bluuebunny/crossref_metadata_2025 huggingface dataset:
Are Statistically Non-Significant Findings Necessarily Negative? A Review of All Phase III Randomized Controlled Trials in Hematology Conducted by NCI Sponsored Cooperative Groups.
Authors: Kumar Ambuj , Soares Heloisa , Djulbegovic Benjamin
Published: October 2019

Background: Randomized clinical trials (RCT) remain the primary mean for advancement of new treatments for the prevention and cure of hematological malignancies. However, if a RCT fails to show a significant difference between the treatment and the control group, is it because treatment is not effective (evidence of absence of treatment effect) or the results were inconclusive (absence of evidence of treatment effect).

Methods: We identified 143 studies related to hematological malignancies out of 566 trials conducted by 6 NCI sponsored cooperative groups (RTOG, COG, GOG, ECOG, NSABP and NCCTG). Data were extracted from the original research articles and protocols. Quality of trials was high. To determine whether non-significant trial findings were true negatives or inconclusive, we compared the a priori effect size used in the power calculation for the trial’s pre-defined primary outcome to the confidence intervals (CIs) of the non-significant trial results. Trial results were construed as true negative if the effect size and the 95%CIs were entirely outside the limits of equivalence and inconclusive if the 95%CIs crossed the line of no effect and one or both limits of equivalence (BMJ2004;328:476–477).

Results: 71% of studies resulted in non significant outcome (102/143, missing data for 11 studies). Comparing the calculated CI to the a priori specified effect size 48% (44/91) of studies resulted in true negative results (evidence of no effect) and 52% (47/91) were inconclusive (no evidence of effect). Most of the studies (73%, 66/91) had conducted a power analysis before the commencement of trials. The range of expected effect size for experimental treatments in this set of trials ranged from as low as 9% to high as 200% (improvement in survival).

Conclusion: High-quality RCTs conducted by the most prestigious institutions often result in inconclusive or negative findings. Unrealistic expectation in treatment effect size appears to be a major culprit for continuing conduct of underpowered trials.
Oral Mucositis (OM) and Outcomes of Allogeneic (AL) Hematopoietic Stem Cell Transplantation (HSCT) in Patients with Hematologic Malignancies.
Authors: Vera-Llonch Montserrat , Oster Gerry , Ford Colleen , Lu John , Khazanov Irina , Sonis Stephen
Published: October 2019

Background. OM is a common toxicity of conditioning regimens for HSCT that has been associated with adverse clinical and economic outcomes.

Methods. A retrospective chart review study of ~400 consecutive HSCT recipients with hematologic malignancies was undertaken at a single academic center. OM severity was assessed across eight oropharyngeal sites using a validated scale which was scored as follows: no erythema/ulceration = 0; erythema only = I; ulceration one site = II; two sites = III; three sites= IV; and four or more sites = V. OM assessments began on the day of conditioning and continued bi-weekly until hospital discharge. Outcomes of interest included number of days of fever, total parenteral nutrition (TPN), and parenteral narcotic therapy, incidence of Grade III or IV (Common Toxicity Criteria) infection, and inpatient days and charges. We report data here for the 281 AL HSCT recipients in the study cohort.

Results. Mean age was 41 yrs. Most common diagnoses were acute (33%) and chronic (19%) myeloid leukemia. Most patients (96%) received total body irradiation. Fifty-one percent of grafts were from unrelated donors. Seventy-six percent of patients experienced OM ≥ grade II. The relationship between worst recorded grade of OM and each of the outcomes of interest is reported below.

Conclusions. OM is associated with worse clinical and economic outcomes in patients with hematologic malignancies undergoing AL HSCT.

Outcomes of AL HSCT in patients with hematologic malignancies, by worst recorded grade of OM Worst Grade of OM (N=281) 0 I II III IV V Outcome n=34 n=33 n=70 n=60 n=43 n=41 -value p *mean, SD; **test for linear trend; Cochran-Armitage for dichotomous measures, GLM for continuous measures Fever days* 3.1 (3.0) 4.8 (4.4) 5.3 (4.9) 5.7 (4.7) 5.9 (4.7) 7.3 (4.6) <0.001 TPN days* 6.7 (7.2) 7.7 (7.8) 8.8 (7.8) 10.2 (7.8) 13.2 (7.2) 14.0 (6.2) <0.001 Narcotic days* 7.6 (6.3) 10.8 (5.0) 10.7 (7.4) 11.5 (5.1) 14.0 (6.8) 17.2 (5.0) <0.001 Infection (%) 85.3 90.9 92.9 93.3 100.0 100.0 0.002 Mortality (%) 8.8 3.0 8.6 15.0 7.0 22.0 0.042 Inpatient days* 28.1 (8.8) 30.3 (10.6) 34.5 (15.1) 32.7 (14.5) 35.9 (10.4) 42.4 (15.1) <0.001 Inpatient charges* (x 1,000 USD) 214 (102) 252 (156) 313 (251) 280 (198) 305 (140) 437 (233) <0.001
Quantitiative Analysis of Roughness of Catheters by Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM).
Authors: Chan Anthony , Parmar Nagina , Jega Anusha , Yip Christopher , Farrokhyar Forough , Walton Mark J.
Published: October 2019

