bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Scoring, the PF4 ENHANCED Assay, and the 14 C-Serotinin Release Assay for the Diagnosis of Heparin Induced Thrombocytopenia (HIT) in Complex Medical and Surgical Patients.
Authors: Shaheed Gurvinder , Malkovska Vera , Horne McDonald , Mendoza Jose , Patel Mehool , Rees John , Wesley Robert , Merryman Paula 
Published: April 2020

Abstract
BACKGROUND: Heparin-induced thrombocytopenia (HIT) is a clinico-pathological entity that presents a diagnostic challenge, especially in medically complex patients.
PATIENTS AND METHODS: In this observational and prospective study, 34 consecutive patients with suspected HIT were enrolled. They were clinically scored using two different clinical scoring systems and underwent testing for PVS-ELISA and 14C-SRA. 14C-SRA results were also available from 21 medical and surgical patients who had previously been tested.
PURPOSE: The objective of our study was to evaluate the sensitivity and specificity of the PF4 ENHANCED (GTI Diagnostics) ELISA for HIT performed at two institutions using the serotonin-release assay (14C-SRA) and clinical scoring systems as reference methods.
RESULTS: With the 14C-SRA as the reference method, the sensitivities and specificities of the ELISA were 100% and 57% respectively at one institution and 93% and 65% at the other. There was only one false-negative ELISA with the 14C-SRA as the reference standard. When compared with the clinical scoring by Greinacher et al., the sensitivity and specificity for PVS-ELISA was 81.82% and 50% and for 14C-SRA 40% and 100% respectively. Five of eleven patients who scored ’Very Likely’ by the Greinacher scoring system were either ’Unlikely’ or ’Possible’ with the Paulpard scoring system. Seven of 11patients with ’Very likely’ HIT and 7 of 8 with ’Possible’ HIT by Greinacher clinical criteria were 14C-SRA negative.
CONCLUSIONS: Our study did confirm the high specificity of 14C-SRA, however the sensitivity was low with clinical scoring as reference standard. Clinical scoring was discordant in more than 50% of patients with ’Very Likely’ HIT by Greinacher score when compared with Paulpard scoring. Thus neither the clinical score nor the 14C-SRA appears suitable as the ’gold standard’ for diagnosis of HIT. Because of its high sensitivity with the 14C-SRA as the reference method, we believe the PF4 ENHANCED ELISA should be used to identify HIT in patients with multiple potential causes of thrombocytopenia, although false-positive results will not be uncommon.
Low Platelet Counts: Diagnosis Using Flow Cytometry and Anti Platelet Antibody.
Authors: Guerra Joao Carlos C. , Kanayama Ruth H. , Nozawa Sonia S. , Ioshida Marcia R. , Takihi Irina Y. , Lazaro Robson J. , Hamerschlak Nelson , Rosenfeld Luiz Gastao M. , Guerra Celso Carlos C. , Bacal Nydia S. 
Published: April 2020

Abstract
In our service of hematology 9,1 % of all consults are related to low platelet number finding in the CBC. Pseudothormbocytopenia is a rather unusual phenomenon, induced by agglutination of platelets by auto-antibodies acting on EDTA collected blood.
Objective: Classify, diagnose and analyze low platelet counts as obtained by automatic blood counters.
Methods: From January 1997 to May 2005; 16761 patients were attended in our service. Among them, 1174 cases (7,0%), 429 males and 745 females (1 to 75 years), were there because of low platelet counts alone. Platelet counts were done by Coulter T-890, with blood collected in EDTA K3 and sodium citrate. Counts were repeated after letting blood at 4o C for 24 hours; also Fonio technique and Neubauer camera counts were done. Normal value was defined as platelet count between 150,000 to 450,000. Bone marrow was obtained when necessary and stained by conventional techniques. Serological and biochemical tests were done in a Cobas Core (Roche) machine. Immuno-phenotyping was done in 115 cases, to identify anti-platelet-antibody (CD41 PE Immunotech and anti human IgG FTIC conjugate Sigma)-direct method, by flow citometry (Coulter Epics XL-MCL).
Results: Among the 1158 cases, platelets counts between 120 000 to 150 000, were found in 15.8 %; laboratory error in 6,3 % and spurious low platelet counts in 1.3 %. In addition, the follow up for four years of those patients with “lower normal” platelet count (120 000 to 150 000) did not show any morbid process. Immunologycal idiopatic purpura (ITP) was diagnosed in 49,1 % of the cases, being as expected, more frequent in females, children and young adults. Hepatitis C as cause of low platelet counts was responsible for 6,9 % and HIV infection for 1.3 % of the cases Anti-platelet antibodies were present in 76.9 % of ITP cases and was not found in 88.3 % of normal platelet values.
Conclusions: In an hematological reference service should be considered a lower range for normal platelet values (120 000 to 450 000). Additionally, low platelet counts as diagnosed by automated machines should be confirmed by slide exam and anti platelet antibody is useful for asserting ITP diagnosis and to rule out spurious thrombocytopenia or normal values.
Treatment of Immune-Mediated Thrombocytopenia Purpura with Concurrent IVIg and Platelet Transfusion: A Retrospective Review of 40 Patients.
Authors: Spahr Joseph E. , Agarwal Neeraj , Rodgers George M. 
Published: April 2020

