bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Lipid Rafts in GPVI Agonist-Induced Platelet Signaling.
Authors: Quinter Patricia G. , Quinton Todd M. , Dangelmaier Carol A. , Kunapuli Satya P. , Daniel James L. 
Published: October 2019

Abstract
The collagen receptor glycoprotein VI (GPVI), plays an essential role in platelet activation and the regulation of hemostasis. Microdomains within the plasma membrane, called lipid rafts, have been implicated in GPVI signaling. The GPVI receptor has been shown to associate with the lipid rafts in both resting and activated platelets. It has been reported that there is a reduction in GPVI signaling in raft-disrupted platelets following activation with various GPVI agonists, especially at low to moderate agonist concentrations. Since platelet aggregation is potentiated by secreted adenosine 5′-diphosphate (ADP) at low concentrations of convulxin and at all concentrations of collagen and collagen-related peptide (CRP), we wanted to determine whether the decrease in GPVI signaling found in platelets with disrupted rafts was due to the loss of agonist potentiation by ADP. We compared platelet aggregation, protein phosphorylation, and calcium mobilization in platelets with intact and disrupted lipid rafts following activation with the GPVI agonists, collagen, convulxin and CRP. We show that lipid raft disruption inhibits aggregation induced by collagen and convulxin, but this inhibition is no longer apparent in the presence of ADP feedback inhibitors. Furthermore, raft-disrupted platelets had the same level of phosphorylation of proteins involved in GPVI signaling (i.e. Syk, LAT, and PLCγ2) and the same ability to mobilize calcium following activation with collagen or convulxin. Therefore, the effects of lipid raft disruption on aggregation can be attributed to the loss of ADP feedback. Interestingly, however, raft disruption directly inhibited aggregation and Syk phosphorylation induced by CRP in the presence and absence of ADP feedback. We propose that these differences are due to the fact that CRP is a relatively small, synthesized peptide of 37 amino acids, while collagen and convulxin are large ligands. These agonists are all able to bind the GPVI receptor, but they may not have the same ability to simultaneously cluster multiple receptors due to their size differential. The lipid rafts may be important for CRP stimulation, but not for collagen or convulxin, because they may have a higher density of the GPVI receptor than nonraft membrane regions, allowing CRP to cluster multiple receptors and activate the GPVI signaling cascade. When we disrupt the lipid rafts, we are reducing the effective concentration of GPVI available for activation by CRP but not by collagen or convulxin.
Validation of T2* Technique in Ex-Vivo Myocardial Tissue.
Authors: Kirk Paul , Firmin David N. , Sampson Barry , Porter John B. , Walker Malcolm J. , Pennell Dudley J. 
Published: October 2019

Abstract
Background
The T2* technique has been validated as a non-invasive measurement of myocardial iron load. However, calibration of the technique against myocardial tissue remains to be determined. Since such calibration is easiest to perform using post-mortem tissue, it is useful to know the effects of storage of the tissue in formalin, and in particular whether iron leaks from the heart during storage, and whether fixation alters the myocardial T2* significantly.
Method
A heart was donated after death of a patient with sideroblastic anemia who had been poorly compliant with deferoxamine treatment and died in heart failure. The pre-mortem myocardial T2* was 5.2 ms indicating severe myocardial iron loading. The majority of the heart was assigned to a calibration study of myocardial iron, but 2 samples from the heart were available for this study.
Sample 1 was an apical short-axis slice of the left ventricle. This was preserved in formalin for 3 days prior to measurement of the initial post-mortem myocardial T2* which was 5.2 ms (Siemens Sonata 1.5T MR scanner). The T2* of the slice was then measured weekly for 4 weeks; then 2 weekly for 8 weeks; and finally 4 weekly.
Sample 2 was a 7.35 g piece of apical myocardium which was stored in 280ml formaldehyde. A 1ml sample of formaldehyde solution was taken for iron analysis from sample 2 initially and at 2 days, and then weekly for 4 weeks; two-weekly for 6 weeks; and then 4-weekly for 12 weeks. The iron concentration in the formalin was measured using Inductive Coupled Plasma with Mass Spectometry (ICPMS).
Results
The results from sample 1 are shown as myocardial T2* against days elapsed. The T2* rose from a pre-mortem and post-mortem (equivalent values) of 5.2ms to a mean of 6ms over the first 20 days, and after this was stable (figure 1).
The results from sample 2, show an increase from 16.6 microgram to 74.9 microgram over the initial 3–4 weeks after which little significant additional accumulation occurred (figure 2). Estimation of the total iron content of the myocardial sample (prior to destructive iron measurement at the end of the study) suggested that the total iron leaked into the formalin was 0.44% of the iron in the myocardial sample.
Conclusion
There was a small increase in myocardial T2* over the first 20 days of formalin fixation, but little significant change after that. This was associated with a loss of approximately 75 micrograms of iron into the formalin fixation solution, which was estimated as only 0.44% of the iron in the myocardial sample. This suggests that formalin fixation has a small but measurable effect on myocardial T2* over time, and this is not explained by loss of iron into the fixation fluid.
Figure 1 Figure 1. Figure 2 Figure 2.
Is Primary Biliary Cirrhosis a Rare Cause of Eosinophilia? A Case Report.
Authors: Tsesmeli Niki E. , Savopoulos Christos G. , Kaiafa Georgia D. , Hatzitolios Apostolos I. , Vretou Eleni E. , Papadopoulos Athanasios , Koliouskas Dimitrios P. 
Published: October 2019

