bluuebunny/crossref_metadata_2025 huggingface dataset:
Is Homozygosity for the MTHFR C677T Mutation a Risk Factor for Miscarriage? — Likely.
Authors: Altomare Ivy , Aledort Louis M.
Published: April 2020

Various hereditary thrombophilias such as activated protein C resistance, the factor V leiden mutation, the prothrombin G20210A mutation, protein S deficiency, hyperhomocysteinemia, or combinations of the above disorders have been linked to pregnancy loss at varying stages of gestation (Robertson L, et al. Thrombophilia in pregnancy: a systematic review. Br J Haematol2006;132(2):171–96; Seligsohn U, et al. Genetic susceptibility to venous thrombosis. N Engl J Med2001;344(16):1222–31). The literature regarding methylenetetrahydrofolate reductase (MTHFR) mutations is confusing. Much of the evidence supporting a causal relationship between MTHFR mutations and early fetal loss comes from individual case reports with little outcome data. Three recent meta-analyses of thrombophilia in pregnancy concluded that although there may be a trend toward higher risk, the relationship between MTHFR mutations and spontaneous abortions is not truly significant (Robertson L, et al. Thrombophilia in pregnancy: a systematic review. Br J Haematol2006;132(2):171–96; Nelen WL, et al. Hyperhomocysteinemia and recurrent early pregnancy loss: a meta-analysis. Fertil Steril2000;74(6):1196–9; Rey E, et al. Thrombophilic disorders and fetal loss: a meta-analysis. Lancet2003;361(9361):901–8). In contrast, a study supporting the role of MTHFR mutations in recurrent unexplained abortions found a 2–3 fold increased risk of early fetal loss among Caucasian females homozygous for the C677T mutation versus normal controls (Nelen WL, et al. Genetic risk factor for unexplained recurrent early pregnancy loss. Lancet1997;350(9081):861). We present 5 patients with this genetic defect who presented consecutively to our clinic for decision making for future pregnancies. Each had strong histories of miscarriages after the 8th week of gestation. The clinical features of these patients are reported in the table below.

Each woman is Caucasian, and is homozygous for the MTHFR C677T mutation with no other acquired or hereditary thrombophilias. Homocysteine levels were also in normal range for each patient. Our case series of 5 Caucasian women gives support to the theory of a causal relationship between MTHFR mutations and fetal loss. We recommended anticoagulation with low-molecular weight or unfractionated heparin for each patient as a therapeutic intervention to reduce the risk of recurrent abortion in future pregnancies.
A Novel Heterozygous Point Mutation Causing Coagulation Factor XII Deficiency as the Molecular Mechanism of Fatal Thrombosis Event in a Chinese Pedigree.
Authors: Feng Ying , Xu Ye , Pang Ying , Dai Jing , Xie Wei , Zhou Xuhong , Ding Qiulan , Wang Xuefeng
Published: April 2020

Objective: To study on the laboratory diagnosis and molecular mechanism of a pedigree with fatal cerebral thrombosis event caused by hereditary coagulation factor XII deficiency.

Methods: Activated partial thromboplastin time (APTT), prothrombin time (PT), coagulation factor VIII activity (FVIII:C), factor IX activity (FIX:C), factor XI and factor XII activity (FXI:C and FXII:C) were determined in the propositus and the other kindred member for phenotypic analysis. All the 14 exons and their flank sequences of factor XII 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 member of the kindred were amplified and sequenced.

Results: The APTT of the propositus was significantly elongated (>120 seconds), while her PT was normal. Her FVIII:C,FIX:C and FXI:C were all normal. But her FXII:C was only 1 %, lower than normal. A heterozygous G3774C missense mutation in exon 4 was identified, which has led to the substitution of serine (TCT) 73 for cysteine (TGT). The sequencing results from the pedigree suggested that the other asymptomatic member of the kindred also had the same heterozygous mutation.

Conclusion: The Propositus was diagnosed with inherited coagulation factor XII deficiency. The novel gene mutation found in this pedigree is the molecular mechanism leading to the fatal thrombosis events in the propositus.
Weight Adjusted Low-Dose Warfarin Decreases the Incidence of Thalidomide (T) Associated Venous Thromboembolism (VTE) in Patients (pts) with Multiple Myeloma (MM) and Waldenstroms Macroglobulinemia (WM).
Authors: Miller Kena C. , Manochakian Rami , Padmanabhan Swaminathan , Coignet Jean-Gabriel , Musial Laurie , DePaolo Dawn , Chanan-Khan Asher Alban
Published: April 2020

Introduction: T is an effective antimyeloma therapy. An important and problematic side effect of T is development of VTE. The underlying etiology of T associated VTE remains unknown, though a definite rise in its incidence is recorded when combined with other antimyeloma therapies such as dexamethasone (15%) and doxorubicin (26%). Defining an optimal way to prevent T associated VTE is important to improve patient’s quality of life and safety of T. We prospectively investigated the use of weight-adjusted, low-dose warfarin to decrease the incidence of VTE among pts treated with T based regimens.

Methods: All pts treated with T-based therapies, at our institute were given VTE prophylaxis with 1 vs. 2mg warfarin for actual body weight <70 kg vs. ≥ 70 kg, respectively. Concurrent aspirin or other anticoagulants were not given. Pts with underlying pre-existing hypercoaguable disorders were fully anticoagulated with warfarin and were excluded from this analysis. Warfarin was continued throughout the duration of T therapy.

Results: A total of 80 pts, 60 years (range 43–82), 35M and 45F are included in this analysis. Sixty three had stage III and 14 had ≤ stage II MM while 3 pts had WM. T was given with dex in 58 (72%) with dox in 57(71%) and with bortezomib in 32 (40%). Table 1 outlines the T based regimens used. IgG MM was noted in 39, IgA in 20 and 31 had other types of MM. Median T dose was 200 mg (range 50–300) and the median duration 4 months (range 1–6 months). Thirty two pts received 1mg and 48 received 2mg warfarin. Overall incidence of VTE was 6.25% (n=5). There were no bleeding complications noted with the use of warfarin in this pt population.

