bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prospective Evaluation Of Risk Factors For Clostridium Difficile Colonization Among Allogenic Hematopoietic Stem Cell Transplant Recipients, 2010-2012
Authors: Jain Tania , Croswell Christopher , Reddy Banavasi Harsha Vardhan , Urday-Cornejo Varinia , Liubakka Alyssa , Cutright Jessica , Salimnia Hossein , Lephart Paul , Revankar Sanjay , Chandrasekar Pranatharthi , Chopra Teena , Deol Abhinav , Alangaden George 
Published: October 2019

Background
Clostridium difficile (CD) is the leading cause of nosocomial diarrhea. After acquisition of CD, some patients (pts) remain colonized whereas others develop symptomatic CD infection (CDI). Hematopoietic stem cell transplant (HSCT) recipients are at higher risk of CDI than the general population. Rates of colonization and risk factors for CD colonization in the HSCT population are unknown. The goals of this study were to determine the rates and predictors of colonization with CD in HSCT pts at our institution.

Methods
We conducted a 16-month prospective study starting in December 2010 in the HSCT unit at the Karmanos Cancer Center. Pts who signed informed consent had stool samples collected within 72 hours of admission and weekly thereafter until discharge. Pts were followed up for 90 days after enrollment. Stool samples were cultured for CD and culture positive samples were tested by PCR for confirmation of toxigenic CD. Demographic information, risk factors and outcomes were examined.

Results
154 patients who underwent allogeneic HSCT were enrolled in the study. Mean age for both CD colonized and non-colonized pts was 51 ±12.8 years, 59% were males. Overall, 47/154 (30%) pts were colonized with CD at admission; 23/154 (15%) with toxigenic CD. 8/23 (34%) pts colonized with toxigenic CD developed symptomatic CDI. Of the pts colonized with non-toxigenic CD, 7/24 (29%) acquired CD during the hospital stay. The median time to acquisition was week 2 of hospitalization. On bivariate analysis, underlying comorbid conditions and traditional risk factors for CDI like prior hospitalization, length-of-stay, antibiotic use, nasogastric tube and gastrointestinal tract procedures were found to be equally prevalent in both colonized and non-colonized population. The outcome in terms of graft versus host disease (GVHD) was also similar in both groups. The use of proton pump inhibitors was more prevalent in non-colonizers (56%) vs. colonizers (43%). However, it was not statistically significant (p=0.16). Similarly, lymphopenia seemed common amongst colonizers vs. non-colonizers (67% vs. 58%) but was not statistically significant (p=0.2).

Conclusion
Colonization with toxigenic CD is high in our HSCT recipients at hospital admission. Symptomatic CDI occurs in a third of pts colonized with toxigenic CD. Unlike in the general population, traditional risk factors for CD colonization are not predictive of colonization in our allogeneic HSCT pts. Novel approaches such as active surveillance to detect colonized patients to guide infection control measures and to direct “pre-emptive” therapy should be evaluated in this population.

Disclosures:
No relevant conflicts of interest to declare.
Role Of miRNA Expression As Novel Biomarker Of GvHD After Allogeneic Stem Cell Transplantation
Authors: Koldehoff Michael , Steckel Nina K , Maniera Christina , Elmaagacli Ahmet H. , Beelen Dietrich W. 
Published: October 2019

Background
MicroRNAs (miRNAs) are a family of 19-24 nucleotide noncoding RNAs, which affect the regulation of gene expression in eukaryotic cells by binding to a 3´-untranslated region which target messenger RNAs. MiRNAs plays important roles in many cellular processes such as development, stem cell division, apoptosis and cancer. We profiled miRNAs expressed on patients (pts) who underwent allogeneic hematopoietic stem cell transplantation (HSCT).

Methods
Here we analyzed in a retrospective study 258 pts (male 124 pts, female 134 pts) for miRNAs expression (miRNA-21; -128; -146; -155; -181; and –let7) in whole blood at different time-points that underwent allogeneic HSCT and analyzed their outcome. MiRNA expressions were performed by miRNA-specific real time RT-PCR.

