bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional Analysis of the Amino- and Carboxyl-Termini of Streptokinase
Authors: Fay William P. , Bokka Lakshmi V. 
Published: May 2019

SummaryStreptokinase (SK) is a 414 amino acid bacterial protein that activates human plasminogen. Streptokinase fragments derived from the central portion of the protein bind plasminogen, but are inactive, indicating that the amino- and/or carboxyl-termini are required for normal plasminogen activator activity. To better define the function of the N- and C-termini of SK we generated and characterized 21 N-terminal and 20 C-terminal deletion mutants. All mutants lacking ≥18 N-terminal or ≥51 C-terminal amino acids exhibited markedly reduced plasminogen activator activity, while mutants lacking ≤12 N-terminal or ≤40 C-terminal residues were fully active. The decrease in SK activity with N-terminal deletion appeared to result not from loss of plasminogen binding capacity, but rather from increased susceptibility of deletion mutants to degradation by plasmin. Point mutations at positions 13 (SK V13D) or 20 (SK V20D) produced functional abnormalities similar to those observed in N-terminal deletion mutants, with SK V13D exhibiting delayed amidolytic activity and SK V20D exhibiting only 1% plasminogen activator activity and marked sensitivity to degradation by plasmin. C-terminal deletion mutants lacking ≥51 amino acids also bound plasminogen, but did not induce significant amidolytic activity in plasminogen or activator activity in plasmin. Prevention of cleavage at position 59 of SK had no effect on plasminogen activator activity, suggesting that the rapid hydrolysis of this bond that occurs after SK-plasminogen complex formation is not required for normal function of the N-terminus. These results suggest that residues within or near positions 13-20 of SK are important determinants of its capacity to generate amidolytic activity and are a critical determinant of the stability of SK, while residues within or near position 364-374 are required for generating amidolytic activity and for conferring plasminogen activator activity to plasmin(ogen). These results also suggest that SK fragments significantly smaller than SK 13-374 are unlikely to be effective thrombolytic agents.
Preparation of a Novel Streptokinase Mutant with Improved Stability
Authors: Shi Guey-Yueh , Chang Bi-Ing , Su Su-Wen , Young Kung-Chia , Wu Dung-Ho , Chang Li-Ching , Tsai Yau-Sheng , Wu Hua-Lin 
Published: May 2019

SummaryThe novel mutant streptokinase, SK-K59E, can activate human plasminogen as efficiently as the purified commercially available streptokinase. Several peptide bonds including Lys59-Ser60 in native streptokinase were hydrolyzed in reaction with plasmin and peptides of small molecular masses were generated. The plasminogen activator activity of native streptokinase in reaction with human plasmin declined to 25% of the original activity in a 120-min incubation. On the other hand, the NH2-terminal peptide of SK-K59E remained intact in reaction with plasmin and the activator activity of streptokinase decreased to 75% of the original activity in 120 min. The major degraded peptide fragments of native streptokinase in reaction with plasmin had molecular masses of 36 and 30 kDa. However, two major peptide fragments of 42 and 34 kDa were observed in the reaction of SK-K59E with human plasmin. The 42 kDa peptide fragment, which contained NH2-terminal of streptokinase, could activate human plasminogen as efficiently as the native streptokinase. SK-K59E can induce greater degree of caseinolysis and fibrinolysis than the native streptokinase. In conclusion, the results demonstrate that the prevention of cleavage at Lys59 of streptokinase prolongs the half-life of streptokinase in complex with plasmin and that the NH2-terminal of streptokinase (Ile1-Lys59) plays an important role in maintaining its stability.
Maturation of Megakaryoblastic Cells Is Accompanied by Upregulation of GSα-L Subtype and Increased cAMP Accumulation
Authors: Vuurst Hans van der , Hendriks Maaike , Lapetina Eduardo G. , Willigen Gijsbert van , Akkerman Jan-Willem N. 
Published: May 2019

