Miracle-dz/UniFAR-FTUs huggingface dataset:
Zymosan suppresses leukotriene C4 synthase activity in differentiating monocytes: antagonism by aspirin and protein kinase inhibitors

Cysteinyl leukotrienes (cysLTs) are potent proinflammatory mediators with particular relevance for asthma. However, control of cysLT biosynthesis in the time period after onset of acute inflammation has not been extensively studied. As a model for later phases of inflammation, we investigated regulation of leukotriene (LT) C4 synthase (LTC4S) in differentiating monocytes, exposed for several days to fungal zymosan. Incubations with LTA4 revealed 20‐fold increased LTC4S activity during differentiation of monocytic Mono Mac 6 (MM6) cells, which was reduced by 80% in the presence of zymosan (25 µg/ml, 96 h). Zymosan (48 h) similarly attenuated LTC4S activity of primary human monocyte‐derived macrophages and dendritic cells. Several findings indicate phosphoregulation of LTC4S: increased activity during MM6 cell differentiation correlated with reduced phosphorylation of 70‐kDa ribosomal protein S6 kinase (p70S6K), which could phosphorylate purified LTC4S; the p70S6K inhibitor rapamycin (20 nM) doubled LTC4S activity of undifferentiated MM6 cells, and protein kinase A and C inhibitors (H‐89, CGP‐53353, and staurosporine) reversed the zymosan‐induced suppression of LTC4S activity. Finally, zymosan (48 h) up‐regulated PGE2 biosynthesis, and aspirin (10 µM) or prostaglandin E2 (PGE2) receptor antagonists counteracted the zymosan effect. Our results suggest a late PGE2‐mediated phosphoregulation of LTC4S during microbial exposure, which may contribute to resolution of inflammation, with implications for aspirin hypersensitivity. —Esser, J., Gehrmann, U. , Salvado, M. D. , Wetterholm, A. , Haeggstrom,J. Z. , Samuelsson, B. , Gabrielsson, S. , Scheynius, A. , Râdmark, O. Zymosan suppresses leukotriene C4 synthase activity in differentiating monocytes: antagonism by aspirin and protein kinase inhibitors. FASEBJ. 25, 1417–1427 (2011). www.fasebj. org
1-1-2013 Cyclin-Dependent Kinase 2 Regulates Foxp 3 and Regulatory T Cell Function

Foxp3 is a transcription factor required for the development and function of regulatory T cells (Treg). Humans lacking functional Foxp3 are afflicted with uncontrolled systemic autoimmunity. How the Foxp3 protein is regulated post-translationally is unclear. Our previous studies demonstrate cyclin-dependent kinase 2 (CDK2) controls Foxp3+Treg function, but the mechanism by which this occurred was not identified. The CDKs are primarily thought to control cell cycle progression. However, recent studies suggest only CDK1 is required for normal mammalian cell cycle, raising questions about the biological role of the other CDKs. Specifically, mice genetically deficient in CDK2 are viable with no significant defects in cell cycle. We probed the Foxp3 sequence for the presence of CDK motifs, finding four such sites in the amino terminus. We confirmed that Foxp3 is phosphorylated by CDK2 using an in vitro kinase assay and mass spectrometry, as well as a phospho-specific antibody that recognizes one of the phosphorylated Foxp3-CDK motifs. We generated a mutant of Foxp3 lacking all four CDK motifs, which has increased half-life compared to wild-type Foxp3. CD4+ T cells transduced with a Foxp3-CDK motif mutant have increased function compared to cells transduced with wild-type Foxp3 as measured by induction and repression of canonical Foxp3 target genes, as well as the ability to suppress conventional T cell proliferation. These data suggest CDKs negatively regulate Foxp3 protein stability, which has an impact on Foxp3 function. To determine when the CDK cascade was actively regulating Foxp3 in vivo we investigated the role of the CDK2 inhibitor, p27kip1. Recent data shows TGFβ signaling drives expression of p27kip1 in B cells. TGFβ is also required for extrathymic induction of Foxp3 in conventional CD4+ T cells. We show that conventional T cells, which have high CDK2 activity and minimal p27kip1 expression, induce large amounts of p27kip1 along with Foxp3 in the presence of TGFβ. Additionally, T cells lacking p27kip1 have defective TGFβ-dependent Foxp3 induction. We hypothesize that TGFβ signaling is required to activate p27kip1 and stabilize Foxp3 protein levels in developing iTreg by repressing CDK2. Degree Type Dissertation Degree Name Doctor of Philosophy (PhD) Graduate Group Immunology First Advisor Andrew Wells Second Advisor Chris Hunter This dissertation is available at ScholarlyCommons: http://repository.upenn. edu/edissertations/900
Phosphate metabolite concentrations and ATP hydrolysis potential in normal and ischaemic hearts