Abstract
INTRODUCTION: Evaluation of eosinophilia involves various medical sybspecialties and the list of diseases, syndromes and inflammatory processes associated with peripheral blood (>300 eosinophils/mm3) and/or tissue eosinophilia, is quite extensive. Little attention, however, has been given to its possible association with primary biliary cirrhosis (PBC).
AIMS & METHODS: To identify PBC as a rare cause of eosinophilia
RESULTS: A 59 female patient, complaining of fatigue, referred to us for abnormal liver function tests. She had no prior history of liver disease, alcohol abuse or medication use. The haematological parameters were normal except for eosinophilia: WBC: 7.5 x 109 /lit, eosinophils 1125/mm3 -15% (normal range: 3–5%) and the biochemical abnormalities included an alkaline phosphatase (ALP) of 314 U/L, γ-gt 146 U/L, total bilirubin 2.2 mg/dl (direct 1.6 mg/dl), AST 62 U/L, ALT 88 U/L and cholesterol 302 mg/dl. Eosinophilia was first detected during an initial evaluation for fatigue 20 months before. She had shown a normal serum biochemistry and negative stool examinations for intestinal parasites and ova at that time. Other causes of eosinophilia, such as skin, allergic, pulmonary, collagen, vascular or immunodeficiency disorders and malignancies, had been excluded by history, physical examination, imaging procedures (chest X-ray and abdominal CT scan) and laboratory investigation, including serum protein electrophoresis, quantitative immunoglobulins, serum immunoelectrophoresis, tumor markers and bone marrow aspirate. On admission, both abdominal and ultrasonography examination revealed a mild hepatomegaly. AMA were detected and liver biopsy was performed. After the serological and histological identification of PBC, ursodeoxycholic acid (15 mg/kg daily) was administered. Three months later, her haematological and biochemical profiles were improved (eosinophils 6%, ALP 234 U/L, γ-gt 80 U/L, AST 59 U/L, ALT 72 U/L)
CONCLUSIONS: Our case suggests that primary biliary cirrhosis should be contemplated in the differential diagnosis of eosinophilia, when all common causes are excluded. In particular, clinicians should evaluate AMA in cases of otherwise unexplained eosinophilia, even in the absence of symptoms or laboratory findings compatible to PBC.
Differential Expression of Granzymes A and K in Subsets of Human T-Cells and NK-Cells.
Authors: Zeberg Lennart , Olofsson Tor 
Published: October 2019

