , Cló Emiliano , Gothelf Kurt V. , Mokhir Andriy
Published: November 2009

Only two nucleic acid directed chemical reactions that are compatible with live cells have been reported to date. Neither of these processes generate toxic species from nontoxic starting materials. Reactions of the latter type could be applied as gene‐specific drugs, for example, in the treatment of cancer. We report here the first example of a chemical reaction that generates a cytotoxic drug from a nontoxic prodrug in the presence of a specific endogeneous ribonucleic acid in live mammalian cells. In this case, the prodrug is triplet oxygen and the drug is singlet oxygen. The key component of this reaction is an inert molecule (InP–2′‐OMe‐RNA/Q–2′‐OMe‐RNA; P: photosensitizer; Q: quencher), which becomes an active photosensitizer (InP–2′‐OMe‐RNA) in the presence of single‐stranded nucleic acid targets. Upon irradiation with red light, the photosensitizer produces over 6000 equivalents of toxic singlet oxygen per nucleic acid target. This reaction is highly sequence specific. To detect the generation of singlet oxygen in live cells, we prepared a membrane‐permeable and water‐soluble fluorescent scavenger, a derivative of 2,5‐diphenylisobenzofurane. The scavenger decomposes upon reaction with singlet oxygen and this is manifested in a decrease in the fluorescence intensity. This effect can be conveniently monitored by flow cytometry.
Cytomorphological diagnosis of tendinous xanthomatosis: A case report
Authors: Aggarwal Seema , Gandhi Akansha , Arora Vinod Kumar
Published: November 2009

Xanthoma is a reactive histiocytic proliferation in response to altered lipid levels. These develop usually in skin and subcutis but may also develop in deep soft tissue as tendon sheath or synovium. We report a rare case of tendinous xanthomatosis in a 23‐year‐old female, diagnosed on aspiration cytology. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.