Conclusion: Our study demonstrates for the first time that a weight adjusted low-dose warfarin approach in MM pts decreases the incidence of T associated VTE and can be safely instituted in pts treated with this important anti-myeloma therapy. Interestingly, none of the pts who received T with Doxil experienced any episode of VTE.

Regimen No. of Patients Thalidomide Alone 1 Dexamethasone + Thalidomide 28 Bortezomib + Doxil + Thalidomide 24 VAD + Thalidomide 22 Bortezomib + Cyclophosphmaide + Dexamethasone + Thalidomide 5
High Incidence of Venous Thromboembolism in Patients with Lymphoma Receiving Chemotherapy: Assessment of Risk Factors.
Authors: Teegala Shyam , Zhou Xiao , Huen Auris , Ji Yuan , Fayad Luis , Romaguera Jorge Enrique , Hagemeister F.B. , Vadhan-Raj Saroj
Published: April 2020

Background: Venous thromboembolism (VTE) is a significant cause of cancer morbidity and mortality. Lymphoma patients (pts) are at increased risk of VTE, however, the exact incidence and risk factors are unknown.

Methods: Of the 1050 newly referred Lymphoma pts to MDACC in 2003 identified, medical records (MR) of 538 consecutive pts were reviewed for demographics, tumor histology, staging, laboratory values, type of chemotherapy (CT) regimens, risk factors for VTE, incidence of VTE and management over a follow up of 2 years.

Results: 207 out of 538 pts received at least one cycle of CT at MDACC (total CT cycles 1125). The median age was 56 years (range 17–82); there were 81 females and 126 males. Majority of pts (61.8%) were newly diagnosed and the most common histologies were Large Cell Lymphoma (31.88%), followed by Hodgkin’s Disease (16.9%) and Follicular Lymphoma (14.98%). Thirteen out of 207 (6.28%) pts had history of VTE prior to CT and 37 (17.9 %) pts out of 207 had 41 new episodes of VTE; 29 Deep Vein Thrombosis (DVT) (12 upper and 12 lower extremity) and 12 Pulmonary Embolism (PE); 2 pts had both DVT and PE. All VTE episodes were confirmed by imaging except in 3 pts. The mean baseline hemoglobin (Hb) in VTE pts was 12.8 g/dL. The median cycle number for VTE occurrence was cycle 3 with 24/37 (64.86%) pts experiencing VTE by cycle 3 and 6/37 (16.2%) pts had VTE in cycle 1. Two out of the 37 (5.4%) pts had recurrent VTE. Among those with new VTE, 31/37 (83.78%) pts were of age greater than 40, 25/37 (67.56%) pts had BMI > 25, 32/37 (86.4%) pts had aggressive or highly aggressive histology and 29/37 (78.37%) pts had stage 3 or 4 disease. Twenty-three out of 31 (74.2%) pts had received erythropoietin before or during the cycle of VTE. Fourteen of 207 (6.79%) pts were on thromboprophylaxis before the chemotherapy. Only 1 of these 14 pts experienced VTE, approximately 2 months after the discontinuation of prophylaxis. Central venous catheter (CVC) thrombosis occurred in 6/174 (3.44%) patients with CVC. The most common systemic treatment for VTE was Enoxaparin (12/33). By multivariate logistic regression analysis of many of the previously described risk factors and other variables, Doxorubicin and/or Methotrexate based CT regimen was found to be a significant independent risk factor for VTE (OR 5.58, 95%CI 1.62 to 19.13. p = 0.0062).

Conclusion: VTE is a frequent and underestimated complication in Lymphoma pts. These findings underscore the importance of prospective clinical trials of anticoagulation prophylaxis in the high risk patients receiving CT.
Influence of ADP in the Course of Thrombin Receptors Activation.
Authors: Lu Xiaoxu , Wu De Pei , Han Yue , Dai Lan , Sheng Wenhong
Published: April 2020

Objective In order to compare the functions of ADP in platelet aggregation and platelet membrane surface glycoproteins expression after thrombin receptors activation, then to investigate the role of ADP in thrombin signal transmission.

Methods Peptide SFLLRN (PAR1-AP, TRAP) and AYPGKF(PAR4-AP)were used for stimulating platelet at different time points (0~30minute) in 12 normal subjects, then the alterations of platelet aggregation and GPIbα, P-selectin were analyzed in the involvement of ADP inhibitor, ApyraseVII, which was observed by aggregometer and flowcytometry respectively.

Results Either PAR1 or PAR4 peptide can lead to platelet activation, inducing absolute platelet aggregation, together with a persistent increase of P-selectin and reversible interlisation of GPIb. Platelet aggregation was partly inhibited by ApyraseVII and showed a reversible curve upon PAR1 activation, and no change upon PAR4 activation. In addition, ApyraseVII accelerated the return of GPIb to platelet surface in PAR1 pathway, produced weak effect on GPIbα internalisation, … ltogether with little effect on GPIb alteration in PAR4 pathway. Meanwhile, no change of P-selectin was obtained in the involvement of Apyrase for both peptides.

