Transient Anemia Research Articles

Efficacy and safety outcomes in heavily pretreated patients (pts) with relapsed ovarian cancer (ROC) after single-agent trabectedin.

e16538 Background: Trabectedin is a marine-derived antineoplastic agent, initially isolated from the tunicate Ecteinascidia turbinata and currently produced synthetically. In combination with pegylated liposomal doxorubicin, trabectedin is approved in Europe for the treatment of pts with platinum-sensitive (PS) ROC. The aim of this multicenter, retrospective study was to evaluate the efficacy and safety of single agent trabectedin in the palliative treatment of heavily pretreated patients with ROC. Methods: Patients with measurable ROC and at least 2 prior treatments were eligible. Patients received single agent trabectedin (1.3 mg/m2, n = 56; 1.1 mg/m2, n=42) as a 3-hour i.v. infusion every 3 weeks. An analysis of the overall response rate (ORR; primary end point) as per RECIST, time to progression (TTP) and overall survival (OS) were performed. Results: Overall, 98 pts were enrolled: median age at diagnosis of recurrence was 53 years (range: 29-79). Forty-four pts (44.9%) were fully PS, while 23 (23.5%) were partially PS and 31 (31.6%) were platinum-resistant. Median number of previous chemotherapy regimens was 4 (range: 2-6). The ORR was 28.6% (5 complete and 23 partial responses); 32 pts (32.6%) experienced stabilization of disease, which lasted ≥6 months in 2 pts, while 38 (38.8%) pts progressed during treatment. The ORR was higher in PS pts (38.6%) compared to partially PS (26.1%) and platinum resistant cases (16.1%), although the statistical significance was not reached (p=0.071). No difference in the ORR was found according to number of prior treatments or trabectedin dose. After a median follow-up of 8 months, median TTP and OS were 5 and 13 months, respectively. The most common grade 3/4 toxicities were transient and non-cumulative anemia and neutropenia in 6.1% and 17.5% of cases, respectively. AST and ALT were increased in 7.1% and 13.3% of pts. Cardiac toxicity was documented in 4 anthracycline pretreated pts, of whom one died due to acute arrhythmia. Conclusions: Single agent trabectedin represents a valid approach in the palliative treatment of pts with heavily pretreated ROC with meaningful clinical benefit and acceptable and manageable safety profile.

Rapidly involuting congenital hemangioma associated with transient anemia and thrombocytopenia

Rapidly involuting congenital hemangioma associated with transient anemia and thrombocytopenia

Population dynamics and host reactions in young foxes following experimental infection with the minute intestinal fluke, Haplorchis pumilio

BackgroundInfections with fish-borne zoonotic trematodes (FZT) including the minute intestinal fluke, Haplorchis pumilio, are highly prevalent in Southeast Asia. However, little is known about the infection dynamics and clinical symptoms in the final hosts which include a range of animal species and man. We aimed to generate such information using an experimental model with H. pumilio in foxes.MethodEight commercially bred foxes were each orally infected with 2000 H. pumilio metacercariae. Another three foxes served as uninfected controls. Faecal examination for eggs was performed twice weekly. The body weight was measured, standard haematological and biochemical analysis were performed regularly. All foxes were euthanized at day 56 post infection (p.i.). Adult worms were quantified and location in the small intestine noted.ResultsAnorexia was observed in all infected foxes starting day 12 p.i. and lasting for approximately a week. A weight loss was noticed in the infected group in weeks 3–6 p.i. Five of eight infected foxes excreted H. pumilio eggs day 9 p.i. onwards, the remaining three started on day 13 p.i. Mean (± SD) faecal egg counts showed an initial peak at day 16–20 with a maximum of 1443 ± 1176 eggs per gram of faeces (epg), where after a stable egg output around 4–500 epg was seen. Worm burdens ranged between 116–2070 adult flukes with a mean (± SD) worm recovery of 948 ± 666. The majority of worms were found in the lower part of the jejunum. Total white blood cell and lymphocyte counts were significant lower in the infected group from first week p.i. onwards and throughout the study period. A significantly lower level of eosinophils was found in week 2 p.i. and transient anaemia was seen in week 2 and 4 p.i.ConclusionThis study showed a short prepatency period, an initial peak in egg excretion, establishment of infection in all animals with predilection site in the lower jejunum and a marked but transient clinical effect of the infection. The findings on egg output and prepatency should be taken into consideration when control programs targeting dogs and other reservoir hosts are to be established.