Abstract
Granzymes are highly specific proteases stored in secretory granule in T-cells and NK-cells. Five different granzymes are known in human (A, B, H, K and M). A and B are the best studied and both can induce apotosis when delivered to target cells. The role for granzyme H, K and M are principally unknown. We have studied the expression of granzymes in subsets of T-cells and NK-cells both in fresh circulating cells and cultured cells stimulated with IL-2, IL-15 and OKT3/antiCD28. Protein expression (biosynthesis/immunoprecipitation) and mRNA level (RT-PCR) was studied in parallel. This report focus on Granzyme A and K which are close relatives, both tryptases located 70 kb apart on chromosome 5.
Granzyme A have the highest expression in NK-cells were granzyme K are barely detected. Granzyme K on the other hand have a particular high expression in CD8+ T cells. Stimulation with IL-2 or IL-15 give a strong upregulation of both enzymes in NK-cells. Stimulation of CD8+ T cells gives mainly upregulation of granzyme A. Fresh CD4+ T cells show very weak synthesis of granzymes but treatment with IL-2 or IL-15 give a clear upregulation of both granzyme A and K. T-cells (both CD4+ and CD8+) are highly responsive to stimulation with OKT3/anti CD28 producing granzyme B but not A and K. This rather discordant pattern of expression for granzyme A and K talks in favour of different modes of regulation and most likely different functions.
A striking difference between granzyme A and K concern the propensity for granzyme A to be secreted into the culture medium after stimulation while this is not the case for granzyme K. The secreted form of granzyme A has a molecular weight slightly higher than the intracellular form indicating the presence of a proform. Secretion of granzymes to the extracellular medium is probably not only an in vitro phenomena since both granzyme A and B can be detected in serum from healthy individuals in picomolar concentrations. Moreover, significant elevation of granzyme levels in serum can bee seen in different diseases with activation of cellular immunity and in T- and NK-cells malignancies. We have previously reported that proforms of certain serine proteases including the five human granzymes have a downregulating activity on myeloid proliferation, a mechanism probably implicated in certain forms of neutropenia. We propose that secreted granzymes (especially A, B and H) constitute a part of T regulatory function working both locally at the site of inflammation and distantly via the circulation.
The Effects of Glycoprotein IIb/IIIa Antagonists and Chloride Channel Blockers on Platelet Cytoplasmic Free Calcium.
Authors: Yin Songmei , Li Yiqing , Xie Shuangfeng , Nie Danian , Li Haiming , Ma Liping , Wang Xiuju , Wu Yudan , Feng Jianghong 
Published: October 2019

Abstract
Objective
To explore the effects and interactions of GPIIb/IIIa antagonists and chloride channel blockers on the platelet cytoplasmic free calcium ([Ca2+]i ).
Methods
We washed and suspended fresh platelets with Hepes buffer containing 0.1% bovine serum albumin (BSA), then loaded platelets with 5μmol/L Fura-3/AM. Then RGDS, the GPIIb/IIIa antagonists, and the chloride channel blockers 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene(DIDS) or niflumic acid(NFA) were added to the platelet suspension. After 2 minutes incubating, we observed the effects and interactions of GPIIb/IIIa antagonists and chloride channel blockers on platelet [Ca2+]i by measuring the Fura-3 fluorescence intensity.
Results
1. Effects of GPIIb/IIIa antagonists and chloride channel blockers on platelet [Ca2+]i.
The fluorescence intensity of resting platelet [Ca2+]i were 369.6±62.2, 381.9±72.4, 392.8 ±69.9 after adding RGDS(250 μmol/L), DIDS(100 μmol/L) or NFA( 100 μmol/L) respectively. every agent had no significant effect on resting [Ca2+]i (p>0.05). After thrombin(0.03 U/ml) stimulating and adding RGDS(250 μmol/L), DIDS(100 μmol/L) or NFA( 100 μmol/L), the platelet [Ca2+]i were 883.9±107.0, 789.8±99.8, 564.1±79.4. Compare with the control(977.9±108.8), the three agents could inhibit the elevation of [Ca2+]i stimulated by thrombin (p<0.05). The inhibiting rates were (9.37±7.5)%, (18.7±10.4)% and (41.8±10.1)% respectively.
2. Combined effects of GPIIb/IIIa antagonists and chloride channel blockers
The fluorescence intensity of resting platelet [Ca2+]i was 383.9±67.9 after incubated with RGDS and DIDS. That had no significant effects. When platelets were stimulated by thrombin (0.03 U/ml), the combined inhibition rate was (24.4±10.8)%, RGDS and DIDS couldn’t weaken or enhance each other on thrombin-induced elevation of [Ca2+]i (p>0.05). Neither RGDS nor NFA had significant combined effects on resting [Ca2+]i(p>0.05). The combined inhibition rate was (46.0±7.3)%, they had no interactions too(p>0.05).
Conclusion
The GPIIb/IIIa antagonists RGDS and the chloride channel blockers DIDS or NFA have no effect on resting platelet [Ca2+]i. All of them can inhibit the elevation of platelet [Ca2+]i induced by thrombin. There are no interactions between GPIIb/IIIa antagonists RGDS and chloride channel blockers (DIDS or NFA) in resting platelet [Ca2+]i and elevation of platelet [Ca2+]i induced by thrombin, and their effects were independent.
Circulating Platelet-Leukocyte Aggregates as a Marker of Microvascular Lesions in Diabetic Patients.
Authors: Elalamy Ismail , Chakroun Tahar , Robert Francoise , Lecrubier Chantal , Elgrably Fabienne , Samama Meyer M. , Gerotziafas Grigoris T. , Hatmi Mohamed T. 
Published: October 2019