SummaryIn platelets and megakaryoblasts Gs, the trimeric G-protein that stimulates adenylyl cyclase, is present in a short, 45 kDa, and a long, 52 kDa isoform termed Gsα-S and Gsα-L, respectively. To assess the relative contribution of these isoforms in the cellular synthesis of cAMP, the ratio Gsα-S/Gsα-L was changed in the megakaryoblastic cell line DAMI by inducing cell maturation with recombinant human thrombopoietin (TPO) or the phorbol ester PMA. Flow cytometric analysis confirmed that this treatment induced a moderate (TPO) and extensive (PMA) increase in nuclear ploidy and expression of the glycoproteins-IIIa and -Ib. Northern blot analysis revealed down-regulation of total GS-mRNA after treatment of DAMI-cells with TPO and PMA. Western blot analysis showed significant (P <0.05) upregulation of Gs-L with respective amounts of 27 ± 4% of total G s in untreated cells, 35 ± 1% in TPO- and 41 ± 3% in PMA-treated DAMI cells (n = 3-4). DAMI cells contained 6 ± 1 pmol cAMP/106 cells, which was not changed by treatment with TPO or PMA. In untreated cells this level increased to 70 ± 9 pmol cAMP/106 cells after 10 min stimulation with 1 μmol/l of the stable prostacyclin analog iloprost. The same stimulation with iloprost resulted in 165 ± 32 pmol cAMP/106 in TPO-treated cells and in 588 ± 100 pmol cAMP/106 in cells treated with PMA. Thus, a shift from Gsα-S to Gsα-L during megakaryoblast maturation strongly potentiates the production of cAMP. A similar shift may occur during normal megakaryocyte maturation and may explain the extreme sensitivity to prostacyclin of platelets, which contain Gsα-S and Gsα-L in approximately equal amounts.
Human Platelet CD36 (GPIIIb, GPIV) Binds to Cholesteryl-Hemisuccinate and Can Be Purified by a Simple Two-Step Method Making Use of this Property
Authors: Kronenberg A. , Grahl H. , Kehrel B. 
Published: May 2019

SummaryCD36, also known as GPIIIb or GPIV, is a main protein of the platelet membrane. Accumulating evidence implicates CD36 in the pathogenesis of atherosclerosis. CD36 binds to cholesteryl-hemisuccinate coupled to agarose. This binding may be of physiologic relevance since CD36 has a long extracellular hydrophobic region at its N-terminus and belongs to a family of proteins involved in lipid metabolism. This newly discovered binding property of CD36 was used to develop a rapid, efficient and gentle two-step purification method for CD36 from human platelets.
CD40 Engagement on Endothelial Cells Promotes Tissue Factor-dependent Procoagulant Activity
Authors: Zhou Ling , Stordeur Patrick , Lavareille Aurore de , Thielemans Kris , Capel Paul , Goldman Michel , Pradier Olivier 
Published: May 2019

SummaryThe CD40 molecule expressed on endothelial cells has been shown to transduce activation signals resulting in upregulation of adhesion molecules. Herein, we studied the impact of CD40 engagement on the induction of tissue factor (TF)-dependent procoagulant activity (PCA) at the surface of human umbilical vein endothelial cells (HUVECs). First, we found that co-incubation of HUVECs with 3T6 fibroblasts transfected with the CD40L gene (3T6-CD40L) resulted in a clear induction of PCA which was not observed with control untransfected fibroblasts. The specificity of this finding was established by inhibition experiments using monoclonal antibodies (mAbs) blocking CD40 or CD40L. PCA induced by CD40 ligation was TF-related as it was not observed in factor VII-deficient plasma and was associated with the accumulation of TF mRNA. To investigate the role of CD40/CD40L interactions in the induction of endothelial cell PCA by lymphocytes, interferon (IFN)-γ-stimulated EC were incubated with T cells in the absence or presence of anti-CD40 or anti-CD40L mAb. The 60-70% inhibition of PCA induced by these mAbs but not their isotype-matched control indicated that the CD40 pathway is involved in the induction of PCA resulting from interactions between activated HUVECs and T cells. We conclude that activation signals elicited by CD40 engagement on endothelial cells result in the induction of TF-dependent PCA. The CD40/CD40L pathway might therefore be involved in the development of prothrombic states during diseases associated with endothelial cell and T cell activation.
The Effects of Sex Steroids on Plasma Levels of Marker Proteins of Endothelial Cell Functioning
Authors: van Kesteren P. J. M. , Kooistra T. , Lansink M. , van Kamp G. J. , Asscheman H. , Gooren L. J. G. , Emeis J. J. , Vischer U. M. , Stehouwer C. D. A. 
Published: May 2019

