Location, location, location : Fibrin, cells, and fibrinolytic factors in thrombi

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Location, location, location: Fibrin, cells, and fibrinolytic factors in thrombi

Thrombi are heterogenous in nature with composition and structure being dictated by the site of formation, initiating stimuli, shear stress, and cellular influences. Arterial thrombi are historically associated with high platelet content and more tightly packed fibrin, reflecting the shear stress in these vessels. In contrast, venous thrombi are generally erythrocyte and fibrin-rich with reduced platelet contribution. However, these conventional views on the composition of thrombi in divergent vascular beds have shifted in recent years, largely due to recent advances in thromboectomy and high-resolution imaging. Interestingly, the distribution of fibrinolytic proteins within thrombi is directly influenced by the cellular composition and vascular bed. This in turn influences the susceptibility of thrombi to proteolytic degradation. Our current knowledge of thrombus composition and its impact on resistance to thrombolytic therapy and success of thrombectomy is advancing, but nonetheless in its infancy. We require a deeper understanding of thrombus architecture and the downstream influence on fibrinolytic susceptibility. Ultimately, this will aid in a stratified and targeted approach to tailored antithrombotic strategies in patients with various thromboembolic diseases

Fibrinogenolysis and fibrinolysis in Vaccine-induced Immune Thrombocytopenia and Thrombosis (VITT)

Acknowledgements The authors would like to thank all the patients whose samples were used as part of this study, and all the NHS Scotland staff who collected patient samples and looked after these patients. We thank Aberdeen Royal Infirmary Haematology laboratory for conducting the anti-platelet factor 4 antibody testing and Dr Sue Pavord, Consultant Haematologist at Oxford Teaching Hospitals for help in gathering clinical data on the patients. Funding This research was supported by The University of Aberdeen Development Trust (RG16009). CSW and NJM are supported by the British Heart Foundation (PG/15/82/31721; PG/20/17/35050).Peer reviewedPublisher PD

Stability indicating method development and validation for simultaneous estimation of atorvastatin calcium and celecoxib in bulk and niosomal formulation by RP-HPLC

The present work describes development and validation of a specific, sensitive, precise and stability-indicating high-performance liquid chromatographic method of analysis of atorvastatin calcium and celecoxib, both as a bulk drug and in niosomal formulation. The analysis has been performed by using Cosmosil-C18 column (4.6 mm´250 mm, 5 m) at 25 °C using acetonitrile: ammonium acetate buffer pH 5.0: methanol (50:25:25 v/v/v) as mobile phase. The detection was carried out at 277nm with a flow rate of 1.0mL/min. The retention times of Atorvastatin calcium and Celecoxib were 6.195 and 3.989min, respectively. The method was validated according to ICH guidelines, for specificity, precision, linearity, accuracy and robustness. Atorvastatin calcium and Celecoxib were subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal degradation. The degradation was observed in oxidation and acid hydrolysis. The linearity for atorvastatin calcium and celecoxib were in the range of 100-500 µg/mL. The recovery study of atorvastatin and celecoxib were found to be in the range of 98.96 - 99.92% and 98.90-100%, respectively. The proposed method was validated and successfully applied to the estimation of Atorvastatin calcium and Celecoxib in combined in-house niosomal formulation.O presente trabalho descreve o desenvolvimento e a validação de método de análise por cromatografia de alta eficiência específico, sensível, preciso e indicador de estabilidade de atorvastatina cálcica e celecoxibe, ambos como fármaco e como formulação niosômica. A análise foi realizada utilizando coluna Cosmosil-C18 (4,6 mm´250 mm, 5 m) a 25 °C, e acetonitrila: tampão acetato de amônio pH 5,0: metanol (50:25:25 v/v/v) como fase móvel. A detecção foi realizada a 277 nm, com fluxo de 1,0 mL/min. Os tempos de retenção de atorvastatina cálcica e de celecoxibe foram 6,195 e 3,989 min, respectivamente. O método foi validado de acordo com as regras da ICH para especificidade, precisão, exatidão e robustez. A atorvastatina cálcica e o celecoxibe foram submetidos a condições de estresse por hidrólise, oxidação, fotólise e degradação térmica. A degradação foi observada por oxidação e hidrólise ácida. Observou-se a linearidade da atorvastatina cálcica e do celecoxibe na faixa de 100-500 µg/mL. A recuperação da atorvastatina e do celecoxibe foi observada na faixa de 98,96-99,92% e 98,90-100%, respectivamente. O método proposto foi validado e aplicado com sucesso para a determinação de atorvastatina cálcica e celecoxibe em formulação niosômica caseira combinada

Bioavailability in soils

The consumption of locally-produced vegetables by humans may be an important exposure pathway for soil contaminants in many urban settings and for agricultural land use. Hence, prediction of metal and metalloid uptake by vegetables from contaminated soils is an important part of the Human Health Risk Assessment procedure. The behaviour of metals (cadmium, chromium, cobalt, copper, mercury, molybdenum, nickel, lead and zinc) and metalloids (arsenic, boron and selenium) in contaminated soils depends to a large extent on the intrinsic charge, valence and speciation of the contaminant ion, and soil properties such as pH, redox status and contents of clay and/or organic matter. However, chemistry and behaviour of the contaminant in soil alone cannot predict soil-to-plant transfer. Root uptake, root selectivity, ion interactions, rhizosphere processes, leaf uptake from the atmosphere, and plant partitioning are important processes that ultimately govern the accumulation ofmetals and metalloids in edible vegetable tissues. Mechanistic models to accurately describe all these processes have not yet been developed, let alone validated under field conditions. Hence, to estimate risks by vegetable consumption, empirical models have been used to correlate concentrations of metals and metalloids in contaminated soils, soil physico-chemical characteristics, and concentrations of elements in vegetable tissues. These models should only be used within the bounds of their calibration, and often need to be re-calibrated or validated using local soil and environmental conditions on a regional or site-specific basis.Mike J. McLaughlin, Erik Smolders, Fien Degryse, and Rene Rietr