To understand how cardiac ATP and CrP remain stable with changes in work rate – a phenomenon that has eluded mechanistic explanation for decades – data from 31phosphate‐magnetic resonance spectroscopy (31P‐MRS) are analysed to estimate cytoplasmic and mitochondrial phosphate metabolite concentrations in the normal state, during high cardiac workstates, during acute ischaemia and reactive hyperaemic recovery. Analysis is based on simulating distributed heterogeneous oxygen transport in the myocardium integrated with a detailed model of cardiac energy metabolism. The model predicts that baseline myocardial free inorganic phosphate (Pi) concentration in the canine myocyte cytoplasm – a variable not accessible to direct non‐invasive measurement – is approximately 0.29 mm and increases to 2.3 mm near maximal cardiac oxygen consumption. During acute ischaemia (from ligation of the left anterior descending artery) Pi increases to approximately 3.1 mm and ATP consumption in the ischaemic tissue is reduced quickly to less than half its baseline value before the creatine phosphate (CrP) pool is 18% depleted. It is determined from these experiments that the maximal rate of oxygen consumption of the heart is an emergent property and is limited not simply by the maximal rate of ATP synthesis, but by the maximal rate at which ATP can be synthesized at a potential at which it can be utilized. The critical free energy of ATP hydrolysis for cardiac contraction that is consistent with these findings is approximately −63.5 kJ mol−1. Based on theoretical findings, we hypothesize that inorganic phosphate is both the primary feedback signal for stimulating oxidative phosphorylation in vivo and also the most significant product of ATP hydrolysis in limiting the capacity of the heart to hydrolyse ATP in vivo. Due to the lack of precise quantification of Piin vivo, these hypotheses and associated model predictions remain to be carefully tested experimentally.
Differences in the methyl ester distribution of homogalacturonans from near-isogenic wheat lines resistant and susceptible to the wheat stem rust fungus.

Plants possess an efficient nonself surveillance system triggering induced disease resistance mechanisms upon molecular recognition of microbial invaders. Successful pathogens have evolved strategies to evade or counteract these mechanisms, e.g., by the generation of suppressors. Pectic fragments produced during host cell wall degradation can act as endogenous suppressors of the hypersensitive response in wheat leaves. We have isolated and characterized homogalacturonans from cell walls of two wheat cultivars susceptible to the stem rust fungus, Puccinia graminis f. sp. tritici, namely cvs. Prelude and Marquis, and from near-isogenic lines of both cultivars containing the Sr5-gene for hypersensitive rust resistance. Two independent approaches were used to compare their methyl esterification: i) immunochemistry using the monoclonal antibodies JIM5, JIM7, PAM1, and LM7 and ii) chromatography of oligogalacturonides representing stretches of contiguous nonmethyl-esterified GalA residues. The results clearly indicate a significant difference in the homogalacturonans from susceptible and resistant wheat lines. The difference can best be explained by assuming a nonrandom and more blockwise distribution of the methyl esters in the homogalacturonans of susceptible wheat cultivars as compared with a presumably more random distribution in the near-isogenic resistant lines. Possible consequences of this difference for the enzymatic generation of endogenous suppressors are discussed.
Function of identified motoneurones and co‐ordination of primary and secondary motor systems during zebra fish swimming.