SummaryWe studied thirteen male-to-female (M→F) and ten female-to-male (F→M) transsexuals who, for four months, received cross-sex treatment with, respectively, ethinylestradiol and cyproterone acetate, and with testosterone esters. We assessed the effects of treatment on plasma levels of tissue-type plasminogen activator (tPA), von Willebrand factor (vWF), vWF-propeptide (vWF:AgII) and bigendothelin-1 (big-ET-1), four proteins that are markers of endothelial cell functioning. We also measured urokinase-type PA (uPA) and plasminogen activator inhibitor-type 1 (PAI-1), which may not be endothelium-derived but share major clearance pathways with tPA.In M→F transsexuals, mean plasma levels of tPA (minus 4.4 ng/ml), big-ET-1 (minus 0.8 pg/ml), uPA (minus 0.5 ng/ml) and PAI-1 (minus 26 ng/ml) decreased (all Ps ≤0.02). The level of vWF increased (plus 24%; P = 0.005), while vWF:AgII did not change (P = 0.49).In F→M transsexuals, levels of big-ET-1 increased (plus 0.4 pg/ml; P = 0.02), while tPA, uPA and PAI-1 did not change (all Ps >0.25). In this group vWF decreased (minus 14%; P = 0.06), but vWF:AgII did not change (P = 0.38).Estrogens and androgens have clear effects on plasma levels of endothelial marker proteins. The mechanisms behind these effects are complex and appear to involve both altered secretion (big-ET-1) and processing and/or clearance (vWF and possibly tPA). Therefore, effects of hormones on the levels of endothelial marker proteins do not necessarily reflect changes in endothelial cell functioning, at least with regard to changes in vWF level associated with the oral administration of high doses of ethinylestradiol and cyproterone acetate to healthy men and the parenteral administration of testosterone to healthy women.
Coagulation Signalling and Metabolic Disorders: Lessons Learned from Animal Models
Authors: Madhusudhan Thati , Ruf Wolfram 
Published: May 2019

AbstractNutrient excess in obesity drives metabolic reprogramming in multiple tissues involving extensive interorgan and intercellular crosstalk. Experimental and clinical studies show that prolonged nutrient excess often compromises metabolic adaptation propagating proobesogenic and proinflammatory responses. Chronic inflammation further promotes insulin resistance and associated comorbidities. Obesity and type 2 diabetes are characterized by a hypercoagulable state and clinical studies show a strong correlation of markers of coagulation activation in metabolic disorders. Coagulation protease-dependent signalling via protease-activated receptors is intimately associated with inflammation. The experimental evidence supports roles of tissue factor and G protein coupled protease-activated receptor-2 signalling in the regulation of insulin resistance and metabolic inflammation in diet-induced obesity. Likewise, increases in plasminogen activator inhibitor-1 levels and fibrin-driven inflammation promote insulin resistance in obesity. Additionally, impaired thrombomodulin-dependent protein C activation is mechanistically linked to diabetic kidney disease. Given the increased usage of direct oral anticoagulants, understanding the role of specific coagulation proteases in regulation of metabolic inflammation is highly relevant and might provide insights into the design of novel treatment regimens for patients suffering from thromboinflammatory and cardiometabolic disorders.
Splinting after Distal Radius Fracture Fixation: A Prospective Cohort Analysis of Postoperative Plaster Splint versus Soft Dressing
Authors: Poiset Spencer , Abboudi Jack , Gallant Gregory , Jones Christopher , Kirkpatrick William , Kwok Moody , Liss Frederic , Rivlin Michael , Takei T. Robert , Wang Mark , Ilyas Asif M. 
Published: May 2019

