bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of T‐cell receptor signaling by the actin cytoskeleton and poroelastic cytoplasm
Authors: Beemiller Peter , Krummel Matthew F.
Published: August 2014

The actin cytoskeleton plays essential roles in modulating T‐cell activation. Most models of T‐cell receptor (TCR) triggering signalosome assembly and immune synapse formation invoke actin‐dependent mechanisms. As T cells are constitutively motile cells, TCR triggering and signaling occur against a cytoskeletal backdrop that is constantly remodeling. While the interplay between actin dynamics and TCR signaling have been the focus of research for many years, much of the work in T cells has considered actin largely for its ‘scaffolding’ function. We examine the roles of the actin cytoskeleton in TCR signaling and immune synapse formation with an emphasis on how poroelasticity, an ensemble feature of actin dynamics with the cytosol, relates to how T cells respond to stimulation.
Mechanical regulation of T‐cell functions
Authors: Chen Wei , Zhu Cheng
Published: August 2014

T cells are key players of the mammalian adaptive immune system. They experience different mechanical microenvironments during their life cycle, from the thymus, secondary lymph organs, and peripheral tissues that are free of externally applied force, but display variable substrate rigidities to the blood and lymphatic circulation systems, where complicated hydrodynamic forces are present. Regardless of whether T cells are subject to external forces or generate their own internal forces, they respond and adapt to different biomechanical cues to modulate their adhesion, migration, trafficking, and triggering of immune functions through mechanical regulation of various molecules that bear force. These include adhesive receptors, immunoreceptors, motor proteins, cytoskeletal proteins, and their associated molecules. Here, we discuss the forces acting on various surface and cytoplasmic proteins of a T cell in different mechanical milieus. We review existing data on how force regulates protein conformational changes and interactions with counter molecules, including integrins, actin, and the T‐cell receptor, and how each relates to T‐cell functions.
Coordinate control of cytoskeletal remodeling and calcium mobilization during T‐cell activation
Authors: Babich Alexander , Burkhardt Janis K.
Published: August 2014

Ca2+ mobilization and cytoskeletal reorganization are key hallmarks of T‐cell activation, and their interdependence has long been recognized. Recent advances in the field have elucidated the molecular pathways that underlie these events and have revealed several points of intersection. Ca2+ signaling can be divided into two phases: initial events leading to release of Ca2+ from endoplasmic reticulum stores, and a second phase involving STIM 1 (stromal interaction molecule 1) clustering and CRAC (calcium release activated calcium) channel activation. Cytoskeletal dynamics promote both phases. During the first phase, the actin cytoskeleton promotes mechanotransduction and serves as a dynamic scaffold for microcluster assembly. Proteins that drive actin polymerization such as WASp (Wiskott–Aldrich syndrome protein) and HS1 (hematopoietic lineage cell‐specific protein 1) promote signaling through PLCγ1 (phospholipase Cγ1) and release of Ca2+ from endoplasmic reticulum stores. During the second phase, the WAVE (WASP‐family verprolin homologous protein) complex and the microtubule cytoskeleton promote STIM 1 clustering at sites of plasma membrane apposition, opening Orai channels. In addition, gross cell shape changes and organelle movements buffer local Ca2+ levels, leading to sustained Ca2+ mobilization. Conversely, elevated intracellular Ca2+ activates cytoskeletal remodeling. This can occur indirectly, via calpain activity, and directly, via Ca2+‐dependent cytoskeletal regulatory proteins such as myosin II and L‐plastin. While it is true that the cytoskeleton regulates Ca2+ responses and vice versa, interdependence between Ca2+ and the cytoskeleton also encompasses signaling events that occur in parallel, downstream of shared intermediates. Inositol cleavage by PLCγ1 simultaneously triggers both endoplasmic reticulum store release and diacylglycerol‐dependent microtubule organizing center reorientation, while depleting the pool of phosphatidylinositol‐4,5‐bisphosphate, an activator of multiple actin‐regulatory proteins. The close interdependence of Ca2+ signaling and cytoskeletal dynamics in T cells provides positive feedback mechanisms for T‐cell activation and allows for finely tuned responses to extracellular cues.
Neutrophil migration: moving from zebrafish models to human autoimmunity
Authors: Shelef Miriam A. , Tauzin Sebastien , Huttenlocher Anna
Published: August 2014

There has been a resurgence of interest in the neutrophil's role in autoimmune disease. Classically considered an early responder that dies at the site of inflammation, new findings using live imaging of embryonic zebrafish and other modalities suggest that neutrophils can reverse migrate away from sites of inflammation. These ‘inflammation‐sensitized’ neutrophils, as well as the neutrophil extracellular traps and other products made by neutrophils in general, may have many implications for autoimmunity. Here, we review what is known about the role of neutrophils in three different autoimmune diseases: rheumatoid arthritis, systemic lupus erythematosus, and small vessel vasculitis. We then highlight recent findings related to several cytoskeletal regulators that guide neutrophil recruitment including Lyn, Rac2, and SHIP. Finally, we discuss how our improved understanding of the molecules that control neutrophil chemotaxis may impact our knowledge of autoimmunity.
Genetic associations between temperament and performance traits in Nellore beef cattle
Authors: Sant'Anna A.C. , Baldi F. , Valente T.S. , Albuquerque L.G. , Menezes L.M. , Boligon A.A. , Paranhos da Costa M.J.R.
Published: August 2014