Thrombosis is a major complication of central venous access devices, and incidence depends on material, diameter, tip position, and tip surface. Catheters are usually cut to the appropriate length for accurate positioning. However, cutting is not recommended as rough surfaces can serve as a nidus for thrombi. This study was done to assess the roughness of the catheter tips as provided by various manufacturers, versus the roughness once cut and handled. Four types of catheters (Groschong, Hickman, Port-a-Cath, and Per Q Cath) were cut by scissors, iris scissors, or scalpel, and handled with debakey forceps, a needle driver, and adsons with or without teeth, to determine the damage created on the catheter. The manufactured tip was compared as a control. Scanning electron microscopy (SEM) was used to do imaging for all samples and roughness and section analysis was quantified using atomic force microscopy (AFM) for the cutting methods. SEM showed that scalpel-cut and manufactured ends appeared smoother relative to those cut with scissors or iris scissors (Fig 1). This complemented the roughness and section analysis by AFM (Fig 2). Catheters handled by debakey equipment and adsons with teeth showed the most roughness, visible as deep holes or a grainy surface when observed by high magnification SEM. Decreasing smoothness of catheters is in the following order: uncut surface, followed by surfaces cut by scalpel, scissors or iris scissors. Handling should be minimized and use of adsons with teeth, needle drivers and debakey forceps avoided, which can leave permanent damage. The least damage appeared to be adsons without teeth. Thus, the cutting of catheters is not recommended since rough surfaces can serve as a thrombotic nidus.

Figure Figure
The Effect of beta-Thalassemia Minor on the Optic Nerve Head Analysis.
Authors: Cetin Turker , Mutlu Fatih M. , Avcu Ferit , Mumcuoglu Tarkan , Nevruz Oral , Ural Ali U.
Published: October 2019

Background: A variety of ophthalmologic findings have been reported in patients with anemia.

Aim: To determine the effect of beta-thalassemia minor on the optic nerve head topographic analysis.

Methods: A total of 39 beta-thalassemia minor patients were divided into 2 groups. Patients with iron, folic acid, or vitamin B12 deficiency were ruled out. Group 1 comprised 20 patients with anemia, and group 2 comprised 19 patients without anemia. One eye of each patient was included into the study. All subjects underwent complete ocular examination. Optic nerve head topographic analysis was performed by using a confocal scanning laser ophthalmoscope type Heidelberg retina tomograph (HRT). The following stereometric parameters were evaluated: disc area, area and volume of cup, area and volume of neuroretinal rim, measure of cup shape, and mean retinal nerve fiber layer thickness.

Results: The mean age of group 1 and 2 were 26.8±7.6 and 25.6±4.5 years, respectively (P=0.91). Their mean disc areas were 2.01±0.3 mm2 and 2.53±0.6 mm2, respectively (P =0.009). The differences between groups for area and volume of cup, area and volume of neuroretinal rim, cup shape measure, and mean retinal nerve fiber layer thickness were insignificant (p>0.05). There was no significant difference between mean intraocular pressure of both groups (p=0.93).

Conclusion: In beta-thalassemia minor, patients with anemia, optic disc area showed a statistically significant reduction compared to the patients without anemia. Further clinical trials on ocular blood flow and optic nerve oxygenation changes may highlight the role anemia in the optic nerve head of the beta-thalassemia minor patients.
Correlation between Entothelial Microparticle Binding to Monocytes and Monocyte Nitric Oxide Production in Prothrombotic and Inflammatory Disorders.
Authors: Chirinos Julio A. , Jy Wenche , Del Carpio Freddy , Arteaga Roque , Gustavo Heresi A. , Valvidia Martin , Horstman Lawrence L. , Jimenez Joaquin J. , Ahn Yeon S.
Published: October 2019

Introduction: Endothelial microparticle binding to monocytes has been shown to induce monocyte activation in vitro. In this study, we examined the correlations between EMP binding to monocytes and monocyte levels of nitric oxide (NO), as a marker of monocyte activation in different clinical conditions.

Methods: We studied 186 subjects with acute venous thromboembolism (n=25), atrial fibrillation (n=48), metabolic syndrome (n=37), congestive heart failure (n=44), and normal controls (n=32). Using flow cytometry, we measured monocyte levels of NO by flow cytometry after loading monocytes with the membrane permeable NO-selective fluorescent indicator DAF-DA. Two different populations of EMP-monocyte conjugates were also measured. EMP62E+-monocyte and EMP54+-monocyte conjugates were measured based on the detection of E-selectin (CD62E) or CD54, respectively, coexpressed with CD45 in monocytes.

Results: Pearson correlation coefficients between monocyte NO levels and EMP-monocyte conjugates are shown in the Table. A highly significant correlation was found between EMP62E+-monocyte conjugates and monocyte NO levels in patients with congestive heart failure (r=0.43; p=0.003). In contrast, EMP54+-monocyte conjugates strongly correlated with monocyte NO in patients with venous thromboembolism (r=0.58; p=0.009) and metabolic syndrome (r=0.49; p=0.002). No correlation was found between these conjugates and monocyte NO levels in atrial fibrillation or normal controls.

Conclusions: The binding of different species of EMP to monocytes correlates with monocyte NO levels in clinical states such as congestive heart failure, metabolic syndrome, and venous thromboembolism. However, this correlation was not found in atrial fibrillation or normal controls. Our findings support the concept that the binding of different species of EMP exert different biological effects and/or reflect different biologic processes in specific disease states. Further research is needed to determine whether the binding of EMP species to monocytes regulates nitric oxide production by monocytes and whether monocytes themselves regulate the binding of different species of EMP in different disease states.

