Urate crystals induce macrophage PAF‑AH secretion which is differentially regulated by TGFβ1 and hydrocortisone

The aim of the present study was to establish the role of platelet‑activating factor acetyl hydrolase (PAF‑AH) in the resolution phase of gout using an established in vitro mononuclear cell model. The effects of signalling pathway inhibitors on PAF‑AH secretion, as well as the effects of the common treatments hydrocortisone and colchicine, an antibody against the anti‑inflammatory cytokine transforming growth factor β1 (TGFβ1), and the transcriptional inhibitor actinomycin D, were also investigated. The effect of recombinant PAF‑AH on cytokine secretion by these cells was also determined. Human peripheral blood‑derived monocytes were isolated and differentiated into macrophages. Monocytes and macrophages were stimulated with monosodium monohydrate urate (MSU) crystals or lipopolysaccharide in the presence or absence of AEG3482 [a c‑Jun N‑terminal kinase (JNK) inhibitor], MG132 (a proteasome inhibitor), hydrocortisone or colchicine. Cultures were then analysed for PAF‑AH secretion using ELISA. A 6‑fold upregulation of PAF‑AH secretion was observed following macrophage exposure to MSU crystals for 24 h (29.3±6 vs. 5.4±0.3 ng/ml unstimulated; P<0.05). Following 72 h, PAF‑AH levels decreased significantly (11.1±1.8; P<0.01). Secretion was further enhanced following pre‑treatment with the JNK protein kinase inhibitor AEG3482 prior to MSU crystal stimulation (P<0.05) and was abrogated when cells were preincubated with actinomycin D or the proteasome inhibitor MG132 (50, 100 and 200 µM). The addition of recombinant PAF‑AH (2.5‑10 ng/ml) to MSU crystal‑stimulated immature monocyte cultures significantly decreased pro‑inflammatory interleukin (IL)‑1β (unstimulated 687±124 vs. stimulated 113±30 pg/ml) and IL‑6 secretion (unstimulated 590±50 vs. stimulated 182±19 pg/ml). Treatment of MSU crystal‑stimulated macrophages with hydrocortisone (2 µM) also significantly decreased PAF‑AH release (P<0.05). Neutralising anti‑TGFβ1 addition decreased PAF‑AH dose‑dependently with the highest inhibition observed at 1 µg/ml (P<0.05). The results implicated that PAF‑AH may have an anti‑inflammatory role in the resolution phase of gout

Current-Voltage Characteristics of Weyl Semimetal Semiconducting Devices, Veselago Lenses and Hyperbolic Dirac Phase

The current-voltage characteristics of a new range of devices built around Weyl semimetals has been predicted using the Landauer formalism. The potential step and barrier have been reconsidered for a three-dimensional Weyl semimetals, with analogies to the two-dimensional material graphene and to optics. With the use of our results we also show how a Veselago lens can be made from Weyl semimetals, e.g. from NbAs and NbP. Such a lens may have many practical applications and can be used as a probing tip in a scanning tunneling microscope (STM). The ballistic character of Weyl fermion transport inside the semimetal tip, combined with the ideal focusing of the Weyl fermions (by Veselago lens) on the surface of the tip may create a very narrow electron beam from the tip to the surface of the studied material. With a Weyl semimetal probing tip the resolution of the present STMs can be improved significantly, and one may image not only individual atoms but also individual electron orbitals or chemical bonding and therewith to resolve the long-term issue of chemical and hydrogen bond formation. We show that applying a pressure to the Weyl semimental, having no centre of spacial inversion one may model matter at extreme conditions such as those arising in the vicinity of a black hole. As the materials Cd3As2 and Na3Bi show an asymmetry in their Dirac cones, a scaling factor was used to model this asymmetry. The scaling factor created additional regions of no propagation and condensed the appearance of resonances. We argue that under an external pressure there may arise a topological phase transition in Weyl semimetals, where the electron transport changes character and becomes anisotropic. There a hyperbolic Dirac phases occurs where there is a strong light absorption and photo-current generation

'Special K' and a loss of cell-to-cell adhesion in proximal tubule-derived epithelial cells: modulation of the adherens junction complex by ketamine