Conclusion All the results confirm a critical role of ADP in thrombin signal transmission through the process of PAR1 pathway. ADP induces platelet aggregation and slows down the restoration of GPIb to platelet surface.
Timing and Safety of Postpartum Enoxparin.
Authors: Freedman Rachel A. , Zwicker Jeffrey , Bauer Kenneth Alan
Published: April 2020

The appropriate time to restart anticoagulation in the postpartum period is not known. Both the American College of Obstetrics and Gynecology as well as American College of Chest Physicians (ACCP) have issued guidelines regarding the use of anticoagulants during pregnancy but neither have generated recommendations regarding the timing of the first dose of low-molecular-weight heparin (LMWH) in the postpartum period. The incidence of postpartum hemorrhage was assessed in a retrospective cohort study of 95 women treated with enoxaparin and compared with 303 consecutive deliveries where anticoagulation was not administered. The rate of severe postpartum hemorrhage did not differ significantly for women treated with peripartum enoxaparin versus a control group of women undergoing vaginal delivery (3.6% versus 1.4%, P=0.72) or cesarean section (5.1% versus 3.4%, P=0.98). There were no severe postpartum hemorrhages following the re-initiation of enoxaparin postpartum. In 75% of vaginal deliveries and 49% of cesarean sections, enoxaparin was restarted within 24 hours. Two incisional hematomas were observed in the group of women who received enoxaparin within 24 hours following cesarean section. We conclude that severe postpartum hemorrhage is an infrequent complication following the administration of enoxaparin postpartum. In support of current clinical practice, enoxaparin can be safely administered 0 to 24 hours following vaginal delivery and 12 to 36 hours following cesarean section.
Anti-Platelet Properties of Salivary Secretions of the Avian Leech.
Authors: Wakeman Lisa J. , Munro Roger C. , Al-Ismail Saad
Published: April 2020

Introduction

Haematophages, animals evolved to a bloodsucking lifestyle as their exclusive mode of feeding secrete compounds capable of arresting haemostasis in the host. It is clear that exploitation of host haemostasis is an absolute requirement for the survival of these species. Since the discovery and with the subsequent characterisation and engineering of Hirudin a potent thrombin inhibitor from the European medicinal leech Hirudo medicinalis, attention has been focused on the potential anticoagulant and platelet aggregation inhibitors derived from an array of different species of leech from both the Rhynchobdellid and Arhynchobdellid orders. Haematophagous leeches of the genus Theromyzon sp. of the Rhynchobdellid order, also termed duck leeches, feed directly on the nasal passages, trachea and nictating membranes of migratory birds. We present the novel observation of inhibition of aggregation of human platelets by the salivary secretion extracts of the avian leech Theromyzon tessulatum.

Methods

Twelve adult leeches of the species T. tessulatum (total weight 2.828g) were anaesthetised with ethanol vapour. The leeches were severed at the anterior end and a homogenate produced containing salivary gland secretions. The posterior two thirds of the leeches were treated identically to serve as control material. Platelet rich plasma (PRP) was prepared from blood from a normal individual (free from known platelet modifying medicines) mixed 9:1 (v:v) with 0.105 M trisodium citrate in siliconised glass vacutainers. Platelet numbers were adjusted with autologous platelet poor plasma (PPP) to obtain concentrations of approximately 300 × 109/L. Leech extracts (anterior or posterior control) were added to PRP at a ratio of 1:4. Aggregation studies were performed using thrombin (10units/ml), collagen 10μg/ml, Ristocetin (1.5mg/ml) and ADP (5μm/ml).

Results

Data from this study shows that platelet aggregation was completely inhibited when stimulated by thrombin, collagen and ADP and partially inhibited (40%) on the addition of ristocetin.

Conclusion

Our observations contradict the belief that the anti-thrombocyte properties of this species of haematophagous leech are restricted to duck thrombocytes. We suggest the presence of one or more inhibitory molecules acting by various mechanisms including inhibition of vWF and platelet integrin mediated collagen interactions, inhibition of ristocetin mediated vWF and platelet GPIb receptor binding and salivary secretion derived apyrase inhibition of arachidonic acid mediated platelet aggregation.

These findings provide conclusive evidence that this blood sucking bird leech has the ability to overcome thrombocyte function in higher vertebrates.
Is Age Related Fibrinogen Affected by Warfarin?.
Authors: Munro Roger C. , Wakeman Lisa J. , Al-Ismail Saad
Published: April 2020

Introduction: There is published evidence which indicates that advancing age may be associated with higher plasma concentrations of fibrinogen. There is also evidence that derived fibrinogen values are significantly higher than Clauss measurements and that this discrepancy is greater in patients receiving warfarin. The purpose of this study was to determine whether age related derived fibrinogen levels are similar in both warfarin and non-warfarin groups.

Methods: Venous samples were collected into siliconised glass B-D Vacutainers containing tri-sodium citrate (Ref: 367691) from 1000 patients receiving long term warfarin treatment and an equal number of age-matched patients not receiving warfarin. Genders were equally represented in both groups. Patients in both groups were categorized into 5 years age bands as follows: <40 n=23: 40–44 n=20: 45–49 n=43; 50–54 n=74: 55–59 n=113: 60–64 n=155: 65–69 n=178: 70–74 n=191: 75–79 n=124; 80–84 n=56: 85–89 n=23. Derived fibrinogen was measured in each patient on an ACL300R coagulometer (I L) within 1 hour of collection using IL PT-FIB HS Plus reagent and following the manufacturer’s protocol. Appropriate CLSI guidelines were followed throughout. A normal probability plot of the data was performed to confirm that it did not deviate too much from the normal distribution.

Results: The T-test for independent samples using the separate variance estimate showed that there was a statistically significant difference in the mean fibrinogen between patients on warfarin and those not on warfarin (p<0.05) in each group except for the last (85–89 years). There was a statistically significant difference (ANOVA) in the fibrinogen levels of patients of different age in both warfarinised and non-warfarinised groups (p<0.05). The modified least significance procedure in the ANOVA test showed that in the non-warfarin group, most of significant difference in fibrinogen between the different age groups is contributed by the difference between patients under 50 years of age. In the non-warfarin group, it requires an age gap of at least 20 years for the difference in fibrinogen to be statistically significant but in the warfarin group, it only requires an age gap of ten years (p<0.05). Both Linear Regression and Cross Tabulation indicate that the relationship between fibrinogen and age does not vary whether or not the patient is on warfarin. These also show that the effect of age on fibrinogen is not affected by warfarin treatment.

