bluuebunny/crossref_metadata_2025_split huggingface dataset:
Induced CD8+FoxP3+ Treg Cells in Rheumatoid Arthritis Are Modulated by p38 Phosphorylation and Monocytes Expressing Membrane Tumor Necrosis Factor α and CD86
Authors: Ellis Shawn D. P. , McGovern Jenny L. , van Maurik André , Howe David , Ehrenstein Michael R. , Notley Clare A. 
Published: July 2014

ObjectiveLimiting the severity of inflammation and promoting its eventual resolution are vital for protecting host tissues both in autoimmunity and chronic infection. The aim of this study was to determine the suitability of repurposing anti‐CD3 monoclonal antibody (mAb) therapy for rheumatoid arthritis (RA) by analyzing its ability to induce CD8+FoxP3+ Treg cells from peripheral blood mononuclear cells (PBMCs).MethodsAnti‐CD3 mAb was cultured with RA PBMCs to induce CD8+FoxP3+ Treg cells, which were analyzed by flow cytometry to determine their phenotype. Treg cell induction was investigated via neutralization or blocking antibodies, cellular depletion, or ImageStream technology. Blotting was used to determine the signaling pathways involved in CD8+FoxP3+ Treg cell induction. Suppression of CD4+ T cell effector responses was assessed by Treg cell suppression assays and Mosaic enzyme‐linked immunosorbent assay.ResultsPotent CD8+FoxP3+ Treg cells were induced from RA PBMCs by anti‐CD3 mAb. Unlike their CD4+ counterparts, CD8+FoxP3+ Treg cells inhibited Th17 responses in a contact‐dependent manner, thereby functioning to limit a wider range of inflammatory pathways. CD8+FoxP3+ Treg cell induction was supported both by p38 phosphorylation intrinsic to naive CD8+ T cells and by monocytes via CD86 and membrane tumor necrosis factor α (TNFα). Artificially increasing monocyte membrane TNFα or inhibiting CD8+ T cell p38 phosphorylation drove FoxP3 expression in a subset of initially unresponsive CD8+ T cells.ConclusionThese data define an unknown mechanism of CD8+FoxP3+ Treg cell induction by anti‐CD3 mAb, which could be combined with a p38 inhibitor to improve therapeutic efficacy in RA patients and resolve chronic inflammation via the restoration of tolerance.
Role of Protein Phosphatase Magnesium‐Dependent 1A and Anti–Protein Phosphatase Magnesium‐Dependent 1A Autoantibodies in Ankylosing Spondylitis
Authors: Kim Yong‐Gil , Sohn Dong Hyun , Zhao Xiaoyan , Sokolove Jeremy , Lindstrom Tamsin M. , Yoo Bin , Lee Chang‐Keun , Reveille John D. , Taurog Joel D. , Robinson William H. 
Published: July 2014

