bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Novel Assay for Tissue Factor Pathway Inhibitor (TFPI) Anticoagulant Activity in Plasma.
Authors: Dahm Anders E.A. , Andersen Trine O. , Rosendaal Frits , Sandset Per Morten 
Published: October 2019

Abstract
Introduction. Several studies have shown that full-length free TFPI is a much stronger anticoagulant in diluted prothrombin time (dPT) assays than truncated and lipoprotein associated TFPI. Although it has been suggested that TFPI anticoagulant activity is of importance for the effect of TFPI in vivo, no assays have so far been established and validated to assay TFPI anticoagulant activity in patient samples.
Aims. To develop and validate an assay of TFPI anticoagulant activity and study the relationship between TFPI anticoagulant activity and TFPI free and total antigen and TFPI chromogenic substrate activity.
Materials and Methods. TFPI anticoagulant assay: 32 μL of plasma was first incubated for 5 min with either 8 μL TBS or anti-TFPI immunoglobulin, which completely blocked TFPI activity. Then, 30 μL recombinant, relipidated tissue factor (Innovin®) at 1/200 fold dilution was added and incubated for 3 min. Clotting was initiated by the addition of 30 μL CaCl2 (35 mM). Results were expressed as a ratio, with clotting times in the presence of anti-TFPI as the denominator. This ratio was normalized against the ratio obtained with a reference plasma within each run to minimize day to day variation in clotting times. After normalization, high ratios reflect high functional TFPI anticoagulant activity, whereas low ratios reflect low activity.
TFPI antigen assays: TFPI free and total antigen were assayed with commercial kits from Stago, France. TFPI free antigen is specific for circulating full-length TFPI, whereas TFPI total antigen detects both full-length and truncated forms of TFPI.
Study population: Control subjects (n=436) of the Leiden Thrombophila Study (LETS).
Results. Mean TFPI anticoagulant ratio was 1.03 (SD 0.0066, range 0.87–1.22). The assay coefficient of variation was 2.8% between days and from 0.6% to 1.5% within days. Linear regression showed that the n-TFPIac ratio was strongly associated to the levels of TFPI free antigen, but weakly associated to bound TFPI (table 1).
Conclusions. We have established a novel and reproducible assay for detection of TFPI anticoagulant activity in plasma. TFPI anticoagulant ratio was strongly associated to free antigen, and may be considered an activity assay of full-length TFPI in plasma.
Table 1 - The association of normalized TFPI anticoagulant ratio to other TFPI assays
TFPI assay Normalized TFPI anticoagulant ratio- b (95% CI) Correlation coefficient- r Standardized regression coefficient (b), 95% confidence interval (CI) and correlation coefficient (r). (b) shows the change in n-TFPIac ratio per increase of one standard deviation in the TFPI assay studied. TFPI free antigen 0.049 (0.044 to 0.053) 0.73 Total TFPI 0.033 (0.027 to 0.038) 0.48 TFPI chrom. subst. activity 0.031 (0.025 to 0.037) 0.46 Bound TFPI 0.020 (0.014 to 0.026) 0.30
Muscle Cells in Fibrin Matrix Actively Collapse the Network Leading to Rapid Allignment and Formation of Muscle Bundles.
Authors: Carr Marcus E. , Lam Phillipe , Simpson David G. , Wnek Gary E. , Bowlin Gary L. 
Published: October 2019

Abstract
The goal of in vitro organogenesis has been hampered by cell culture techniques that promote two dimensional cell proliferation. Since cell growth stops once confluence is achieved, three-dimensional organization does not occur. Possibly due to a lack of normal spatial orientation and cell-to-cell interactions, cells grown in such systems often assume morphologies significantly different from those assumed in normal biological tissues. We have recently developed a simple technique for growing cells in fibrin gels. This is accomplished by suspending cells in fibrinogen solutions and subsequently mixing the fibrinogen-cell suspension with buffered thrombin. The fibrinogen solution can be prepared from purified protein or from plasma diluted with growth media. The mixture rapidly forms a fibrin gel to which the cells are attached. When muscle cells are cultured in fibrin matrices, they initially form a latticework of proliferating cells. Over the next forty-eight hours, the cells begin to collapse the edge of the structure in a process similar to platelet induced clot retraction. As the fibrin network collapses, muscle cells are brought into linear alignment to form structures closely resembling muscle bundles. This process is relatively rapid. Within forty-eight to seventy-two hours, three-dimensional structures morphologically similar to muscle tissue are formed. By manipulating the gel structure through the imposition of outside forces, the alignment of muscle cells can be caused to occur even more rapidly. This system is simple to use and rapidly results in a muscle culture structure that resembles muscle tissue. This technique holds promise to significantly advance the goal of true in vitro “tissue” formation.
The Care of Immigrant Children with Haemophilia.
Authors: Barnes Chris , Hilliard Pam , Stain Ann Marie , Floros Georgina , Blanchette Victor , Carcao Manuel 
Published: October 2019