Conclusion: Differences or correlations detected in this analysis are of statistical significance but not necessarily clinically significant. Placing age and warfarin treatment in the same model shows that variations in fibrinogen have to be explained by other factors (e.g. technical) not included in the study as only 12% of the error in predicting fibrinogen levels can be reduced by knowing both the age and status of warfarin treatment in individual patients.
High Incidence of Venous Thromboembolism (VTE) in Patients with Cancer Hospitalized at a University Hospital in Chile. Efficacy of Pharmacological Thromboprophylaxis.
Authors: Conte Guillermo F. , Figueroa Gaston L. , Altmann Arie D. , Donaire Luisa A.
Published: April 2020

Inpatients are at increased risk of VTE due to multiple factors. Cancer diagnosis is an important risk factor determined through prospective studies. The aims of this study are to know the incidence of VTE in cancer patients hospitalized due to acute medical conditions and describe the use of pharmacological thromboprophylaxis and its efficacy.

Methods: Retrospective analysis of cancer patients admitted to the University of Chile Clinical Hospital due to acute medical conditions between 2003 and 2004. Exclusion criteria: VTE diagnosed at admission, oral anticoagulant therapy at admission, age <18 years, admission to Intensive Care Unit (ICU). It was necessary to confirm VTE diagnosis by ultrasonography or angio-CT scan.

Results: Data of 366 patients was retrieved. The cancer origin was: gastric (38%), lung (19%), colorectal (15%), breast (10%), hepatocarcinoma (5%), others (13%). Seventy-seven percent of the cases presented an advanced disease (stage TNM III-IV). The main diagnoses at admission were: pneumonia (17%), vomits/dehydratation (16%), urinary tract infection (7%), decompensated diabetes mellitus (7%), digestive hemorrhage (7%). In 125 cases (34%) no type of pharmacological thromboprophylaxis was used; unfractionated heparin was used in 120 (33%) (5000 U sc c/8–12 hr) and low-molecular-weight heparin (dalteparin or enoxaparin) in 121 (33%). The VTE incidence was 3% (11 cases). In patients who did not receive thromboprophylaxis, the VTE incidence was 6.4% (8/125) versus 1.2% (3/241) in those who were administered heparin or LMWH (Odds Ratio=0.18 CI 95% 0.05–0.65). The group of patients who did not receive thromboprophylaxis was younger (median 60 vs. 65 years), had a higher frequency of thrombocytopenia (<150.000; 39% vs. 15%) and hypoprothrombinemia (INR>1.5; 35% vs. 14%), and a lower frequency of recent oncologic surgery (3% vs.19%).

Conclusions: Cancer patients with acute medical conditions showed a high incidence of symptomatic VTE (3% in this series). One third of patients were not administered pharmacological thromboprophylaxis. The use of thromboprophylaxis in these high-risk patients was associated to a significative reduction of the VTE frequency.
A Randomized, Double-Blind Trial of the Interaction between Cranberry Juice and Warfarin.
Authors: Ansell Jack , McDonough Maryellen , Harmatz Jerold S. , Greenblatt David J.
Published: April 2020

Therapeutic warfarin therapy is highly influenced by concomitant drug therapy or dietary factors by effecting warfarin metabolism or vitamin K status. Cranberry juice (CJ) was recently implicated to influence the INR in patients on warfarin based on a number of brief anecdotal or case reports. This study was designed to determine what influence, if any, daily cranberry juice consumption has on INR levels and plasma warfarin concentrations in patients stably anticoagulated with warfarin. Patients on warfarin for a variety of indications from a population of anticoagulation clinic patients were selected because of their past history of stable anticoagulation and willingness to participate in the study. Major inclusion criteria required a therapeutic INR range of 2.0 – 3.0, no hepatic or renal dysfunction, a stable INR (1.7 – 3.3) on at least 2 measurements within 8 weeks prior to study entry, and informed consent. The study consisted of a 2 week lead-in phase (with weekly INRs in range); a 2 week intervention phase (double-blind daily consumption of an 8 oz glass of cranberry juice or placebo); and a one week follow-up phase (35 day total). Any patient with an out of range INR during the intervention phase ceased the intervention but continued through follow-up. 30 patients were randomized, 14 to CJ and 16 to placebo. 28.6% (4/14) in the CJ group developed a minimally elevated INR and 7.1%(1/14) developed a low INR during the intervention phase. 25% (4/16) in the control group developed a minimally elevated INR during the intervention. There was no significant difference in the mean INRs from each group on each day of measurement except for day 11 of the intervention phase when the CJ group was significantly higher than the placebo group (p<0.02), although on day 15 (end of intervention) the mean INRs from both groups were identical. Plasma warfarin levels are currently pending and should be helpful in identifying any effect on warfarin clearance or warfarin plasma protein binding. A daily 8 oz glass of cranberry juice appears to have no consistent effect on the INR in patients on warfarin therapy compared to placebo.
Cost-Consequences of Fondaparinux Use for Treatment of Heparin-Induced Thrombocytopenia.
Authors: Kanaan Abir O. , Donovan Jennifer L. , Belliveau Paul P. , Roy Tara , Al-Homsi Ahmad-Samer
Published: April 2020