1. The activity of the two classes of motoneurones, primary and secondary, which innervate myotomal muscle fibres in the zebra fish, was monitored with electromyographic and intracellular techniques. 2. Simultaneous EMG and intracellular recordings from muscle fibres showed that the activity of the two motor systems and of individual primary motoneurones can be distinguished by recording EMG spikes during swimming. 3. Measurements of EMG spikes demonstrated that primary and secondary motoneurones are co‐ordinately activated over a wide range of conditions during normal swimming. 4. During swimming the primary motoneurones within a given segment are usually co‐activated although they sometimes fire independently. 5. When different primary motoneurones within a given segment are co‐activated, they fire nearly synchronously. 6. We conclude that the primary motoneurones are used principally, although not exclusively, during fast swimming, struggling and the startle response, whereas secondary motoneurones function primarily during slower swimming.
A visual study of computers on doctors' desks.

BACKGROUND General practice has rapidly computerised over the past ten years, thereby changing the nature of general practice rooms. Most general practice consulting rooms were designed and created in an era without computer hardware, establishing a pattern of work around maximising the doctor-patient relationship. General practitioners (GPs) and patients have had to integrate the computer into this environment. METHODS Twenty GPs allowed access to their rooms and consultations as part of a larger study. The results are based on an analysis of still shots of the consulting rooms. Analysis used dramaturgical methodology; thus the room is described as though it is the setting for a play. RESULTS First, several desk areas were identified: a shared or patient area, a working area, a clinical area and an administrative area. Then, within that framework, we were able to identify two broad categories of setting, one inclusive of the patient and one exclusive. CONCLUSION With the increasing significance of the computer in the three-way doctor-patient-computer relationship, an understanding of the social milieu in which the three players in the consultation interact (the staging) will inform further analysis of the interaction, and allow a framework for assessing the effects of different computer placements.
Continued decay in the cyclotron line energy in Hercules X-1

The centroid energy Ecyc of the cyclotron line in the spectrum of the binary X-ray pulsar Her X-1 had been found to decrease with time on a time scale of a few tens of years - surprisingly short astrophysically. This was found for the pulse phase averaged line centroid energy using observational data from various X-ray satellites over the time frame 1996 to 2012, establishing a reduction of about 4 keV. Here we report on the result of a new observation by NuSTAR performed in August 2015. The earlier results are confirmed and strengthened, with respect to both, the dependence of Ecyc on flux (it is still present after 2006) and the dependence on time: the long-term decay continued with the same rate, corresponding to a reduction of about 5 keV in 20 years.
The T cell receptor begins to reveal its many facets

lished work, Baker says he has been able to use them to make dendrimer-DNA complexes that have now successfully transferred genes to more than 30 different types of cells with an efficiency up to 1000 times higher than that found by Szoka. Those results have aroused some interest from gene therapy researchers. "I believe dendrimers have a serious future as vectors for gene therapy," says Claude Nicolau, a professor of medicine at Harvard Medical School, who has spent years working on liposome vectors. Thus far, however, dendrimers have not made the jump from success in the petri dish to success in live animals, he adds. Indeed, it's far from certain that dendrimers will ever make that jump, says James Wilson, director of the Institute for Human Gene Therapy at the University ofPennsylvania in Philadelphia. Polylysines, another DNAbinding synthetic vector, also looked good at transfecting cells in culture, he notes, but they failed miserably when it came to tests in live animals. Researchers may be more successful in attempts to use dendrimers to move electrons around rather than genes. Larry Miller and his colleagues at the University of Minnesota, for example, have added electronically active groups called imides to the outer branches ofPAMAM dendrimers in hopes of engineering a new class of 3D conducting polymers that may have unique electrical properties. The conducting polymers that are currently available, such as polyacetylene, are linear molecules. But in recent years researchers have found that 3D conducting molecules have unique properties not found in their linear counterparts. The soccerball-shaped carbon molecules known as fullerenes, for example, superconduct (conduct electricity without resistance) at a temperature higher than that of any other molecular conductor. To make dendrimers with the right properties for this job, Miller's group starts with spherical PAMAM dendrimers that have 192 branches to which they attach pancakeshaped imide groups. Miller suspects this will allow electrons to jump from imide to imide around the surface of the sphere (although to demonstrate this he must first fashion the dendrimers into a larger material). Others think the work holds promise. Formulating the spheres "sounds like a significant advance in terms of controlling the surface chemistry" of dendrimers, according to Martin Bryce, professor ofchemistry at the University of Durham in England, who is working toward the same goal but using different conductive molecules as dendrimer add-ons. However, "you shouldn't get too excited about applications yet," he cautions. But a little excitement, the dendrimer-makers feel, is finally warranted. -Robert F. Service IMMUNOLOGY
Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups.

