bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Children with Coagulation Test Abnormality in Pre-Surgical Evaluation
Authors: Kwon Seung Yeon , Han Jung Woo , Won Sung Chul , Song Jaewoo , Lyu Chuhl Joo 
Published: April 2020

Abstract
Prothrombine time (PT) and activated prothrombine time (aPTT) are common tests used for screening of coagulation function before surgical procedures. We analyzed underlying causes of abnormal coagulation test results which were incidentally found during pre-surgical evaluation in healthy patients without definite bleeding history. Total 58 children referred to pediatric hematoloy service for abnormal PT and aPTT results in pre-surgical evaluation between June 2006 and May 2008 were analyzed retrospectively by review of medical records. 50 patients showed aPTT prolongation, 5 patients PT prolongation, 2 patients PT and aPTT prolongation and another three patients showed normal PT and aPTT. Among 55 patients with abnormal results, 25 patients (43%) were recovered spontaneously during their follow up tests, 17 patients (29%) showed lower level of certain coagulation factor than reference range and the other 13 patients were lost during follow up despite of recommendation for further evaluation. Mean value of international normalized ratio (INR) for PT and aPTT of the patients recovered spontaneously were 1.05±0.11, 44.53±5.01seconds(s), and 1.12±0.11, 47.0±5.36s in patients with lower level of coagulation factor, showing significant increase of PTT in patients with lower factor levels (p&lt;0.05). Median time required for spontaneous recovery was four weeks and 18 patients (72%) were recovered within this time. Among 17 patients with lower level of certain coagulation factor then reference level, there were 11 patients with low factor XII level, three patients with low factor VIII level, three patients with low von Willebrand factor, two patients each for low factor VII and factor XI and one patient with low factor V level. Among them three patients with low level in von Willebrand factor, one patient with low factor VII and two patients with low factor XI showed deficient level of coagulation factors requiring factor replacement for the surgical procedures. From this analysis of patients with incidentally found PT or aPTT prolongation, 43% of patients were spontaneously recovered during follow up period within 4 weeks in median. However, we also found that 29% of patients had relatively lower level of coagulation factor than reference range. Even though most of them were factor XII decrease which is not closely related with bleeding tendency, six patients had significant deficiencies of coagulation factors requiring factor replacement during surgical procedures. These results suggest that we should keep following up and undergo adequate evaluation for underlying coagulation factor deficiencies in patients who have sustaining PT and aPTT prolongation abnormalities despite of absence of any bleeding history.
Involvement of Heparanase in Early Pregnancy Losses
Authors: Nadir Yona , Henig Israel , Vlodavsky Israel , Brenner Benjamin 
Published: April 2020

Abstract
Heparanase activity is implicated in cell invasion, tumor metastasis and angiogenesis. Recently, we have reported that heparanase stimulates tissue factor (TF) expression in endothelial and cancer cells, resulting in elevation of coagulation activity (Nadir et al. JTH, 2006). We also found that heparanase releases tissue factor pathway inhibitor (TFPI) from endothelial and tumor cells, contributing to the local procoagulant balance (Nadir et al. TH, 2008). Heparanase was cloned from and is abundant in the placenta. We investigated the role of heparanase in the placenta, focusing on its effect on TF, TFPI, TFPI-2, and vascular endothelial growth factor-A (VEGF-A). Twenty formalin embedded samples of early pregnancy losses (weeks 5–10) were studied. Ten cases were miscarriages of women with thrombophilia and recurrent fetal losses, and ten control cases were pregnancy terminations. Applying real time RT-PCR and immunostaining for heparanase, TF, TFPI, TFPI-2, and VEGF-A it was found that in fetal losses related to thrombophilia the levels of heparanase, VEGF-A and TFPI-2 were significantly increased (2–3 fold) compared to placentas from controls. Next, JAR (human choriocarcinoma throphoblast) cells were transfected to overexpress the full length (65 kDa) heparanase or incubated with active (50+8 kDa) recombinant heparanase. Immunoblotting and real time RT-PCR showed that throphoblasts overexpressing or incubated with exogenous heparanase exhibited a 2–3 fold increase in TFPI and TFPI-2 in cell lysates both at the protein and mRNA levels, with no detectable effect on VEGF-A and TF. Accumulation of TFPI and TFPI-2 in the cell culture medium was increased 5–6 fold, exceeding the observed induction of TFPI and TFPI-2 gene transcription. Extracellular accumulation of TFPI and TFPI-2 was already evident 60 min following heparanase addition, prior to protein induction, indicating an active release of preformed proteins. These results indicate a regulatory effect of heparanase on TFPI and TFPI-2 in throphoblasts, suggesting a potential involvement in early miscarriages.
Coagulation Induced by Dilute Tissue Factor Depends More Critically on Factor II and X Concentration Than on FVII or FIX
Authors: Gudmundsdottir Brynja R. , Bjornsdottir Alexia M , Onundarson Pall T. 
Published: April 2020