Correlation between Monocyte NO levels and EMP-Monocyte Conjugates in different conditions. EMP54+-Monocyte Conjugates EMP62E+-Monocyte Conjugates Pearson r p value Pearson r p value Metabolic Syndrome 0.49 0.002 0.13 0.54 Venous Thromboembolism 0.51 0.009 0.12 0.57 Congestive Heart Failure 0.22 0.14 0.43 0.003 Atrial Fibrillation 0.22 0.14 0.17 0.24 Normal Controls 0.03 0.85 0.27 0.13
Tumor Necrosis Factor-a Enhances Human Leukemia Cell Differentiation Induced by DMSO, but Not Retinoic Acid.
Authors: Yu Hong-Nu , Lee Young-Rae , Shim Hyun-Jaung , Han Myung-Kwan , Song Eun-Kyung , Park Jinny , Kim Jong-Suk
Published: October 2019

One of the human leukemia treatment methods is to differentiate leukemia cells into mature cells. Because differentiated cells lose their proliferative and tumor-forming abilities, differentiation inducers may be useful for the treatment of leukemia. Differentiation of leukemia cells has been studied using HL60 cells, a human promyelocytic leukemia cell line, which can be differentiated into granulocyte-like or monocyte/macrophage-like cells by various pharmacological agents such as dimethyl sulfoxide (DMSO), retinoic acid and phorbol myristic acetate (PMA). We previously reported that nuclear factor - kB (NF-kB) activation plays the important role in DMSO-induced differentiation of HL60 cells. Thus, we hypothesized that NF-kB activators could enhance DMSO-induced differentiation of HL60 cells. Here we examine whether tumor necrosis factor-a (TNF-a), a potent NF-kB inducer, enhance DMSO-induced differentiation of HL60 cells. TNF-a was found to enhance HL60 cell differentiation induced by DMSO. CD11b, a differentiation marker, was increased in 0.5 % DMSO-treated cells compared to control cells. When TNF-a was added to the same condition, CD11b expression was further enhanced in a dose and a time dependent manners. We also found that nitro blue tetrazolium (NBT) reducing activity, a marker for granulocytic differentiation, was further increased in DMSO plus TNF-a treated cells compared to only DMSO- treated cells. However, TNF-a alone had no effect on CD11b expression and NBT reducing activity. The enhancement of DMSO-induced HL60 differentiation by TNF-a was offset by NF-kB inhibition. Interestingly, retinoic acid- induced differentiation of HL60 cells showed no enhancing effect of TNF--a on the differentiation. These findings indicate that TNF--a might affect only NF-kB dependent differentation of HL60 cells. Taken together, we demonstrated that TNF-a enhances DMSO-induced differentiation of HL60 cells by stimulating NF-kB activation. Our results suggest that NF-kB inducers such as TNF-a are useful for the treatment of leukemia in combination with DMSO.
Sub-Marginal Endothelial Cells Dynamically Regulate Endothelial Sheet Migration.
Authors: Ofori-Acquah Solomon F.
Published: October 2019

Migration of endothelial cells as a sheet in fully differentiated blood vessels is essential for reducing vascular permeability during wound healing. Indeed, loss of collective endothelial sheet migration contributes to increased vascular permeability in tumor angiogenesis and several vascular proliferative disorders. Despite this significance, mechanisms responsible for keeping migrating endothelial cells in a monolayer, sheet or tube are poorly understood. To unravel the basis for collective endothelial cell migration, we used time-lapse video microscopy to study early events of wound closure in confluent monolayers of primary microvascular endothelial cells in a live cell chamber. Immediately after wounding (0– 20 min), endothelial cells at the margin of the wound (marginal cells) retracted away from the wounded area and showed no visible lamellipodia extensions. The next phase of early wound healing (20 min – 6 hours) revealed extensive lamellipodia formation and migration of marginal cells into the wounded region. Remarkably, sub-marginal endothelial cells that were several microns away from the wound edge protruded lamellipodia that formed dynamic cell-cell contacts with the substratum of marginal cells at the wound edge. In several instances sub-marginal cells physically and coordinately pulled back endothelial cells at wound edge to maintain regularity of the endothelial sheet front. Cell-tracking measurements revealed autonomous and yet coordinated migration of marginal and sub-marginal endothelial cells culminating in net protrusion of the endothelial sheet into the wound. This study provides in real-time evidence of retraction of endothelial cells at the wound edge by several microns prior to the initiation of forward migration. In addition, we show for the first time that endothelial cells several microns away from the wound edge actively participate in sheet migration through the extention of lamellipodia into the substratum of cells at the wound edge. These findings highlight an important role in endothelial sheet migration for the Rho family of GTPases given their intimate control of cell retraction and lamelipodia extensions. Future studies will directly evaluate the influence of Rac, Cdc42 and RhoA in retraction of marginal cells and formation of lamellipodia by sub-marginal cells in endothelial sheet migration.
NIH-Funded Research Using Human Embryonic Stem Cells.
Authors: Blau C. Anthony , Nelson Angelique , Ware Carol B.
Published: October 2019

In October 2003, the National Institutes of Health (NIH) established three extramural “Exploratory Centers for Human Embryonic Stem Cell Research.” Our Center’s experience in acquiring and manipulating NIH-approved human embryonic stem (hES) cell lines may be useful for other institutions interested in pursuing NIH-funded hES cell research. We acquired 14 of the 22 NIH-approved cell lines. Modifications to the proposed Material Transfer Agreements (MTAs) with hES cell suppliers were sought to improve accessibility to these hES cell lines by local researchers. Lines were characterized for survival following cryopreservation and for their ability to adapt to a uniform set of culture conditions while maintaining a normal karyotype. Each of the 4 suppliers contacted eventually agreed to terms that improved access to their hES cell lines. Eleven hES cell lines were received frozen, and in 10 cases very few cells survived cryopreservation. Ten hES cell lines were successfully converted to simplified culture conditions that enhanced their ability to be maintained and expanded in culture. One hES cell line had an unstable karyotype at an early passage. Current MTA provisions continue to present significant obstacles to NIH-funded hES cell research. Many hES cell lines can be maintained using culture conditions less onerous than those recommended by their suppliers.
The South East England Thrombotic Thrombocytopenic Purpura Registry.
Authors: Scully Marie , Yarranton Helen , Liesner Ri , Cavenagh Jamie , Hunt Beverley , Benjamin Sylvia , Laffan Michael , Bevan David , Mackie Ian , Machin Samuel J.
Published: October 2019