The aim of this study was to estimate genetic associations between temperament and performance traits. Temperament was evaluated in yearling male and female Nellore cattle, using four traits: temperament score (TS), for assessing animals’ reactions in a corral pen (n = 25 691); movement score (MOV), for animals’ movements recorded inside the crush; crush score (CS), for animal's general reactivity inside the crush; and flight speed (FS), for the speed (in m/s) at which the animals exited the crush (n = 11 697, for the last three methods); for all the temperament traits, lower scores indicate animals with calmer temperament. Performance traits were visual scores for conformation (C), finishing precocity (P) and muscling (M) evaluated at yearlings, and average daily gain (ADG) was estimated from weaning to yearling. Bayesian inference using Gibbs sampling was applied to estimate (co)variance components and genetic and phenotypic parameters. Heritability estimates for the temperament traits ranged from 0.07 (CS) to 0.28 (FS). Genetic correlations of the temperament traits with ADG and C, P and M were negative and ranged from −0.02 to −0.31. Phenotypic correlations were negative and consistently lower than the genetic, ranging from −0.08 to −0.02. It was concluded that the temperament traits assessed had favourable genetic correlation estimates with the performance traits studied. However, indirect responses in temperament when selecting for higher ADG and visual scoring system of C, P and M, will be low.
MLK3 is a novel target of dehydroglyasperin D for the reduction in UVB‐induced COX‐2 expression in vitro and in vivo
Authors: Jung Sung Keun , Ha Su Jeong , Kim Yeong A , Lee Jihoon , Lim Tae‐Gyu , Kim Yun Tai , Lee Nam Hyouck , Park Jun Seong , Yeom Myeong‐Hun , Lee Hyong Joo , Lee Ki Won
Published: August 2014

Dehydroglyasperin D (DHGA‐D), a compound present in licorice, has been found to exhibit anti‐obesity, antioxidant and anti‐aldose reductase effects. However, the direct molecular mechanism and molecular targets of DHGA‐D during skin inflammation remain unknown. In the present study, we investigated the effect of DHGA‐D on inflammation and its mechanism of action on UVB‐induced skin inflammation in HaCaT human keratinocytes and SKH‐1 hairless mice. DHGA‐D treatment strongly suppressed UVB‐induced COX‐2 expression, PGE2 generation and AP‐1 transactivity in HaCaT cells without affecting cell viability. DHGA‐D also inhibited phosphorylation of the mitogen‐activated protein kinase kinase (MKK) 3/6/p38, MAPK/Elk‐1, MKK4/c‐Jun N‐terminal kinase (JNK) 1/2/c‐Jun/mitogen, and stress‐activated protein kinase (MSK), whereas phosphorylation of the mixed‐lineage kinase (MLK) 3 remained unaffected. Kinase and co‐precipitation assays with DHGA‐D Sepharose 4B beads showed that DHGA‐D significantly suppressed MLK3 activity through direct binding to MLK3. Knockdown of MLK3 suppressed COX‐2 expression as well as phosphorylation of MKK4/p38 and MKK3/6/JNK1/2 in HaCaT cells. Furthermore, Western blot assay and immunohistochemistry results showed that DHGA‐D pre‐treatment significantly inhibits UVB‐induced COX‐2 expression in vivo. Taken together, these results indicate that DHGA‐D may be a promising anti‐inflammatory agent that mediates suppression of both COX‐2 expression and the MLK3 signalling pathway through direct binding and inhibition of MLK3.
Periodontal disease and Helicobacter pylori infection: a community‐based study using serology and rapid urease test
Authors: Nisha Krishnavilasom J. , Nandakumar Krishnankutty , Shenoy Kottacherry T. , Janam Presanthila
Published: August 2014

Aims

The aims of the present study were to assess the prevalence of periodontal disease and Helicobacter pylori (H. pylori) infection and their associations within a predefined Indian population.

Methods

A community‐based cross‐sectional study of 500 selected individuals using a questionnaire, oral examination, rapid urease testing of dental plaque, and serological examination for immunoglobulin G antibody to H. pylori was carried out.

Results

Periodontal disease and H. pylori infection were prevalent in more than 50% of the population. Age, smoking, and diabetic status of the individuals were risk factors for periodontal disease after multivariate analysis, and a lack of proper sewage and waste disposal facilities were found to increase the risk of H. pylori infection. Although there was no association between periodontal disease and H. pylori seropositivity in the community, a highly‐significant association was found between periodontal disease and colonization of H. pylori in dental plaque.

Conclusions

Because periodontal disease is associated with the increased colonization of H. pylori, new treatment modalities, such as plaque control measures, should be employed for the complete management of H. pylori‐associated gastric disease.
Continuous infusion of substance P into rat striatum alleviates nociceptive behavior via phosphorylation of extracellular signal‐regulated kinase 1/2
Authors: Nakamura Yoki , Izumi Hiroki , Fukushige Ryo , Shimizu Takumi , Watanabe Kyohei , Morioka Norimitsu , Hama Aldric , Takamatsu Hiroyuki , Nakata Yoshihiro
Published: August 2014

Intraplantar injection of 0.4% formalin into the rat hind paw leads to a biphasic nociceptive response; an ‘acute’ phase (0–15 min) and ‘tonic’ phase (16–120 min), which is accompanied by significant phosphorylation of extracellular signal‐regulated kinase (ERK)1/2 in the contralateral striatum at 120 min post‐formalin injection. To uncover a possible relationship between the slow‐onset substance P (SP) release and increased ERK1/2 phosphorylation in the striatum, continuous infusion of SP into the striatum by reverse microdialysis (0.4 μg/mL in microdialysis fiber, 1 μL/min) was performed to mimic volume neurotransmission of SP. Continuous infusion for 3 h of SP reduced the duration of ‘tonic’ phase nociception, and this SP effect was mediated by neurokinin 1 (NK1) receptors since pre‐treatment with NK1 receptor antagonist CP96345 (10 μM) blocked the effect of SP infusion. However, formalin‐induced ‘tonic’ phase nociception was significantly prolonged following acute injection of the MAP/ERK kinase 1/2 inhibitor PD0325901 (100 pmol) by microinjection. The coinfusion of SP and PD0325901 significantly increased the ‘tonic’ phase of nociception. These data demonstrate that volume transmission of striatal SP triggered by peripheral nociceptive stimulation does not lead to pain facilitation but a significant decrease of tonic nociception by the activation of the SP‐NK1 receptor–ERK1/2 system.