Abstract
Canada, like many industrialised countries, has a history of accepting immigrants and a significant proportion of the population of major cities like Toronto is comprised of visible minority groups. The provision of health care to recently arrived immigrant families presents a number of challenges including difficulties in communication, economic inequalities experienced by migrating families and complications of underlying conditions not often seen in countries with well-resourced health care systems. Over the last decade the Hospital for Sick Children (HSC) has been referred a number of children with haemophilia who have recently immigrated. We reviewed the presenting features and subsequent management of recently immigrated children with haemophilia at HSC. We compared objective joint assessment scores of these patients at presentation to follow-up joint assessment scores. Joint assessment scores were performed according to published guidelines with a normal joint score being graded as zero and the maximum joint score (total scores of both elbows, knees and ankles) being 162. Seventeen patients with haemophilia (haemophilia A N=15, haemophilia B N=2) were identified representing 9.2% of all patients with haemophilia at HSC and is comparable to the proportion of people who have recently immigrated to the Toronto area from other countries over the years 1995 to 2001 according to census data. The patients were seen initially at HSC at an average age of 7.3 years (range 0.6 years to 12.2 years) and have been managed at HSC for an average of 3.1 years (range 0.5 to 9.3 years). Fifteen patients have severe haemophilia and of these patients, 7 had no definite previous treatment with clotting factor concentrate. Two patients had been exposed to infection with the hepatitis C virus. None of the patients were HIV virus positive. Joint assessment scores at initial presentation to HSC were available for 13 patients with a mean score of 9.4 (range 0 to 24). Two of the patients were not weight-bearing at presentation. The mean repeat joint assessment scores for ten of the recently immigrated patients was 7.8 (range 0 to 27). With specific treatment and rehabilitation the joints scores improved in the majority of patients. Addressing the challenges of providing health care to recently immigrated children with haemophilia in a culturally sensitive manner can lead to effective medical management and greatly improve joint function in these children.
Short-Term Use of Phenobarbital in the Treatment of “Superwarfarin” Poisoning.
Authors: Oliveri Cecilia V. , Stubbs James R. , Chernova Tatyana , Carnahan Gary Ellis , Evans Tara N. , Gammill Andrew , Siegel Jonathan 
Published: October 2019

Abstract
Background: “Superwarfarin” was developed in the 1970s as an anticoagulant rodenticide. The half-life of the “superwarfarin”, brodifacoum, is four times longer than warfarin. Patients with significant brodifacoum ingestions have often required long-term treatment with vitamin K1 (e.g., 6 months), occasionally with adjunctive phenobarbital therapy.
Case: A 52-year-old male was admitted to the hospital after being found unconscious at home. He was pale and tachycardic at presentation with an oozing laceration on his right leg and bright red blood per rectum. Two liters of maroon fluid were recovered via nasogastric lavage. Admission laboratory values included: PT > 300 seconds, aPTT > 300 seconds, hemoglobin 2.8 g/dL, hematocrit 8.8%, and platelet count 400,000 per μL. Metabolic acidosis with an increased anion gap was present. A drug screen (Toxi-Lab®) was negative. Alcohol, salicylate, and acetaminophen assays yielded negative results. The following day, the patient admitted ingesting with a total of 170 grams of brodifacoum with suicidal intent. Subcutaneous vitamin K1 (10 mg QID), octreotide acetate (50 mg PO), and pantoprazole sodium (40 mg PO) were administered. He improved clinically, but his PT (range: 22.4–47.4 seconds), INR (range: 2.1–5.5), and aPTT (range: 40–70 seconds) remained abnormal. On hospital day 5 his vitamin K1 dose was increased to 25 mg QID and oral phenobarbital (30 mg TID) was started. A total of thirty-eight units of plasma and 8 red cell units were transfused during the admission. He was transferred to an inpatient psychiatric facility on day 8, and he was discharged from that facility after 5 days. Vitamin K1 and phenobarbital were discontinued at this time. Three and one-half months later, the patient was asymptomatic. His laboratory values included: PT 13.2 seconds, INR 1.1, aPTT 25 seconds, hemoglobin 14.3 g/dL, and hematocrit 42%. A serum brodifacoum level yielded negative results.
Conclusion: Superwarfarin has been noted to be an increasingly common cause of poisoning in the past 20 years, especially in intentional ingestions. Many cases have been successfully treated with long-term vitamin K1 — with adjunctive phenobarbital given to some patients. The rationale for using phenobarbital in this setting is that it increases warfarin metabolism by inducing cytochrome P450 isozymes known to be major catabolic enzymes for warfarin (CYP2C9/19) in the liver. We report the successful treatment of significant brodifacoum toxicity with a short course of vitamin K1 (13 days) in combination with phenobarbital (6 days).
Figure Figure
Compare Two Arsenic Trioxide Administration Methods in APL Therapy.
Authors: Zhou Jin , Meng Ran , Yang Baofeng 
Published: October 2019