Thrombotic thrombocytopenic purpura (TTP) is an acute, rare, life threatening disorder. We present data from the South East (SE) England registry for TTP, from April 2002–December 2005. Included are 156 patients and 179 acute episodes. Seventy four percent of cases were female and 26% males, median age at presentation, 41 and 47.5 years respectively. Mortality to December 2003 was 12%, and from January 2004 to December 2005 was 5%; 5/12 died before treatment was instigated. The main ethnic groups were Caucasian (63%), Afro Caribbean (22%) and Asian (12%). In 61%, this was their first TTP episode; 39% represented with acute relapse. Precipitating events (n=123); idiopathic (56%), a defined infection (11%), pregnancy (6%), HIV disease (7%), congenital (5%) and associated autoimmune disease (6%). Features at presentation include: neurological disease (39%), temperature (15%), abdominal symptoms (12%), renal impairment (10%), symptomatic cardiac disease (7%), symptoms related to low platelets (17%) and no symptoms (2%). From April 2002–December 2003, the median number of plasma exchanges until complete remission (CR) recorded in 36 patients was 20 (5–79). One patient received plasma infusion (PI) in conjunction with PEX. 33 patients received 3 doses of pulsed intravenous methylprednisolone (IV MP) and 9 orally. Other therapies used were Rituximab (1), defibrotide (5), cyclosporine (9) and vincristine (15). In the second cohort (January 2004–December 2005), the median number of PEX to CR in 88 episodes was 8 (3–80). PI was used in 5 cases, 48 patients received 3 doses of pulsed IV MP on admission and 6 patients oral prednisolone. Other therapies used were Rituximab (26), defibrotide (2), cyclosporine (3), vincristine (5) and BPL 8Y (9). The latter is an intermediate purity Factor VIII concentrate used in congenital TTP. The median Haemoglobin (Hb) was 9.9g/dL at presentation and platelet count 18 ×109/L. The 2002–2003 group had 48% cases with platelet counts <150 ×109/L at 7 days, but only 25% of cases in 2004–2005. ADAMTS 13 activity, measured by collagen binding assay (CBA) (normal range 66–126%), in 136 episodes was <5% in 60%, 5–40% in 21%, 40–66 in 12% and >66% in 7% at presentation. Antibody to ADAMTS 13 (determined by mixing studies using CBA and/or anti-ADAMTS 13 IgG antibodies) was present in 61% of all cases. In those patients with acute idiopathic TTP, 93% had associated antibody. In conclusion: Improved recognition and prompt diagnosis of TTP may be associated with reduction in mortality and changes in treatment protocols reduced the number of PEX to achieve CR. Acute idiopathic TTP is the most common presentation, but improved identification of subgroups such as HIV and congenital TTP have resulted in appropriate tailoring of therapy.
TDAG51 Deficiency Promotes Migration and Proliferation of Mouse Embryonic Fibroblasts.
Authors: Hossain Gazi S. , Dickhout Jeffrey G. , McCulloch Christopher A. , Zhou Ji , May Linda , Al-Bayati Hiam , Rho J. , Austin Richard C.
Published: October 2019

T-cell death associated gene 51 (TDAG51) is a member of the pleckstrin homology-related domain family having pro-apoptotic characteristics. Indirect immunofluorescence studies demonstrate that endogenous TDAG51 co-localizes with focal adhesion kinase (FAK), a molecule found within focal adhesion complexes (FACs). FACs are believed to be the structural connection between the extracellular matrix (ECM) and actin cytoskeleton. Given that overexpression of TDAG51 promotes detachment-induced programmed cell death (PCD) and that TDAG51 co-localizes with FAK, it was proposed that deficiency of TDAG51 may stabilize FAC assembly, thereby affecting actin cytoskeletal organization and/or cell migration. To better understand the role of TDAG51 in cell migration, mouse embryonic fibroblasts (MEFs) deficient in TDAG51 were derived from TDAG51-deficient mice and compared to wild type MEFs. TDAG51-deficient MEFs showed increased migration following monolayer disruption or in response to chemotaxis on fibronectin-coated Boyden Chambers. In addition, loss of TDAG51 promotes the proliferation of MEFs. Collagen gel contraction experiments were performed to confirm the aggregate effect of the differences in migration and proliferation observed in TDAG51-deficient MEFs. Interestingly, migration and proliferation can be blocked by active b-integrin inhibitory antibody. In terms of a cellular phenotype, migratory TDAG51-deficient MEFs have distinct filopodial and lamellipodial extentions, compared to migratory wild type MEFs. Laser scanning confocal microscope demonstrated a significant decrease in F-actin stress fibers as well as increased distribution of vinculin within the lamellipodial protrusions of TDAG51-deficient MEFs. Importantly, reintroduction of TDAG51 into TDAG51-deficient MEFs reversed these phenotypic changes. Taken together, our findings suggest that loss of TDAG51 affects cell migration and proliferation through modifying focal adhesion complex assembly and cytoskeletal rearrangements.
Clinical Usefulness of the Simple Technique To Diagnose Thrombocytopenia Using Immature Platelet Fraction (IPF).
Authors: Cho Yong Gon , Lee Jae Hyun , Kim Dal Sik , Lee Hye Soo , Choi Sam Im
Published: October 2019

Background : It is defined that immature platelet fraction (IPF) is the percentage of reticulated platelet (RP) of total platelet count. We have measured an IPF reference range using XE-2100 blood cell counter with upgraded software (Sysmex, Kobe, Japan) and evaluated the clinical utility of this parameter for the laboratory diagnosis of thrombocytopenia due to increased peripheral platelet destruction.

