Dengue fever, which is transmitted by the mosquito Aedes aegypti is a common acute viral febrile illness, affecting 390 million people worldwide every year. It is endemic in several countries. Although the usual presentation is that of a self-limiting illness, its complications are protean. Infection Associated with Hemophagocytic Syndrome (IAHS) is a rare and fatal complication of dengue fever. It should be suspected in a patient with fever beyond seven days, associated with hepatosplenomegaly, hyperferritinemia, worsening cytopenias and development of multiorgan dysfunction(MOD). We report a 19-year-old female, who presented with an acute febrile illness and was diagnosed with primary dengue fever. Despite appropriate supportive therapy, she worsened clinically during the course of hospitalisation. A disproportionately high ferritin level and persistent bicytopenia prompted investigations for hemophagocytic lymphohistiocytosis (HLH). Further evaluation revealed features of HLH, as per the diagnostic criteria laid down by the Histiocyte Society. She was successfully treated with glucocorticoids, etoposide and intravenous immunoglobin and other supportive therapy. She had severe cytopenia and cardiac dysfunction during the course of her illness and its management. This case adds to the limited adult cases of dengue-associated hemophagocytic syndrome and shows its difficulty in management due to associated extra complications.
Rho GTPases are involved in homing and mobilization of hematopoietic stem and progenitor cells due to their impact on cytoskeleton remodeling. We have previously shown that inhibition of Rho, Rac and Cdc42 clearly impairs adhesion of normal and leukemic hematopoietic progenitor cells (HPC) to fibronectin and migration in a three-dimensional stromal cell model. Here, we identified the Ras GTPase-Activating Protein SH3 Domain-Binding Protein (G3BP) as a target gene of Rho GTPases and analysed its role in regulating HPC motility. Overexpression of G3BP significantly enhanced adhesion of murine 32D HPC to fibronectin and human umbilical vein endothelial cells, increased the proportion of adherent cells in a flow chamber assay and promoted cell migration in a transwell assay and a three-dimensional stromal cell model suggesting a strong impact on the cytoskeleton. Immunofluorescent staining of G3BP-overexpressing fibroblasts revealed a Rho-like phenotype characterized by formation of actin stress fibers in contrast to the Rac-like phenotype of control fibroblasts. This is the first report implicating a role for G3BP in Rho GTPase-mediated signalling towards adhesion and migration of HPC. Our results may be of clinical importance, since G3BP was found overexpressed in human cancers.
Intravenous immunoglobulin (IVIG) has been manufactured from pooled plasma of 10,000 or more units from healthy donors.Recently, we reported that the IVIG manufactured even before the 2009 influenza pandemic contained antibodies reactive to seasonal H1N1 and pandemic H1N1 2009 (H1N1 pdm) viruses.In this study, we used an animal model to evaluate the efficacy of IVIG against influenza infections.A seasonal influenza H1N1 strain (New Caledonia, A/NC/20/99) and an H1N1 pdm strain (A/Osaka/168/2009) were used.The BALB/c and severe combined immunodeficiency mice (SCID; C.B-17/lcr-scid/scid) were also used.Mice inoculated with A/NC/20/99 or A/Osaka/168/2009 were administrated IVIG and monitored for 3 weeks.The administration of IVIG 48 h before and after inoculation with a mouse-adapted seasonal H1N1 virus, resulted in survival rates of 80 and 88%, respectively.The rate among control mice was 30%.In addition, infectivity in lungs from IVIG-treated mice also decreased significantly.Similar effects of IVIG on the survival rate were obtained with H1N1 pdm.Thus, IVIG was shown to be effective against both viruses in mice.
Background: Red blood cells and platelet concentrates are frequently used in paediatric oncology.There is little literature on the indications and costs of this adjuvant therapy.Aim: To retrospectively evaluate the indications, amounts and costs of transfusing blood products in 2008 in the Department of Paediatric Oncology at Tygerberg Children's Hospital, Cape Town. Material and Method:The patient records were analyzed for age, sex, disease and stage, indications for transfusion and type of blood product as well as amount.The costs were obtained from the Blood Bank for each item released for every patient.Results: Thirty-nine children with cancer were transfused, between one unit and 34 units of blood products per patient, during their treatment in 2008.The total cost of this therapy in 2008 was ZAR 941,966 = USD 125,595 = EUR 89,711.The maximum cost per patient amounted to ZAR 70,682 = USD 9,424 = EUR 6,731 and the minimum ZAR 914 = USD 121 = EUR 87.The average expenditure per patient was ZAR 24,125 = USD 3,216 = EUR 2,297.The management of leukemia required the highest usage of blood products per patient. Conclusion:The use of blood products is indispensable during the treatment of numerous haematology -oncology diseases.Their indications should be specified in internal protocols and their actual use should be audited frequently due to the considerable costs.