Results
In this cohort 84 pts received grafts from HLA-identical siblings (32%), 126 pts from matched (49%) and 48 pts from mismatched (19%) unrelated donors. Transplant consisted of unmanipulated peripheral blood stem cells (n=225, 87%) or bone marrow (n=33, 13%). Of all pts, 64 (25%) had relapsed and 41 (16%) died of June 2013. There was no significant correlation between miRNA expressions from siblings and unrelated donors. For miRNA-181, we found a significant up-regulation in mismatched unrelated donors versus matched donors (489 vs. 330%, p<0.005). Analysis of each miRNAs for relapse showed no statistically differences. Among all pts, 210 (81%) developed acute GVHD (93 pts had an aGvHD of grade ≥2). There was a significant differences between low level of miRNA-146 (529 ± 516% vs. 804 ± 1335%; p<0.04), and high level of miRNA-let7 (642 ± 419% vs. 546 ± 335%, p<0.05) of pts with aGvHD ≥ 2. In pts with severe aGVHD (grade >3) comparing all other aGvHD pts., we found a high significant reductions of miRNA-146 (333% vs. 720%, p<0.001), and miRNA-21 (528% vs. 2356%, p<0.02). In regard to chronic GvHD, we found a down-regulation of miRNA-let7 expressions in pts with severe cGVHD (384% vs. 922%, p<0.002) comparing other cGVHD in the first year. The probability of severe aGvHD with low level of miRNA-146 (< 215%, p<0.001) is 20 ± 7% versus 4 ± 2% with higher levels of miRNA-146 levels. However, an estimate for death in remission or overall survival was not significantly associated with miRNAs.

Conclusions
These results suggest that pts with different expression of miRNA-146, miRNA-21 and miRNA-let7 confirms a relevant association of the development of aGVHD and miRNA profiling could be an early indicator of severe aGvHD.

Disclosures:
Off Label Use: HCG will be discussed as new therapy for chronic GVHD.
Update On Phase 2 Clinical Trial To Induce Tolerance In Mismatched Living Donor Renal Transplant Recipients
Authors: Leventhal Joseph , Abecassis Michael , Miller Joshua , Gallon Lorenzo , Tollerud David J , Ildstad Suzanne T. 
Published: October 2019

Nineteen subjects have been enrolled in a phase 2 protocol (FDA IDE 13947) to induce donor-specific tolerance in HLA-mismatched related and unrelated recipients of living donor renal allografts (KTx). The protocol is based upon tolerogenic CD8+/TCR- facilitating cells (FC) and nonmyeloablative conditioning. Recipients are conditioned with fludarabine (30 mg/kg/dose, days -5, -4, -3), cyclophosphamide (50 mg/kg/dose, day -3 and +3), 200 cGy TBI (day -1) followed by KTx (day 0). A G-CSF mobilized peripheral blood mononuclear cell product is apheresed from the donor > 2 weeks prior to the KTx, processed to remove GVHD-producing cells yet retain CD34+ cells and FC (FCRx), and cryopreserved until infusion on day +1 post-KTx. Subjects ranged in age from 18 to 65 years. All were HLA-disparate from their donors, ranging from 5 of 6 matched related to 0 of 6 unrelated. 18 of 19 subjects exhibited engraftment at 1 month; one highly sensitized subject (PRA > 50%) did not engraft. Subjects were discharged on post-operative day 2 and managed as outpatients. Mycophenolate mofetil and tacrolimus-based immunosuppression were administered for 6 months. At 6 months the MMF was discontinued if the protocol biopsy was normal and stable renal function present. Tacrolimus was weaned at month 9 (trough levels 3-6) and discontinued at 1 year if chimerism, normal renal function, and normal kidney biopsy were noted. No subjects have experienced GVHD or engraftment syndrome. Two early subjects who received a suboptimal cell dose and one other highly sensitized subject were only transiently chimeric (< 6 months); all transiently chimeric subjects resumed endogenous hematopoiesis and are maintained on low-dose tacrolimus monotherapy with stable renal function. The protocol was subsequently modified to exclude subjects with a PRA > 20%. One chimeric subject developed viral sepsis at month 3, thrombosed his kidney, and was successfully re-transplanted off study. A second subject developed CNI-induced hemolytic uremic syndrome (HUS) which resulted in graft loss. He is currently off all immunosuppression and his HUS has resolved. The remaining subjects are either off all immunosuppression (n = 9; 1 to 36 months) or are in the process of tapering. None of the chimeric subjects have developed rejection on protocol biopsy while 3 of the 5 who were not durably chimeric had Banff 1A rejection on protocol biopsy. Recurrence of autoimmune disease has occurred in 2 of 4 non-chimeric subjects, but not in any of the durable chimeric subjects. In summary, high levels of durable chimerism and tolerance without GVHD have been achieved in mismatched related and unrelated recipients of living KTx.