Materials and methods : Peripheral blood samples collected into K2EDTA (Beckton Dickinson, Franklin Lakes, NJ, USA) were analysed at Chonbuk National University Hospital. 142 samples from apparently healthy adults (all routine full blood count parameters including platelets within the healthy reference range) were used to establish a normal reference range for the IPF%. The patients are classified 3 group including hypoplastic (consisted of 22 patients undergoing chemotherapy with falling platelet counts and 14 with aplastic anemia), destructive (14 with ITP and 7 with infection), and cirrhotic (40 with cirrhosis of liver).

Results : An IPF reference range in healthy individuals was established as 0.4~5.4%, with a mean of 1.7%. The significant increase in IPF values was found in destructive patient group. The cut-off value of IPF is 6.0%, its sensitivity and specificity are 95.2%, 82.9% respectively. Reproducibility was good.

Conclusion : A rapid, inexpensive automated method for measuring the IPF% is feasible and should become a standard parameter in evaluating the thrombocytopenic patient.
Use of Activated Recombinant Factor VII for the Control of Post Partum Hemorrhage (PPH).
Authors: Hossain Nazli H. , Farzana Tasneem F. , Shamsi Tahir S. , Soomro Nargis S.
Published: October 2019

Objective: To evaluate the efficacy and safety of activated recombinant factor VII (rFVIIa) in the control of PPH

Patients and Methods: All patients with massive PPH who failed medical and surgical treatment were eligible for the investigation. Massive PPH was defined as loss of > 1000 ml of blood within 24 h after delivery. Patients were considered for treatment with rFVIIa after failure of conventional measures such administration of myotonic agents including oxytocin, ergotamine, misoprostol and failure of other interventions aimed at controlling blood loss such as ligation of uterine and ovarian vessels, or ligation of internal iliac artery. The primary outcome measures were control of the bleeding episode and reduction in the number of administered blood products.

Results: A total of 16 patients with massive PPH were eligible for the current study. Vaginal and vacuum delivery was used in 6 patients while 10 patients underwent C-section. The mean dose of rFVIIa was 68 mcg/kg administered intravenously as a single bolus injection. Mean time of diagnosing PPH and administration was 5.5 hours (+/−2). The mean number of blood products used prior the administration of rFVIIa was 14 units RBC and 12 units of FFP. Cessation of blood loss was achieved in 14 patients within an average of 20 minutes after the administration of rFVIIa. The blood products utlilized after the administration of rFVIIa in these patients were 2 URBC and 2UFFP. The uterus was conserved in 7 patients who received rFVIIa prior to hysterectomy. While in 9 patients rFVIIa was administered after failure of hysterectomy to control blood loss. Of the 16 patients, 3 died because of multiorgan failure following severe blood loss. No adverse events including thromboembolic phenomena were observed.

Conclusion: Massive PPH is the most common cause of maternal mortality in developing countries. The use of rFVIIa in patients with massive PPH can achieve hemostasis with a single dose only. The administration of rFVIIa in patients with massive PPH may avoid the need for hysterectomy. There were no safety issues in the current study. Further trials should address the dose and timing of administration of rFVIIa in patients with massive PPH.
The Impact of Rebleeds in Cost Modelling of Treatment Strategies in Patients with Hemophilia A and Inhibitors.
Authors: Huth-Kuehne Angela , Lages Peter , Zimmermann Rainer
Published: October 2019

Introduction: To investigate the cost-effectinvess of the treatment in patients with hemophilia and inhibitor, the expected number of rebleeds in home treatment is an important factor if one compares different treatment options e.g. APCC or rFVIIa. To evaluate the economic impact in mild to moderate bleeds in home treatment we used a robust established model to compare 1st, 2nd and 3rdline treatment either with rFVIIa or APCC. For the rate of rebleeds we refer to published data on a broad basis.

Methods: To compare the economic impact of different treatment regimes, this model refers to an on-demand home treatment of bleeding episodes in adult hemophiliacs (mean body weight 75 kg) and is based on a previously published flow-diagram and calculation formula (Knight 2003). Dosage per bolus: rFVIIa 90μg/kg, aPCC 60 U/kg. Costs for German market were fixed at 1.21 €/IU for APCC, 0.73 €/μg for rFVIIa and 308.98 €/day in hospital. 3 treatment strategies (TS) were calculated:

TS 1:

APCC at home, followed by in hospital APCC (2nd-line) and rFVIIa (3rd-line)

TS 2:

APCC at home, followed by in hospital rFVIIa (2nd-line) and rFVIIa (3rd-line)

TS 3:

rFVIIa at home, followed by in hospital rFVIIa (2nd-line) and rFVIIa (3rd-line)

The model was based on published data on dosing and outcome of therapy in each treatment phase. Probability of failure to control bleeding or rebleeding after first-line home treatment was set at 23.5% with APCC and 9.1% with rFVIIa.

Results: Median costs for TS 1 are 23,521 Euro per bleeding episode as compared to 21,963 Euro for TS 2 and 14,328 Euro for TS 3. The difference is caused by costs for first attempt to control bleeding at home (APCC 16,335 Euro as compared to rFVIIa 11,333 Euro) and costs at later stages of treatment. Further analyses show, that favorable result for TS 3 is robust to changes of different parameters such as patients’ weight.