The BCR/ABL fusion protein is the hallmark of Philadelphia Chromosome positive (Ph+) leukemia. The constitutive activation of the ABL-kinase in BCR/ABL cells induces the leukemic phenotype. Targeted inhibition of BCR/ABL by small molecule inhibitors reverses the transformation potential of BCR/ABL. Recently, we definitively proved that targeting the tetramerization of BCR/ABL mediated by the N-terminal coiled-coil domain (CC) using competitive peptides, representing the helix-2 of the CC, represents a valid therapeutic approach for treating Ph+ leukemia. To further develop competitive peptides for targeting BCR/ABL, we created a membrane permeable helix-2 peptide (MPH-2) by fusing the helix-2 peptide with a peptide transduction tag. In this study, we report that the MPH-2: (i) interacted with BCR/ABL in vivo; (ii) efficiently inhibited the autophosphorylation of BCR/ABL; (iii) suppressed the growth and viability of Ph+ leukemic cells; and (iv) was efficiently transduced into mononuclear cells (MNC) in an in vivo mouse model. This study provides the first evidence that an efficient peptide transduction system facilitates the employment of competitive peptides to target the oligomerization interface of BCR/ABL in vivo .
A chronic undefined illness characterized by infectious mononucleosis (IM)-like symptoms and signs, possibly associated with Epstein-Barr virus (EBV) infection, designated as so-called chronic active EBV infection (CAEBV), is focused and discussed in this mini-review. Patients with CAEBV often develop T cell lymphoproliferative disorder (LPD)/lymphoma or NK cell LPD/lymphoma. Unique manifestations with generally poor prognosis of the disease prompt us to understand in particular the entity, diagnosis and treatment.
We studied the geographical distribution of all cases of HM that were diagnosed among residents of Sardinia during the 20-year period from 1974 to 1993.Cases, grouped into three categories (lymphoproliferative diseases, myeloproliferative diseases, and total haematological malignancies), were assigned to the 356 municipalities of the island according to the reported residence of each patient.In each municipality, the relative risk (RR) of diagnosis was calculated for the three disease groups, for females, for males, and for the total population.The RR of diagnosis of HM was significantly different from 1 (p<0.05)for a limited number of estimates (5.4%), with almost as many instances of elevated RR as of reduced RR.Interested municipalities (87) were distributed unevenly across Sardinia.Nevertheless, we did observe the following: 1) Municipalities with increased risk were mostly situated in central and northern Sardinia, especially for lymphoproliferative diseases in females;2) One municipality in north-central Sardinia featured numerous estimates of elevated RR;3) A group of municipalities near the east coast of the island had 1-5 estimates of reduced RR, particularly for lymphoproliferative diseases in females; 4) Only a few municipalities located in areas harbouring potentially polluting activities exhibited increased RRs.Observed differences in RR, if not due to chance, might be linked to unknown environmental or genetic factors.
NPM1 belongs to a new category of genes that function both as oncogenes and tumor suppressor genes, depending on gene dosage, expression levels, interacting partners, and compartmentalization. Nucleophosmin mutations within exon 12 have been described as the most frequent acquired molecular abnormalities in adult and pediatric acute myeloid leukaemia (AML), mutation can be observed in nearly half of patients with a normal karyotype and is associated with a favorable outcome. NPM1 mutations are characterized by the aberrant cytoplasmic localization of NPM, the absence of CD34, involvement of several cell lineages myeloid, monocytic, erythroid and megakaryocytic but not lymphoid and a high frequency of FLT3-ITD mutation. We aimed to study the prevalence, association with Flt3 mutations, and prognostic impact of NPM1 exon-12 mutations in 71 AML patients with normal karyotype. We studied NPM1 and FLT3 by RT PCR. NPM1 gene mutation was detected among 34 patients (47.9%) and was associated with a high white blood cell count, involvement of the monocytic lineage, CD34 negativity, and high frequency of FLT3ITD. DFS and OS did not differ between mutated and unmutated NPM patients. Prospective studies are needed to confirm the definitive place of NPM mutation among patients with normal karyotype.