Heparin-induced thrombocytopenia (HIT) is a severe complication of therapy with heparin and low molecular-weight heparins and is associated with increased morbidity and mortality. Furthermore, HIT constitutes an economic burden on health care systems. Direct thrombin inhibitors (DTIs) lepirudin and argatroban; the approved agents for HIT treatment; are expensive and require intravenous administration with substantial use of resources and increased duration of hospital stay. Indeed, the cost of management of HIT utilizing DTIs in heart surgery has been estimated to range from over $100 to $300 million dollars nationwide. Published case reports, abstracts, and our own data support the use of fondaparinux, a pentasaccharide, in the management of HIT as an effective and safe alternative to DTIs. We evaluated the cost associated with fondaparinux use in a series of patients with HIT treated at our tertiary care teaching institution. We performed a retrospective review of 18 patients with HIT receiving fondaparinux during study period from January 2002 through June 2006. HIT diagnosis was found in 77.7% of patients during their hospital stay and in 22.2% at admission. Ten patients were initially treated with argatroban (9) or lepirudin (1) before switching to fondaparinux due to concerns of increased risk of bleeding mandating the use of an anticoagulant with short half-life or to renal failure prohibiting the use of fondaparinux. The total cost for the group including actual direct variable (room, pharmacy, lab, and supplies), actual direct fixed, and indirect costs was $1,484,297 and the mean was $82,462±70,153. The actual direct variable cost was $888,461 with a mean of $49,359±43,047 per patient. The actual direct fixed cost was $344,563 with a mean of $19,142±15,257. The indirect cost was $251,273 with a mean of $13,960±12,094. In this series, the daily cost of anticoagulant per patient based on average wholesale price (AWP) $407. This compared favorably to a daily cost per patient of $1093.76 for argatroban and $1732.8 for lepirudin. The total cost of drug for all patients was $70,414 as opposed to an estimated cost of $189,220 or $299,774 should argatroban or lepirudin have been used, respectively. This translated into an estimated savings from $129,000 to $229,000. In summary, the utilization of fondaparinux in HIT treatment at our institution resulted in a dramatic drug cost reduction. Coupled with its subcutaneous route of administration and the absence of need for laboratory monitoring, allowing reduction in hospital stay and spared resources, fondaparinux would represent an attractive alternative to DTIs in the management of HIT pending further clinical studies to confirm its safety and efficacy.
Irradiation of Cellular Blood Components with Cobalt 60 Is Very Efficient and Safe in the Prevention of Transfusion Associated Graft Versus Host Disease (TA-GVHD) in the Allogeneic Transplant Setting.
Authors: Karduss Amado J. , Leon Eduardo , Buitrago Lina , Gomez Luis R. , Jimenez Alejo , Reyes Pedro , Coll Yadira , De Ossa Hilda
Published: April 2020

For the prevention of TA-GHVH in patients who received a allogeneic stem cell transplant is mandatory the gamma irradiation of the all cellular blood components. This irradiation is usually done with Cesium 137 and with a special blood bank irradiators. However these devices are expensive; because that, in developing countries, is frequent the utilization of Cobalt 60 and the same device that is used in the radiotherapy department, instead of blood bank irradiators. We present our experience with this technique.

From Dec 2002 to Dec 2005 thirty patients received a allogeneic stem cell transplant and 28 were analysed. The stem cells source was: peripheral blood 25, unrelated cord blood 2, and bone marrow 1. The irradiation of the blood was performed with Cobalt 60–1.24 Mev- (theratron 780 C); the irradiation field was calculated for covering all of the bag surface and a dose of 3.5 Gy was administered to the mild plane of the bag.

158 blood concentrates were transfused, 68 red cell (X:2.5 per patient), and 90 platelets (3.2 per patient). The pre transfusion median hemoglobin and platelet levels were 7.63 g/dl and 12.000/ul; after transfusion was a median increase of 2.3 gm/dl (0.6–4.7) in hemoglobin and 18.000/ul (0–140.000) in platelets.

There was no any case of TA-GVHD. Four patients developed pos transplant aGVHD, in all of the cases the disease began 50 days or more after the last transfusion, there were no pancytopenia and the aGVHD was resolved completely with the treatment.

Conclusion

In receptors of allogeneic stem cell transplant the gamma irradiation of blood components with Cobalt 60 and with the same device which is used for patients radiotherapy is 100% effective and safe in the prevention of TA-GVHD. This is a good alternative in centers without blood bank irradiator
Review of Serologically Confirmed Cases of Transfusion-Related Acute Lung Injury Reported to a Blood Center.
Authors: Houseworth Julie L. , Rhamy Jennifer , Yarnell Vicki , Parks Jodi A. , Smith Daniel S. , White Linda , Waxman Dan A.
Published: April 2020

We reviewed 13 cases of serologically confirmed Transfusion-Related Acute Lung Injury (TRALI) reported to our blood center over a five-year period. Cognate antigen-antibody pairs were noted, as well as product transfused, indication for transfusion, onset of symptoms related to transfusion and time to resolution. All patients demonstrated bilateral pulmonary edema on CXR. Onset of symptoms for all cases occurred during transfusion of implicated product, or within one hour post-transfusion. All donors of implicated products were multiparous females. The majority of implicated products (77%) were FFP. There were 3 TRALI deaths, all resulted from transfusion of FFP. Anti-neutrophil, HLA Class I and/or Class II antibodies were identified in all fatal cases. Platelets comprised 15% of implicated products, and RBC’s only a single case (7 %). Among patients with TRALI receiving FFP, the most common indication was reversal of anticoagulation (50%). Chart review in two cases indicated Vitamin K in lieu of FFP may have been appropriate.