image

Noxious stimulation induces a slow‐onset substance P (SP) release as a volume transmitter, activating extra‐synaptic NK1 receptors, and evokes phosphorylation of extracellular signal‐regulated kinase (ERK) 1/2. The SP‐NK1‐ERK1/2 system in the striatum decreases tonic nociception.
Cardioprotective Effects of Oral Nicorandil Use in Diabetic Patients Undergoing Elective Percutaneous Coronary Intervention
Authors: SHEHATA MOHAMED
Published: August 2014

Objective

This study sought to evaluate the cardioprotective role of oral nicorandil (a hybrid between the nitrates and adenosine triphosphate [ATP] sensitive potassium channel activators) in diabetic patients undergoing elective percutaneous coronary intervention (PCI).

Background

Myocardial injury commonly occurs during PCI. Several agents that mimic ischemic preconditioning could help minimize this phenomenon.

Methods

One hundred diabetic patients with stable angina were prospectively enrolled, then randomly assigned to receive oral nicorandil: 20 mg once daily (group A, 50 patients) or placebo (group B, 50 patients) 1 week before and 6 months after PCI. Cardiac troponin I (cTnI) and creatine kinase‐MB (CK‐MB) levels were measured before and 6, 12, and 24 hours post‐PCI.

Results

Mean age of the study cohort was 59.8 ± 5.8 years (males = 68%). cTnI level was significantly lower in group A (6 hours: 7.3 ± 0.3 vs. 14.5 ± 0.4 pg/mL; 12 hours: 12.7 ± 0.7 vs. 25.3 ± 0.5 pg/mL; and 24 hours: 7.7 ± 0.5 vs. 15.0 ± 0.4 pg/mL, P < 0.001). After 6 months, the same group showed significantly higher left ventricle ejection fraction (LVEF%), that is, 63.5 ± 7.7% versus 56.5 ± 8.3% (P < 0.05), with a trend toward lower incidence of major adverse cardiac events (MACE).

Conclusion

In diabetic patients undergoing elective PCI, nicorandil intake was associated with decreased incidence of PCI‐related myocardial injury and improvement of LVEF% after 6 months. (J Interven Cardiol 2014;27:472–481)
Cobalt–Chromium KAname™ coRonary stEnt System in the Treatment of Patients With Coronary Artery Disease (KARE Study)
Authors: CARRIE DIDIER , SCHÄCHINGER VOLKER , DANZI GIAN BATTISTA , MACAYA CARLOS , ZEYMER UWE , PUTNIKOVIC BILJANA , INIGUEZ ANDRES , MORENO RAUL , MEHMEDBEGOVIC ZLATKO , BELESLIN BRANKO ,   ON BEHALF OF KARE INVESTIGATORS
Published: August 2014

Objectives

To evaluate the safety and effectiveness of the Kaname™ cobalt–chromium (Co–Cr), thin strut, bare metal stent (BMS) system for the treatment of coronary artery lesions.

Background

Despite widespread use of drug‐eluting stents, a certain percentage of patients with coronary artery disease are still treated with BMS. Therefore, it is essential to evaluate their clinical performance.

Methods

Two hundred eighty‐two patients were enrolled in this prospective, single‐arm study including a predefined subset of 79 patients with small vessels. The primary end‐point was freedom from target vessel failure (TVF) at 6 months. Key angiographic and intravascular ultrasound (IVUS) end‐points were late loss, diameter stenosis, binary restenosis, and neointimal hyperplasia volume.

Results

Freedom from TVF at 6 months was 93.3% and at 1 year 90.8% in total population, and 92.4% and 87.3% in small vessels, respectively. Clinically driven target lesion revascularization (TLR) rates at 6 and 12 months were 4.3% and 6.4% in total population, and 3.8% and 7.6% in small vessels, respectively. At 6 months in‐stent late loss was 0.75 ± 0.43 mm and binary restenosis rate was 16.9% in total population, and 0.64 ± 0.40 mm and 26.1% in small vessels, while IVUS assessed neointimal hyperplasia volume at 6 months was 128.9 ± 42.6 mm3 for total population. There were no definite and probable stent thromboses up to 12 months.

Conclusions

Results indicate good safety and effectiveness of the Kaname™ stent with clinically equivalent results in small and larger vessels, as such providing useful treatment option for patients with ischemic heart disease considered for BMS implantation. (J Interven Cardiol 2014;27:491–499)
Smartphone use and work–home interference: The moderating role of social norms and employee work engagement
Authors: Derks Daantje , van Duin Desiree , Tims Maria , Bakker Arnold B.
Published: August 2014

Work in our modern society that is facilitated by communication technology involves connectivity, immediacy, and a blurring of boundaries between work and non‐work domains. This 4‐day quantitative diary study (N = 100 employees, N = 367–400 data points) aims to shed light on the relationship between daily smartphone use and daily work–home interference (WHI). Two potential moderators of this relationship are examined: (1) (strong) social norms represented by the influence of colleagues and supervisors regarding availability after work hours and (2) work engagement. Overall, the results of multilevel analyses were in line with the hypotheses. The findings suggest that supervisors should be clear about their expectations regarding smartphone use in private hours in that they should not expect employees to be always available. In addition we conclude that engaged workers can prevent work from interfering too much with their private lives, even when they use their smartphones during evening hours.