Abstract
Background The effect of postoperative dressing and splinting after distal radius fracture (DRF) open reduction internal fixation (ORIF) is not well understood. A prospective cohort analysis was performed to assess differences in functional and radiographic outcomes with the use of plaster splinting or soft dressing following DRF ORIF.
Methods All patients undergoing DRF ORIF with locking volar plates were consecutively enrolled. Preoperative demographic and postoperative radiographic and functional outcome data were collected at 2 weeks and 3 months postoperatively. Functional data included range of motion (ROM), pain on visual analog scale (VAS), Patient-Rated Wrist Evaluation (PRWE), and quick Disabilities of the Arm, Shoulder and Hand (DASH) scores. Radiographic data included loss of fracture reduction.
Results A total of 139 patients were enrolled (79 plaster splinting, 60 soft dressing). By the first postoperative visit (POV), there was one case of loss of reduction with plaster splinting and one case with soft dressing with no hardware failure or revision surgery in either group, and no difference in DASH, PRWE, or VAS pain scores. By the final POV, the soft dressing group showed greater ROM in extension by 9.6, flexion by 10.9, and supination by 4.8 degrees over plaster splinting. Additionally, the soft dressing group demonstrated statistically significant improvement in PRWE and DASH scores, as well as VAS pain scores as compared with plaster splinting.
Conclusions Applying only soft dressing following DRF ORIF demonstrated improvements in ROM, VAS, and functional outcomes by final follow-up, with no significant differences in radiographic outcomes. No benefit of applying a plaster splint was identified.
Long-Term Outcomes of All-Arthroscopic Pre-Tied Suture Device Triangular Fibrocartilage Complex Repair
Authors: Sarkissian Eric J. , Burn Matthew B. , Yao Jeffrey 
Published: May 2019

Background A pre-tied suture device has been utilized for all-arthroscopic peripheral triangular fibrocartilage complex (TFCC) repairs with promising early clinical results.
Purpose The purpose of this study was to evaluate long-term functional outcomes of these repairs.
Patients and Methods A retrospective review of patients undergoing arthroscopic TFCC repair was performed. Inclusion criteria were the Palmer type 1B TFCC tears diagnosed on arthroscopy and repaired using the all-arthroscopic pre-tied suture device. Patients with any evidence of concomitant wrist injury at the time of surgery, history of prior wrist surgery, or nonrepairable and nonperipheral TFCC tears were excluded. Postoperative complications, range of motion, grip strength, and outcome assessments were recorded for each patient.
Results Eleven patients (mean age, 36 years; range, 20–64 years) satisfied our inclusion criteria and comprised the study cohort. The mean follow-up period was 7.0 years (range, 4.3–10.9 years). Mean range of motion of the wrist revealed flexion of 76 ± 11 degrees and extension of 73 ± 12 degrees. Mean grip strength was 98 ± 15% of the nonsurgical extremity. QuickDASH, modified Mayo, and patient-rated wrist evaluation (PRWE) average scores were 9 ± 8, 80 ± 6, and 12 ± 12, respectively. No surgical complications were observed and no patient required any further surgical intervention.
Conclusions Our cohort of patients following all-arthroscopic pre-tied suture device repair of isolated Palmer type 1B TFCC tears demonstrated excellent clinical function at a mean follow-up of 7 years. These findings indicate that the pre-tied suture device is a reliable, safe, effective, and most importantly, durable treatment option for repair of peripheral TFCC tears.
Level of Evidence This is a Level IV, therapeutic study.
Experimentalism and cognition
Authors: Farsi Roghayeh 
Published: May 2019

AbstractThis study approaches experimental literary texts from a cognitive perspective. It investigates if a constructivist modeling of cognition can be applied to such texts, and contends that there is a two-way relation between memory and literary experimentations. It suggests a fresh look at literary experimentalism – from the perspective of the cognitive processes involved in challenging (text, language, and world) schemata to varying degrees. There exists a vast body of knowledge on experimental texts, but the study of cognitive processing of such texts has until now been a less studied area of cognitive research.This study defines two main types of experimental texts based on their closeness to or divergence from the schematic parameters of world, text, and language: proximal and distal. The study shows how distal experimentations are conventionalized over the course of time and call for re-innovation.
Weaknesses of wickedness: a critical perspective on wickedness theory
Authors: Noordegraaf Mirko , Douglas Scott , Geuijen Karin , Van Der Steen Martijn 
Published: May 2019