Conclusions: First line treatment with rFVIIa in case of on demand treatment seems to be the most cost effective option in inhibitor treatment in a home treatment setting or in-hospital treatment. Local prices have to be taken into consideration.
Prognostic Value of Non-Overt DIC Criteria in Patients with Sepsis.
Authors: Jang Moon Ju , Chong So Young , Kang Myung Seo , Lee Sun Ju , Oh Doyeun
Published: October 2019

DIC is a frequent complication of sepsis resulted in high mortality. The early diagnosis of DIC in septic patients is necessary to reduce the mortality by the prevention of irreversible organ damage through earlier therapeutic intervention. We investigated the performance of the ISTH non-overt DIC criteria and its modification by adding antithrombin (AT), protein C and Organ Failure Scoring (OFS) system for mortality prediction were evaluated in 100 septic patients prospectively enrolled between July 2004 and March 2006. Platelet count, PT, aPTT, fibrinogen, D-dimer, AT, protein C on day 0,1,2,3,4, and 7 were measured in each patient. ISTH overt and non-overt DIC scoring with 28-day mortality, OFS was evaluated to assess the diagnostic value of each criterion in the prediction of clinical outcome. Twenty-five, 14 and 61 had overt DIC, non-overt DIC and non-DIC respectively. Platelet count, PT, AT, protein C levels were significantly different between DIC and non-DIC. D-dimer and protein C levels were significantly different between overt and non-overt DIC. The 28-day mortality rate of overt DIC, non-overt DIC and non-DIC was 44, 50 and 11.5% respectively. By adding AT and protein C on non-overt DIC criteria, the 28-day mortality rate of overt DIC, non-overt DIC and non-DIC became changed to 44, 31.4 and 7.5% respectively, but there was no difference between overt and non-overt DIC patients. Addition of AT, protein C or OFS to ISTH non-overt DIC criteria did not improve the diagnostic power significantly. In conclusion, ISTH non-overt DIC criteria is feasible to predict the poor outcome of septic patients.
Platelet and Coagulation Activation in Myeloproliferative Diseases (MPD), and Relationship to JAK2(V617F) Mutation Status and Clonality.
Authors: Robertson Beverley J. , Urquhart Colin , Ford Isobel , Watson Henry G. , Vickers Mark , Greaves Michael
Published: October 2019

Patients with MPD have an increased risk of thrombosis. Previous reports suggest an association between clonality and thrombosis in ET. The contribution of the JAK2 mutation to thrombotic risk is unclear. In our cohort of patients with MPD (n=122) (PRV n = 54, ET n = 64, IMF n= 4), we compared coagulation and platelet activation markers (D-Dimers, thrombin antithrombin complexes (TAT), prothrombin fragments 1+2 (F1+2), soluble E-selectin(sE-selectin) and soluble P-selectin(sP-selectin)) between MPD patients and hypertensive controls. sP-selectin was significantly increased in patients with MPD (p=<0.001). The JAK2 mutation status was determined in our cohort by ARMS PCR. Of the total MPD cohort, 59% were JAK2 positive, (76% of PRV, 45% of ET, and 50% of IMF). Coagulation activation markers were compared in JAK2 positive and JAK2 negative patients. sP-selectin levels were highly significantly elevated in JAK2 positive patients compared to JAK2 negative (p= 0.002), or controls (p<0.001).

There was no significant difference in platelet count between JAK2 positive and negative patients (p=0.19). The clonality of the MPD was determined in 54 female patients using an x-chromosome inactivation pattern (XCIP) assay (HUMARA). A significant proportion of “polyclonal” patients, as defined by XCIPs were positive for the JAK2 mutation. We therefore calculated the proportion clonality of samples and found no correlation between proportion clonality and any coagulation or platelet activation marker.

Our results show an increase in platelet activation, as determined by sP-selectin levels, in patients with MPD compared to controls. Furthermore, platelet activation was a feature of patients positive for the JAK2 mutation when compared with wild type patients and controls. The role of the JAK2 mutation as a risk factor for thrombosis in MPD is still unclear but our study indicates that the presence of the mutation may be linked to platelet activation in MPD. Whether this translates into a higher clinical thrombosis risk requires further evaluation in a large prospective study.
Human Cell Expressed Cytokines for Expansion of CD34+ Hematpoietic Stem Cells.
Authors: Rede Teresa , Domagala Teresa , Liddell Kate , Crofts Linda , Pilkington Glenn , Marks Denese
Published: October 2019

Many cytokines and growth factors are heavily glycosylated, with up to 75% of their mass consisting of carbohydrate moieties. Glycosylation is important for secretion, solubility, resistance to proteolysis, immunogenicity, biological recognition, biological activity, in vivo stability and clearance of glycoproteins including cytokines and growth factors. This study was designed to compare the ability of human cell expressed cytokines to induce CD34+ hematopoietic stem cell expansion with human cytokines expressed in E. coli. CD34+ stem cells were cultured with either human cell expressed G-CSF and stem cell factor (SCF), or with human G-CSF and SCF expressed in E. coli. CD34+ stem cell expansion was determined by counting total nucleated cells (TNC), and colony forming unit (CFU) potential of the expanded cells was determined by colony number, size and cell morphology. During early cell proliferation and expansion, the TNC count of cells treated with the human cell expressed cytokines was markedly higher than cells treated with E. coli expressed human cytokines. In CFU assays, the size and mean number of colonies were greater with the human expressed G-CSF and SCF cytokine combination. In addition, the proportion of colonies with macrophage-like morphology was greater. Human cell expressed cytokines also induced greater myeloid differentiation as indicated by higher CD33 expression compared to cytokines expressed in E. coli.

We are currently investigating the ability of human cell expressed G-CSF/SCF to activate signaling pathways including the MEK/ERK and STAT pathways, which are known to be involved in G-CSF mediated proliferation and differentiation. Differences in receptor-ligand binding using ligands from various expression systems are also being examined.