Abstract
Prothrombin time based tests used to monitor coumarin anticoagulation measure the initiation phase of fibrin formation (clotting time, CT) in platelet poor plasma. Additional information can be obtained using computerized rotational thromboelastometry (ROTEM) which also measures the consequent propagation phase (measured as maximum velocity, Vmax), and the stabilization phase (maximum clot firmness, MCF).
We used ROTEM to study the effect of individual vitamin K dependent (VK) coagulation factor (F) concentration on clotting induced by dilute thromboplastin in platelet poor and platelet rich plasma (PPP and PRP).
As expected, FII, FVII and FX in PPP equally affected the prothrombin time whereas FIX did not. In PPP the ROTEM CT, however, was affected more by the concentration of FII and FX than by FVII (data not shown). To imitate physiological clotting better, factor deficient platelet poor plasma was repleted by adding frozen platelets in an optimal concentration corresponding to 100 × 109/L.. In the PRP the ROTEM CT was more dependent on the concentration of FII than of FVII, more dependent on FX than on FII and not dependent on FIX at all (left figure). The Vmax (reflecting the propagation phase) was also more dependent on FII and factor X than on the concentration of FVII or FIX which both may demonstrate a threshold effect (right figure). The stabilization phase or maximum clot firmness (MCF, not shown) was influenced similarly by FII and FX but was not influenced by FVII or FIX.
Figure Figure
Conclusion: During deficiency of vitamin K dependent coagulation factors the concentration of FII and FX may be more critical for hemostasis than FVII or FIX concentration. This may have practical implications for the appropriate choice of monitoring assays during coumarin administration.
Human Hepatic Stellate Cells Carry a New Splice Variant of the Human ADAMTS13
Authors: Kimchi-Sarfaty Chava , Blaisdell Adam , Marple Andrew , Roma Isabella , Newell Jordan , Dobkin Alexandra , Allen Courtni , Friedman Scott , Burge Christopher , Shomron Noam 
Published: April 2020

Abstract
The ADAMTS13 gene plays an integral role in vascular homeostasis. A severe deficiency in ADAMTS13 triggers a rare disorder known as thrombotic thrombocytopenic purpura (TTP). Various missense and nonsense point mutations have been described in exonic regions of ADAMTS13 along with several frameshift deletions and one frameshift insertion. Additionally, several intronic mutations have been discovered in USA patients that result in aberrant mRNA splicing forms. A tissue based screen of ADAMTS13 alternative splicing has never been assessed. Besides the known mRNA splice isoforms, others are evident from the Expressed Sequence Tag (EST) databases. We analyzed alternative splicing of ADAMTS13 in several tissues in order to assess whether alternative splicing might play a role during its gene expression. We identified a novel isoform of ADAMTS13 that retains intron 25, evident in human, but not in mouse, hepatic cells. Moreover, the retained intron is not detected in mice possibly because they do not possess regulatory sites inserted into the human intron at a later evolutionary stage. This might suggest that ADAMTS13 is post-transcriptionally regulated at the splicing level. It also shows the need for future analysis of transcript levels of this new isoform in TTP patients. We hypothesize that irregular balance between the two splice isoforms might affect ADAMTS13 functionality, thus, it is possible that our findings might be of clinical significance.
“The findings and conclusions in this presentation have not been formally disseminated by the Food and Drug Administration and should not be construed to represent any Agency determination or policy.”
Use of Amphipathic Helical Peptides as An Anticoagulant
Authors: Charbonneau Sophie , Peng Henry , Shek Pang N , Blostein Mark 
Published: April 2020