Exploration of the nicotinamide-binding site of the tankyrases, identifying 3-arylisoquinolin-1-ones as potent and selective inhibitors <em>in vitro</em>

Tankyrases-1 and -2 (TNKS-1 and TNKS-2) have three cellular roles which make them important targets in cancer. Using NAD(+) as a substrate, they poly(ADP-ribosyl)ate TRF1 (regulating lengths of telomeres), NuMA (facilitating mitosis) and axin (in wnt/β-catenin signalling). Using molecular modelling and the structure of the weak inhibitor 5-aminoiso quinolin-1-one, 3-aryl-5-substituted-isoquinolin-1-ones were designed as inhibitors to explore the structure-activity relationships (SARs) for binding and to define the shape of a hydrophobic cavity in the active site. 5-Amino-3-arylisoquinolinones were synthesised by Suzuki-Miyaura coupling of arylboronic acids to 3-bromo-1-methoxy-5-nitro-isoquinoline, reduction and O-demethylation. 3-Aryl-5-methylisoquinolin-1-ones, 3-aryl-5-fluoroisoquinolin-1-ones and 3-aryl-5-methoxyisoquinolin-1-ones were accessed by deprotonation of 3-substituted-N,N,2-trimethylbenzamides and quench with an appropriate benzonitrile. SAR around the isoquinolinone core showed that aryl was required at the 3-position, optimally with a para-substituent. Small meta-substituents were tolerated but groups in the ortho-positions reduced or abolished activity. This was not due to lack of coplanarity of the rings, as shown by the potency of 4,5-dimethyl-3-phenylisoquinolin-1-one. Methyl and methoxy were optimal at the 5-position. SAR was rationalised by modelling and by crystal structures of examples with TNKS-2. The 3-aryl unit was located in a large hydrophobic cavity and the para-substituents projected into a tunnel leading to the exterior. Potency against TNKS-1 paralleled potency against TNKS-2. Most inhibitors were highly selective for TNKSs over PARP-1 and PARP-2. A range of highly potent and selective inhibitors is now available for cellular studies.</p

Residual effects of natural Zn chelates on navy bean response, Zn leaching and soil status

greenhouse experiment was conducted on weakly acidic and calcareous soils to evaluate the aging and residual effects of three natural organic Zn chelates [Zn-ethylenediaminedisuccinate (Zn-EDDS), Zn-polyhydroxyphenylcarboxylate and Zn-aminelignosulfonate] each administered in a single application to a first navy bean (Phaseolus vulgaris L.) crop at several different Zn application rates. In a second navy bean crop, we determined the following parameters: the extent of Zn leaching, the amount of available Zn remaining in soils, the amount of easily leachable Zn, the size of Zn fractions in soils, the pH and redox potential, the dry matter yield, and the soluble and total Zn concentrations in plants. The residual effect after 2 years of Zn fertilization mainly depended on the aging effect of Zn chelates and losses due to Zn leaching. The data relating to the evolution from the first to the second crop showed that the aging effect was noticeable in the calcareous soil. In the latter soil, the Zn-S,S-EDDS treatments showed greater decreases in the Zn uptake by plants than the other Zn treatments and the greatest Zn uptake by plants occurred when Zn was applied as Zn-aminelignosulfonate (10 mg Zn kg−1 rate, 6.85 mg Zn per lysimeter; 5 mg Zn kg−1 rate, 3.36 mg Zn per lysimeter). In contrast, in the calcareous soil, the maximum amount of Zn uptake, for the three chelates was 0.82 mg Zn per lysimeter. Consequently, a further application of Zn would be needed to prevent Zn deficiencies in the plants of a subsequent crop. The behaviour of the pH and Eh parameters in the soils and leachates did not depend on the natural Zn sources applied. In this study, the easily leachable Zn estimated by BaCl2 extraction was not adequate to predict Zn leaching from the soils in subsequent crops

First measurement of the Michel parameter ξ′\xi^\prime in the τ−→μ−νˉμντ\tau^-\to\mu^-\bar{\nu}_\mu\nu_\tau decay at Belle

We report the first measurement of the Michel parameter $\xi^\prime$ in the $\tau^-\to\mu^-\bar{\nu}_\mu\nu_\tau$ decay with a new method proposed just recently. The measurement is based on the reconstruction of the $\tau^-\to\mu^-\bar{\nu}_\mu\nu_\tau$ events with subsequent muon decay-in-flight in the Belle central drift chamber. The analyzed data sample of $988\,\text{fb}^{-1}$ collected by the Belle detector corresponds to approximately $912\times10^6$ $\tau^+ \tau^-$ pairs. We measure $\xi^\prime=0.22\pm0.94(\text{stat})\pm0.42(\text{syst})$, which is in agreement with the Standard Model prediction of $\xi^\prime=1$. Statistical uncertainty dominates in this study, being a limiting factor, while systematic uncertainty is well under control. Our analysis proved the practicability of this promising method and its prospects for further precise measurement in future experiments.Comment: 6 pages, 4 figures, submitted to Phys. Rev. Let