Acadesine for patients with relapsed/refractory chronic lymphocytic leukemia (CLL): a multicenter phase I/II study

PurposeAcadesine has shown in vitro to selectively induce apoptosis in B cells from chronic lymphocytic leukemia (CLL) patients. We conducted a phase I/II open-label clinical study, to determine the safety and tolerability of acadesine given intravenously as a 4-h infusion to CLL patients.MethodsPatient population included CLL patients with relapsed/refractory disease who had received one or more prior lines of treatment including either a fludarabine or an alkylator-based regimen. Twenty-four patients were included: eighteen in Part I treated at single doses of 50–315 mg/kg, and six in Part II, three with two doses at 210 mg/kg and three with five doses at 210 mg/kg.ResultsA manageable and predictable safety profile was demonstrated for acadesine at single doses between 50 and 210 mg/kg; 210 mg/kg was the maximum tolerated dose (MTD) and optimal biological dose (OBD). Grade ≥2 hyperuricemia occurred commonly but was not clinically significant and resolved with the administration of prophylactic allopurinol. Other adverse events included transient anemia and/or thrombocytopenia (not clinically significant), renal impairment, and transient infusion-related hypotension (clinically significant). Trends of efficacy such as a reduction of peripheral CLL cells and reduction in lymphadenopathy were observed; however, the results were variable due to the small population and the range of doses tested.ConclusionsA MTD of 210 mg/kg was established with single acadesine dose. Multiple dose administrations at the OBD were tested with an acceptable safety profile, showing that acadesine might be a valuable agent for the treatment of relapsed/refractory CLL patients.

Acute megakaryoblastic leukemia and severe pulmonary fibrosis in a child with down syndrome: Successful treatment with ultra low-dose cytarabine using GATA1 mutation to monitor minimal residual disease

Acute megakaryoblastic leukemia and severe pulmonary fibrosis in a child with down syndrome: Successful treatment with ultra low-dose cytarabine using GATA1 mutation to monitor minimal residual disease

Baseline Anemia Is Not a Predictor of All-Cause Mortality in Outpatients With Advanced Heart Failure or Severe Renal Dysfunction: Results From the Norwegian Heart Failure Registry

The aim of this study was to evaluate the prognostic impact of anemia in outpatients with chronic heart failure attending specialized heart failure clinics and specifically to investigate its prognostic utility in patients with severe renal dysfunction or advanced heart failure. Anemia is an independent prognostic marker in patients with heart failure. The effect of anemia on mortality decreases with increasing creatinine levels. Multivariate Cox regression analyses were used to investigate the prognostic effect of anemia in 4,144 patients with heart failure from 21 outpatient heart failure clinics in Norway. Severe renal failure was defined as estimated glomerular filtration rate ≤45 ml/min/1.73 m2 and advanced heart failure as New York Heart Association functional classes IIIb and IV. Baseline anemia was present in 24% and was a strong predictor of all-cause mortality (adjusted hazard ratio [HR]: 1.30, 95% CI: 1.09 to 1.56, p = 0.004). Baseline anemia did not predict mortality in the 752 patients with severe renal dysfunction (adjusted HR: 1.08, 95 % CI: 0.77 to 1.51, p = 0.662) and the 528 patients with advanced heart failure (adjusted HR: 0.87, 95% CI: 0.56 to 1.34, p = 0.542). In the 1,743 patients who attended subsequent visits, sustained anemia independently predicted worse prognosis (adjusted HR: 1.47, 95% CI: 1.10 to 1.94, p = 0.008), whereas transient and new-onset anemia did not. According to our study, baseline anemia was not an independent predictor of all-cause mortality in outpatients with heart failure and accompanied severe renal dysfunction or advanced heart disease. Sustained anemia after optimizing heart failure treatment might imply worse prognosis independently of renal function and New York Heart Association functional class.

The Xla (X-linked anemia) Mouse: A Transient Neonatal Anemia Caused by a Gata1 Splicing Mutation,