Abstract
We have previously shown that an ideal amphipathic helical peptide of K7L15 composition (IAP) accelerates factor IXa-mediated factor X turnover and factor Xamediated prothrombin turnover in a phospholipid free system (Biochem J., 2008, 412:545). Under these conditions, IAP behaves as a phospholipid membrane allowing coagulation factors to bind and exert their actions. However, when IAP is used with in vitro assays that employ phospholipids such as an active partial thromboplastin time (aPTT), IAP paradoxically behaves as an anticoagulant by prolonging clotting times. We hypothesize that this anticoagulant effect occurs by blocking binding sites for coagulation factors on phospholipids membranes. To test this hypothesis, we employed three phopholipid-dependant coagulation assays, the aPTT, dilute PT and dilute RVV, with both low and high concentrations of phospholipids. We show that these coagulation times are prolonged by IAP in a concentration dependent manner and that this prolongation is abrogated by adding excess phospholipid, demonstrating phospholpid dependence for this inhibition. In purified tenase and prothrombinase assays, in the presence of phospholipids, IAP inhibits substrate turnover consistent with our hypothesis. To show direct binding between IAP and phospholipids, we conducted fluorescence spectroscopy experiments and show direct binding between IAP and phospholipid membranes. In summary, the above data demonstrate that IAP acts as an anticoagulant by blocking the interaction of coagulation factors with phospholipids membranes.
Effect of Human Recombinant VWF Administration on Plasma ADAMTS13 Activities in Non-Human Primates and Rabbits
Authors: Varadi Katalin , Rottensteiner Hanspeter , Muchitsch Eva M , Weber Alfred , Gritsch Herbert , Turecek Peter L. , Ehrlich Hartmut J , Schwarz Hans Peter 
Published: April 2020

Abstract
Von Willebrand factor (VWF) is composed of a series of multimers, the sizes of which are regulated by the plasma metalloprotease ADAMTS13. Reports suggest that a transient increase in VWF levels, triggered for instance by DDAVP treatment, will result in a decrease in ADAMTS13 activity. This phenomenon is widely believed to be due to ADAMTS13 being exhausted when confronted with an excess of substrate. Treating patients who have type 3 von Willebrand disease with a recombinant human von Willebrand factor (rVWF) that has not yet been exposed to ADAMTS13 might thus cause a drop in ADAMTS13 levels. We therefore tested whether a rise in VWF concentrations to supraphysiological levels caused by administration of rVWF could influence endogenous ADAMTS13 levels in animal models. Various doses of human rVWF (300, 600, and 1200 RCo IU/kg BW) were injected into rabbits and cynomolgus monkeys and plasma samples were collected at a range of time points. As expected, VWF antigen rose sharply in a dose-dependent manner (~25 IU/ml VWF:Ag for the highest dose, 15 min after injection) and then declined gradually (~7 IU/ml VWF:Ag for the highest dose, 18 hours after injection). When these samples were tested for ADAMTS13 activity using the FRETS assay, no relevant changes were observed throughout the entire test period in the rabbit or in the monkey samples, indicating that rVWF, even at high doses, did not compromise ADAMTS13 activity. Both rabbit and cynomolgus ADAMTS13 recognized human rVWF as the specific cleavage products were detectable by electrophoresis at all doses administered. These animal studies thus indicate that an excess of intravenously administered rVWF leading to supraphysiological levels does not exhaust ADAMTS13.
Aplastic Anemia, Pesticide Exposure and Response to Cyclosporine Monotherapy: A Single Institution Case Controlled Study
Authors: Malhotra Hemant , Singh Ajay Pal , Rishi Jai Prakash , Khandelwal Purshottam D 
Published: April 2020