ObjectiveAlthough ankylosing spondylitis (AS) is driven by immune‐mediated processes, little is known about the presence and role of autoantibodies in this disease. This study was undertaken to investigate whether autoantibodies occur in and are involved in AS.MethodsWe performed human protein microarray analysis of sera derived from patients with AS or other autoimmune disorders to identify autoantibodies associated specifically with AS, and identified autoantibody targeting of protein phosphatase magnesium‐dependent 1A (PPM1A) in AS. We performed enzyme‐linked immunosorbent assay (ELISA) analysis of sera from 2 independent AS cohorts to confirm autoantibody targeting of PPM1A, and to assess associations between levels of anti‐PPM1A antibodies and AS disease severity or response to anti–tumor necrosis factor (anti‐TNF) therapy (as measured by Bath AS Disease Activity Index [BASDAI] score). Levels of anti‐PPM1A antibodies were also evaluated in sera from rats transgenic for HLA–B27 and human β2‐microglobulin. The expression of PPM1A was assessed by immunohistochemistry in synovial tissue samples from patients with AS, rheumatoid arthritis, or osteoarthritis. The role of PPM1A in osteoblast differentiation was investigated by gene knockdown and overexpression.ResultsAS was associated with autoantibody targeting of PPM1A, and levels of anti‐PPM1A autoantibodies were significantly higher in patients with more advanced sacroiliitis and correlated positively with BASDAI score after treatment with anti‐TNF agents. The levels of anti‐PPM1A autoantibodies were also higher in the sera of transgenic rats that are prone to develop spondyloarthritis than in those that are not. PPM1A was expressed in AS synovial tissue, and PPM1A overexpression promoted osteoblast differentiation, whereas PPM1A knockdown suppressed it.ConclusionAnti‐PPM1A autoantibodies are present in AS, and our findings suggest that PPM1A may contribute to the pathogenic bone ankylosis characteristic of AS.
Evaluation of Developmental Toxicity of Propylthiouracil and Methimazole
Authors: Mallela Murali K. , Strobl Marie , Poulsen Ryan R. , Wendler Christopher C. , Booth Carmen J. , Rivkees Scott A. 
Published: July 2014

BACKGROUNDPropylthiouracil (PTU) and methimazole (MMI) are antithyroid drugs used to treat hyperthyroidism. Despite the widespread use of PTU and MMI during pregnancy, modest clinical data and less animal data are available on the teratogenic potential of these drugs.METHODSWe evaluated the teratogenicity of in utero exposure to PTU or MMI in mice and rats. First, pregnant C57Bl/6 mice were treated daily with PTU (10 or 100 mg/kg), MMI (2 or 20 mg/kg), or vehicle from gestation day (GD) 6 to 16. GD 18 fetuses were evaluated for gross and histopathological abnormalities. Next, pregnant Sprague‐Dawley rats were treated daily with PTU (50 or 100 mg/kg), MMI (10 or 20 mg/kg), or vehicle from GD 6 to 19, followed by evaluation for gross and histopathological abnormalities at GD 20.RESULTSIn mice treated with PTU or MMI, no significant histopathological abnormalities or external gross malformations, and no adverse effects on placental weight, litter size, resorption rates, or fetal weight were observed at GD 18. In rats, no adverse effects on litter size, placental weights, or maternal body weights were observed with either PTU or MMI treatment. PTU treatment (50 and 100 mg/kg) and MMI (10 mg/kg) treatment resulted in a decrease in crown‐rump length in rat fetuses but no external gross malformations or histopathological abnormalities were observed.CONCLUSIONWe did not observe either gross external malformations or histopathological malformations in mice or rats treated long‐term with high doses of PTU or MMI during pregnancy
Short-term complications and long-term morbidity of laparoscopic and open appendicectomy in a national cohort
Authors: Andersson R E 
Published: July 2014

Abstract

Background
Laparoscopic appendicectomy has been proposed as the standard for surgical treatment of acute appendicitis, based on controversial evidence. This study compared outcomes after open and laparoscopic appendicectomy in a national, population-based cohort.

Methods
All patients who underwent open or intended laparoscopic appendicectomy in Sweden between 1992 and 2008 were identified from the Swedish National Patient Register. The outcomes were analysed according to intention to treat with multivariable adjustment for confounding factors and survival analytical techniques where appropriate.