Abstract 3162The Xla (X-linked anemia) mutant mouse was generated by N-ethyl-N-nitrosourea (ENU) mutagenesis and results in a severe and transient neonatal anemia. Xla/+ females exhibit severe anemia with 50% the level of red blood cell number, hematocrit and hemoglobin. Male Xla mice die in utero at 10.5 days gestation. The neonatal anemia observed in Xla/+ female pups is resolved by weaning age at 3 weeks by which time the mice present with a normal hematological phenotype. It is unknown how the neonatal anemia in Xla/+ females is alleviated. Previously, we mapped the Xla locus to the proximal end of the X chromosome near candidate gene Gata1 which showed no change in the coding sequence of GATA1 protein. Now we report the identification of a Gata1 mutation in Xla mice that results in an mRNA splicing defect. A nucleotide change (G to A) was identified 5 base pairs downstream of Exon 1E in intron 1 of the Xla Gata1 gene and results in the lack of incorporation of Exon 1E in the Gata1 mRNA expressed from the mutant locus. Therefore, in some erythroid lineage cells in Xla/+ mice, the normal 1E exon of Gata1 mRNA is replaced by Exon 1Eb/c which is known not to impact erythropoeisis since no GATA1 protein is made by this mRNA due to its inability to bind to ribosomes. These data show the Xla mouse results from a single nucleotide change impacting the normal splicing of the Gata1 gene. A second goal of this study was to understand why Xla/+ mice exhibit the neonatal transient anemia. A contributing factor is X chromosome inactivation which occurs in female mice during development. The short-term anemia in Xla mice was thought to be due to clonal selection of erythroid lineage cells characterized by the expression of GATA1 protein from the active X chromosome expressing only from the wild type Gata1 locus. Using an X-linked gene expressed in red blood cells (Pgk1, phosphoglycerate kinase 1) that varies between Xla mice and a wild derived strain, CAST/Ei, we examined the active state of the X chromosomes based on the expression of Pgk1 RNA in reticulocytes from hybrid Xla mice generated by breeding of these different strains. Examining expression of the X-linked Pgk1 SNP variant in the RNA of reticulocytes from hybrid Xla/+ mice reveals red blood cells are generated from two types of erythroid lineage cells. Pgk1 SNP RT-PCR analysis reveals that red blood cells not only derive from erythroid progenitors with the active X chromosome carrying the wild type Gata1 gene but also red blood cells are produced by erythroid lineage cells expressing the Xla mutant Gata1 mRNA on the active X chromosome (which does not make GATA1 protein). Therefore, some Xla erythroid cells derive from progenitors which express Gata1 transcripts using Exon 1Eb/c that does not stimulate erythropoiesis due to lack of GATA1 protein. The question is how these erythroid precursors generate normal red blood cells without the production of GATA1 protein. We hypothesize there is a developmentally expressed compensatory gene or pathway replacing GATA1 expression in GATA1-lacking erythroid precursors and required for the production of red blood cells in Xla mice. Analysis is underway to identify a potential novel gene or pathway impacting erythropoiesis in these mutant mice. Disclosures:No relevant conflicts of interest to declare.

Mtor Complex 1 Plays Critical Roles in Hematopoiesis and Pten-Loss-Evoked Leukemogenesis