Ketamine, a mild hallucinogenic class C drug, is the fastest growing ‘party drug’ used by 16–24 year olds in the UK. As the recreational use of Ketamine increases we are beginning to see the signs of major renal and bladder complications. To date however, we know nothing of a role for Ketamine in modulating both structure and function of the human renal proximal tubule. In the current study we have used an established model cell line for human epithelial cells of the proximal tubule (HK2) to demonstrate that Ketamine evokes early changes in expression of proteins central to the adherens junction complex. Furthermore we use AFM single-cell force spectroscopy to assess if these changes functionally uncouple cells of the proximal tubule ahead of any overt loss in epithelial cell function. Our data suggests that Ketamine (24–48 hrs) produces gross changes in cell morphology and cytoskeletal architecture towards a fibrotic phenotype. These physical changes matched the concentration-dependent (0.1–1 mg/mL) cytotoxic effect of Ketamine and reflect a loss in expression of the key adherens junction proteins epithelial (E)- and neural (N)-cadherin and β-catenin. Down-regulation of protein expression does not involve the pro-fibrotic cytokine TGFβ, nor is it regulated by the usual increase in expression of Slug or Snail, the transcriptional regulators for E-cadherin. However, the loss in E-cadherin can be partially rescued pharmacologically by blocking p38 MAPK using SB203580. These data provide compelling evidence that Ketamine alters epithelial cell-to-cell adhesion and cell-coupling in the proximal kidney via a non-classical pro-fibrotic mechanism and the data provides the first indication that this illicit substance can have major implications on renal function. Understanding Ketamine-induced renal pathology may identify targets for future therapeutic intervention

Mining time-series data using discriminative subsequences

Time-series data is abundant, and must be analysed to extract usable knowledge. Local-shape-based methods offer improved performance for many problems, and a comprehensible method of understanding both data and models. For time-series classification, we transform the data into a local-shape space using a shapelet transform. A shapelet is a time-series subsequence that is discriminative of the class of the original series. We use a heterogeneous ensemble classifier on the transformed data. The accuracy of our method is significantly better than the time-series classification benchmark (1-nearest-neighbour with dynamic time-warping distance), and significantly better than the previous best shapelet-based classifiers. We use two methods to increase interpretability: First, we cluster the shapelets using a novel, parameterless clustering method based on Minimum Description Length, reducing dimensionality and removing duplicate shapelets. Second, we transform the shapelet data into binary data reflecting the presence or absence of particular shapelets, a representation that is straightforward to interpret and understand. We supplement the ensemble classifier with partial classifocation. We generate rule sets on the binary-shapelet data, improving performance on certain classes, and revealing the relationship between the shapelets and the class label. To aid interpretability, we use a novel algorithm, BruteSuppression, that can substantially reduce the size of a rule set without negatively affecting performance, leading to a more compact, comprehensible model. Finally, we propose three novel algorithms for unsupervised mining of approximately repeated patterns in time-series data, testing their performance in terms of speed and accuracy on synthetic data, and on a real-world electricity-consumption device-disambiguation problem. We show that individual devices can be found automatically and in an unsupervised manner using a local-shape-based approach

Differential proteolysis of insulin-like growth factor binding protein-1 (IGFBP-1) in pregnancy

The insulin-like growth factors and their binding proteins are important for placental and foetal growth. In this study, we have investigated the presence of proteolytic activity directed against insulin-like growth factor binding protein-1 (IGFBP-1) in pregnancy. In addition, the effect of protease activity on IGFBP-1 immunoreactivity and IGF binding was characterised. 125I-IGFBP-1 was incubated with maternal and foetal serum, amniotic fluid and placental extracts. Breakdown of 125I-IGFBP-1 was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and autoradiography. The size distribution of endogenous IGFBP-1 was determined by Western immunoblotting. Protease inhibitor studies characterised the proteolytic activity, and Western ligand blotting with 125I-IGF-I was used to determine IGF binding capacity of proteolysed IGFBP-1. Amniotic fluid samples collected after labour onset contained proteolytic activity that generated 12- and 19-kDa IGFBP-1 fragments that did not bind to 125I-IGF-I. This activity was not detected in amniotic fluid collected prior to labour onset or in other tissues. Activity was blocked by aprotinin, leupeptin, phenyl methyl sulphonyl fluoride, and Kunitz soybean trypsin inhibitor but not by ethylene diamine tetraacetic acid or pepstatin. Incubation of IGFBP-1 with trypsin generated fragments of a similar size to the amniotic fluid protease. In conclusion, we have demonstrated the presence in vivo of a trypsin-like proteolytic activity that alters the IGF-binding function of IGFBP-1 in pregnancy