Results
A total of 169 896 patients underwent open (136 754) or intended laparoscopic (33 142) appendicectomy. The rate of intended laparoscopic appendicectomy increased from 3·8 per cent (425 of 11 175) in 1992 to 32·9 per cent (3066 of 9329) in 2008. Laparoscopy was used most frequently in middle-aged patients, women and patients with no co-morbidity. The rate of conversion from laparoscopy to open appendicectomy decreased from 75·3 per cent (320 of 425) in 1992 to 19·7 per cent (603 of 3066) in 2008. Conversion was more frequent in women and those with perforated appendicitis, and the rate increased with age and increasing co-morbidity. After adjustment for co-variables, compared with open appendicectomy, laparoscopy was associated with a shorter length of hospital stay (by 0·06 days), a lower frequency of negative appendicectomy (adjusted odds ratio (OR) 0·59; P &lt; 0·001), wound infection (adjusted OR 0·54; P = 0·004) and wound rupture (adjusted OR 0·44; P = 0·010), but higher rates of intestinal injury (adjusted OR 1·32; P = 0·042), readmission (adjusted OR 1·10; P &lt; 0·001), postoperative abdominal abscess (adjusted OR 1·58; P &lt; 0·001) and urinary infection (adjusted OR 1·39; P = 0·020). Laparoscopy had a lower risk of postoperative small bowel obstruction during the first 2 years after surgery, but not thereafter.

Conclusion
The outcomes of laparoscopic and open appendicectomy showed a complex and contrasting pattern and small differences of limited clinical importance. The choice of surgical method therefore depends on the local situation, the surgeon's experience and the patient's preference.
Randomized clinical trial of endovenous laser ablation versus steam ablation (LAST trial) for great saphenous varicose veins
Authors: van den Bos R R , Malskat W S J , De Maeseneer M G R , de Roos K P , Groeneweg D A G , Kockaert M A , Neumann H A M , Nijsten T 
Published: July 2014

Abstract

Background
The aim was to compare endovenous laser ablation (EVLA) and endovenous steam ablation (EVSA) for great saphenous varicose veins in a non-inferiority study.

Methods
Patients with primary great saphenous vein reflux were randomized to EVLA (940 nm) or EVSA (SVS™). Primary outcomes were treatment success (vein obliteration) at 52 weeks, and Venous Clinical Severity Score (VCSS) at 12 weeks. Secondary outcomes were pain, satisfaction with treatment, duration of analgesia use and days lost from daily activities, changes in Aberdeen Varicose Vein Questionnaire (AVVQ) and EQ-5D™ scores after 12 weeks, and complications at 2 and 12 weeks.

Results
A total of 227 legs were treated (EVSA, 117; EVLA, 110); 36 legs treated with EVSA received a low dose and the remaining 81 a higher dose. At 1 year, the treatment success rate after high-dose EVSA was not inferior to that of EVLA: 92 (95 per cent confidence interval (c.i.) 86 to 98) versus 96 (92 to 100) per cent respectively. Changes in VCSS after 12 weeks were similar: −2·69 (95 per cent c.i. −2·34 to −3·04) and −2·51 (−2·10 to −2·93). AVVQ, EQ-5D™ and EQ VAS scores improved equally 12 weeks after both treatments. Patients treated with EVSA reported less postprocedural pain, fewer days of analgesia use, were more satisfied with therapy, and had a shorter convalescence. Complication rates were comparable.

Conclusion
The 1-year treatment success of high-dose EVSA was not inferior to that of EVLA. Several secondary outcomes were in favour of EVSA. Registration number NCT02046967 (http://www.clinicaltrials.gov).
The use of whole‐mount in situ hybridization to illustrate gene expression regulation
Authors: Llamusí Beatriz , Muñoz‐Soriano Verónica , Paricio Nuria , Artero Rubén 
Published: July 2014

AbstractIn situ hybridization is a widely used technique for studying gene expression. Here, we describe two experiments addressed to postgraduate genetics students in which the effect of transcription factors on gene expression is analyzed in Drosophila embryos of different genotypes by whole‐mount in situ hybridization. In one of the experiments, students analyzed the repressive effect of Snail over rhomboid expression using reporter lines containing different constructs of the rhomboid neuroectodermal enhancer fused to the lacZ gene. In the second experiment, the epistatic relationship between the cabut and decapentaplegic genes was analyzed. These simple experiments allowed students to (1) understand the role of transcription factors and cis‐regulatory elements over gene expression regulation and (2) practice a widespread laboratory technique, in situ hybridization with nonradioactive labeled probes, to detect gene expression patterns. These experiments required 12 hr and were organized into four daily sessions that included the discussion of the results with students. Examples of the results obtained and their relevance are shown and discussed herein. The methods described in these laboratory exercises can be easily adapted to model organisms other than Drosophila. © 2014 by The International Union of Biochemistry and Molecular Biology, 42(4):339–347, 2014.
Carbonaceous Hydrogen‐Evolution Catalyst Containing Cobalt Surrounded by a Tuned Local Structure
Authors: Maruyama Jun , Ioroi Tsutomu , Hasegawa Takahiro , Mori Takuya , Orikasa Yuki , Uchimoto Yoshiharu 
Published: July 2014