Abstract
Introduction: In January 2000, two patients with severe Immune-Mediated Thrombocytopenia (ITP) at our institution were successfully treated with prolonged infusions of IVIg and platelets. The dose of IVIg was 1 g/kg given by continuous infusion over 24 hours with concurrent platelets (1 pheresis unit every 8 hours). Based on these preliminary results, we evaluated this protocol in a larger series of 40 ITP patients.
Methods: We performed an IRB-approved retrospective review of adult hospitalized patients with ITP treated with this regimen from January 2000 - December 2005. Patients with clinically significant thrombocytopenia and either active bleeding, need for anticoagulation, or requirement for a surgical procedure received the combined treatment. The subjects received IVIg and platelets as described above. Additional treatments, such as steroids, immunosupressives, or rituximab, as well as splenectomy were utilized at the discretion of the hematologist overseeing their care.
Results: The average age of patients treated was 52 years. The majority of patients ranged from 20–80 years old, but 12.5% were older than 80 years. The average pretreatment platelet count was 10,000/μl, with an increase to 55,000/μl after 24 hours, and 69,000/μl after 48 hours. By 72 hours, the average platelet count had begun to decline, although the platelet count remained at an acceptable level (58,200/μl). After 24 hours, 62.7% of patients had a platelet count &gt; 50,000/μl. Bleeding was controlled initially in all patients, and those requiring a procedure experienced no bleeding complications. Over half of the patients (52.5%) required additional treatments for recurrent or refractory ITP, and 32.5% of the patients underwent splenectomy. Six of the 21 patients requiring later retreatment (29%) received IVIg and platelets again in a similar fashion. The average retreatment platelet counts after 24 and 48 hours were 53,000/μl and 49,000/μl respectively, with clinical improvement in bleeding in all patients. No side effects of the combined treatment were noted. The response rates for the 3 IVIg products used were similar.
Discussion: For ITP, IVIg and platelets are considered to be first line treatment for patients with very low platelet counts, active bleeding, or those requiring urgent procedures. There is limited literature on the optimal dose and schedule for administration of IVIg and platelets. Our approach for administration of IVIg and platelets concurrently was associated with minimal side effects, resolution of bleeding, ability to safely undergo procedures, and rapid restoration of adequate platelet counts. Additionally, elderly patients had equivalent benefit with no increased side effects, indicating that this regimen is appropriate and safe for patients of all age groups.
Rituximab in Refractory Idiopathic Thrombocytopenic Purpura (ITP).
Authors: Santoro Cristina , Biondo Francesca , De Angelis Gioia , De Propris Mariastefania , De Vellis Annalisa , Guarini Anna , Malandruccolo Luigi , Rago Angela , Foa Robert , Mazzucconi Mariagabriella 
Published: April 2020