Practitioner points

Employees working in an ‘always‐on’ culture experience more WHI.

Important role models, such as supervisors, should be aware that the emails they send during evening hours and weekends also have recipients.

Supervisors should be careful in creating expectations regarding availability when they decide to provide smartphones to their employees.
Insecticidal and repellent effects of tea tree and andiroba oils on flies associated with livestock
Authors: KLAUCK V. , PAZINATO R. , STEFANI L. M. , SANTOS R. C. , VAUCHER R. A. , BALDISSERA M. D. , RAFFIN R. , BOLIGON A. , ATHAYDE M. , BARETTA D. , MACHADO G. , DA SILVA A. S.
Published: August 2014

This study aimed to evaluate the insecticidal and repellent effects of tea tree, Melaleuca alternifolia (Myrtales: Myrtaceae), and andiroba, Carapa guianensis (Sapindales: Meliaceae), essential oils on two species of fly. For in vitro studies, free‐living adult flies were captured and reared in the laboratory. To evaluate the insecticidal effects of the oils, adult flies of Haematobia irritans (L.) and Musca domestica L. (both: Diptera: Muscidae) were separated by species in test cages (n = 10 per group), and subsequently tested with oils at concentrations of 1.0% and 5.0% using a negative control to validate the test. Both oils showed insecticidal activity. Tea tree oil at a concentration of 5.0% was able to kill M. domestica with 100.0% efficacy after 12 h of exposure. However, the effectiveness of andiroba oil at a concentration of 5.0% was only 67.0%. The insecticidal efficacy (100.0%) of both oils against H. irritans was observed at both concentrations for up to 4 h. The repellency effects of the oils at concentrations of 5.0% were tested in vivo on Holstein cows naturally infested by H. irritans. Both oils demonstrated repellency at 24 h, when the numbers of flies on cows treated with tea tree and andiroba oil were 61.6% and 57.7%, respectively, lower than the number of flies on control animals. It is possible to conclude that these essential oils have insecticidal and repellent effects against the species of fly used in this study.
A pathogenic trace of Tannerella forsythia – shedding of soluble fully active tumor necrosis factor α from the macrophage surface by karilysin
Authors: Bryzek D. , Ksiazek M. , Bielecka E. , Karim A.Y. , Potempa B. , Staniec D. , Koziel J. , Potempa J.
Published: August 2014

Tannerella forsythia is implicated as a pathogen causing chronic and aggressive periodontitis. However, its virulence factors, including numerous putative proteases, are mostly uncharacterized. Karilysin is a newly described matrix metalloprotease‐like enzyme of T. forsythia. Since pathogen‐derived proteases may affect the host defense system via modulation of the cytokine network, the aim of this study was to determine the influence of karilysin on tumor necrosis factor‐α (TNF‐α). The results showed that karilysin cleaved the membrane form of TNF‐α on the surface of macrophages, and that this led to an increased concentration of soluble TNF‐α in the conditioned medium. Importantly, despite partial degradation of soluble TNF‐α by karilysin, the released cytokine retained its biological activity, inducing apoptosis and stimulating autocrine pathway of pro‐inflammatory gene expression. Notably, the observed effect required proteolytic activity by karilysin, since a catalytically inactive mutant of the enzyme did not affect TNF‐α secretion. The shedding was independent of the activity of ADAM17, a major endogenous TNF‐α converting enzyme. Karilysin‐dependent TNF‐α release from the cell surface is likely to occur in vivo because human plasma, the main constituent of gingival crevicular fluid, only slightly affected the sheddase activity of karilysin. Taken together, these results indicate that karilysin modulates the host immune response through regulation of TNF‐α secretion, and should therefore be considered as a new virulence factor of T. forsythia.
Peptidylarginine deiminase from Porphyromonas gingivalis contributes to infection of gingival fibroblasts and induction of prostaglandin E2‐signaling pathway
Authors: Gawron K. , Bereta G. , Nowakowska Z. , Łazarz–Bartyzel K. , Łazarz M. , Szmigielski B. , Mizgalska D. , Buda A. , Koziel J. , Oruba Z. , Chomyszyn‐Gajewska M. , Potempa J.
Published: August 2014

Porphyromonas gingivalis (P. gingivalis) expres‐ses the enzyme peptidylarginine deiminase (PPAD), which has a strong preference for C‐terminal arginines. Due to the combined activity of PPAD and Arg‐specific gingipains, P. gingivalis on the cell surface is highly citrullinated. To investigate the contribution of PPAD to the interaction of P. gingivalis with primary human gingival fibroblasts (PHGF) and P. gingivalis‐induced synthesis of prostaglandin E2 (PGE2), PHGF were infected with wild‐type P. gingivalis ATCC 33277, an isogenic PPAD‐knockout strain (∆ppad) or a mutated strain (C351A) expressing an inactive enzyme in which the catalytic cysteine has been mutated to alanine (PPADC351A). Cells were infected in medium containing the mutants alone or in medium supplemented with purified, active PPAD. PHGF infection was assessed by colony‐forming assay, microscopic analysis and flow cytometry. Expression of cyclo‐oxygenase 2 (COX‐2) and microsomal PGE synthase‐1 (mPGES‐1), key factors in the prostaglandin synthesis pathway, was examined by quantitative reverse transcription polymerase chain reaction (qRT‐PCR), while PGE2 synthesis was evaluated by enzyme immunoassay. PHGF were infected more efficiently by wild‐type P. gingivalis than by the ∆ppad strain, which correlated with strong induction of COX‐2 and mPGES‐1 expression by wild‐type P. gingivalis, but not by the PPAD activity‐null mutant strains (Δppad and C351A). The impaired ability of the Δppad strain to adhere to and/or invade PHGF and both Δppad and C351A to stimulate the PGE2‐synthesis pathway was fully restored by the addition of purified PPAD. The latter effect was strongly inhibited by aspirin. Collectively, our results implicate PPAD activity, but not PPAD itself, as an important factor for gingival fibroblast infection and activation of PGE2 synthesis, the latter of which may strongly contribute to bone resorption and eventual tooth loss.
The effectiveness of ambulatory continuous peripheral nerve blocks for postoperative pain management in children and adolescents
Authors: Visoiu Mihaela , Joy Lendi N. , Grudziak Jan S. , Chelly Jacques E.
Published: August 2014