Abstract
Background As2O3, on the one hand is successful in the treatment of acute promyelocytic leukemia (APL), on the other hand many side effects may occur during the early arsenic treatment. We assessed the effectiveness and security of two As2O3 administration methods in acute promyelocytic leukaemia treatment.
Methods seventy-four cases of APL treated using As2O3 in two administration methods have been studied randomly. Thirty-two cases were administrated As2O3 with routine method as control, the other 42 cases were given As2O3 with a ‘continuous and slowing intravenous infusion’ method. Firstly, we calculated the total quantity of As2O3 needed every day of every patient according to the criteria of 0.16mg/kg, then gave half of the total amount of As2O3 in the morning and the other half in the afternoon, diluted each half of total amount of As2O3 with 5% glucose 250ml and infused at the speed of 8 drops/min, collected the blood serums of the patients treated with the two methods respectively at different time points, and finally we assayed arsenic concentration in blood serums.
Results Most of the APL treated by As2O3 gained complete remission at the time of 4th week with the two administrated methods. The total effective rate in ‘continuous and slowing intravenous infusion ‘ group was 93.75%, while that in control group was 83.3%. The effective arsenic level in blood circulation in ‘continuous and slowing intravenous infusion’ group was lasted at least 18.38±3.27 hours, while in control group was lasted 9.38 ±1.58 hours, the average hospitalization duration in ‘continuous and slowing intravenous infusion ‘ group was26.4±2.04 days, while that in control group was 35.7±4.78 days.
Conclusions The ‘continuous and slowing intravenous infusion’ method can relieve leukocytosis, increase complete remission (CR) rate of APL, and accelerate to CR.
Table1 arsenic concentration, its lasting time and the average time reaching CR (mean± SD)
Group (n) The peakconcentration in 24h μmol/L) ( The lasting time above μmol/L(h / 24h) 1 The average time reaching CR(d) Experiment 42 3.32±0.55 18.38±3.27 26.4±2.04 control 32 5.84±1.05 9.38 ±1.58 35.7±4.78 t-value 6.08 3.64 4.78 P -value < 0.01 < 0.01 < 0.01
Table2 The incidence of side effects in As2O3 therapy ( %)
peripheral WBC↑ DIC QTc prolongation nerve conductspeed ↓ cerebral vasospasm liver disfunction experiment (n=42) 31.3(15) 16.7(8) 8.1(4) 8.1(4) 14.6(7) 14.6(7) control (n=32) 71.2(34) 35.4(17) 25.0(12) 6.3(3) 33.3(16) 22.9(11) X2 - value 17.64 4.38 4.80 0.14 4.62 1.08 P - value <0.001 < 0.05 < 0.05 > 0.1 < 0.05 > 0.05
Peripheral Blood HPC Counts Are Augmented in CML Patients with Cytogenetic Findings of Disease as Compared to Patients in Cytogenetic Remission.
Authors: de Meis Ernesto , Morais Rita C.B. , Carvalho Luize O. , Moellmann-Coelho Arthur , Martinez Maria M.K. , Costa Ana J.U.B. , Barros Ailce S. , Castilho Ilda A.M. 
Published: October 2019