Abstract
Introduction:The present study is a prospective, hospital-based, case-controlled study of 19 patients of acquired Aplastic Anaemia (AA) from the SMS Medical College Hospital, Jaipur, India.
Patients & Methods:All newly diagnosed patients of Aplastic Anaemia over a one year period starting from Jan 2005 were included in the study. Detailed demographic parameters including a careful evaluation for pesticide exposure, haematological profile, serum organochlorine levels and response to oral Cyclosporine-A (Cyc-A) were recorded as per a preset Performa. Patients were followed for a minimum of 12 months. Patients receiving stem cell transplantation (SCT) and anti-thymocyte globulin (ATG) were excluded from the study. Serum quantitative and qualitative estimation of organochlorine pesticide levels by gas chromatography were done in all patients at study entry and compared with age and sex matched normal control.
Observations & Results:
Of the 19 patients included in the study, 15 (78%) were males and the mean age was 23 years (range 7 to 64 years) in the study. Thirteen of the 19 patients (68.4%) were from rural versus urban habitat and 9/19 (47.4%) gave a history of pesticide exposure. All these were from rural background. Six patients (31.6%) had non-severe AA, 9/19 (47.4%) had severe AA and 4/19 (21%) had very severe AA. Overall response to Cyc-A at 3 months was 47.4% [complete response (CR) 10.6%] and at 6 months was 47.4% (CR 21.1%). Non-severe AA patients had a response rate of 83.3% versus 33.3% in the severe AA and 25% in the very severe AA group. Cyc-A was generally well tolerated with mild toxicity. No patient was taken off the drug because of toxicity. No patient required serum Cyc-A level monitoring. Serum organocholorine levels were significantly higher (p&lt;0.005) in the AA patients (mean 0.35 ppm) versus controls (0.22 ppm). Mean serum organoclorine levels appear to correlate with the severity of AA. Patients with higher serum organochlorine levels were less likely to respond to Cyc-A therapy.
Conclusions:Even though this is a relatively small, single-institution study, the following conclusions can be drawn:
Single agent Cyc-A can produce a response in close to half of AA patients within 3 months of starting treatment. Considering the ease of administration and the low toxicity, Cyc-A could be considered as initial therapy in all patients of AA in a resource-limited setting (as compared to ATG and SCT). Pesticide exposure may play a role in the aetiopathogenesis of AA, particularly in patients with occupational exposure to the agent. Pesticide exposure and elevated serum pesticide levels may identify patients of AA who will not respond to immunotherapy (Cyc-A).
Is the Early Cyclosporine a Level Predictive to the Outcome of Immunosuppressive Therapy in Severe Aplastic Anemia?
Authors: Song Moo-kon , Chung Jooseop , Choi Young-Jin , Seol Young-Mi 
Published: April 2020

Abstract
Although there are many recent studies of efficacy of antithymoglobulin (ATG) combined with cyclosporine A (CsA), there has been no study to clarify the correlation between variability of CsA level and the response rate of immunosuppressive therapy (IST). Therefore, we assessed whether the early CsA level would be important or not for IST in acquired severe aplastic anemia (SAA), retrospectively. Thirty one patients were treated with ATG combined with CsA for 6 months. The patient group that achieved higher mean CsA level (≥200 ng/ml, higher CsA group) during first 2 weeks, was shown to more higher response rate than the group, lower mean CsA level (&lt; 200ng/ml, lower CsA group)(response rate of 82.3% in higher CsA group, 21.4% in lower CsA group, p=0.0023). There was also a significant difference in survival rate during median 785 days between higher CsA group and lower CsA group (survival rate of 64.7% in higher CsA group, 28.6% in lower CsA group, p=0.0115). Therefore, we suggest that early higher CsA level during first 2 weeks would be an important factor to achieve the overall response of IST in severe aplastic anemia.
Long-Term Clinical Outcome in Children with Very Severe Aplastic Anemia
Authors: Chung Nack Gyun , Jeong Dae-Chul , Lee Jae-Wook , Jang Pil-Sang , Cho Bin , Kim Hack-Ki 
Published: April 2020

Abstract
Introduction: The clinical course of pediatric patients with severe aplastic anemia (SAA) treated with immunosuppressive therapy (IST) or hematopoietic cell transplantation (HCT) is well known. However, reports on the course and prognosis of patients with very severe aplastic anemia (vSAA) are few. Here, we report on the characteristics and outcome of children with vSAA.
Methods: We reviewed the medical records of patients diagnosed with vSAA at the Department of Pediatrics, St. Mary’s Hospital, The Catholic University of Korea between January, 1995 to December, 2007. We analyzed the clinical characteristics of the patients at diagnosis, and the effects of IST, HSCT and other supportive treatment measures on patient outcome.
Results: Forty three patients were diagnosed with vSAA during the study period, of whom 9 received supportive treatment measures only, 21 received IST consisting of cyclosporine A and either ATG or ALG, and 15 underwent HCT (including 2 patients who showed no response to initial IST). Of the 9 children who received supportive measures only, one patient recovered spontaneously and is surviving event-free for 22 months, 6 patients died from either severe infection or bleeding from 1 to 10 months after diagnosis (median 5 months), and the other two children are continuing to receive supportive transfusions. Of the patients with IST, two children showed a complete response (10.5%), 6 patients showed a partial response (31.6%), and eleven failed to respond (57.9%). Of the 11 patients who did not respond, 8 children died from infection or bleeding within 9 months after IST respectively, 2 patients were lost to follow-up, and 1 patient is receiving supportive transfusions. Of the 8 patients who showed a response, 2 have shown relapse and are being maintained with transfusions. Of the 15 HCT’s, 12 were from HLA-matched sibling donors and 3 were from matched unrelated donors. Except for the 2 patients who died from serious infections at 5 and 9 months post-HSCT respectively, all are currently surviving disease-free. The 10-year survival of the patients who received supportive measures only, IST, or HCT is 33.3±15.7%, 47.4±14.0 and 86.7±8.8% respectively (P=0.009).
Conclusion: As with SAA, HCT is the best treatment option for vSAA. If IST is undertaken, measures should be undertaken to prevent death from early infection. With only supportive care, median survival approximates only 6 months, calling for more intensive treatment strategies such as IST or HCT in the care of vSAA patients.
Viability of CD34+ Cells in Cryopreserved Cord Blood
Authors: Eom Young-woo , Jeong Seong Hyun , Choi Jin-Hyuk , Kang Seok Yun , Lee Hyun Woo , Yang Mal Sook , Kim Hyo Eun , Jang In Keun , Lee Jong Eun , Kim Yeong Jin , Kim Hugh Chul , Hwang Yoon Ho , Ahn Mi Sun , Park Joon Seong 
Published: April 2020