Background

Ambulatory continuous peripheral nerve blocks (CPNBs) are feasible for pediatric patients. We sought to evaluate the efficacy of CPNBs in a pediatric population.

Methods

This retrospective report of 33‐month prospectively collected data investigates patient, parent, and nurse pain control satisfaction score (PCSS), the incidence and severity of pain, daily analgesic consumption following discharge home with various CPNBs and On‐Q pumps, and any complications and side effects related to CPNBs.

Results

Four hundred and three patients (403; aged 5–22) were discharged home with 410 CPNBs (brachial and lumbar plexus, femoral, sciatic, and paravertebral); 76.7% on the day of surgery. The median/interquartile range (IQR) ropivacaine continuous infusion via On‐Q pump was 0.24 (0.20–0.30) mg·kg−1·hr−1, and the median/IQR duration was 72 (48–72) h. The median/IQR home PCSS was 10 (9–10). Median Postoperative Ambulatory Care Unit (PACU)/IQR pain control satisfaction scores were 10 (8–10) for the patient, 10 (9–10) for the parent, and 10 (9–10) for the nurse. Thirty‐three (10.0%) patients did not report any pain at home, and median maximum home/IQR pain score was 4 (2–6). In the PACU, 126 (31.3%) patients did not report any pain and median/IQR pain score was 1 (0–3). No opioids were administered at home for 12 (4.3%) patients and in the PACU for 150 (37.4%). Sixty‐three (14.4%) complications and side effects for 58 patients were reported. We report 93.1% ambulatory efficacy of CPNBs.

Conclusion

Our patients and their caregivers were very satisfied with ambulatory CPNBs. When combined with oral analgesics, CPNBs provided effective home postoperative analgesia.
Treatment of BK virus‐associated nephropathy with CMX001 after kidney transplantation in a young child
Authors: Reisman Lewis , Habib Sabeen , McClure Gloria B. , Latiolais Lisa S. , Vanchiere John A.
Published: August 2014

NC, with renal failure secondary to bilateral dysplastic kidneys, received an LRD renal transplant (tx) at 17 months of age. Her early post‐tx course was complicated by persistently elevated blood polyoma BK virus DNA loads. A protocol biopsy at six months post‐transplant revealed BKVAN. Blood viral loads did not respond to decreased immunosuppression or treatment with ciprofloxacin and leflunomide. Six months post‐tx, her serum creatinine began to rise and we sought experimental therapy to prevent the loss of her graft. At seven months post‐tx, with FDA approval under an eIND, the patient was started on a 36‐wk course of treatment with the investigational drug. The patient is now more than 24 months after stopping treatment with CMX. BKV viral DNA loads remain at low, but still detectable levels. Urine viral loads have declined, but remain elevated. EBV DNA loads become undetectable. The patient's serum creatinine has declined back to a baseline of 0.5–0.7 mg/dL and has been stable for two yr. Renal function was preserved in association with the use of CMX001 to treat BKV nephropathy in a young pediatric kidney transplant recipient. There were no serious adverse events associated with the use of CMX001. This novel medication may be of value in the treatment of BKVAN in pediatric renal transplant recipients.
I Was a Teenage Vegan: Motivation and Maintenance of Lifestyle Movements
Authors: Cherry Elizabeth
Published: August 2014

Recent research has pointed to the rise of socially conscious consumption and of lifestyle movements or social movements that focus on changing one's everyday lifestyle choices as a form of protest. Much of this research addresses how adults maintain socially conscious consumption practices. Using interviews with youths who are vegan—strict vegetarians who exclude all animal products from their diet and lifestyle—I isolate the factors influencing recruitment into and retention of veganism as a lifestyle movement. I show that initial recruitment requires learning, reflection, and identity work, and that subsequent retention requires two factors: social support from friends and family, and cultural tools that provide the skills and motivation to maintain lifestyle activism. I also show how participation in the punk subculture further facilitated these processes. This work contributes to studies of youth subcultures and social movements by showing how the two intersect in lifestyle movement activism.
“Your Honor” is a Female: A Multistage Electoral Analysis of Women's Successes at Securing State Trial Court Judgeships*
Authors: Bullock Charles S. , MacManus Susan A. , Owen Karen Padgett , Penberthy Corttney C. , Reid Ralph O. , McPhee Brian
Published: August 2014

Objectives

Using a multistage gender‐focused analysis of candidates for trial court judgeships, we seek answers to these leading questions: (1) Are women as strategic as men in choosing the conditions under which they run for an open seat or against an incumbent and against a male opponent or against another female? (2) Are women winning at the same rates as men under the same conditions at the primary election stage and at the runoff stage? (3) Are women as likely as men to move into a vacant trial court judgeship via a gubernatorial appointment? (4) Are women more likely to be appointed than to win election to the bench?