Effect of glycosaminoglycans on growth factor-stimulated trophoblast invasion

Objectives: To determine the effect of glycosaminoglycans and a series of growth factors on the viability and invasion of the extravillous trophoblast cell line SGHPL4. Methods: Cells were cultured in Hams F10 media supplemented with fetal bovine serum and L-glutamine. For viability studies cells were seeded into 96-well culture plates (104 cells/well), maintained in serum free medium for 24h and then incubated with glycosaminoglycans (heparin, heparin sulphate and hyaluronic acid; each 100ng/ml) ± growth factors (VEGF, FGFand HB-EGF). Cell viability was measured in cells using the MTS assay. Cellular invasion was assessed using the FluoroBlok invasion assay. Cells were serum-starved for 24 h, incubated with the fluorescent dye DiIC12(3) (10mg/ml) for 1 hour prior to seeding onto an artificial extracellular matrix coated 8 mm FluoroBlok porous membrane inserts (2.5 x 105 cells per insert). Growth factors ± GAGs were added to the cell suspension and the inserts were lowered into a 96-well plate containing 10% fetal calf serum. Plates were incubated at 37°C for 24h. Invasion was determined by measurement of fluorescence of invaded cells using a fluorescent plate reader (Ex549/Em565 nm). Results: Cell numbers were significantly increased following incubation with VEGF, FGF and HB-EGF. Cell number was also increased after incubation with each of the glycosaminoglycans tested. The largest increase was observed following incubation with heparin sulphate. Cell numbers were further increased when the GFs were incubated with HS and heparin, but not with hyaluronic acid. Invasion was increased following incubation with VEGF, HBEGF and HGF. Heparan sulphate and heparin increased invasiveness in a dose-dependent manner. In contrast, hyaluronic acid had no significant effect. Conclusion: This study demonstrates a role for glycosaminoglycans in key features of trophoblast function

Establishment and characterization of single and triple‐agent resistant osteosarcoma cell lines

Two human osteosarcoma cell lines (MG-63 and HOS-143B) are developed into drug-resistant models using a short-term drug exposure and recovery in drug-free media. Cisplatin, doxorubicin, and methotrexate are used as single agents and in triple combination. The highest level of resistance to cisplatin is observed in MG-63/CISR8, doxorubicin in HOS-143B/DOXR8, and methotrexate in HOS-143B/MTXR8. The MG-63/TRIR8 and HOS-143B/TRIR8 tripleresistance models show lower levels of resistance to combination treatment and are not resistant to the drugs individually. Apoptosis assays suggest that the resistance in MG-63/TRIR8 isfrom cisplatin and methotrexate and not doxorubicin. In contrast, the resistance in HOS-143B/TRIR8 is from doxorubicin and methotrexate instead of cisplatin. Upregulation of P-glycoprotein is seen in all resistant models except those developed with single-agent methotrexate. However, P-glycoprotein is not causing resistance in all cell lines as the inhibitor elacridar only reverses the resistance of doxorubicin on MG-63/ DOXR8 and HOS-143B/TRIR8. The migration of the MG-63 resistant models is significantly increased, their invasion rate tends to increase, and RT-PCR shows a switch from epithelial to mesenchymal gene signaling. In contrast, a significant decrease in migration is seen in HOS-143B resistant models with their invasion rate tending to decrease and a switch from mesenchymal to epithelial gene signaling

1/n expansions for two-electron Coulomb matrix elements

The study of 1/n expansions for various atomic matrix elements, where n is the principal quantum number, plays an important role in the theoretical foundations of the quantum defect method. The paper develops an expansion in powers of 1/n 2 for hydrogenic boundstate wavefunctions which can be used to calculate 1/n expansions of matrix elements. The 1/n expansions of the two-electron direct and exchange Coulomb integrals are evaluated as an example. © 1993 IOP Publishing Ltd

Whole body cryotherapy, cold water immersion, or a placebo following resistance exercise: a case of mind over matter?

PURPOSE: The use of cryotherapy as a recovery intervention is prevalent amongst athletes. Performance of high volume, heavy load resistance exercise is known to result in disturbances of muscle function, perceptual responses and blood borne parameters. Therefore, this study investigated the influence of cold water immersion (CWI), whole body cryotherapy (WBC) or a placebo (PL) intervention on markers of recovery following an acute resistance training session. METHODS: 24 resistance trained males were matched into a CWI (10 min at 10 °C), WBC (3- and 4 min at - 85 °C) or PL group before completing a lower body resistance training session. Perceptions of soreness and training stress, markers of muscle function, inflammation and efflux of intracellular proteins were assessed before, and up to 72 h post exercise. RESULTS: The training session resulted in increased soreness, disturbances of muscle function, and increased inflammation and efflux of intracellular proteins. Although WBC attenuated soreness at 24 h, and positively influenced peak force at 48 h compared to CWI and PL, many of the remaining outcomes were trivial, unclear or favoured the PL condition. With the exception of CRP at 24 h, neither cryotherapy intervention attenuated the inflammatory response compared to PL. CONCLUSION: There was some evidence to suggest that WBC is more effective than CWI at attenuating select perceptual and functional responses following resistance training. However, neither cryotherapy intervention was more effective than the placebo treatment at accelerating recovery. The implications of these findings should be carefully considered by individuals employing cryotherapy as a recovery strategy following heavy load resistance training