The ability of human cell expressed cytokines to induce greater in vitro differentiation and proliferation of hematopoietic stem cells may translate to better ex vivo expansion protocols.
A High-Density MicroRNA Array for Analyzing the Expression of 15,090 Verified and Predicted Small, Non-Coding RNAs.
Authors: Wilson Michael , Leng Nan , Sah Sachin , Sekinger Edward , Fletcher Frederick , Johnson Charles
Published: October 2019

We have designed and evaluated the performance of a custom Affymetrix microRNA (miRNA) microarray that enables the analysis of 15,090 unique mature miRNAs that are predicted or biologically verified. The comprehensive probe content of the array was selected from multiple sources including the Sanger miRBase sequence database (version 8.0), published reports, and a proprietary database of predicted miRNAs. Most of the ~31,000 25-mer probes on the array were designed by selecting two overlapping oligonucleotide sequences that span the mature miRNA sequences and a portion of the flanking precursors (pre-miRs). We also included antisense control probes as well as probes that target the predicted hairpin loop region of the pre-miR for a subset of well-characterized human miRNAs. We evaluated the array performance using a model system of two total RNA reference standards isolated from different tissues that were mixed in calibrated ratios. The custom array performance was compared to an enhanced mirVana™ miRNA Bioarray, using the same calibrated mixtures of reference RNAs. We assessed the reproducibility, relative sensitivity, dynamic range and concordance between the results from the two platforms. The performance of the two platforms was similar, with both platforms reliably detecting differential miRNA expression. For those miRNAs that are represented on both platforms, there was a strong correlation of the log2 ratios of differential expression levels between the RNA reference standards. Many of the miRNAs with detectable expression signals were novel predicted miRNAs derived from bioinformatic exploration of human genomic sequence data. This provides initial evidence that some of the predicted miRNAs are expressed in tissues and, therefore, may function as regulators by mechanisms consistent with other well-characterized miRNAs. In summary, our data illustrate the technological and biological feasibility of the custom high-density miRNA microarray for the comprehensive identification and global expression profiling of novel miRNAs.
Osmotic Fragility of Human Blood Platelets
Authors: GUREVITCH J. , NELKEN D.
Published: October 2019

The osmotic fragility of 50 samples of platelets from normal healthy donors has been studied. It has been found that fragility of platelets starts at a concentration of about 0.44 per cent NaCl and is complete at a concentration of 0.34 per cent. The serotonine released from the platelets increased with the gradual disintegration as the saline concentration fell.
Ultrastructure of Erythrocyte Membranes in Thalassemia Major and Minor
Authors: HOFFMAN J. F. , WOLMAN I. J. , HILLIER J. , PARPART A. K.
Published: October 2019

The plasma membranes of erythrocytes derived from Thalassemia major (five cases) and minor (seven cases) were isolated and studied by electron microscopy. It was found that the surface texture of ghosts from all T. minor bloods appeared similar and indistinguishable from ghosts of normal humans blood. The surface texture of ghosts from all T. major bloods likewise were quite similar but distinctly different from T. minor and normal ghosts. These differences were observable whether the specimens for comparison were prepared at the same time or on different days. The results indicate that the morphological characteristics observed in T. major ghosts constitute an expression of an alteration in the molecular structure of their plasma membranes.
Sickle Cell Anemia (Homozygous S) with Aseptic Necrosis of Femoral Head
Authors: TANAKA KOUICHI R. , CLIFFORD GEORGE O. , AXELROD ARNOLD R.
Published: October 2019

1. Six patients with sickle cell anemia who exhibit aseptic necrosis of the femoral head are presented.

2. All patients were proven homozygous S by paper electrophoresis.

3. In our material, aseptic necrosis of the femoral capital epiphysis in patients with sickle cell anemia is not an uncommon finding, 12% of 51 cases demonstrating this lesion.

4. The incidence of this lesion may be even greater than we report, inasmuch as some of our patients were asymptomatic, although typical roentgenographic changes of aseptic necrosis of the femoral head were present.
The Hemostatic Defect of Uremia
Authors: LARRAIN CAMILO , LANGDELL ROBERT D.
Published: October 2019

Acute urinary retention was produced in 15 adult mongrel dogs by ligation of the ureters. Studies of the hemostatic mechanism were made as the uremia progressed. All animals developed a coagulation defect characterized by abnormally long clotting time of whole blood in siliconized glassware. Other indices of coagulation efficiency were found to be normal or only slightly impaired.

It is not clear whether the basic defect is a lack of an essential clotting factor or an excess of an inhibitor. Urea does not seem to be the cause of the defect. The data suggest that the duration of urinary retention is as important as the degree of azotemia.
Immunologic Features of Erythrocyte Sensitization
Authors: VAUGHAN JOHN H.
Published: October 2019

By the use of specifically absorbed Coombs sera, it has been possible to show that cell sensitization in acquired hemolytic disease may be due either to γ-globulin or to non-γ-globulin serum components. The latter may be either warm reacting or cold reacting (cold agglutinin).

In certain instances acquired hemolytic disease is associated with autoimmunization. In other instances, it is possible that nonspecific adsorptive processes are involved, though definitive evidence for this is lacking.
The "Morular Cell" and the "Grape Cell" in Bone Marrow and Peripheral Blood
Authors: ZLOTNICK AVINOAM
Published: October 2019

Bone marrow aspirates of 39 patients with different diseases were examined. In 8 of these patients, who showed hyperglobulinemia of nonmyelomatous origin, three kinds of cells were found:

1) Plasmocytoid reticulum cells whose cytoplasm was filled with crystal-like configurations.