Methods

This research represents the first use of an “all‐stage” analysis of the relative success of female versus male candidates for trial court judge in nonpartisan settings. The three stages analyzed are (1) candidacy, (2) the initial primary round (outright victory or progression to the runoff), and (3) the end stage—victory in a runoff, if necessary. The study is based on longitudinal data from Florida and Georgia—two of the nation's 10 largest states, each with higher contestation rates than the national average for nonpartisan trial court judicial races—and gubernatorial appointments to new or vacant judgeships.

Results

Women behave strategically, more often seeking open seats than challenging incumbents and more often competing against men than women. Women participated in the candidate pools for most seats having contested primaries and when women run, they win more often than they lose, usually by defeating men. While women fare less well in runoffs than initial primaries, they win more runoffs than they lose. Women more often come to the bench by winning an election than securing a gubernatorial appointment, with female appointees disproportionately serving in newly created judgeships.

Conclusions

Women frequently compete for trial court judgeships and when they run, they usually succeed. Open seats rather than challenging incumbents or hoping for a gubernatorial appointment provide the best way to increase gender equality on the bench.
Immunoagglutination test to diagnose Chagas disease: comparison of different latex–antigen complexes
Authors: Garcia Valeria S. , Gonzalez Verónica D. G. , Marcipar Ivan S. , Gugliotta Luis M.
Published: August 2014

Objective

To evaluate the diagnostic performance of novel latex–protein complexes obtained from different antigens of Trypanosoma cruzi through immunoagglutination test using a panel of T. cruzi‐positive sera, leishmaniasis‐positive sera and negative sera for both parasites.

Methods

Complexes’ behaviour using total parasite homogenate (TPH), two simple recombinant proteins (RP1 and RP5) and two chimeric recombinant proteins (CP1 and CP2) was comparatively evaluated. The area under ROC curves was used as an index of accuracy. Sensitivity, specificity and discrimination efficiency were assessed.

Results

All recombinant antigens showed higher specificity than TPH. The lower specificity of TPH was mainly due to cross‐reacting peptides between T. cruzi and Leishmania spp. In turn, all performance indicators were higher for CP1 and CP2 than for RP1 and RP5. The carboxylated latex–CP2 (C2–CP2) complex was able to detect antibodies against T. cruzi. The values of area under ROC curve (0.96), sensitivity (92.3%, 95% CI: 79.4–100.0%) and specificity (84.0%, 95% CI: 67.6–100.0%) indicate that the assay could be used as a screening test.

Conclusion

The C2–CP2 complex could be an important tool to carry out sero‐epidemiological studies.
Presumed canine trigemino‐abducens synkinesis in a dog
Authors: Eminaga Salih , Williams David , Cherubini Giunio Bruto
Published: August 2014

A ten‐year‐old male neutered Rhodesian ridgeback cross dog was presented for the investigation of abnormal bilateral protrusion of the third eyelid when chewing. Physical, ophthalmological, and neurological examinations were unremarkable. Thoracic radiographs, abdominal ultrasound, and magnetic resonance of the brain and orbits failed to reveal any abnormalities. Cerebrospinal fluid analysis revealed elevated protein, but the nucleated cell count was normal. trigemino‐abducens synkinesis was presumptively diagnosed. Aetiopathogenesis of this condition is discussed. To the authors' knowledge, this is the first report of presumed trigemino‐abducens synkinesis in a dog.
Isolation and cultivation of canine uveal melanocytes
Authors: Dawson‐Baglien Ethan M. , Winkler Paige A. , Bruewer Ashlee R. , Petersen‐Jones Simon M. , Bartoe Joshua T.
Published: August 2014

Objectives

To establish a method for isolation and culture of canine uveal melanocytes.

Animals studied

Uveal explants from five mixed‐breed dogs.

Procedures

Donor globes were dissected, and the anterior uvea removed. The uveal explants were placed in trypsin solution for enzymatic digestion. Extracted cells were cultured in modified F12 media. Immunocytochemistry was performed to confirm the identity of the extracted cells.

Results

Melanocytes were successfully isolated from uveal explants. Contaminating cell types were not observed. Repeated passaging of the melanocytes resulted in a gradual decrease in intracellular pigment. Melanocyte cell lines could be cryopreserved, thawed, and cultures successfully reestablished.

Conclusions

This extraction technique allows for generation of large populations of canine uveal melanocytes in a relatively short period of time. This technique could be a useful tool for future studies investigating both normal cellular characteristics and alterations found in melanocytes from dogs with ocular melanocytic disorders.
A Scoping Review of the Role of Wildlife in the Transmission of Bacterial Pathogens and Antimicrobial Resistance to the Food Chain
Authors: Greig J. , Rajić A. , Young I. , Mascarenhas M. , Waddell L. , LeJeune J.
Published: August 2014