Abstract 391The mammalian Target of Rapamycin (mTor) is an evolutionary-conserved serine/threonine protein kinase that plays critical roles in energy homeostasis, and cell-decision processes, such as cell growth and proliferation. Genetic alterations that lead to increased mTor activity in humans and mouse models have suggested a critical role for this kinase and its signal-transduction pathway in hematopoietic-stem cell (HSC) biology as well leukemogenesis, and has garnered much attention as a therapeutic target in human hematopoietic diseases. Further confirmation for a critical function of mTor in hematopoiesis has been suggested through pharmacological inhibition with rapamycin. However, recent evidence has shown that rapamycin is an incomplete inhibitor of mTor and its two associated complexes (mTorc1 and mTorc2). We thus sought to definitively discern the role of mTorc1 in hematopoiesis and leukemogenesis by utilizing mice containing inducible “knock-out” alleles of the essential mTorc1 component Raptor.To conditionally ablate the Raptor gene, mice with flanking loxP sites (floxed) of exon 6 of raptor were crossed with mice expressing Cre under the interferon-inducible Mx promoter (MxCre), and deletion induced in 4–6 week old offspring with injections of polyIpolyC. Induced MxCre:raptor homozoygous mice (KO) and controls (WT) were then analyzed for hematopoietic pertubations. Deletion of raptor leads to a complete ablation of mTorc1 biochemical activity in HSC and progenitors (HSPC) as assessed by phosphoflow analysis of key mTorc1 substrates in response to stem-cell factor. Furthermore, cell-type specific pertubations of a major negative feedback loop from mTor to the Akt pathway were uncovered by this analysis, with increased pAkt levels apparent only in early monocytic cells. Loss of mTorc1 activity produced a non-lethal hematopoietic disorder characterized by leucopenia, transient hypochromic anemia, and thrombocytopenia. Bone marrow (BM) of KO mice was characterized by an apparent lack of normal hematopoiesis and a striking expansion of monoyctic cells at the expense of neutrophils and B lymphocytes. These data suggest a critical role for mTorc1 in the execution of myelomonocytic development. Splenomegaly was prevalent in KO animals, reflecting extramedullary hematopoiesis. As many of these hematopoietic alterations could result from defective HSPC function, we examined the HSPC compartment in more detail. Raptor loss leads to an accumulation of mostly G0 immunophenotypically-defined HSC and multi-potent progenitors in the BM. Metabolic and transcriptional profiling of HSC/MPP indicated pertubations in major metabolic pathways in the KO cells, including decreases in cholesterol and steroid biosynthesis, and alterations in transamination reactions, among others. Strikingly, loss of mTorc1 in HSC/MPP leads to alterations in expression of genes that play critical roles in HSC. To examine the relevancy of these findings to HSC function we performed a series of competitive transplantation experiments. Results from these experiments indicate that mTorc1 is required in a cell-autonomous manner for HSC fitness and proper myelomonocytic differentiation, likely through the control of key metabolic and gene-expression programs.While mTorc1/Raptor loss resulted in profound hematopoietic dysfunction, KO mice survived for greater than 1 year after induced deletion of the raptor gene and maintained hematopoiesis in the absence of Raptor. We next asked whether loss of mTorc1 could be a reasonable therapeutic target in leukemia by generating mice with compound floxed alleles of both Raptor and the Pten gene (deleted again with Cre under the Mx promoter). While homozygous Pten deletion results in a rapidly-fatal myeloproliferative disease in both WT and induced Raptor heterozygous mice, homozygous deletion of Raptor leads to a significantly prolonged survival. Furthermore, through transplantation experiments, we show that mTorc1 is required in a cell-autonomous manner for Pten-loss-evoked leukemogenesis. These studies have demonstrated critical functions of mTorc1 in benign hematopoiesis, suggest rapamycin-independent functions of mTorc1 in this system, and imply that mTorc1 ablation can have beneficial effects in mouse models of leukemogenesis. Disclosures:Scadden:Fate Therapeutics: Equity Ownership. Armstrong:Epizyme: Consultancy.

Analysis of Ribosomal Protein Genes Associated with Diamond Blackfan Anemia (DBA) In German DBA Patients and Their Relatives

Abstract 729On behalf of the German DBA registry. The first two authors contributed equally.Mutations of ribosomal protein (RP) genes cause diamond blackfan anemia (DBA), a congenital bone marrow failure syndrome characterized by erythroid failure associated with diverse malformations of multiple organ systems. Familial dominant inheritance has been established in at least one third of all cases. Some affected family members present only with history of transient anemia during childhood without additonal clinical abnormalities. Laboratory parameters supporting the diagnosis of DBA and pointing towards a non-penetrant carrier are macrocytosis, elevated hemoglobin F (HbF) and adenosine deaminase (ADA). The presence of a more severe disease phenotype in DBA offsprings affected by the disease raised the question of potential genetic anticipation. However, the insufficient longitudinal datasets, the non-prospective nature of investigations and other complex issues such as fecundity bias make it difficult to evaluate this problem in DBA patients. The purpose of the current study was to investigate the incidence of mutations in ribosomal protein genes in family members of DBA patients, irrespective of family history, specific symptoms or laboratory parameters.Based on the results from the ongoing DBA re-sequencing study we determined the mutational status of German DBA patients for the following 12 RP genes: RPS7, RPS10, RPS17, RPS19, RPS24, RPS26, RPL5, RPL9, RPL11, RPL19, RPL26, RPL35a. Out of 179 index patients with DBA, 94 patients carried an exclusive mutation or deletion of the following 9 genes: RPS19, n=46 (25,7%); RPL5, n=15 (8,4%); RPS26, n=12 (6,7%); RPL11, n=8(4,5%), RPL35a, n=6 (3,4%), RPS17, n=3 (1,7%), RPS24, n=2 (1,1%), RPS10, n=1 (0,6%), RPS7, n=1 (0,6%). In two patients, large genomic deletions were discovered using Affymetrix 6.0 SNP-array karyotyping, which encompass RPL5 and both copies of RPS17, respectively. No aberrations were found in RPL9, RPL19 and RPL26. A total of 85 patients (47%) were negative for all 12 tested genes. In the next step, RP genes with nonsynonymous changes identified in index patients were interrogated in patients` relatives. The investigation of parent-offspring trios revealed de novo gene aberrations in 32/55 (58%) of families (RPS19, n=14; RPL5, n=8; RPS26, n=4; RPL11, n=3, RPL35a, n=2; RPS17, n=1). Maternal inheritance pattern was identified in a total of 17/55 families (RPS19, n=10; RPS24, n=2; RPL5, n=2; RPS10, n=1; RPS26, n=1; RPL11, n=1), whereas a paternal transmission of the mutation was observed in 6/55 families (RPS19, n=3; RPL5, n=1; RPL35a, n=1 and RPS26, n=1). Among parents with identified mutation, 43% were silent carriers (presenting with normal phenotype and no history of anemia). Strikingly, 4/10 silent carriers with mutation (RPS10 Lys24Arg; RPS26 Arg87X; RPS19 Val30SerfsX46; RPS19 Arg94X) had normal HbF and ADA values. Taken together, spontaneous remission and silent clinical phenotype accounted for 87% of parental cases with identified RP mutations as opposed to 24% of offsprings with respective RP mutations.In summary, in German DBA cohort, RP genetic defects arise de novo at a frequency of 58% and are inherited over at least one generation in 42% of cases, with no significant clustering of RP gene mutations. Maternal inheritance pattern predominates at 74% of all inherited cases. The unequal cumulative frequency of spontaneous remission and silent phenotype among parental subjects versus offsprings with mutated RP genes might indicate increasing severity of the disease in each generation. This observation and the unexpectedly high frequency of silent carriers with normal HbF and ADA values warrants further studies with a large number of longitudinal datasets and known RP gene status. Disclosures:No relevant conflicts of interest to declare.