Abstract
Purpose: On performing umbilical cord blood (UCB) transplantation, faster engraftment may lead better clinical outcome. Because transplanted viable cell count in UCB is related to the engraftment, we examined cryopreserved UCB cells with several methods after thawing.
Methods: Viability of cryopreserved cells were examined with trypan blue, DNA contents analysis, caspase-3 activation test, intracellular esterase activity and Annexin-V/PI staining.
Results: A total of 60 samples were used in this study. After thawing, 89% of the total MNCs and 84% of CD34+ cells were viable as identified by trypan blue exclusion assay. In the CD34+ cell population, the cell death rate was found to be 47 % by Annexin-V/PI staining and less than 5 % by DNA contents analysis. Caspase-3 activity failed to document apoptosis. The intracellular esterase activity test also showed a cell death rate of about 10–20 % at 2, 4, and 6 hours after thawing.
Conclusion: Viable cells in UCB should be measured by several compensatory techniques rather than a single method. Discordance among Annexin-V/PI staining versus trypan blue exclusion, DNA contents analysis, and the caspase-3 activation test or intracellular esterase activity should be clarified in order to apply these techniques for actual cord blood transplantation.
Peripheral Blood Stem Cell Collection in Elderly Patients
Authors: Tempescul Adrian , Ianotto Jean-Christophe , Eveillard Jean-Richard , Hardy Elisabeth , Guillerm Gaelle , Dalbies Florence , Berthou Christian 
Published: April 2020

Abstract
Introduction Intensive treatments like autologous stem cells transplantations are final standard treatment in lymphoma and myeloma for young people (1,2). In the context of population’s ageing, those treatments should be eligible for older patients. One of the major causes which did not allowed these procedures in elderly patients is the reports of the impact of the age on the process of mobilisation of peripheral stem cell (3). We proceed a retrospective study to explore the feasibility of stem cells collection in patients older than 65 years, compared to younger people. Material and patients During the period of 1999–2006, we identified all the patients older than 65 years, with myeloma and lymphoma, who performed peripheral stem cells collection in Brest’s blood collection centre. We excluded patients for bone marrow collection. Patients were eligible if they achieved more than 2.106 CD34/kg. Then we compared the results obtained in older patients with the results obtained in younger people. Results We identified 100 patients older than 65 years between the 359 patients who underwent peripheral stem cell harvest in Brest’s harvest centre. We excluded 37 others patients for non-conformity. The elderly patients were significantly older (age 68.9 vs. 49.3 y, p&lt;10−6). No differences were seen for the number of collection between the older and the younger patients (1.59 vs. 1.50, p&gt;0.05). The number of CD34/kg per patient was sufficient to realise more than one autologous transplantation in both groups of patients (5.3 vs. 6.67, p=0.004). Samples were sufficiently rich for CD34/kg (4.36 vs. 5.54, p=0.02). Patients were harvested in one (60), two (25) or more than 2 collections (15). Among the older patients, 71% achieved a sufficient collection of more than 2.106 CD34/kg in one sample only. Conclusion We identified one hundred patients older than 65 years, during a period of 8 years, who performed peripheral stem cells collection, with sufficient wealth (&gt;2.106 CD34/kg). The majority of patients, 60%, need only one stem cells harvesting. The study confirmed that old patients could be harvested like young people, and eligible for stem cells autologous transplantation.