Wildlife can contribute to environmental contamination with bacterial pathogens and their transfer to the human food chain. Global usage and frequent misuse of antimicrobials contribute to emergence of new antimicrobial resistant (AMR) strains of foodborne pathogens. We conducted a scoping review of published research to identify and characterize the evidence on wildlife's role in transmission of AMR and/or bacterial pathogens to the food chain. An advisory group (AG) of 13 North American experts from diverse disciplines was surveyed to solicit insight in the review scope, priority topics and research characteristics. A pre‐tested search strategy was implemented in seven bibliographic databases (1990 to January 2013). Citations were relevance screened, and key characteristics on priority topics extracted independently by two reviewers. Analysis identified topic areas with solid evidence and main knowledge gaps. North America reported 30% of 866 relevant articles. The prevalence of five targeted bacterial pathogens and/or AMR in any pathogen in wildlife was reported in 582 articles. Transmission risk factors for selected bacteria or AMR in any bacteria were reported in 300. Interventions to control transmission were discussed in 124 articles and formally evaluated in 50. The majority of primary research investigated birds, cervids, rodents, feral pigs, opossums, E. coli (n = 329), Salmonella (n = 293) and Campylobacter (n = 124). An association between wildlife and transmission of bacterial pathogens and/or AMR to the food chain was supported in 122 studies. The scoping review identified a significant body of research on the role of wild birds in the prevalence and transmission of E. coli, Salmonella and Campylobacter. There was little research employing molecular methods contributing to the evidence concerning the importance and direction of transmission of wildlife/pathogen combinations. Given the advancements of these methods, future research should focus in this area to help prioritize future intervention studies and risk mitigation strategies.
Increased blood‐oxygen binding affinity in Tibetan and Han Chinese residents at 4200 m
Authors: Simonson T. S. , Wei G. , Wagner H. E. , Wuren T. , Bui A. , Fine J. M. , Qin G. , Beltrami F. G. , Yan M. , Wagner P. D. , Ge Ri Li
Published: August 2014

New Findings

What is the central question of this study?

Is Tibetan and Chinese highlanders’ blood oxygen‐binding affinity (P50) different from that of other populations (at altitude or sea level), and does Tibetan P50 relate to haemoglobin concentration and/or exercise capacity at altitude?

What is the main finding and its importance?

Tibetans and Chinese at 4200 m have slightly lower P50 than sea‐level residents.

During maximal exercise at 4200 m, reduced P50 does not enhance pulmonary gas exchange, impair systemic oxygen extraction or affect peak exercise capacity.

Oxygen saturation measurements based upon forehead oximetry are sufficiently reduced during exercise at altitude (and accurate compared with those obtained from arterial blood by co‐oximetry) to determine P50 reliably.

High‐altitude natives are challenged by hypoxia, and a potential compensatory mechanism could be reduced blood oxygen‐binding affinity (P50), as seen in several high‐altitude mammalian species. In 21 Qinghai Tibetan and nine Han Chinese men, all resident at 4200 m, standard P50 was calculated from measurements of arterial and forehead oximeter oxygen saturation, which was validated in a separate examination of 13 healthy subjects residing at sea level. In both Tibetans and Han Chinese, standard P50 was 24.5 ± 1.4 and 24.5 ± 2.0 mmHg, respectively, and was lower than in the sea‐level subjects (26.2 ± 0.6 mmHg, P < 0.01). There was no relationship between P50 and haemoglobin concentration (the latter ranging from 15.2 to 22.9 g dl−1 in Tibetans). During peak exercise, P50 was not associated with alveolar–arterial difference or peak O2 uptake per kilogram. There appears to be no apparent benefit of a lower P50 in this adult high‐altitude Tibetan population.
Cerebral vasomotor reactivity: steady‐state versus transient changes in carbon dioxide tension
Authors: Brothers R. Matthew , Lucas Rebekah A. I. , Zhu Yong‐Sheng , Crandall Craig G. , Zhang Rong
Published: August 2014

New Findings

What is the central question of this study?

The relationship between changes in cerebral blood flow and arterial carbon dioxide tension is used to assess cerebrovascular function. Hypercapnia is generally evoked by two methods, i.e. steady‐state and transient increases in carbon dioxide tension. In some cases, the hypercapnia is immediately preceded by a period of hypocapnia. It is unknown whether the cerebrovascular response differs between these methods and whether a period of hypocapnia blunts the subsequent response to hypercapnia.

What is the main finding and its importance?

The cerebrovascular response is similar between steady‐state and transient hypercapnia. However, hyperventilation‐induced hypocapnia attenuates the cerebral vasodilatory responses during a subsequent period of rebreathing‐induced hypercapnia.

Cerebral vasomotor reactivity (CVMR) to changes in arterial carbon dioxide tension () is assessed during steady‐state or transient changes in . This study tested the following two hypotheses: (i) that CVMR during steady‐state changes differs from that during transient changes in ; and (ii) that CVMR during rebreathing‐induced hypercapnia would be blunted when preceded by a period of hyperventilation. For each hypothesis, end‐tidal carbon dioxide tension () middle cerebral artery blood velocity (CBFV), cerebrovascular conductance index (CVCI; CBFV/mean arterial pressure) and CVMR (slope of the linear regression between changes in CBFV and CVCI versus ) were assessed in eight individuals. To address the first hypothesis, measurements were made during the following two conditions (randomized): (i) steady‐state increases in of 5 and 10 Torr above baseline; and (ii) rebreathing‐induced transient breath‐by‐breath increases in . The linear regression for CBFV versus (P = 0.65) and CVCI versus (P = 0.44) was similar between methods; however, individual variability in CBFV or CVCI responses existed among subjects. To address the second hypothesis, the same measurements were made during the following two conditions (randomized): (i) immediately following a brief period of hypocapnia induced by hyperventilation for 1 min followed by rebreathing; and (ii) during rebreathing only. The slope of the linear regression for CBFV versus (P < 0.01) and CVCI versus (P < 0.01) was reduced during hyperventilation plus rebreathing relative to rebreathing only. These results indicate that cerebral vasomotor reactivity to changes in is similar regardless of the employed methodology to induce changes in and that hyperventilation‐induced hypocapnia attenuates the cerebral vasodilatory responses during a subsequent period of rebreathing‐induced hypercapnia.
Modulation of non‐NMDA receptor gating by auxiliary subunits
Authors: Howe James R.
Published: August 2014