AbstractThe low activity and durability of noble‐metal‐free catalysts in acidic media are major problems that hinder their practical use. This study reports significantly enhanced activity of a cobalt‐containing carbonaceous catalyst used for the hydrogen‐evolution reaction in a water electrolyzer by using a proton‐exchange membrane (PEM), which is an efficient energy‐conversion device that is suitable for intermittent renewable energy sources. On the basis of X‐ray absorption near‐edge structure, the enhancement in activity was attributed to a change in the local structure surrounding the Co atom. The PEM water electrolyzer formed by using the catalyst with an optimized structure and Co surface concentration showed an enthalpy efficiency of 80 % at 1 A cm−2.
Purification of Lysozyme from Protein Mixtures by Solvent‐Freeze‐Out Technology
Authors: Yu Xiaoxi , Wang Jingkang , Ulrich Joachim 
Published: July 2014

AbstractThe purification of hen egg white lysozyme from a lysozyme/crude extract mixture was carried out by the solvent‐freeze‐out (SFO) technology on the laboratory scale. Compared to previous works, this investigation focuses on the crystallization of lysozyme from a complicated system. The crystallization conditions were determined by screening tests. Based on these results, the crystallization process was performed in basic buffer solution in which the formation of tetragonal lysozyme crystals is considered to be difficult. The protein composition and purity of the crystals were evaluated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The enzymatic activity was also measured by the use of a spectrophotometer. The results show that lysozyme retained its original enzyme activity.
Formation and Properties of a Bicyclic Silylated Digermene
Authors: Hlina Johann , Baumgartner Judith , Marschner Christoph , Albers Lena , Müller Thomas , Jouikov Viatcheslav V. 
Published: July 2014

AbstractIn the presence of PMe3 or N‐heterocyclic carbenes, the reaction of oligosilanylene dianions with GeCl2⋅dioxane gives germylene–base adducts. After base abstraction, the free germylenes can dimerize by formation of a digermene. An electrochemical and theoretical study of a bicyclic tetrasilylated digermene revealed formation of a comparably stable radical anion and a more reactive radical cation, which were characterized further by UV/Vis and ESR spectroscopy.
The Discovery and Development of the N‐Substituted trans‐3,4‐Dimethyl‐4‐(3′‐hydroxyphenyl)piperidine Class of Pure Opioid Receptor Antagonists
Authors: Carroll F. Ivy , Dolle Roland E. 
Published: July 2014

AbstractN‐Substituted trans‐3,4‐dimethyl‐4‐(3‐hydroxyphenyl)piperidines are a class of pure opioid receptor antagonists with a novel pharmacophore. This opioid receptor antagonist pharmacophore was used as a lead structure to design and develop several interesting and useful opioid receptor antagonists. In this review we describe: 1) early SAR studies that led to the discovery of LY255582 and analogues that are nonselective opioid receptor antagonists developed for the treatment of obesity; 2) the discovery and commercialization of LY246736 (alvimopan; ENTEREG®), a peripherally selective opioid receptor antagonist that accelerates the time to upper and lower GI recovery following surgeries that include partial bowel resection with primary anastomosis; and 3) the discovery and development of the potent and selective κ opioid receptor antagonist JDTic and analogues as potential pharmacotherapies for treating depression, anxiety, and substance abuse (nicotine, alcohol, and cocaine). In addition, the use of JDTic for obtaining the X‐ray structure of the human κ opioid receptor is discussed.
Noninvasive radiofrequency treatment effect on mitochondria in pancreatic cancer cells
Authors: Curley Steven A. , Palalon Flavio , Lu Xiaolin , Koshkina Nadezhda V. 
Published: July 2014