Prevalence and determinants of anemia in the immediate postkidney transplant period

At the time of renal transplantation, erythropoiesis-stimulating agents and iron supplementation are routinely discontinued in the prospect of recovery of renal function. This recovery, however, is often delayed and suboptimal. In addition, blood loss because of frequent diagnostic phlebotomies may be substantial. Renal transplant recipients may thus be considered at high risk of anemia in the immediate post-transplant period. We performed a single-center observational study, including 391 recipients of a single kidney. Hemoglobin levels and parameters of iron metabolism were monitored during the immediate post-transplant period, i.e., the first 3 months after transplantation. Hemoglobin levels decreased by 3.8 ± 1.5 g/dl to reach a nadir of 9.1 ± 1.2 g/dl at day 7. Transient severe anemia was observed in 91.3% of the patients. Donor age, gender, renal diagnosis of polycystic disease, pretransplant hemoglobin and ferritin level, estimated glomerular filtration rate at month 3, and duration of initial hospitalization were observed to be independently associated with the hemoglobin level at month 3. Transient severe anemia is an almost universal observation in incident renal transplant recipients. Poor graft function, high donor age, and low iron stores are independently associated with low hemoglobin levels at month 3.

Gene Expression Profiling in rMd5- and rMd5Δmeq-Infected Chickens

Marek's disease (MD) is a lymphoproliferative disorder of domestic chickens caused by a highly contagious and oncogenic alpha-herpesvirus, Marek's disease virus (MDV). MD is characterized by bursal-thymic atrophy and rapid onset of T-cell lymphomas that infiltrate lymphoid tissues, visceral organs, and peripheral nerves with severe clinical signs that include transient paralysis, anemia, weight loss, and neurologic disorders. Using overlapping cosmids- and BAC-cloned MDV, it has been shown that MDV-encoded vIL-8, pp38, vTR, vLIP, RLORF4, and meq are among the many essential genes that play critical roles in viral pathogenesis. Of all the genes investigated so far, only meq has been shown to be consistently expressed in all MDV-derived tumors and lymphoblastoid cell lines. Meq is a basic leucine-zipper protein that shares homology with the jun/fos family of transcriptional factors. There are two copies of meq gene within the MDV genome that are only present in the serotype-1 strains. It has been shown conclusively that deletion of meq results in loss of transformation of T cells in chickens, with no effect on the early cytolytic phase of infection in lymphoid organs, which is essential for induction of innate and adaptive immunity. The goal of this study was to investigate 1) the effect of the meq oncogene on the expression pattern of select chicken immune and nonimmune-related genes, and 2) its potential role in MDV-induced apoptosis. We used real-time reverse transcriptase-polymerase chain reaction to evaluate the expression profiling of a panel of chicken genes in rMd5- and rMd5deltameq-infected chickens at 5, 14, 21, and 35 days postinfection (dpi). Although the transcriptional activities of several immune-related genes, including IL-6, IL-10, cMGF, GM-CSF, iNOS, IFNbeta, and INFgamma, were higher in rMd5deltameq-infected chickens at 5 dpi when compared to the rMd5-infected birds, the differences in expression levels of the tested genes between the two viral constructs were not significant. In addition, a reduction in the transcriptional activity of Bdcl2 in recombinant fowlpox virus (rFPV)+meq-infected chicken embryonic fibroblasts suggested that meq alone did not impede FPV-induced apoptosis. The likely suppressive nature and anti-inflammatory function of the meq oncogene and its possible role in virus-induced cell death is discussed.