Abstract
It is attractive to frame societal challenges such as climate change, terrorism and migration as ‘wicked issues’. Wickedness theory in its various guises has become popular as it connects the uncertain and political nature of issues with hopeful strategies for strengthening networks, trust and learning. In this paper we take a critical approach towards wickedness, advancing three criticisms: (1) the daily experiences of people and their practices are missing from the grand narratives about wickedness, (2) the potential of collaborations and learning to address these problems is romanticized, (3) the implications for managerial and professional perspectives are unclear. We argue that the wickedness literature can be strengthened by further emphasizing situated relations, routines and rituals, adopting the perspective of situated wickedness. This would link insight into grand wickedness to insights into daily ambiguity. We illustrate this argument with two specific cases, (counter)terrorism and forced migration/refugees.
Use of the anaerobic speed reserve to normalize the prescription of high‐intensity interval exercise intensity
Authors: Julio Ursula F. , Panissa Valéria L. G. , Paludo Ana C. , Alves Elaine D. , Campos Fábio A. D. , Franchini Emerson 
Published: May 2019

AbstractThe aim of this study was to compare the perceptual and physiological responses and time‐to‐exhaustion in high intensity interval exercise (HIIE) protocols that are prescribed based on the relative anaerobic speed reserve (ASR) or maximal aerobic speed (MAS) in athletes with different ASR values, as well as the coefficient of variation (CV) of the abovementioned variables. Eleven long‐distance runners and ten rugby players were submitted to five experimental sessions on different days; the first and second session were intended for the determination of the anthropometry, MAS and maximal sprint (MSS). In the subsequent sessions, three HIIE15:15s protocols were performed until exhaustion (110%MAS, Δ25%ASR, and Δ50%ASR) in random order. The anthropometric characteristics and variables obtained from the MAS and MSS tests in the different groups were compared by Student's unpaired t‐test. The analysis of mixed models for repeated measures (groups and protocols) was used to compare the speed, delta blood lactate, rating of perceived exertion, and time‐to‐exhaustion. Rugby players presented higher ASR (13.6 ± 0.9 km h−1) compared to long‐distance runners (12.6 ± 0.9 km h−1) (P = .049). For the HIIE15:15s protocols, there were no protocol and group interaction effects. However, lower CV values were observed for time‐to‐exhaustion (a mean reduction of 52%) and delta blood lactate (a mean reduction of 48%) in Δ25%ASR and Δ50%ASR when compared to 110%MAS. Furthermore, the rating of perceived exertion CV was similar in all HIIE15:15s protocols. The prescription of intensity of HIIE based on the ASR was able to reduce the inter‐subject variability of lactate and time‐to‐exhaustion in rugby players and long‐distance runners.
Effectiveness of sensor monitoring in a rehabilitation programme for older patients after hip fracture: a three-arm stepped wedge randomised trial
Authors: Pol Margriet C , ter Riet Gerben , van Hartingsveldt Margo , Kröse Ben , Buurman Bianca M 
Published: May 2019

AbstractObjectivesto test the effects of an intervention involving sensor monitoring-informed occupational therapy on top of a cognitive behavioural treatment (CBT)-based coaching therapy on daily functioning in older patients after hip fracture.Design, setting and patientsthree-armed randomised stepped wedge trial in six skilled nursing facilities, with assessments at baseline (during admission) and after 1, 4 and 6 months (at home). Eligible participants were hip fracture patients ≥ 65 years old.Interventionspatients received care as usual, CBT-based occupational therapy or CBT-based occupational therapy with sensor monitoring. Interventions comprised a weekly session during institutionalisation, followed by four home visits and four telephone consultations over three months.Main outcomes and measuresthe primary outcome was patient-reported daily functioning at 6 months, assessed with the Canadian Occupational Performance Measure.Resultsa total of 240 patients (mean[SD] age, 83.8[6.9] years were enrolled. At baseline, the mean Canadian Occupational Performance Measure scores (range 1–10) were 2.92 (SE 0.20) and 3.09 (SE 0.21) for the care as usual and CBT-based occupational therapy with sensor monitoring groups, respectively. At six months, these values were 6.42 (SE 0.47) and 7.59 (SE 0.50). The mean patient-reported daily functioning in the CBT-based occupational therapy with sensor monitoring group was larger than that in the care as usual group (difference 1.17 [95% CI (0.47-1.87) P = 0.001]. We found no significant differences in daily functioning between CBT-based occupational therapy and care as usual.Conclusions and relevanceamong older patients recovering from hip fracture, a rehabilitation programme of sensor monitoring-informed occupational therapy was more effective in improving patient-reported daily functioning at six months than to care as usual.Trial registrationDutch National Trial Register, NTR 5716.