BACKGROUNDThe development of novel therapeutic approaches for cancer therapy is important, especially for tumors that have poor response or develop resistance to standard chemotherapy and radiation. We discovered that noninvasive radiofrequency (RF) fields can affect cancer cells but not normal cells, inhibit progression of tumors in mice, and enhance the anticancer effects of chemotherapy. However, it remains unclear what physiological and molecular mechanisms this treatment induces inside cells. Here, we studied the effect of RF treatment on mitochondria in human pancreatic cancer cells.METHODSThe morphology of mitochondria in cells was studied via electron microscopy. The alteration of mitochondrial membrane potential (Δψ) was accessed using a Mitotracker probe. The respiratory activity of mitochondria was evaluated by analyzing changes in oxygen consumption rates determined with a Mito Stress Test Kit. The production of intracellular reactive oxygen species was performed using flow cytometry. The colocalization of mitochondria and autophagosome markers in cells was performed using fluorescence immunostaining and confocal microscopy analysis.RESULTSRF fields treatment changed the morphology of mitochondria in cancer cells, altered polarization of the mitochondrial membrane, substantially impaired mitochondrial respiration, and increased reactive oxygen species production, indicating RF‐induced stress on the mitochondria. We also observed frequent colocalization of the autophagosome marker LC3B with the mitochondrial marker Tom20 inside cancer cells after RF exposure, indicating the presence of mitochondria in the autophagosomes. This suggests that RF‐induced stress can damage mitochondria and induce elimination of damaged organelles via autophagy.CONCLUSIONRF treatment impaired the function of mitochondria in cancer cells. Therefore, mitochondria can represent one of the targets of the RF treatment. Cancer 2014;120:3418–3425. © 2014 American Cancer Society.
The orexinergic neurons receive synaptic input from C1 cells in rats
Authors: Bochorishvili Genrieta , Nguyen Thanh , Coates Melissa B. , Viar Kenneth E. , Stornetta Ruth L. , Guyenet Patrice G. 
Published: July 2014

ABSTRACTThe C1 cells, located in the rostral ventrolateral medulla (RVLM), are activated by pain, hypoxia, hypoglycemia, infection, and hypotension and elicit cardiorespiratory stimulation, adrenaline and adrenocorticotropic hormone (ACTH) release, and arousal. The orexin neurons contribute to the autonomic responses to acute psychological stress. Here, using an anatomical approach, we consider whether the orexin neurons could also be contributing to the autonomic effects elicited by C1 neuron activation. Phenylethanolamine N‐methyl transferase‐immunoreactive (PNMT‐ir) axons were detected among orexin‐ir somata, and close appositions between PNMT‐ir axonal varicosities and orexin‐ir profiles were observed. The existence of synapses between PNMT‐ir boutons labeled with diaminobenzidine and orexinergic neurons labeled with immunogold was confirmed by electron microscopy. We labeled RVLM neurons with a lentiviral vector that expresses the fusion protein ChR2‐mCherry under the control of the catecholaminergic neuron‐selective promoter PRSx8 and obtained light and ultrastructural evidence that these neurons innervate the orexin cells. By using a Cre‐dependent adeno‐associated vector and TH‐Cre rats, we confirmed that the projection from RVLM catecholaminergic neurons to the orexinergic neurons originates predominantly from PNMT‐ir catecholaminergic (i.e., C1 cells). The C1 neurons were found to establish predominantly asymmetric synapses with orexin‐ir cell bodies or dendrites. These synapses were packed with small clear vesicles and also contained dense‐core vesicles. In summary, the orexin neurons are among the hypothalamic neurons contacted and presumably excited by the C1 cells. The C1–orexin neuronal connection is probably one of several suprabulbar pathways through which the C1 neurons activate breathing and the circulation, raise blood glucose, and facilitate arousal from sleep. J. Comp. Neurol. 522:3834–3846, 2014. © 2014 Wiley Periodicals, Inc.
Semi‐quantitative estimation of cellular SiO2nanoparticles using flow cytometry combined with X‐ray fluorescence measurements
Authors: Choi Seo Yeon , Yang Nuri , Jeon Soo Kyung , Yoon Tae Hyun 
Published: July 2014