Recovery From Hospital-Acquired Anemia After Acute Myocardial Infarction and Effect on Outcomes

New-onset, hospital-acquired anemia (HAA) during acute myocardial infarction (AMI) is independently associated with poor outcomes. The patterns of recovery from HAA after AMI and their association with mortality and health status are unknown. In the prospective 24-center Translational Research Investigating Underlying disparities in acute myocardial infarction Patients' Health Status (TRIUMPH) registry, we identified 530 patients with AMI and HAA (defined as normal hemoglobin at admission with the development of anemia by discharge) who had a repeat, protocol-driven hemoglobin measurement at 1 month after discharge. The 1-month measures were used to define persistent (persistent anemia) and transient (anemia resolved) HAA. The patients' health status was assessed at 1, 6, and 12 months after AMI using the Short-Form 12 Physical Component Summary, and the health status of patients with persistent and transient HAA was compared using multivariate repeated measures regression analysis. Mortality was compared using the log-rank test and proportional hazards regression analysis. Overall, 165 patients (31%) developed persistent HAA. The adjusted mean Short-Form 12 Physical Component Summary scores at the follow-up visit were significantly lower in those with persistent HAA than in those with transient HAA (-2.0 points, 95% confidence interval -3.6 to -0.3; p = 0.02). During a median follow-up of 36 months, the crude mortality (13% vs 5%, p = 0.002) and multivariate-adjusted mortality (hazard ratio 2.08, 95% confidence interval 1.02 to 4.21, p = 0.04) was greater in patients with persistent HAA. In conclusion, HAA persists 1 month after discharge in nearly 1 of 3 patients and is associated with worse health status and greater mortality. Additional investigation is needed to understand whether HAA prevention, recognition, and treatment, particularly among those with persistent HAA, will improve outcomes.

Cytotoxic Effect of Poly-Dispersed Single Walled Carbon Nanotubes on Erythrocytes In Vitro and In Vivo

Single wall Carbon Nanotubes (SWCNTs) are hydrophobic and do not disperse in aqueous solvents. Acid functionalization of SWCNTs results in attachment of carboxy and sulfonate groups to carbon atoms and the resulting acid functionalized product (AF-SWCNTs) is negatively charged and disperses easily in water and buffers. In the present study, effect of AF-SWCNTs on blood erythrocytes was examined. Incubation of mouse erythrocytes with AF-SWCNTs and not with control SWCNTs, resulted in a dose and time dependent lysis of erythrocyte. Using fluorescence tagged AF-SWCNTs, binding of AF-SWCNTs with erythrocytes could be demonstrated. Confocal microscopy results indicated that AF-SWCNTs could enter the erythrocytes. Treatment with AF-SWCNTs resulted in exposure of hydrophobic patches on erythrocyte membrane that is indicative of membrane damage. A time and dose dependent increase in externalization of phosphatidylserine on erythrocyte membrane bilayer was also found. Administration of AF-SWCNTs through intravenous route resulted in a transient anemia as seen by a sharp decline in blood erythrocyte count accompanied with a significant drop in blood haemoglobin level. Administration of AF-SWCNTs through intratracheal administration also showed significant decline in RBC count while administration through other routes (gavage and intra-peritoneal) was not effective. By using a recently developed technique of a two step in vivo biotinylation of erythrocytes that enables simultaneous enumeration of young (age <10 days) and old (age>40 days) erythrocytes in mouse blood, it was found that the in vivo toxic effect of AF-SWCNTs was more pronounced on older subpopulation of erythrocytes. Subpopulation of old erythrocytes fell after treatment with AF-SWCNTs but recovered by third day after the intravenous administration of AF-SWCNTs. Taken together our results indicate that treatment with AF-SWCNTs results in acute membrane damage and eventual lysis of erythrocytes. Intravenous administration of AF-SWCNTs resulted in a transient anemia in which older erythrocytes are preferably lysed.

A safety evaluation of DAS181, a sialidase fusion protein, in rodents.