Disclosures:
Tollerud: Regenerex, LLC: Equity Ownership. Ildstad:Regenerex, LLC: Equity Ownership.
Health Consequences Of Benzene Exposure Among Children Following a Flaring Incident At Petroleum Refinery In Texas City
Authors: D'Andrea Mark , Mitter Ajay , Reddy G. Kesava 
Published: October 2019

Objective
Human exposure to benzene is associated with multiple adverse health effects leading to hematological malignancies. The objective of this retrospective study was to evaluate the health consequences of benzene and other toxic chemical exposure in children following a flaring incident at the British petroleum refinery in the Texas City, Texas.

Methods
The study included children aged < 17 years who had been exposed and unexposed to benzene due to a flaring incident at the British petroleum refinery facility. Using medical charts, clinical data including white blood cell (WBC) counts, platelets counts, hemoglobin, hematocrit, blood urea nitrogen (BUN,) creatinine, alkaline phosphatase (ALP), aspartate amino transferase (AST), alanine amino transferase (ALT), and somatic symptom complaints by the children exposed to benzene were reviewed and analyzed.

Results
A total of 312 subjects (benzene exposed, n=157 and unexposed, n=155) were included. Hematologic analysis showed that WBC counts were significantly decreased in benzene exposed children compared with the unexposed children (6.8 ± 2.1 versus 7.3 ± 1.7, P=0.022). Conversely, platelet (X 103 per µL) counts were increased significantly in the benzene exposed group compared with the unexposed group (278.4 ± 59.9 versus 261.6 ± 51.7, P=0.005). Similarly, benzene exposed children had significantly higher levels of ALP (183.7± 95.6 versus 165 ± 70.3 IU/L, P=0.04), AST (23.6 ± 15.3versus 20.5 ± 5.5 IU/L, P = 0.015), and ALT (19.2 ± 7.8 versus 16.9 ± 6.9 IU/L, P=0.005) compared with the unexposed children.

Conclusion
Together, the results of the study reveal that children exposed to benzene experienced significantly altered blood profiles, liver enzymes and somatic symptoms indicating that children exposed to benzene are at a higher risk of developing hepatic or blood related disorders.

Disclosures:
No relevant conflicts of interest to declare.
Citrate Synthase Activity Is Increased In Children With Sickle Cell Disease (SCD) On Hydroxyurea (HU) Therapy
Authors: Morris Claudia R. , Latronica Cornelia , Kuypers Frans A , Rosenfeld Howard , Larkin Sandra K , Doniger Stephanie , Vichinsky Elliott , Shiva Sruti 
Published: October 2019

Background
Mitochondrial dysfunction is associated with abnormal cell function, oxidative stress and cell death. A progressive impairment of mitochondrial function and/or increased oxidative damage likely contribute to the pathophysiology of SCD, however little is known about bioenergetics in SCD. While previous work demonstrated mitochondrial dysfunction in the platelets of adult patients with SCD characterized by deficient complex IV and V activity which was associated with hemolysis, no data has yet been described in children with SCD.

Objective
To initiate investigation into mitochondrial function in children with SCD.

Methods
Children with a diagnosis of SCD requiring parenteral opioids for vaso-occlusive pain episodes (VOE) were prospectively enrolled within 24 hours of presentation to an urban emergency department (ED). Blood was obtained for biomarkers of hemolytic rate, and estimated pulmonary pressures were noninvasively measured by a point-of-care Doppler echocardiography (echo). Platelet rich plasma was isolated from patient whole blood by differential centrifugation and flash frozen for mitochondrial studies. The activities of Complexes V and IV were measured since these enzymes are the sites of oxygen consumption (Cx IV) and ATP production (CxV). The activity of citrate synthase, a mitochondrial matrix protein, was also determined as an indicator of mitochondrial number.

Results
11 children with SCD seen in the ED or admitted for VOE were included in this pilot study. Mean age was 17±3 years (range: 9-21 years), 64% were male, and 45.5% had HbSS, 45.5% HbSC and 9% HbSbThal. Mean Hb was 9.9±2 g/dL, reticulocyte (retic) count was 7.6±5%, and 45.5% were on HU. Mean tricuspid regurgitant jet velocity (TRV) = 2.0±0.4 m/s. Of interest patients on HU had significantly higher citrate synthase activity (Figure 1), although Complex IV and V activities were similar in both groups. Citrate synthase activity inversely correlated with TRV (r=-0.63, p=0.04). Similar to the previously reported adult data, complex IV activity inversely correlated with retic count (R2=0.27), although not reaching statistical significance, likely due to small sample size. No difference in mitochondrial studies were found when HbSS patients were compared to those with HbSC.