Abstract
Background. Rituximab, a chimeric anti-CD20 monoclonal antibody effective in B-cell depletion, may be useful in autoimmune disorders by interfering with the production of auto-antibodies.
Aims. To investigate the efficacy of Rituximab in patients with resistant ITP.
Patients and Methods. Fourteen adult ITP patients (3 males, 11 females; median age 44.5 years [21.1–67.6]) were treated with Rituximab (375 mg/m2/weekly for four doses). The median time between diagnosis and start of Rituximab was 2 years (0.2–33.1 months). All patients had already received at least two lines of therapy (median 3; 2–6): prednisone, pulsed high-dose dexamethasone, azathioprine, immunoglobulins, interferon or splenectomy. At the start of Rituximab, the median platelet count was 10 x 109/L (3–20 x 109/L). Response definitions: complete response (CR), platelet count ≥150 x 109/L; partial response (PR), &gt;50 &lt;150 x 109/L; minimal response (MR), &gt;20 ≤50 x 109/L; no response (NR) ≤20 x 109/L. After completing therapy, patients were evaluated for platelet count after 1 and 3 months, and thereafter every 3 months until relapse or start of a different treatment. Peripheral blood B lymphocytes were evaluated by flow-cytometry as CD19+ cells before treatment, 1 and 3 months after stopping therapy, and then every 3 months up to recovery.
Results. One month after Rituximab therapy, 7 responses (2 CR, 4 PR, 1 MR; 50%) and 7 NR (50%) were observed. Two relapses occurred 5 and 18 months after response. The median follow-up of all treated patients is 6 months (1.8–34.6), while the median follow-up of all responsive patients is 6.1 months (2–18.7).
Before starting therapy, 11/14 cases were evaluable for flow-cytometry studies. The median baseline value of peripheral blood CD19+ B cells was 137 x 106/L (58–476). One month after completing therapy, 7/9 evaluable cases showed absence of CD19+ cells and 2/9 showed a count of 9 and 4.4 x 106/L CD19+ cells, respectively. At the last available control (median follow-up of 6.8 months; 1.9–32.5), 11/14 evaluable patients had still not recovered the baseline CD19+ cell count (median value: 5.5 x 106/L; 0–287).
The following side effects were observed: 1 case of papulosquamous dermatitis, 2 cases of fever.
Conclusions. Seven out of 14 (50%) ITP patients had an early response to Rituximab (2 CR, 4 PR, 1 MR), that persisted in 5 cases. No late responses were observed. The response was independent of the post-therapy CD19+ cell numbers. No serious infections were observed during the clinical follow-up. No patient had to stop therapy because of severe side effects.
Contribution of the Heparin Neutralization Step to the Anti-PF4 ELISA.
Authors: Dlott Jeffrey S. , Amen Nadia N. , Frock Ruth A. , Chou Peter P. , Gallivan Monica V. , Sahud Mervyn A. 
Published: April 2020

Abstract
The combination of anti-PF4 ELISA and serotonin release assay (SRA) has a reported sensitivity approximating 100% for diagnosing heparin-induced thrombocytopenia (HIT) - 7th ACCP Chest Supplement. Current therapeutic guidelines are to initiate therapy with a direct thrombin inhibitor (DTI) based on clinical parameters and to use HIT assays for diagnostic confirmation. Consequently, timely lab data may potentially minimize unnecessary drug exposure, reduce cost, and decrease morbidity. Due to test complexity, the ELISA results consistently precede the SRAs - the “gold standard” for HIT testing. This study examines the predictive value of the optical density (OD) reading and heparin neutralization procedure (100 IU/ml) for determining positive SRA. 200 patients with orders for the anti-PF4 ELISA (GTI) were retested with and without 100 IU of unfractionated heparin. Confirmation by heparin neutralization was reported as % inhibition (&gt;50% inhibition = positive) as calculated by manufacturer’s instructions.
Negative SRA Positive SRA Negative, OD/0.4 34 2 Positive, OD/0.4 120 44
The sensitivity and specificity of the ELISA was 95.6% and 21.6% respectively. The heparin neutralization step improved the specificity to 38.6% and did not alter the sensitivity. Figure 1 shows a single variable binary logistic regression analysis demonstrating the relationship between OD and probability for a positive SRA. An additional binary logistic regression analysis showed no statistical significance for the % neutralization with heparin. A receiver operators curve (ROC) (Figure 2)demonstrating an optimal sensitivity/specificity was generated for both the neat and the heparin neutralized samples. The curves were virtually identical with the same OD cut-off of 1.008. The specificity was nominally improved with the neutralization procedure.
Figure 1 Figure 1. Figure 2 Figure 2.
The heparin neutralization step only marginally improves the specificity of the anti-PF4 ELISA and does not correlate with a positive SRA result, and therefore, does not justify the added expense. Higher ELISA OD measurements correlate significantly with the probability of a positive SRA.
Bone Marrow Reticulin in Patients with Immune Thrombocytopenic Purpura.
Authors: Mufti Ghulam , Bagg Adam , Hasserjian Robert , Bain Barbara , Kuter David , Dreiling Lyndah , Nichol Janet 
Published: April 2020