DAS181 is a novel inhaled drug candidate blocking influenza virus (IFV) and parainfluenza virus (PIV) infections through removal of sialic acid receptors from epithelial surface of the respiratory tract. To support clinical development, a 28-day Good Laboratory Practices inhalation toxicology study was conducted in Sprague-Dawley rats. In this study, achieved average daily doses based on exposure concentrations were 0.47, 0.90, 1.55, and 3.00 mg/kg/day of DAS181 in a dry powder formulation. DAS181 was well tolerated at all dose levels, and there were no significant toxicological findings. DAS181 administration did not affect animal body weight, food consumption, clinical signs, ophthalmology, respiratory parameters, or organ weight. Gross pathology evaluations were unremarkable. Histological examination of the lungs was devoid of pulmonary tissue damage, and findings were limited to mild and transient changes indicative of exposure and clearance of a foreign protein. DAS181 did not show any cytotoxic effects on human and animal primary cells, including hepatocytes, skeletal muscle cells, osteoblasts, or respiratory epithelial cells. DAS181 did not cause direct or indirect hemolysis. A laboratory abnormality observed in the 28-day toxicology study was mild and transient anemia in male rats at the 3.00 mg/kg dose, which is an expected outcome of enhanced clearance of desialylated red blood cells resulting from systemic exposure with DAS181. Another laboratory observation was a transient dose-dependent elevation in alkaline phosphatase (ALP), which can be attributed to reduced ALP clearance resulting from increased protein desialylation due to DAS181 systemic exposure. These laboratory parameters returned to normal at the end of the recovery period.

A Hemoglobin Variant Associated with Neonatal Cyanosis and Anemia

Globin-gene mutations are a rare but important cause of cyanosis. We identified a missense mutation in the fetal Gγ-globin gene (HBG2) in a father and daughter with transient neonatal cyanosis and anemia. This new mutation modifies the ligand-binding pocket of fetal hemoglobin by means of two mechanisms. First, the relatively large side chain of methionine decreases both the affinity of oxygen for binding to the mutant hemoglobin subunit and the rate at which it does so. Second, the mutant methionine is converted to aspartic acid post-translationally, probably through oxidative mechanisms. The presence of this polar amino acid in the heme pocket is predicted to enhance hemoglobin denaturation, causing anemia.

Acute Nonimmune Hemolytic Anemia Without Fulminant Hepatitis in Wilson Disease

Owing to the insidious course and variable presentation, Wilson disease is often diagnosed months to years after the initial symptoms. Although fulminant hepatitis with nonimmune hemolytic anemia is frequently reported, chronic mild hepatitis can occur with bouts of transient hemolytic anemia. We report a 16-year-old female who presented with fatigue, dizziness, and new onset jaundice. She had a hemolytic anemia, although diagnosis of Wilson disease was initially confounded by a family history of autoimmunity with a high erythrocyte sedimentation rate and only mildly elevated bilirubin and aspartate aminotransferase. Macrocytosis, poor liver synthetic function, and low serum alkaline phosphatase led to the diagnosis.

Identification of Bartonella henselae in 2 Cats With Pyogranulomatous Myocarditis and Diaphragmatic Myositis

Most cats infected with Bartonella henselae remain outwardly healthy carriers for years; however, self-limiting fever, transient anemia, neurologic dysfunction, lymphadenopathy, reproductive disorders, aortic valvular endocarditis, and neutrophilic myocarditis have been described in experimentally or naturally infected cats. Two cats in a North Carolina shelter died with pyogranulomatous myocarditis and diaphragmatic myositis. Bacteria were visualized in the lesions by Warthin-Starry silver impregnation and by B. henselae immunohistochemistry. B. henselae DNA was amplified and sequenced from the heart of 1 cat and from multiple tissue samples, including heart and diaphragm, from the second cat. This study supports a potential association between B. henselae and what has been historically described as "transmissible myocarditis and diaphragmitis" of undetermined cause in cats.

Transient aplastic anemia in Down’s syndrome – A rare association

The association of Down’s syndrome with aplastic anemia is extremely rare with only six such cases reported in world literature. Herein, we report a child of Down’s syndrome with pancytopenia and hypocellular marrow. There was associated hypothyroidism and the pancytopenia resolved with thyroxine treatment. The child made uneventful recovery.