Conclusions
These data suggest mitochondrial dysfunction and potentially decreased mitochondrial number in SCD children with increased hemolytic rate (TRV). Further, the greater citrate synthase activity in platelets of patients with SCD on HU compared to those not treated with HU may be indicative of increased mitochondrial number with HU therapy. These observations have never been previously described, and may suggest a novel mechanism of action for HU warranting further study.

Disclosures:
No relevant conflicts of interest to declare.
Toll Like Receptors Have mRNA Differentiated Expression In Dendritic Cells In Crisis-State Sickle Cell Anemia Patients, Suggesting a Pivotal Role Of These Molecules and Cell Type In The Maintenance Of Inflammatory Response
Authors: Cajado Cyntia , Fukutani Kyioshi Ferreira , Boas Wendell Vilas , Veloso Cerqueira Bruno Antônio , Tavares Natalia Machado , Leite Ivana , Siqueira Isadora , Lyra Isa , Adorno Elisangela , Goncalves Marilda Souza 
Published: October 2019

Introduction
The vascular alterations play an important role in the SCA chronic inflammatory state with the participation of different cell types, including macrophages and dendritic cells (DCs).

Aim
Here, we evaluated the mRNA expression of TLRs in DCs cells pre and post LPS stimulus and also measured IL-1β, TNF-α and IL-10 cytokines in plasma and supernatant of DCs pre and post LPS challenge, in order to indentify a possible role of this cell and receptor in the inflammatory state described in sickle cell anemia.

Methods
In vitro generation of immature DCs (imDC) monocyte-derived was obtained of monocytes from peripheral blood mononuclear cells (PBMC) of seven hospitalized HbSS patients. Relative mRNA abundance of each transcript was investigated by quantitative real time PCR and cytokines levels were measured by ELISA.

Results
We found that different HbSS patients showed specific DCs TLR expression and a differential levels of IL-1β among HbSS patients with and without pneumonia history (p=0.0253) and with hemoglobin concentration (p=0.0288) (Fig. 1). There was difference of TLR4 mRNA expression between HbSS patients groups without and with leg ulcers history (0.320+0.302 and 1.60+0.014) respectively (p=0.0016); with platelets count between HbSS patients groups with TLR2 expression < 50th (569.7+3.512) and > 50th (304.5+111.7) (p=0.0102) and reticulocytes count between HbSS patients groups with TLR2 expression < 50th (15.97+2.450) and > 50th (2.025+2.417) (p=0.0007) and of TNF-alpha concentration in DCs supernatant culture among HbSS patients with TLR5 expression < 50th (30.57+11.92) and > 50th (7.053+6.918) (p=0.0496) and of IL-10 concentration in DCs supernatant culture among HbSS patients with TLR5 expression < 50th (3.525+0.921) and > 50th (5.790+1.226) (p=0.0374) (Fig.2). It was also found statistic difference of IL-1 beta concentration in DCs supernatant culture post LPS stimulus between HbSS patients without and with pneumonia history (P=0.0253); analysis of TLR4 mRNA expression between HbSS patients without and with leg ulcer history (P=0.0016); analysis of hemoglobin concentration between HbSS patients with different plasma IL-1 beta concentration (P=0.0288)

Conclusion
Ours results show that these molecules and cell type possibly have an important role in maintain the inflammatory state described in this disease.

Disclosures:
No relevant conflicts of interest to declare.
Pyknotic Lymphocytes In Infectious Mononucleosis
Authors: Dittman William A. 
Published: October 2019

Context
Pyknotic lymphocytes were infrequently observed in routine blood film evaluation of patients with clinical infectious mononucleosis. A marked increase in frequency was observed in 1995-1997.

Objective
To characterize the cause of these changes.

Design
Collection of samples from patients (25) with clinical infectious mononucleosis to ascertain the effect of anticoagulant and/or time lapse on lymphocytes and to correlate with monospot or Epstein-Barr virus antibody testing. Control samples (33), mixed hematopoietic disease (29) and lymphoproliferative disorders (36) were obtained also. Additional patients (53), identified by variant lymphocyte morphology during routine CBC’s, were verified by monospot or EBV testing and slides were made at 24 and 48 hours for differentials. Finally, single specimens (86) were obtained to establish predictability or correlation between pyknotic cells and monospot testing

Setting
Inpatient and outpatient hematology laboratory.

Results
Patients with clinical infectious mononucleosis demonstrated 8-10% pyknotic lymphocytes in 24 and 48 hour samples in all three anticoagulants (see Figure 1) compared with controls, “other diseases” (Table 2) and lymphoproliferative disorders (Table 3). An unexpected predictive correlation of pyknotic cells with positive monospot or Epstein-Barr Virus antibodies was demonstrated when pyknotic cells were seen with or without significant atypical lymphocytes during daily routine differential counting.