2) Plasmocytoid reticulum cells with hyaline transparent vesicles of different sizes in the cytoplasm. These cells resembled the "morular cell" of Mott.

3) Plasma cells the cytoplasm of which contained opaque bluish staining globular bodies varying in number and size. These cells were identical with the "grape cell."

The "grape cell" was also found in the buffy coat of the blood from 2 patients with hyperglobulinemia, and the "morular cell" in the buffy coat from a patient suffering from an unusual parasitic disease.

The protein nature of these inclusions is assumed, since they stained with the M.G.G. combination, but did not stain with Sudan black nor with P.A.S.
The Response of Transplanted Chloroleukemia in the Rat to Treatment with Antileukemic Drugs
Authors: SHAY HARRY , HARRIS CHARLES , GRUENSTEIN MARGOT
Published: October 2019

Using a short-term screening program, we have demonstrated a parallelism in response between our myelogenous chloroleukemia in the rat and that of myelogenous leukemia in man when treated with a number of chemotherapeutic agents.
Erythrokinetics: Quantitative Measurements of Red Cell Production and Destruction in Normal Subjects and Patients with Anemia
Authors: GIBLETT ELOISE R. , COLEMAN DANIEL H. , PIRZIO-BIROLI GIACOMO , DONOHUE DENNIS M. , MOTULSKY ARNO G. , FINCH CLEMENT A.
Published: October 2019

Red cell turnover of 19 normal subjects and 25 anemic patients was measured with the following technique: erythroid-myeloid ratio of the marrow, reticulocyte counts, plasma iron turnover, red cell utilization of radioiron, and urobilinogen determinations. Measurements of blood production and destruction were so expressed as to allow comparison between normal and anemic individuals of different size and different red cell mass. The usefulness and disadvantages of each procedure in the study of anemia are discussed.

From studies of various types of anemia, it has become apparent that erythropoiesis must be defined in terms of total quantity of red cells produced and in terms of the portion of red cells produced in the marrow which are delivered to the circulating blood (effective versus ineffective erythropoiesis). A quantitative defect alone exists when a normal ratio is maintained between effective and total erythropoiesis. Here, there are changes of similar magnitude of all erythrokinetic indices, although reticulocyte and urobilinogen values are occasionally disproportionately high. The normal marrow appears to be able to increase its effective red cell production to three times normal in acute anemia and six times normal in chronic anemia. In many disease states this maximal quantitative response is impaired.

Dyspoiesis of the marrow is characterized by a dissociation of erythrokinetic indices. Values which reflect total erythropoiesis (i.e., plasma iron turnover, fecal urobilinogen and erythroid-myeloid ratio of the marrow) are considerably greater than the reticulocyte level and red cell utilization of radioiron which represent effective erythropoiesis. Such defects may result in the pattern of a hemolytic process or aregenerative anemia, depending on their severity.
The Plasma Erythropoietic Stimulating Factor
Authors: LINMAN AMES W. , BETHELL FRANK H. , TASCOTT HELENA K.
Published: October 2019

1. Boiled plasma extracts from anemic rabbits are capable of stimulating erythropoiesis in normal rats as demonstrated by reticulocytosis, erythrocytosis, and increased marrow erythropoietic activity.

2. In the nonanemic rat recipient the increased erythropoiesis is manifested by the production of cells of decreased size without associated increase in the hemoglobin or hematocrit determinations. The cells probably have a greatly shortened life span.

3. The factor in anemic rabbit plasma is not destroyed by prolonged boiling and is not precipitated by perchloric acid. On the basis of these observations it would seem to be nonprotein in nature.

4. Further investigations concerning the role of the erythrocyte stimulating factor in normal and abnormal erythropoiesis should be facilitated by the use of nonprotein plasma extracts which may be administered to small laboratory animals.
Observations on the Hereditary Nature of Hageman Trait
Authors: MARGOLIUS ALVIN , RATNOFF OSCAR D.
Published: October 2019

A study was made of the family tree of two sisters with Hageman trait, an asymptomatic disorder of blood coagulation. In this family the defect behaves genetically as if it is due to the transmission of an autosomal recessive gene.
Study of Leukocyte Dynamics by Means of Cross-Circulation Between Normal and Leukopenic Rats
Authors: HOLLINGSWORTH J. W. , FINCH STUART C. , CHANG CHU H. , CHALTAS MARY
Published: October 2019

(1) Certain aspects of leukocyte dynamics in rats were studied by means of a mechanical cross-circulation technic.

(2) Cross-circulation between two normal rats caused little change in the leukocyte counts.

(3) When irradiated leukopenic animals were cross-circulated with normals, the leukocyte counts of the normal animals were moderately decreased, while the level of the irradiated animals rose. Prolonged mixing of the circulations, however, failed to bring the counts of the two animals into equilibrium.

(4) After cessation of cross-circulation, the leukocyte counts of the normal donors usually rose slowly, while those of the irradiated animals fell progressively with a half-time disappearance of 30-60 minutes.

(5) The possible significance of these observations in terms of normal leukocyte mobilization, viability and destruction was discussed.
Granulomatous Lesions in Bone Marrow
Authors: PEASE GERTRUDE L.
Published: October 2019

A survey has been made of granulomatous lesions found in sections of bone marrow from 150 patients studied at the Mayo Clinic. These lesions have been found in a variety of disorders, some with known etiologic agents and others with unknown causes.

With the exception of tuberculosis, histoplasmosis and brucellosis, in which the causative organisms were seen and identified bacteriologically, the granulomatous lesions were not thought to have any distinctive histopathologic characteristics.

Prominent lesions, Langhans’ giant cells and necrosis were more common in the marrow of patients with disorders known to be associated with a granulomatous inflammatory process than they were in associations with other conditions.