Abstract
Chronic Myeloid Leukemia (CLM) is a stem cell disease characterized by inhibition of apoptosis and increased proliferation of bone marrow (BM) compounds. Evaluation of hematopoetic precursors cells on the peripheral blood (PB) of these patients show a high prevalence of leukemic CD34+ cells. Recently it was demonstrated that measurement of HPC by IMI channel on Sysmex showed an acceptable correlation between Hematopoetic Progenitors CD34+ cells counts. The aim of this work was to compare precursor cells (HPC) numbers in the PB of CML patients with disease and under remission. For this we studied 61 CML patients that had been treated with different protocols. From these 22 were in cytogenetic remission but 39 still presented more than 5% BM cells with Philadelphia chromosome (Ph+). Although the normal levels of CD34+ HPC in PB is considered to be very low, we considered less than 0.05% of leukocytes as normality. HPC in PB were counted using an automated peripheral blood counter (Sysmex XE-2100). We found that Ph+ CML patients had a significant increased number of PB HPC compared to CML patients under remission (with a confidence interval of 95%, and a corrected Yates analyses with X2 16.78 and p<0.0001). The increase in PB HPC was not related with the number of white cells counts at moment of the exam. These findings suggest that the presence of a high number of PB HPC in CML can be used as a non-invasive marker of the disease, more reliable than total and differential leukocyte count.
Constitutive JAK3 Activation as a Mouse Model for Human Peripheral T-Cell Lymphoma.
Authors: Cornejo Melanie G. , Kharas Michael G. , Lee Benjamin , Moore Sandra A. , Gilliland Gary , Mercher Thomas 
Published: October 2019

Abstract
Molecular pathogenesis of human peripheral T-cell malignancies remains poorly understood. We aimed to establish a model for these disorders by using the recently identified JAK3A572V allele. This mutation, which lies in the pseudokinase (JH2) domain of JAK3, transforms Ba/F3 cells to factor-independent growth and causes a striking lymphoproliferative phenotype in a murine bone marrow transplantation assay. Further characterization of JAK3A572V animals revealed that the aberrantly expanded cell population consisted of a mature effector/memory subtype of CD8+T-cells that infiltrated all major lymphoid and several non-lympoid organs and could be transplanted into secondary and tertiary recipients. These JAK3A572V T-cells had increased proliferative capacities and displayed enhanced phosphorylation of common JAK3 targets, such as STAT5 and S6-kinase. Proliferation of primary T-cells transformed by JAK3A572V was effectively inhibited with a small molecule JAK inhibitor that had no effect on the proliferative potential of control cells transduced with a wildtype JAK3 allele. Furthermore, the mutant cells showed increased production of cytotoxic cytokines, such as IFN-γ and TNF-α, compared to wildtype counterparts, which correlated with an increased cellular cytotoxicity towards allogeneic target cells. Of particular interest, JAK3A572V animals presented with skin lesions and histopathologic analysis showed aberrant skin-homing T-cells tagging along the epidermal/dermal junctions. Mice receiving Rag1-deficient donor cells transduced with the JAK3A572V allele also developed a lethal lymphoproliferative disease characterized by the expansion of immature CD3−TCRβ−CD4+/−CD8+ cells, suggesting that the JAK3A572V-dependent lymphoproliferation does not require proper TCR rearrangement. Altogether these results indicate that in this murine model, constitutive activation of JAK3 results in peripheral/cutaneous T-cell lymphoma (PTCL) that closely resembles the human disease. These findings suggest the possibility that the molecular basis of human PTCL could include aberrant JAK3 signaling and might provide a useful platform for deciphering the molecular and cellular mechanisms and requirements for peripheral lymphoid disease development and progression. Furthermore, it provides an opportunity to investigate the therapeutic potential of selective JAK3 inhibitors for this subset of lymphoid disorders, whose treatment remains a challenge.
PIR-B-Deficient Mice Are Susceptible to Salmonella Infection.
Authors: Oka Satoshi , Torii Ikuko , Hotomi Muneki , Benjamin William H. , Takai Toshiyuki , Kearney John F. , Briles David E. , Kubagawa Hiromi 
Published: October 2019