Conclusion
Observation of pyknotic lymphocytes adds a significant and inexpensive element to the diagnosis of infectious mononucleosis and consideration of Cytomegalic Virus infection in older patient

Disclosures:
No relevant conflicts of interest to declare.
The Effect Of Thrombotic Markers On The Severity Of Bleeding Symptoms In Women With Inherited Bleeding Disorders
Authors: Halimeh Susan , Davies Joanna , Kadir Rezan A 
Published: October 2019

Background
Women with inherited bleeding disorders demonstrate heterogenicity in bleeding symptoms that do not always correlate to underlying clotting factor levels. This is problematic when predicting the bleeding risk, especially when haemostatic challenge is anticipated. The effect of thrombotic variables in reducing the overall bleeding tendency, have demonstrated a protective effect in cases of severe haemophilia.

Aims
To assess the correlation between bleeding tendency, haemostatic and thrombotic variables in women with inherited bleeding disorders.

Method
Quantitative measurement of the bleeding tendency was assessed using the standardized ISTH Bleeding Assessment Tool. Pictorial Blood Assessment Chart (PBAC) was used to quantify the subjective assessment of menstrual blood loss. 78 women with inherited bleeding disorders (75 with von Willebrand disease and 3 carriers of haemophilia A) were included. Blood samples were then obtained from individuals for assessment of clotting factors (factor VIII and von Willebrand factor) and thrombophilia screen (lupus anticoagulant, antithrombin, protein C and protein S, factor V Leiden mutation and prothrombin mutation).

Results
Bleeding score was inversely related to von Willebrand factor antigen (vWF:Ag), ristocetin cofactor (vWF:RCo) levels. A positive correlation was demonstrated with antithrombin III activity. Ten women tested positive for thrombotic markers. Multiple regression analysis did not reveal a significant correlation with presence of thrombotic marker on bleeding score.

Conclusion
Bleeding score is a useful assessment for bleeding phenotype and seems to correlate with vWF:Ag and vWF:RCo, which may by useful for assessment of the bleeding risk. Although there was no significant correlation demonstrated by the presence of a thrombotic marker, the levels of antithrombin III appears to affect the bleeding phenotype and this requires further consideration.

Disclosures:
Halimeh: Octapharma AG: Investigator Other, Research Funding.
Changes Of Coagulation Parameters During The Menstrual Cycle
Authors: Halimeh Susan , Rott Hannelore , Kappert Guenther , Siebert Manuela 
Published: October 2019

Introduction
Von Willebrand disease is the most common bleeding disorder with a prevalence of 1-2% of the population. Nevertheless diagnosis of a von Willebrand Syndrom Typ 1 is still challenging. In a newer publication (1), 30 studies about the haemostatic variables during the menstrual cycle were compared. 11 studies were focused on the von Willebrand parameters but only in one study these parameters in patients with von Willebrand disease were observed. We investigated possible cyclic variations in women with menorrhagia, which can lead to a diagnosis of a coagulation disorder.

Samples and Methods
We conducted a laboratory workup in 122 women sent to our lab for menorrhagia. The following tests were conducted: Blood count, VWF:RCo, VWF:Ag, VWF:CB, Fibrinogen (Clauss), activities of FII, FV, FVII, FVIII (clotting and chromogenic), FIX, FX, FXI, FXII, FXIII during the menstrual cycle on predefined time points (day 1-6, day 7-11, day 12-18, day 19-23, day 24-28).

Results
In 51 (40%) patients a von Willebrand disease could be detected, 37% hat other coagulation disorders like p. e. factor-VII-deficieny and factor-XIII-deficiency. 11.5% had an iron deficiency. In 11.5% no coagulation disorder could be found.
In patients with von Willebrand disease we found cyclic variations especially in the VWF:Ag (p = 0.02). They showed the lowest level during the ovulation. For other coagulation parameters no significance for variations during the menstrual cycle were found.

Conclusion
There are cyclic variations in von Willebrand antigen. To investigate women on predefined time points during the menstrual cycle can be useful to diagnose a von Willebrand disease particularly in mild cases in which no other suspicious bleeding symptoms exists.
Literature
Knol H.M., Kemperman R.F.J., Kluin-Nelemans C., Mulder A.B., Meijer K., Haemostatic variables during normal menstrual cycle, Thrombosis and Haemostasis 107.1/2012

Disclosures:
Halimeh: Octapharma AG: Investigator Other, Research Funding.