Renal insufficiency is commonly associated with an erythropoietin (EPO) dependent anemia. CNTO 530 is an EPO-mimetic antibody fusion protein with EPO-receptor agonist activity. The pharmacokinetics and pharmacodynamic activity of CNTO 530 were evaluated in a rat model of chronic renal insufficiency (CRI) induced anemia. Following 5/6 nephrectomy and 13 -17 weeks of stabilization, rats received a single subcutaneous (SC) dose of saline or CNTO 530 (0.03, 0.1, or 0.3 mg/kg). CNTO 530 caused dose-dependent reticulocytosis and long-lived increases in red blood cells, hemoglobin, and hematocrit. After receiving a dose of 0.1 mg/kg, red cell indices increased and remained within normal range for 44 days. Pharmacokinetic analysis showed that clearance of CNTO 530 increased 1.4 fold and terminal t1/2 and AUC decreased 1.6 fold in CRI rats compared to controls. CNTO 530 is active in a CRI rat model of anemia and widely spaced treatments with CNTO 530 may be sufficient to combat anemia.
We developed and tested a compound called AGEM400(HES) that consists of a novel erythropoietin mimetic peptide (EMP) which is produced as a continuous N- to C-linked dimer and is conjugated to biodegradable hydroxyethyl starch (HES). In various in vitro assays, AGEM400(HES) demonstrated excellent efficacy, better than the peptide alone, and comparable to the efficacy of erythropoietin (EPO) and Aranesp (Darbepoietin alpha). The assays included survival assays on EPO-responsive cell lines (EC50 below 1 ng/ml peptide) and clonogenic assays on human bone marrow cells (EC50 1 to 10 ng/ml). AGEM400(HES) caused phosphorylation of STAT5 and ERK signalling proteins in UT7/EPO cells in a similar fashion as EPO. AGEM400(HES) replaced EPO from its receptor and the in vitro activity of AGEM400 (HES) was inhibited by soluble EPO receptor. Antibodies generated in mice and rabbits against EPO did not recognize AGEM400(HES) peptide, and vice versa. A sensitive ELISA was able to detect AGEM400(HES) at low nanogram per ml concentrations which allows for bioanalytics of AGEM400(HES) serum levels in future in vivo studies. As a result, AGEM400(HES) is a promising drug candidate for anemias related to renal insufficiency and/or in oncological settings.
Erythropoietin (EPO) has been shown to have tissue protective properties by binding to its receptor (EPOR) which is also expressed on non-haematopoietic cells.The mechanisms underlying this protection have not yet been elucidated and the renal cell types mediating these effects remain ill-defined.This study aimed to identify the EPOR expression in human tubular epithelial cells (hTECs) and in rat kidney and to investigate the role of EPOR in EPO-mediated renoprotection.Male Wistar rats were treated with saline or EPO (3000 U/kg, i.p.) 24h prior to sham-operation or 30 min bilateral renal ischemia.Renal morphology and function, tubular regeneration, apoptosis and expression of EPOR, hemeoxygenase-1 (HO-1) and hepatocyte growth factor (HGF) were analyzed.Primary cultures of human proximal (PTC) and distal/collecting duct (DTC) tubular cells were incubated with EPO (5-50-500 ng/mL) either or not in the presence of soluble EPOR.Total RNA was extracted and mRNA expression of HO-1 was investigated by quantitative RT-PCR.EPOR mRNA could be demonstrated in hTECs and in cortical tubules of the rat kidney.Furthermore, EPOR protein was expressed at the membrane and as intracellular vesicles in hTECs.In vivo, EPO treatment attenuated histological and functional renal damage, decreased both cell necrosis and apoptotic cell death, enhanced tubular regeneration and resulted in an upregulation of HO-1 and HGF mRNA.In vitro, EPO administration resulted in an early upregulation of HO-1 mRNA which was restricted to PTC and inhibited by simultaneous addition of supra-equivalent amounts of soluble EPOR.These data strongly suggest that the EPO-mediated renoprotection results from direct interaction of EPO with EPOR on tubular cells.