AbstractIn this study, we have demonstrated feasibility of a semi‐quantitative approach for the estimation of cellular SiO2nanoparticles (NPs), which is based on the flow cytometry measurements of their normalized side scattering intensity. In order to improve our understanding on the quantitative aspects of cell–nanoparticle interactions, flow cytometry, transmission electron microscopy, and X‐ray fluorescence experiments were carefully performed for the HeLa cells exposed to SiO2NPs with different core diameters, hydrodynamic sizes, and surface charges. Based on the observed relationships among the experimental data, a semi‐quantitative cellular SiO2NPs estimation method from their normalized side scattering and core diameters was proposed, which can be applied for the determination of cellular SiO2NPs within their size‐dependent linear ranges. © 2014 International Society for Advancement of Cytometry
Mammalian Nkx2.2+ perineurial glia are essential for motor nerve development
Authors: Clark Jessica K. , O'keefe Ashley , Mastracci Teresa L. , Sussel Lori , Matise Michael P. , Kucenas Sarah 
Published: July 2014

ABSTRACTBackground: All vertebrate peripheral nerves connect the central nervous system (CNS) with targets in the periphery and are composed of axons, layers of ensheathing glia and connective tissue. Although the structure of these conduits is well established, very little is known about the origin and developmental roles of some of their elements. One understudied component, the perineurium, ensheaths nerve fascicles and is a component of the blood‐nerve‐barrier. In zebrafish, the motor nerve perineurium is composed of CNS‐derived nkx2.2a+ perineurial glia, which establish the motor exit point (MEP) during development. To determine if mouse perineurial cells also originate within the CNS and perform a similar function, we created a Nkx2.2:EGFP transgenic reporter line. Results: In conjunction with RNA expression analysis and antibody labeling, we observed Nkx2.2+ cells along peripheral motor nerves at all stages of development and in adult tissue. Additionally, in mice lacking Nkx2.2, we demonstrate that Nkx2.2+ perineurial glia are essential for motor nerve development and Schwann cell differentiation. Conclusions: Our studies reveal that a subset of mouse perineurial cells are CNS‐derived, express Nkx2.2, and are essential for motor nerve development. This work highlights an under‐appreciated but essential contribution of CNS‐derived cells to the development of the mammalian peripheral nervous system (PNS). Developmental Dynamics 243:1116–1129, 2014. © 2014 Wiley Periodicals, Inc.
Photosensitizing Properties of Alkynylrhenium(I) Complexes [Re(–C≡C–R)(CO)3(N∩N)] (N∩N = 2,2′‐bipy, phen) for H2 Production
Authors: Oberholzer Miriam , Probst Benjamin , Bernasconi Dominik , Spingler Bernhard , Alberto Roger 
Published: July 2014

AbstractInvited for the cover of this issue is the group of Roger Alberto at the University of Zürich, Switzerland. The cover image shows the reduction of protons by illumination of a catalytic mixture containing a cobalt‐based water‐reduction catalyst (WRC), a rhenium(I)–alkynyl photosensitizer, a proton source (H+), and a sacrificial electron donor (D/D+). The final product, hydrogen gas, stores the energy of light in its chemical bond.