Comparison of Serologically Confirmed TRALI Cases Patient Product Reason for Tx Time to Resolution Neutrophil Antibody HLA I HLA II HLA PRA 1 FFP Reverse anticoag. Death @ 7 hours HNA-3a Neg. Neg. 0 2 Platelet Thrombocytopenia <1 day + Neg. Neg. 0 3 FFP Reverse anticoag. 2 days + Neg. Neg. 0 4 Platelet Pancytopenia <1 day + Neg. Neg. 0 5 FFP Reverse anticoag. 4 days + A2 B3 DR 52 I - 90% II - 56% 6 FFP Not available unknown + Neg. Neg. 0 7 FFP Periop. Bleeding unknown Not tested A2 Neg. 40% 8 FFP Reverse anticoag. 2 weeks Not tested Neg. DR 17 63% 9 Cryopoor plasma TTP 4 days Not tested Neg. DR 52 41% 10 FFP Reverse anticoag. Death @ 4 hours Not tested B 50 DR 16 I - 95% II - 89% 11 RBC Bleeding 2 days Not tested Neg. DR 52 60% 12 FFP GI Bleed Death Not tested Neg. DR 53 DQ 5 60% 13 FFP GI Bleed 2 days Not tested Neg DR 52 60%

In our small series, the majority of implicated products were FFP, and all products were from multiparous females. Production of FFP exclusively from male donors is currently under consideration. Although male donor plasma can be expected to reduce the risk of TRALI associated with transfusion of FFP, it will not eliminate it entirely. Therefore, the risk of TRALI should be factored into the decision to transfuse all products. FFP should be administered cautiously for all indications, including reversal of anticoagulation.
False Positive Pregnancy Test Resulting from Passive Transfusion of beta-Human Chorionic Gonadotropin (bHCG) from Donor Red Blood Cells.
Authors: Shelat Suresh G. , Friedman David F. , Smith-Whitley Kim , Meny Geralyn
Published: April 2020

Blood products may contain donor-derived soluble factors such as allergens or antibodies, which when transfused, may cause pathology in the recipient or unexpected test results.

Case: We describe a 16 year old female with hemoglobin SC disease who was referred for erythrocytapheresis to reduce the level of sickle hemoglobin prior to a surgical procedure that required general anesthesia. The patient was exchanged with five units of packed red blood cells (PRBC). The following day, routine pre-operative laboratory testing revealed an unexpected positive result in assays for beta subunit of human chorionic gonadotropin (bHCG), an early indicator of pregnancy. The patient denied sexually activity. The result caused postponement of the elective surgical procedure and prompted an abdominal ultrasound study to identify the source of bHCG, which was normal.

Methods: Immunoassay was used to determine the bHCG level in the patient (Table 1) and in the residual plasma in each of the transfused units (Table 2).

Conclusion: One blood donor, who was unaware of her pregnancy status at the time of donation, harbored a high bHCG level which caused the positive test results in the recipient patient’s serum and urine. If an unexpected result is detected in a recipient of a blood transfusion, it is important to consider the possibility of passive transfusion of the analyte.

Table 1: bHCG test results from recipient’s blood following transfusion. Test Result Negative cut-off Qualitative bHCG (serum) Positive < 10 mIU/ml Qualitative bHCG (urine) Positive < 20 mIU/ml Quantititative bHCG (serum) 9 mIU/ml < 5 mIU/ml Table 2: bHCG levels in the segments from the PRBC units transfused. Unit number bHCG, mIU/ml* *Negative cut-off, <5 mIU/ml; **Exceeds the dynamic range (0–1000 mIU/ml) of qualitative bHCG assay. 1 0.39 2 0.71 3 >1000** 4 0.18 5 0.24
Fetal Liver Hematopoiesis Becomes Sensitive to Estrogen after Transplantation to Adult Mice.
Authors: Chang Ko-Tung , Pelichovska Tereza , Sefc Ludek , Broulik Petr , Necas Emanuel
Published: April 2020

The hematopoietic stem/progenitor cells from fetal liver (FL) of the mouse do not express estrogen receptors in contrast to the cells from adult bone marrow (BM) [Igarashi H et al: Proc Natl Acad Sci USA (2001) 98, 15131–15136]. Since estrogen had been shown to suppress B-lymphopoiesis in vivo [Medina KL et al: Blood (2000) 95, 2059–2067; Erlandsson MC: Immunology (2003) 108, 346–351], we investigated estrogen action on B-lymphopoiesis derived from FL cells transplanted to adult mice. FL cells from E14.5 embryos of C57Bl/6 mice (Ly5.1), or a mixture of FL cells (Ly5.1) with BM cells (Ly5.2), were transplanted either into sublethally (7 Gy) or into lethally (11 Gy) irradiated male Ly5.2 recipients, respectively. Half of the recipients were kept on a diet containing estradiol (estimated intake of 100 micrograms/day). Peripheral blood obtained from the retrobulbar venous plexus was analyzed for presence of Ly5.1 (FL origin) or Ly5.2 (BM origin) WBCs using flow cytometry (FACS Calibur, Becton Dickinson) from 2 weeks up to 16 weeks after transplantation. The WBCs were also stained with anti-mouse CD45R/B220 antibody to detect B-lymphocytes. Numbers of repopulating B-lymphocytes of FL origin were little affected by estrogen 2 and 4 weeks after transplantation while those of BM origin were significantly suppressed. However, the difference between responses of FL or BM origin B-lymphopoiesis to estrogen became much smaller after 8 weeks and was not evident after 16 weeks. The chimeric bone marrow was further transplanted to secondary lethally irradiated recipients and effect of estrogen on B-lymphopoiesis of either FL or BM origin was studied for another 20 weeks. In this case B-lymphopoiesis of FL origin was suppressed by estrogen immediately after transplantation similarly as that of BM origin. The results provide a functional confirmation of previously reported molecular differences between FL and adult BM stem/progenitor cells dwelling in lack of the expression of estrogen receptors in the FL cells. They also conclusively demonstrate that FL stem/progenitor cells become invariably sensitive to estrogen after being exposed to the environment of the adult organism for a long enough time.
Mortalin-2 (mthsp70/PBP74/Grp75) Mediates the Erythropoietin Signal To Stimulate Growth of Purified Human Erythroid Progenitor Cells.
Authors: Ohtsuka Rie , Abe Yasunobu , Fujii Tomomi , Nishimura Junji , Muta Koichiro
Published: April 2020