Abstract
Introduction: Evolution of diabetes is associated with multiple disorders including metabolic, cellular and blood disturbances leading to various vascular complications (micro-and macro-angiopathies). Increased circulating levels of platelet-leukocyte aggregates (PLA) have been described in several thrombotic diseases.
Material and Methods: In this study, we have evaluated the circulating PLA in diabetic patients and we have investigated whether they may be linked to the vascular complications currently occurring during diabetes evolution. Using flow cytometry assay, we have quantified PLA percentages in 65 diabetics (37 males/28 females, 57 ± 11 years old) including 20 patients with type I and 45 with type II diabetes, and 25 healthy subjects (15 males/10 females, 44 ± 9 years old). Labelling approach using specific monoclonal antibodies permitted us to identify platelet-polymorphonuclear aggregates (PPA) and platelet-monocyte aggregates (PMA).
Results: We have observed a significant increase of PPA and PMA levels in diabetics (22 ± 12% and 45 ± 18%, respectively) compared to control subjects (7 ± 4% and 19 ± 10%, respectively). However, both PPA and PMA values were similar in the two types of diabetes and they were not correlated to the disease duration. Circulating PPA and PMA percentages were significantly enhanced in diabetics with vascular lesions (n=37; PPA: 24 ± 13%; PMA: 50 ± 18%) than in diabetics without vascular lesions (n=28; PPA: 18 ± 8%; PMA: 38 ± 15%). Patients with PPA > 18% and /or PMA > 38% had a more important vascular thrombotic damage (OR: 6; 95%; IC: 1.6; 23). The increased PMA circulating percentage seems to be more specific for micro-retinopathy occurrence (OR: 19, 95% IC: 2.3; 154). The rare patients with elevated PMA percentage and without vascular lesions have a shorter duration of diabetes (7 ± 5 years) than patients presenting retinopathy lesions (16 ± 11 years).
Conclusion: Together our findings established a relationship between increased circulating PLA rate, particularly that of PMA, and the incidence of microvascular complications in diabetic patients. In addition, they reinforce the concept that pro-inflammatory cells are involved in diabetic retinopathy pathogenesis.
Urinary 11-Dehydrothromboxane B2 Levels in Healthy Individuals Following a Single Dose Response to Two Concentrations of Aspirin.
Authors: McGlasson David L. , Chen Michael W. , Knight Zita A. , Dobbs Michael 
Published: October 2019

Abstract
Introduction: The purpose of this protocol was to determine the effect of aspirin on urinary 11-dehydrothromboxane B2 (11-DHT) levels in normal healthy adults who had ingested a single dose of aspirin (ASA).
Methods: We enrolled 50 normal healthy volunteers 18–58 years of age who had not ingested ASA or other NSAIDs for the previous 14 days. A baseline urine specimen was obtained followed by ingestion of a single 81 mg enteric-coated ASA. Twenty-four hours later a post-ASA urine specimen was obtained. Two weeks later another baseline urine specimen was obtained followed by a single 325 mg ASA enteric-coated tablet. Twenty-four hours later another post-ASA urine specimen was obtained. Urines were stored at -70°C until testing. Platelet response to aspirin was assessed using the 11-DHT immunoassay (Cayman Biochemical, Ann Arbor, MI). Aspirin Works® controls (Creative Clinical Concepts, Denver, CO) were utilized to control for lot to lot and run to run variability. Testing was completed by Hemostasis Reference Laboratories (Hamilton, ON). In 2002, HOPE study investigators computed quartiles by comparing clinical outcomes to urinary 11-DHT levels from subjects who were taking ASA to prevent a second adverse cardiovascular event. A level of 298 pg 11-DHT/mg urinary creatinine was the limit for determining platelet response to ASA and for projecting cardiovascular risk While 298 pg/mg was the lower limit of the fourth quartile, this limit may not apply to the 11-DHT effect of a single dose of ASA. Our limit for ASA response was established by determining the 50% difference between baseline and outcome. This approach was applied to both ASA concentration arms.
Results: Twenty-two of 48 subjects in the 81 mg ASA group showed an ASA response (46%). Forty of the 46 subjects (85%) in the 325 mg dosage group responded. The 325 mg ASA dose showed a significantly greater platelet response, based on mean 11-DHT levels.
Discussion: Previous studies have shown that non-responders to ASA have a greater risk of cardio-vascular problems using the quartile distribution. The current study may be used to extend these findings by applying them to single-dose ASA in normal healthy subjects. Further studies may apply these data to primary cardiovascular risk.
A Meta-Analysis on the Use of Intravenous Gamma Globulin and Corticosteroids in Children with Acute Immune Thrombocytopenic Purpura.
Authors: Chua Mary N. , Manuela Tintin , Sarol Jesus N. , Ligsay Antonio D. 
Published: October 2019