Hepatitis C virus (HCV) is an important human pathogen that is associated with transfusion-related non-A, non-B hepatitis. Recently, HCV cDNA was cloned and the nucleotide sequence of approximately three-quarters of the virus genome was determined. A region of the predicted polyprotein sequence was found to share similarity with a nonstructural protein encoded by dengue virus, a member of the flavivirus family. We report here that HCV shares an even greater degree of protein sequence similarity with members of the pestivirus group (i.e., bovine viral diarrhea virus and hog cholera virus), which are thought to be distantly related to the flaviviruses. In addition, we find that HCV shares significant protein sequence similarity with the polyproteins encoded by members of the picornavirus-like and alphavirus-like plant virus supergroups. These data suggest that HCV may be evolutionarily related to both plant and animal viruses.
Technical Contribution: A Cautionary Note on the Use of Behavioural Diversity (H-Index) in Animal Welfare Science

Animal welfare scientists actively seek reliable and practical metrics that can serve as indicators of animal welfare for use with agricultural, laboratory and zoo-housed animals. Behavioural diversity as a welfare concept originated from early welfare scientists linking poor animal welfare with a high proportion of time spent engaged in stereotyped behaviours and little expression of behaviours that would be expected under wild, or natural conditions. Recently, the concept of behavioural diversity as a welfare indicator has been widely adopted and is frequently quantified by the Shannon-Wiener Diversity Index (H-index). However, the H-Index is fraught with theoretical and mathematical limitations when applied to animal welfare. Four key problems with this metric are demonstrated: the metric’s responsivity to the size of the behavioural repertoire and the underlying assumption that larger behavioural repertoires reflect better welfare, the sensitivity of the metric to arbitrary decisions about the resolution of behavioural categories, the fact that the calculation of the metric is agnostic to the valence of behaviours, and the metric’s susceptibility to the common practice of excluding some behavioural categories. Moving forward, we recommend focusing on validated welfare measures that are sensitive to valence when evaluating animal welfare.
Followup radiographic data on patients with rheumatoid arthritis who participated in a two-year trial of prednisone therapy or placebo.

OBJECTIVE In a previous clinical trial of patients with early rheumatoid arthritis (RA), it was determined that patients who received 10 mg of prednisone per day for 2 years had less radiographic joint damage compared with those who received placebo. Our goal was to investigate whether this beneficial effect persisted after the end of the trial. METHODS A blinded assessment of radiographic joint damage was performed approximately 3 years after the end of the original 2-year study. Twenty-four patients from the original prednisone group (60%) and 28 patients from the original placebo group (68%) participated in this followup study. At the end of the original trial, prednisone dosages were tapered down in the prednisone group and stopped, if possible. Patients from the original prednisone group took prednisone during 35% of the followup period (approximately 1 year) at a mean daily dose of approximately 5 mg. Two patients from the original placebo group started taking prednisone during followup. Radiographs of the hands and feet were scored according to the van der Heijde modification of the Sharp method. RESULTS During 3 additional years of followup, radiographic scores showed significantly less progression in the original prednisone group than in the original placebo group. Radiographic damage in the original prednisone group did not show an accelerated rate of progression during the followup period. CONCLUSION The inhibition of radiographic joint damage in patients with early active RA treated with 10 mg of prednisone per day for 2 years seems to persist after the end of prednisone therapy.
Expression of miR-32 in human non-small cell lung cancer and its correlation with tumor progression and patient survival.