Erythropoietin (EPO) stimulates erythroid growth by enhancing the proliferation, maturation and survival of late stage erythroid progenitor cells. Intracellular signaling molecules such as Janus kinase 2 (JAK2) and phosphoinositide-3 kinase (PI-3K)/Akt are considered mediators of the EPO signal; however, the entire process of EPO stimulation remains undetermined. Previously, we used siRNA to show that mortalin-2, which is a member of the heat shock protein 70 family of chaperones, mediates EPO signaling to stimulate the growth of human erythroid colony forming cells (ECFCs). In the present study, we examined the relationship between cell growth and mortalin-2 overexpressed in ECFCs, and analyzed the gene expression pattern of ECFCs treated with mortalin-2 siRNA using a DNA microarray, to further clarify the intracellular mechanism by which EPO and mortalin-2 interact. In the presence of different concentrations of EPO (0–1.0 U/ml), the effect of mortalin-2 overexpression on ECFC growth was determined by MTT assay. There was tendency to better viability of ECFCs treated with mortalin-2 expression vectors than control cells, especially in the EPO 0 U/ml group (p=0.08). BrdU ELISA, used to investigate the effects of mortalin-2 on the DNA synthesis of ECFCs, revealed that when ECFCs were treated with mortalin-2 expression vectors, the cells showed an increase in the amount of BrdU incorporation into DNA without EPO. Next we analyzed the gene expression pattern using mRNA obtained from ECFCs cultured with or without EPO after treatment with mortalin-2 siRNA or control siRNA. When ECFCs were cultured with EPO after treatment with mortalin-2 siRNA, the expression of 19 genes was suppressed to less than 0.6 fold, and these genes included those involved in cell growth, apoptosis or transport, such as interleukin 6 receptor, ATP-binding cassette, Mdm2, BCL2 interacting protein and interleukin 10 receptor alpha. Furthermore, the expression of 8 genes was upregulated to over 1.5 fold, and these genes included transcription or signal transmission related genes, such as ubiquitin A-52 residue ribosomal protein fusion product 1 and serum/glucocorticoid regulated kinase 2. There were also some genes whose expressions overlapped with genes obtained from ECFCs cultured without EPO after treatment with control siRNA. Our data suggests that mortalin-2 expression depends on various pathways, and that one of these pathways mediates EPO signaling to stimulate mortalin-2 expression, which is related to the growth of erythroid progenitor cells.
A Novel Role of Complement Components C1q and C5a in Hematopoietic Stem/Progenitor Cell Migration.
Authors: Jalili Ali , Marquez-Curtis Leah , Montano Jencet , Shirvaikar Neeta , Ratajczak Mariusz Z. , Janowska-Wieczorek Anna
Published: April 2020

The complement protein C1q primarily provides a recognition and activation signal that triggers the classical pathway of complement, but it also has multiple immune functions including acting as a chemoattractant for neutrophils, eosinophils, mast and monocyte-derived dendritic cells. Moreover, hematopoietic stem cells have recently been shown to express the C1q receptor, C1qRp. C5a also strongly chemoattracts monocyte/macrophages to inflammatory sites. Recently we showed that G-CSF mobilization activates complement by a classical IgM-dependent pathway (Blood2005;106:1976a). In this study we examined the possible roles of C1q and C5a in hematopoietic stem/progenitor cells (HSPC) migration. We found that C5aR was expressed at the mRNA level on mobilized peripheral blood (mPB) and cord blood (CB) CD34+ cells but at the protein level only on mPB CD34+ cells. Flow cytometry revealed high C1qRp expression on both mPB and CB CD34+ cells. When the expression of C5aR and C1qRp was examined on the CB CD34+ cells expanded towards myeloid, megakaryocytic and erythroid lineages (at days 0, 3, 6, 11 and 14 of expansion), we found that C5aR expression increased with cell maturation (days 3–14) in both myeloid and megakaryocytic progenitors. In contrast, C1qRp was highly expressed on day 0, stayed constant in myeloid and megakaryocytic cells (days 3–11), and was down-regulated by day 14 in megakaryocytic cells. C1qRp was down-regulated in erythroid precursors during their maturation. We also found that C5a but not C1q is a chemoattractant for mPB CD34+ cells. In chemotaxis assays towards an SDF-1 gradient,

C5a primed chemotactic responses of both mPB and CB CD34+ cells to a low (10 ng/mL) gradient of SDF-1 (up to 80% of their response to a high (200 ng/mL) SDF-1 gradient); C1q primed the chemotactic responses of both types of CD34+ cells (up to 100% of the response to a high SDF-1 gradient), and the priming effect of C1q on SDF-1-induced chemotaxis of expanded myeloid and megakaryocytic precursors decreased, consistent with down-regulation of C1qRp on these cells by day 14.

Hence these results indicate that HSPC and progenitor cells express functional C5a and C1q receptors and that both the C5a-C5aR and C1q-C1qRp axes sensitize the responses of these cells to SDF-1 and thus could play a role in HSPC homing/mobilization to bone marrow. Further studies in animal models are needed to elucidate the roles of C5a and C1q in HSPC trafficking.
Myeloid Lineage Restriction Is Prior to and Independent of Lymphoid Lineage Restriction.
Authors: Matsubara Azusa , Ooehara Jun , Yamazaki Yuji , Takano Hina , Nakauchi Hiromitsu , Ema Hideo
Published: April 2020