Abstract
Immune thrombocytopenic purpura (ITP) is an uncommon disorder, and research that characterizes the bone marrow stroma in ITP patients is lacking. It is known that megakaryocytes are often increased in number in ITP patients. Megakaryocytes are associated with reticulin formation in other conditions, including malignant and nonmalignant diseases, such as systemic lupus erythematosus and other immune disorders. Our objective was to determine if the reactive megakaryocyte proliferations in ITP are associated with increased reticulin deposition. We identified 40 ITP patients from clinical records in a retrospective survey of bone marrow biopsy material. Patients were required to have had a diagnosis of ITP, a bone marrow biopsy with available tissue blocks, and a complete blood count at the time of the biopsy. Paraffin-embedded sections from the bone marrow biopsy specimens were stained for reticulin using standard silver-impregnation methods. Reticulin was quantified using the Bauermeister scale (0 = no reticulin fibers demonstrable; 1 = occasional fine individual fibers and foci of a fine fiber network; 2 = fine fiber network throughout most of the section but no coarse fibers; 3 = diffuse fiber network with scattered thick coarse fibers but no mature collagen; and 4 = diffuse often coarse fiber network with areas of collagen). In addition, bone marrow cellularity and megakaryocyte numbers were evaluated. Of the 40 patients, 13 (33%) were considered to have absent (grade 0) bone marrow reticulin, 1 (2%) had grade 0–1, 20 (50%) had grade 1, 5 (13%) had grade 1–2, and 1 (2%) had grade 2. Thus, reticulin was present in the bone marrow of approximately two-thirds of the patients, with 15% having greater than grade 1. Previous research on bone marrow reticulin in 100 hematologically normal subjects found that 27% had Bauermeister reticulin grade 1 and 4% had grade 2 (Arch Pathol Lab Med1990;114:1241–3). Analysis is ongoing to further characterize the deposition of reticulin in the bone marrow of patients with ITP, the relationship of reticulin deposition to megakaryocyte numbers and location, and correlations with patient clinical findings.
Recombinant Plasmin from Lemna: Purification, Characterization and In Vitro Comparison with Human Plasma-Derived Plasmin.
Authors: Brookhart Gary L. , Tomalski Michael D. , Lounes Karim C. , Meijer Piet , Kluft Cornelis , Radtke Klaus-Peter 
Published: April 2020

Abstract
Background: The fibrinolytic enzyme plasmin has gained renewed interest as a direct thrombolytic agent. Local delivery of plasmin via catheter into thrombi, combined with the efficient neutralization of plasmin that may escape from the clot by its inhibitors α2-antiplasmin (α2-AP) and α2-macroglobulin may provide safe and effective removal of pathologic blood clots.
Objective: To overcome the potential for pathogen contamination associated with plasmin prepared from pooled human plasma, we have generated recombinant plasmin (r-PLM) from transgenic Lemna plants and compared its biological properties with those of plasma-derived plasmin (pd-PLM).
Results: Lemna minor, an aquatic plant, was transfected with the c-DNA encoding human plasminogen, and grown in media under controlled conditions. Lemna-derived plasminogen (r-PLG) was extracted from plant-tissue, separated on lysine-Sepharose and subsequently converted into plasmin (r-PLM), using urokinase-type plasminogen activator (u-PA). r-PLM was further purified to homogeneity using affinity chromatography on benzamidine-Sepharose. r-PLG and r-PLM co-migrate with pd-PLG and pd-PLM on non-reducing SDS-PAGE with the respective molecular weights of about 90,000 kDa and 82,000 kDa. When analyzed on reducing SDS-PAGE, r-PLM, just like pd-PLM, separates into a 56,000 kDa heavy chain and a 26,000 light chain. On Western blots r-PLG and r-PLM cross-react with polyclonal antibodies that were raised against pd-PLG, indicating immunologic identity of r-PLG/PLM and pd-PLG/PLM. All three plasminogen activators (PA), tissue-type PA, u-PA and streptokinase, activate r-PLG. Activity assays using the PLM-specific chromogenic substrate S-2403 show that catalytic rates of r-PLM are indistinguishable from pd-PLM, indicating that in this assay r-PLM and pd-PLM have comparable activity. Purified r-PLM and pd-PLM have similar specific activities. Like pd-PLM, r-PLM activity is inhibited by its physiological inhibitor α2-AP and Kunitz-type inhibitors SBTI and aprotinin. Inhibition of r-PLM by α2-AP results in the formation of a high-molecular weight SDS-stable complex, which migrates on SDS-PAGE with a molecular weight of 150,000 kDa, the cumulative molecular weight of r-PLM (82,000 kDa) and α2-AP (65,000 kDa). In microtiter-based fibrinolysis assays, both pd- and r-plasmin dissolve fibrin clots at the same rate and in a concentration-dependent manner.
Conclusion: This work demonstrates that proteins of the fibrinolytic system can be expressed in Lemna. Lemna-derived r-PLM and pd-PLM have comparable biochemical properties, activity, and are equally sensitive to known plasmin inhibitors. Plant-derived r-PLM may be a valuable alternative to PLM preparations from pooled human plasma.
A Study on the Molecular Mechanism for Hereditary Type II Antithrombin Deficiency in a Chinese Pedigree.
Authors: Xu Ye , Feng Ying , Pang Ying , Jing Peipei , Wei Xie , Zhou Xuhong , Ding Qiulan , Wang Xuefeng 
Published: April 2020