Promising Efficacy with the New Anti-CD20 Antibody GA101 In Heavily Pre-Treated NHL Patients – First Results From a Phase II Study In Patients with Relapsed/Refractory DLBCL and MCL

AbstractAbstract 2878 Background:GA101 is the first type II, glycoengineered and humanized monoclonal anti-CD20 antibody with Phase I (Salles, ASH 2008, 2009; Cartron, EHA 2009; Morschhauser, ASH 2009; Sehn, ASH 2009) and indolent NHL phase II results (Salles, EHA, 2010) previously reported. Methods:Forty eligible patients [25 DLBCL/15 MCL] were randomized to receive GA101 in a low-dose (LD, n=21 [10 DLBCL/11 MCL]) or a high dose (HD, n=19 [15 DLBCL/4 MCL]) cohort. GA101 was given on d1, d8, d22 and q21 days for total of 9 infusions. In the LD cohort, GA101 was given 400mg all infusions; in the HD cohort, d1 and d8 at 1600mg and 800mg thereafter. Primary endpoint was end of treatment response (EOR), assessed 4 weeks after last infusion (44 weeks after treatment start). Secondary objectives were safety, pharmacokinetics and progression free survival (PFS). Results:Patients (Table 1) were heavily pre-treated (median 3 prior therapies), with 63% of patients not responding to or relapsing within six months after a previous rituximab-containing regimen (rituximab refractory) and 45% of patients completed all 9 infusions. For the LD cohort, EOR was 24% 5/21 (DLBCL – 2 PR, 1 CRu; MCL – 2 CR) and 32% (6/19) in the HD cohort (DLBCL – 4 PR; MCL – 2 PR). For DLBCL patients EOR was:28% (7/15) (1 CRu, 6 PR). For MCL patients EOR was: 27% (4/15) (2 CR, 2 PR). Of 13 refractory patients in LD, 1 patient achieved PR (8%) and of 12 refractory patients in HD, 3 patients achieved PR (25%), with two converting to CR during additional follow-up. Median PFS was 78 days [8-356+] and 83 days [7-428+] for the LD and HD cohorts respectively (Hazard ratio 0.88 [95% CI 0.43;1.79]).The most common AEs were G1-2 infusion related reactions (LD 81%, HD 68% of patients). Fourteen patients experienced at least one SAE during treatment period (LD=9, HD=5), with 7 related to GA101 (LD n=5, HD n=2), and 5 of these events associated with the first infusion (IRR=3; TLS=2), one after day 8 infusion (pyrexia), and one after cycle 6 infusion (bradycardia). During treatment, related G3-4 hematological AEs were transient neutropenia (n = 1 in HD), anemia (n = 2 in LD) and thrombocytopenia (n = 3 in LD). Ten patients had at least one G1-2 infection (5 in each cohort) with no G3-4 infections reported. GA101 plasma profiles were explored reflecting that mantle cell lymphoma patients in the HD (n=4) and LD (n=10) appeared to show lower plasma concentrations compared to DLBCL patients. There were 17 patient deaths reported (LD=11, HD=6), with 14 due to PD and 3 due to AE (1 during treatment period; cardio-respiratory arrest; not GA101-related) and two SAEs (LD) in follow-up (gastric haemorrhage, lung infection – both not GA101-related). Conclusion:In this group of heavily pre-treated DLBCL and MCL patients, single-agent GA101 was well tolerated with promising evidence of efficacy.Table 1:Baseline patient characteristics and response dataLD Cohort (400mg)HD Cohort (1600/800mg)Alln211940DLBCL histology (n)101525MCL histology (n)11415Median age (range)70 (43–80)72 (22–85)71 (22–85)Median # of prior treatments (range)4 (1–17)3 (1–6)3 (1–17)Previous rituximab (n)211940Rituximab refractory (n)131225Prior stem cell transplant (n)268Response DataEOR5 (24)%6 (32%)11 (28%)EOR (DLBCL only)3 (30%)4 (27%)7 (29%)EOR (MCL only)2 (18%)2 (50%)4 (27%)EOR Rituximab-refractory1 (8%)3 (25%)4 (16%)Median PFS (days)78 days [8–356]83 days [7–428]Hazard ratio 0.88 [95% CI 0.43;1.79]Median PFS (DLBCL only)57 days [8–351]83 days [7–428]Hazard ratio 0.79 [95% CI 0.32; 1.92] Disclosure:Cartron:Roche: Consultancy, Honoraria; GSK: Honoraria. Morschhhauser:Roche: Honoraria. Birkett:Roche: Employment. Wenger:Roche: Employment. Asikanius:Roche: Employment. Salles:Roche: Honoraria.

E0372 Post-operative observation of the safety and angiogenesis effect of direct current stimulation in a myocardial infarction rabbit model

IntroductionThe purpose of the current study was to evaluate the safety of low voltage direct current (DC) electric stimulation and its angiogenesis effect in a rabbit myocardial infarction (MI) model...