Abstract
To determine the difference in proportion of patients who achieved a rise in platelet count and to determine the risk difference of adverse events in patients with acute immune thrombocytopenic purpura treated with corticosteroids or gamma globulin (IVIG). Medline, EMBASE, Pediatric Hematology and Oncology, Pediatrics and Journal of Pediatrics (all up to 2005) and personal collections of the authors. Systematic search of articles utilizing a search strategy was performed to identify relevant articles according to prior inclusion and exclusion criteria. Fixed effects and random effects models were used to determine summary effect estimates. Difference in proportion of patients who achieved the desired platelet count and risk difference of adverse events between treatment groups. Analysis of 9 randomized controlled trials showed that IVIG produced an additional 12–14% increase in proportion of patients with platelet count >20 x 109/L and 15% increase in proportion of patients with platelet count >50 x 109/L on days 2 to 4. No significant differences were seen between the treatment groups on days 14 and 28 except for platelet count >50 x 109/L. More patients in oral steroid treated group had adverse events compared to IVIG treated group but no differences were noted in the parenteral treated group. IVIG is more effective compared to corticosteroids in raising the platelet count during the early part of treatment. IVIG and parenteral steroids shows less adverse events as compared to oral steroid.
Characterisation of the Novel, High Purity, Double Virus Inactivated Von Willebrand Factor and Factor VIII Concentrate Wilate®.
Authors: Stadler Monika , Gruber Gerhard , Kannicht Christoph , Biesert Lothar , Radomski Kai-Uwe , Suhartono Haryadi , Pock Katharina , Neisser-Svae Andrea , Weinberger Josef , Roemisch Juergen , Svae Tor-Einar 
Published: October 2019

Abstract
Purpose: This study summarizes the characteristics and virus safety profile of Wilate®, a new human plasma-derived, high purity, double virus inactivated VWF/FVIII concentrate. The manufacturing process comprises two chromatographic steps, ensuring high purity and preserving the integrity and functionality of the VWF/FVIII complex. The optimised solvent/detergent treatment (S/D) and terminal dry-heat treatment (PermaHeat) of the lyophilised product provide two effective and robust virus inactivation steps for enveloped (EV) and one step for non-enveloped viruses (NEV).
Methods: Functional and structural properties of the VWF/FVIII concentrate were studied utilising state-of-the-art assays. The reduction factors for a panel of model viruses were determined in down-scale experiments valid for the respective manufacturing steps.
Summary of results: The integrity of the VWF/FVIII concentrate was confirmed by all tests. The ratio between VWF:RCo and FVIII:C was close to that in plasma of blood donors. The VWF multimeric profile demonstrated >10 bands in all batches and the triplet structure was very well preserved. A high specific activity, on average 122 IU FVIII:C/mg total protein, was measured in 15 batches recently produced. SE–HPLC revealed a one-peak-product, representing the VWF/FVIII-complex. All viruses tested were rapidly inactivated by at least 4 log10. The PermaHeat treatment of the lyophilised product at +100°C for 2 hours, at a specified residual moisture of 0.7–1.6%, was efficient for the inactivation of both EV and NEV.
Conclusion: Wilate® is a high purity, double virus inactivated concentrate containing functional VWF and FVIII in a ratio close to plasma of healthy subjects. The product has an excellent viral safety profile. The safe and effective treatment of patients with von Willebrand disease and haemophilia A was demonstrated in multiple clinical trials.