Abstract
Objective: To identify the phenotype and study for the genetic mechanism underlying the recurrence of deep vein thrombosis formation in members of a Chinese pedigree caused by antithrombin (AT) deficiency.
Methods: AT antigen level (AT:Ag) and AT activity (AT:A) were detected with respectively immuno-nephelometry and chromogenic assay. All the seven exons and the flank regions of AT gene form the propositus were amplified by PCR. The purified PCR products were sequenced directly. After mutation was found, the corresponding gene fragments covering the mutated point from the other members of the pedigree were amplified and sequenced.
Results: The plasma level of AT:Ag of the propositus was 29.80mg/dl and those of other 3 members of the kindred ranged from 30.90 to 34.00mg/dl, all were among normal range. The AT:A of the propositus was 56%, and those of the other members were 62% to 68%, all were lower than normal. A heterozygous G13830A missense mutation in exon 6 was identified, which has led to the substitution of histamine (CAT) 393 for arginine (CGT). The sequencing results from the pedigree suggested that all the three other members of the kindred also had the same heterozygous mutation.
Conclusion: The R393H mutation in AT which is similar to the AT Glasgow and AT Avranches can cause reactive site injury in AT so that it is not able to inhibit thrombin anymore. The gene mutation found in this pedigree which was unreported in China is the molecular mechanism for recurrent thrombosis events in two members of the kindred.
Routine Measurement of Endogenous Thrombin Potential (ETP) To Assess Hemostatic Function in Patients with Liver Cirrhosis.
Authors: Gadisseur Alain P. , Schouten Jeoffrey , Franque Sven , Van der Planken Marc , Michielsen Peter , Berneman Zwi N. 
Published: April 2020

Abstract
The maintenance of hemostasis through the production of most of the coagulation proteins is a basic liver function. In patients with cirrhosis of the liver the decrease in these proteins is one of the contributory factors to an increased bleeding tendency. Normally the hemostatic capacity of the liver is measured through routine clotting tests as the activated Partial Thromboplastin Time (aPTT) and Prothrombin Time (PT). In the Child classification for liver cirrhosis the coagulation as expressed by the PT is one of the determinants.
Recently a test has become available which will make it possible to routinely measure the endogenous thrombin generation potential (ETP) which may be a better alternative. In this test according to the method first described by Hemker (1993) thrombin generation is continuously measured by use of a chromogenic substrate. Results are calculated as area under the curve and as a percentage of normal.
We analysed 110 patients with liver cirrhosis classified according to the Child classification, 79 patients with stage A, 19 stage B and 12 stage C, without known pre-existing coagulation abnormalities like inherited bleeding disorders, or anticoagulant drugs. In these patients ETP, APTT, PT/INR, FV, FVII and FXI were measured. We used a fully automated assay for the determination of the endogenous thrombin potential (ETP) on the BCS® System (both Dade Behring, Marburg, Germany).
The results for the mean PT were 81% (CI95 77 - 85) for Child A patients, 53% (CI95 44 - 63) for Child B, and 41% (CI95 33 - 50) for Child C. For the mean INR the results were 1.11 (CI95 1.08 - 1.14) for Child A patients, 1.65 (CI95 1.22 - 2.08) for Child B, and 1.82 (CI95 1.34 - 2.31) for Child C. The results for the mean normalized ETP were 0.87 (CI95 0.84 - 0.90) for Child A patients, 0.78 (CI95 0.68 - 0.88) for Child B, and 0.58 (CI95 0.51 - 0.64) for Child C (p&lt;0.001). While the normalized ETP correlated well with both PT and INR for Child B (P&lt;0.05) and C (p&lt;0.01), there were significant differences within the child A patients where no significant correlation could be identified (Pearson correlation 0.203, R2=0.04, p=0.075.
We conclude that the fully automated assay for the determination of the endogenous thrombin potential (ETP) on the BCS® System (both Dade Behring, Marburg, Germany) is a potentially interesting test to measure the coagulation abilities in patients with liver cirrhosis which in the future may supersede the PT/INR in classification systems for hepatic disease.