Abstract
Paired Immunoglobulin-like receptors of activating (PIR-A) and inhibitory (PIR-B) isoforms are expressed by many blood cell types including B cells, dendritic cells, monocyte/macrophages, granulocytes, mast cells and megakaryocyte/platelets. To determine the role of PIR-B in bacterial infection, PIR-B deficient (PIR-B−/−) and control C57BL/6 mice of the same age and sex were infected intravenously with the attenuated strain of Salmonella typhimurium (WB335). When injected with high doses of bacteria (5 × 104 ∼ 5 × 106 CFU), both types of mice died within 3 to 14 days post infection. When given small doses (103 ∼ 104 CFU), however, PIR-B−/− mice exhibited progressively increasing morbidity beginning at 4-day post-infection over a 4-week period, whereas many control mice could survive (see Figure). PIR-B−/− mice were found to have higher bacterial loads in liver, spleen and lungs than the control mice and bacteremia at days 3 and 7 post-infection. In liver histology typhoid nodules were observed as early as day 1 and their incidence increased during the next six days in both groups of mice. However, quite distinct changes were observed at day 7: in control mice inflammatory cellular lesions were localized and exhibited nodular distribution, whereas in PIR-B−/− mice mononuclear cells infiltrated along the liver sinusoids and exhibited a spreading distribution. There were no significant differences in plasma levels of liver enzymes (ATL, AST and ALK), cytokines (TNFα, IL-10 and IFNγ) and antibodies (IgM, IgG and IgA) against Salmonella between PIR-B−/− and control mice. While splenic lymphoid and myeloid subsets were indistinguishable during the first week of infection, the increases of CD19+ B and CD8+ T cells in infected PIR-B−/− mice were not as marked as those in infected control mice at day 14 post-infection (see Table). For phagocytic cell functions ex vivo, there were no differences in phagocytosis and intracellular killing of bacteria and superoxide anion release between PIR-B−/− and control mice. However, PIR-B−/− macrophages were found to release more nitrite and TNFα upon bacteria stimulation than the control macrophages. These results suggest that PIR-B plays an important regulatory role in bacterial infection.
(Supported by NIH/NIAID grants AI42127 and AI52243).
Mac-1+ MΦ CD4 + T CD8+ T CD19+ B cell numbers (× 107), mean ± 1 S.D. Uninfected Control 1.3 ± 0.6 * 1.4 ± 0.4 0.7 ± 0.3 5.1 ± 1.8 Uninfected PIR-B−/− 1.2 ± 0.3 1.5 ± 0.2 1.2 ± 0.2 4.9 ± 0.3 Infected Control 19.4 ± 7.0 5.5 ± 1.5 2.6 ± 0.7 10.0 ± 2.6 Infected PIR-B−/− 16.5 ± 2.3 4.2 ± 2.1 1.3 ± 0.6 4.5 ± 1.3 Figure Figure
Macrocytosis in COPD and Course of the Disease
Authors: Kontoninas Zisis , Girtovitis Fotios , Kaiafa Georgia , Ntaios George , Saouli Zoi , Hatzitolios Apostolos , Charisopoulos George , Savopoulos Christos , Papadopoulos Athanasios 
Published: October 2019

Abstract
INTRODUCTION: Macrocytosis (MCV>94fl) is frequently observed at patients suffering from chronic obstructive pulmonary disease (COPD) with or without respiratory insufficiency. Recent studies have demonstrated the role of erythropoietin massive excretion subsequent to hypoxemia as a trigger factor in excessive marrow erythropoiesis. Thus, a large number of immature red cells are released in peripheral blood leading to macrocytosis. AIM: To determine possible correlation between macrocytosis and severity of he disease in patients with COPD.
PATIENTS AND METHODS: Thirty eight patients (mean age: 62,16±10,88 years, twenty nine male-Group A and nine female-Group B) with COPD were attended regarding their clinical status for one year, diagnosed according to spirometric classification of COPD based on post-bronchodilator FEV1 (Global Initiative for Chronic Obstructive Lung Disease, 2006). All patients were smokers suffered from mild or moderate COPD (FEV1/FVC <0,70, FEV1≥50% predicted, with PaO2 more than 60mmHg). MCV and hematocrit were measured separately for male and female additionally to the whole sample. Exacerbations considered to be happen when dyspnea or PaO2<60mmHg occurred. Patients with anemia, hematological malignancy, hypothyroidism, heart or renal failure and diabetes mellitus excluded.
RESULTS: Mean value for MCV was 98,84±2,66 fl (range:95–107 fl). Average for Group A estimated at 99,52±2,92 (range: 95–107) and for Group B at 96,87±1,40 (range: 95–99). Mean value of exacerbations for both groups in one’s year period was 0,89±0,61 (range :0–2), that was not significantly different from those concerning Group A and Group B (0,79±0,60, 1,22±0,74 respectively). Inverse correlation between MCV and number of exacerbations was statistically significant (r = − 0,43, p<0,01). That correlation was more apparent for Group A (r = −0,45, p<0,01) than Group B (r = −0,35, p<0,05). Mean value of hematocrit was determined at 45,92 ± 3,77 (range: 40–54). Correlation between MCV and Ht was not significant (r=0,2).
CONCLUSIONS: An unexpected result revealed considering that elevated MCV in patients with mild to moderate COPD correlates with fewer exacerbations of the disease. A game of statistics or something more important for the pathophysiology of mild COPD?