INTRODUCTION miR-32 has recently been found to be implicated in many critical processes in various types of human cancer. However, its clinical significance in human non-small cell lung cancer (NSCLC) has not yet been elucidated. In the present study, we investigated the expression of miR-32 in NSCLC and analyzed its association with clinical features and prognosis of NSCLC patients. METHODS Quantitative real-time PCR (qRT-PCR) was used to measure expression level of miR-32 in lung cancer cell lines, normal bronchial epithelial cells, 90 pairs of tumor samples and adjacent non-tumor tissues. To determine its prognostic value, overall survival was evaluated using the Kaplan-Meier method. Univariate and multivariate analysis were performed using the Cox proportional hazard analysis. RESULTS The expression of miR-32 was significantly decreased in lung cancer cell lines and NSCLC tissues compared with normal bronchial epithelial cells and adjacent non-tumor tissues (P < 0.05). This reduction of miR-32 was associated with tumor stage and lymph node metastasis (P < 0.05). Moreover, Kaplan-Meier analysis demonstrated that patients with low miR-32 expression had shorter overall survival time than those with high miR-32 expression (P < 0.05). Univariate analysis revealed statistically significant correlations between overall survival and miR-32 level, tumor stage and lymph node metastasis (P < 0.05). Furthermore, miR-32 levels, tumor stage and lymph node metastasis were independently associated with overall survival (P < 0.05). CONCLUSIONS Our results provided the first evidence that down-regulation of miR-32 was correlated with NSCLC progression, and miR-32 might be a potential molecular biomarker for predicting the prognosis of patients.
Escherichia coli msbB Gene as a Virulence Factor and a Therapeutic Target

ABSTRACT A mutation in the msbB gene of Escherichia coli results in the synthesis of E. colilipopolysaccharide (LPS) that lacks the myristic acid moiety of lipid A. Although such mutant E. coli cells and their purified LPS have a greatly reduced ability to stimulate human immune cells, a minor reduction in the mouse inflammatory response is observed. When the msbB mutation is transferred into a clinical isolate ofE. coli, there is a significant loss in virulence, as assessed by lethality in BALB/c mice. When a cloned msbBgene is provided to functionally complement the msbBmutant, virulence returns, providing direct evidence that themsbB gene product is an important virulence factor in a murine model of E. coli pathogenicity. In the genetic background of the clinical E. coli isolate, themsbB mutation also results in filamentation of the cells at 37°C but not at 30°C, a reduction in the level of the K1 capsule, an increase in the level of complement C3 deposition, and an increase in both opsonic and nonopsonic phagocytosis of the msbBmutant, phenotypes that can help to explain the loss in virulence. The demonstration that the inhibition of msbB gene function reduces the virulence of E. coli in a mouse infection model warrants further investigation of the msbB gene product as a novel target for antibiotic therapy.
Genetic parameters for faecal egg count following mixed, natural, predominantly Ostertagia circumcincta infection and relationships with live weight in young lambs

Faecal egg counts and live weights were measured on approximately 200 predominantly twin-born Scottish Blackface lambs each year for 3 years, from 1 to 6 1'lonths of age. Measurements lvere made at 4-ti'eek mt/:'ltlals following anthelmintic treatment. Heritability estimates (with s.e.s) of log transformed faecal egg count at each age were 0.01,0.00,0.12 (0.10), 0.14 (0.12), 0.15 (0.07) ard 0.22 (0.13), for ages 1 to 6 months respectively. Therefore, genetic variation exists for acquired but apparentl~ not for innate resistance to infection. Maternal common environmental effects (with s.e.s) were 0.36 (0.11), 0.2b (0.05), 0.27 (0.09),0.06 (0.08), 0.15 (0.09) and 0.16 (0.08), for ages 1 to 6 months respectively. Genetic correlatiqns between faecal egg counts in Iambs older than 3 mpnths u'ere not significantly less than 1.0, indicating that faecal egg counts at different ages are expressions of the same trait. Phenotypic correlations between faecal egg colfnts were generally positive but small. Measurement ,error contributed one-third of the observed variation for indrvidual egg counts. The heritability of mean faecal egg count from.3 to 6 months was 0:33 (s.e. 0.15), . indicatin~ ~hat selection decisions can be ma~e mo~e accurately using multIple egg counts per anImal. PhenotypIc correlatIons between faecal egg counts and live weIght were generally negative but close to zero. However, genetic correlati~ns between faecal egg counts and live weight in lambs older than 3 months were close to -1.0, indicating that r~sistance to gastrointestinal parasites may be an important genetic determinant of growth rate in this environment.