During the past decade, considerable evidence has accumulated that non‐NMDA glutamate receptors (both AMPA and kainate subtypes) are modulated by the association of the core tetrameric receptor with auxiliary proteins that are integral components of native receptor assemblies. This short review focuses on the effect of two types of auxiliary subunits on the biophysical properties and kinetic behaviour of AMPA and kainate receptors at the level of single receptor molecules. Type I transmembrane AMPA receptor proteins increase the number of AMPA receptor openings that result from a single receptor activation as well as the proportion of openings to conductance levels above 30 pS, resulting in larger peak ensemble currents that decay more slowly and bi‑exponentially. Co‐expression of Neto1 and 2 with pore‐forming kainate receptor subunits also increases the duration of bursts and destabilizes desensitized states, resulting in a rapid component of recovery and clusters of bursts that produce a slow component in desensitization decays. The distinct gating seen in the presence of auxiliary subunits reflects slow switching between gating modes with different single‐channel kinetics and open probability. At any given time, the relative proportions of receptors in each gating mode determine both the shape and the amplitude of synaptic currents.
Na+ current properties in islet α‐ and β‐cells reflect cell‐specific Scn3a and Scn9a expression
Authors: Zhang Quan , Chibalina Margarita V. , Bengtsson Martin , Groschner Lukas N. , Ramracheya Reshma , Rorsman Nils J. G. , Leiss Veronika , Nassar Mohammed A. , Welling Andrea , Gribble Fiona M. , Reimann Frank , Hofmann Franz , Wood John N. , Ashcroft Frances M. , Rorsman Patrik
Published: August 2014

Key points

α‐ and β‐cells express both Nav1.3 and Nav1.7 Na+ channels but in different relative amounts.

The differential expression explains the different properties of Na+ currents in α‐ and β‐cells.

Nav1.3 is the functionally important Na+ channel α subunit in both α‐ and β‐cells.

Islet Nav1.7 channels are locked in an inactive state due to an islet cell‐specific factor.

Mouse pancreatic β‐ and α‐cells are equipped with voltage‐gated Na+ currents that inactivate over widely different membrane potentials (half‐maximal inactivation (V0.5) at −100 mV and −50 mV in β‐ and α‐cells, respectively). Single‐cell PCR analyses show that both α‐ and β‐cells have Nav1.3 (Scn3) and Nav1.7 (Scn9a) α subunits, but their relative proportions differ: β‐cells principally express Nav1.7 and α‐cells Nav1.3. In α‐cells, genetically ablating Scn3a reduces the Na+ current by 80%. In β‐cells, knockout of Scn9a lowers the Na+ current by >85%, unveiling a small Scn3a‐dependent component. Glucagon and insulin secretion are inhibited in Scn3a−/− islets but unaffected in Scn9a‐deficient islets. Thus, Nav1.3 is the functionally important Na+ channel α subunit in both α‐ and β‐cells because Nav1.7 is largely inactive at physiological membrane potentials due to its unusually negative voltage dependence of inactivation. Interestingly, the Nav1.7 sequence in brain and islets is identical and yet the V0.5 for inactivation is >30 mV more negative in β‐cells. This may indicate the presence of an intracellular factor that modulates the voltage dependence of inactivation.
LRRC8A protein is indispensable for swelling‐activated and ATP‐induced release of excitatory amino acids in rat astrocytes
Authors: Hyzinski‐García María C. , Rudkouskaya Alena , Mongin Alexander A.
Published: August 2014

Key points

Swelling‐activated release of amino acids in the CNS is thought to be mediated by an unidentified volume‐regulated anion channel.

Two recent studies discovered that LRRC8 family members form a volume‐regulated anion channel in non‐neural cells.

In this work we established a critical contribution of the LRRC8A gene product to swelling‐activated glutamate and taurine release from primary rat astrocytes.

We also found that LRRC8A is indispensable for glutamate and taurine release from non‐swollen astrocytes when they are stimulated with ATP.

These findings suggest that LRRC8A may play a role in physiological release of gliotransmitters, and mediate pathological glutamate release in the CNS disorders associated with cellular swelling.

In mammals, cellular swelling activates release of small organic osmolytes, including the excitatory amino acids (EAA) glutamate and aspartate, via a ubiquitously expressed volume‐regulated chloride/anion channel (VRAC). Pharmacological evidence suggests that VRAC plays plural physiological and pathological roles, including excitotoxic release of glutamate in stroke. However, the molecular identity of this pathway was unknown. Two recent studies discovered that LRRC8 gene family members encode heteromeric VRAC composed of LRRC8A plus LRRC8B‐E, which mediate swelling‐activated Clˉ currents and taurine release in human non‐neural cells (Z. Qiu et al. Cell 157: 447, 2014; F.K. Voss et al. Science 344: 634, 2014). Here, we tested the contribution of LRRC8A to the EAA release in brain glia. We detected and quantified expression levels of LRRC8A‐E in primary rat astrocytes with quantitative RT‐PCR and then downregulated LRRC8A with gene‐specific siRNAs. In astrocytes exposed to hypo‐osmotic media, LRRC8A knockdown dramatically reduced swelling‐activated release of the EAA tracer d‐[3H]aspartate. In parallel HPLC assays, LRRC8A siRNA prevented hypo‐osmotic media‐induced loss of the endogenous intracellular l‐glutamate and taurine. Furthermore, downregulation of LRRC8A completely ablated the ATP‐stimulated release of d‐[3H]aspartate and [14C]taurine from non‐swollen astrocytes. Overall, these data indicate that LRRC8A is an indispensable component of a permeability pathway that mediates both swelling‐activated and agonist‐induced amino acid release in brain glial cells.