It has been widely accepted that hematopoietic stem cells (HSCs) exclusively give rise to either myeloid or lymphoid lineage along their differentiation. Phenotypically defined as FcRloCD34+IL-7R−Sca-1−Kit+Lin− cells, common myeloid progenitors (CMPs) are supposedly at the first branched point for myeloid lineage commitment. Phenotypically defined as Thy1.1−IL-7R+KitloSca-1loLin− cells, common lymphoid progenitors are supposedly at the first branched point for both B- and T-lymphoid lineage commitment. Contradicting this model, we previously made observations of the myeloid lineage restriction in very early stages of HSC differentiation. We therefore decided to compare the myeloid and lymphoid differentiation potentials in long-term HSCs, short-term HSCs, multipotent progenitors (MPPs), CMPs, granulocyte/macrophage progenitors (GMPs), megakaryocyte/erythrocyte progenitors (MEPs), and CLPs at the single cell level. Because any available assays are not so sensitive enough for detection of lymphoid lineage differentiation potential, we primarily focused on myeloid differentiation potentials in this study. Here we provide data indicative of the absence of CMPs in the adult mouse bone marrow. Long-term HSCs (CD34−Kit+Sca-1+Lin− cells), short-term HSCs (Flt-3−CD34+Kit+Sca-1+Lin− cells), MPPs (Flt-3+CD34+Kit+Sca-1+Lin− cells), CMPs (FcRloCD34+IL-7R−Sca-1−Kit+Lin− cells), GMPs (FcRhiCD34+IL-7R−Sca-1−Kit+Lin− cells), MEPs (FcRloCD34−IL-7R−Sca-1−Kit+Lin− cells), and CLPs (IL-7R+KitloSca-1loLin− cells) were purified from adult B6 mice by flow cytometry. Single cell cultures were performed in the presence of SCF+TPO+IL-3+EPO. All colonies made by single cells were subjected to Cytospin preparations, followed by May-Gruenwald-Giemsa staining, for morphological classifications of colony cells: neutrophil (n), macrophage (m), erythroblast (E), or megakaryocyte (M). Cells with all n, m, E, M differentiation potentials (nmEM cells) were detected in on average 31%, 5%, and fewer than 1 % of long-term HSCs, short-term HSCs, and MPPs, repectively. Notably, none of a total of over 800 CMPs showed the full nmEM differentiation potential. These CMPs instead showed potentials belonging to members of GMPs and MEPs, suggesting CMPs are mostly an overlaping population of GMPs and MEPs rather than a distinct population. Single cell transplantation experiments revealed the coexistence of cells with the myleoid and B-lymphoid potentials or cells with the myeloid and T-lymphoid potentials among CD34−Kit+Sca-1+Lin− long-term HSC population. These progentiors are likely to be immediate progeny of HSCs. Together, these data support our view that myeloid lineage restriction takes place prior to and independent of lymphoid lineage restriction.
A High Dose of Erythropoietin Induces the Expansion of CD34+CD133+ Cells in Human Umbilical Cord Blood.
Authors: Noguchi Mitsuho , Tashiro Haruko , Shirasaki Ryosuke , Gotoh Moritaka , Kawasugi Kazuo , Shirafuji Naoki
Published: April 2020

Objectives: We compared the responsiveness to the high dose stimulation of human EPO between the transplant grafts in an in vitro culture system to clarify the characteristics of cord blood and to make clear the repopulation characteristics on human hematopoietic stem/progenitor cells.

Methods: Cells (cord blood, bone marrow and peripheral blood stem/progenitor cells) were cultured for six days with G-CSF (100 ng/ml), EPO (10 U/ml), or without additives, and were sorted with antihuman CD133-1 monoclonal antibody. The obtained fractions were further labeled with antihuman CD34 monoclonal antibody and CD133-2 antibody as well as the lineage markers, and their expressions were analyzed. Cells were also assayed with ordinal colony-formation. For the analysis of long-term culture-initiating cells (LTC-ICs), Dexter’s long-term culture method was carried out. The expression of the specific proteins was also analyzed with RT-PCR method.

Results. The ratio of the CD34+ cell count to the total cultured mononuclear cell count was not changed between CB, BM and PBSC. However, the ratio of the CD133+ cell count to the total cultured mononuclear cell count was 1.4-fold that of when CB cells were cultured with a high dose of EPO. No significance was observed for BM and PBSC. The count of CD34+CD133+ cells was 1.9-fold that of when CB was cultured with EPO. Dexter’s long-term culture system demonstrated that CB cultured with EPO contained more LTC-ICs and colony-forming cells with a high proliferation potential than that cultured without additives.

Discussion: These results indicate that the CB has unique characteristics to respond to a high dose of EPO.
The Role of Telomerase Activity and Telomere Length in Cellular Sensitivity to DNA Damaging Agents.
Authors: Rice Greg T. , Beasley Michael A. , Akabogu Ike I. , Westin Erik R. , Winnike Dale A. , Klingelhutz Aloysius J. , Goldman Frederick D.
Published: April 2020

Dyskeratosis congenita (DC) is a premature aging syndrome characterized by progressive bone marrow failure, abnormal skin pigmentation and nail dystrophy. We have described an autosomal dominant form of DC (AD DC) in a large three-generation kindred that is due to a mutation in the gene encoding human telomerase RNA (hTR). While telomere shortening is a normal consequence of the aging process, DC patients display extremely short telomeres in many somatic cell types, including hematopoietic cells, and they often suffer from bone marrow failure. Allogeneic hematopoietic stem cell transplant (HSCT) remains the only curative therapy for marrow failure in DC. However, HSCT in DC is generally poorly tolerated and associated with significant morbidity, perhaps as a consequence of increased sensitivity of dividing cells to cytotoxic agents during myeloablative therapy. To test this hypothesis, we characterized lymphocytes from various AD DC patients and age matched controls that had been placed in long term culture following in vitro exposure to irradiation (137Cs) and varying doses of Taxol, Adriamycin, and Etoposide. Cell proliferation and viability were quantified by direct visual counting on a hemocytometer, and flow cytometry was employed to assess apoptosis and cell surface expression of senescent markers. In addition to DC lymphocytes having a decreased proliferative capacity and higher basal apoptotic levels, an increased sensitivity to irradiation, Taxol, Adriamycin, and Etoposide was noted. These results suggest that telomere shortening may be an important factor in determining cellular tolerance to cytotoxic therapy and support the concept of reduced intensity HSCT regimens in both aged individuals and DC patients. Further studies have been initiated to determine whether reconstitution of telomere length in DC cells alters response to cytotoxic agents.