Using a mouse monoclonal antibody (mAb) (“2D4”) with high specific reactivity to Hb Bart’s and a rabbit polyclonal antibody (“RPB”) with high reactivity to Hb Bart’s but low reactivity to HbF, an ELISA assay was developed for the quantification of Hb Bart’s in hemolysates of peripheral blood. In the preliminary study, hemoglobin solutions containing 4,000 μg/mL of hemoglobin were analyzed for the concentration of Hb Bart’s in samples collected from the following children and adult subjects of HbH families: 12 children with deletional HbH disease (--/3.7 ) or nondeletional HbH disease (HbH disease with HbCS) (--/ cs ), 12 adults with 0 thalassemia (--/ ), 12 adults with deletional or nondeletional + thalassemia (3.7 / or / cs ) and 12 normal adult subjects ( / ). The mean ± S.D. of Hb Bart’s concentration in those with deletional HbH disease or HbH disease with HbCS, 0 thalassemia, deletional or nondeletional + thalassemia, and normal subjects were 1,374±210 (range 1,164-1,584), 1,118±357 (range 761-1,475), 451 ± 230 (range 221-681), and 0 ng/mL, respectively. When the developed ELISA was further evaluated with additional samples of various types of thalassemia, including: 18 with deletional HbH disease (--/3.7 ); 21 of nondeletional HbH disease (HbH disease with HbCS) (--/ cs ); 33 with 0 thalassemia (--/ ); 19 with nondeletional + thalassemia ( / cs ); 11 with deletional + thalassemia (3.7 / ) and 58 normal subjects ( / ). It was found that the levels of Hb Bart’s in deletional + thalassemia was significantly lower than in 0 thalassemia (p<0.001). The levels of Hb Bart’s in 0 thalassemia was also significantly lower than in nondeletional and deletional HbH diseases (p=0.023 and p<0.001, respectively). When all types of thalassemia were compared with normal subjects, the Hb Bart’s levels in all types of thalassemia were significantly higher (p<0.0001). All of our results indicated that the developed ELISA was highly sensitive and specific for quantitative determination of Hb Bart’s in hemolysates. The ELISA assay might be used as a rapid screening test for the detection of thalassemias in general population.
To assess the prevalence and risk factors of anaemia in pregnancy and recommend a cut-off value for antenatal women in developing countries, a cross sectional study was carried in the University of Port Harcourt Teaching Hospital, Port Harcourt, a 523 bed tertiary health care institution in Nigeria.1371 apparently healthy asymptomatic pregnant women visiting the antenatal clinic for the first time were enrolled for the study.Another 60 age-matched non-pregnant apparently healthy women served as controls.Haemoglobin concentration, malaria, human immunodeficiency virus (HIV)S, haemoglobin electrophoretic pattern and C-reactive protein as marker of infection were investigated using standard haematological and serological procedures.Anaemia in pregnancy (Hb <11.0g/dl) and (Hb<10.0g/dl)were found to be 23.2% and 6.7% respectively.The aetiology of anaemia was found to be multifactorial; 40.2% had anaemia of infection, 20.3% had Plasmodium falciparum alone, 8.5% had HIV alone, 2.5% had HIV and malaria parasite co-infection; 8.9% undetermined infections and 0.6% had sickle cell anaemia.There is moderate prevalence of anaemia in pregnancy in this part of the world.Since the mean Hb value of the pregnant women in this study was 11.62±1.21g/dland the pregnant women with Hb values around 10g/dl are apparently healthy, a cut-off value of 10.0g/dl may be considered ideal for defining anaemia in pregnancy in developing countries.
Hemophilia A & B are congenital bleeding disorders affecting 1:10-20,000 population and 1:20 to 40,000 population respectively.Hemophilia represents the prototype of high cost low volume disease.Eighty persons of world hemophilia population lives in financially poor developing countries, where <2% of GDP is usually spent for total health care.In India, with a population of more than 1 billion and growth rate of around 2% atleast 2000 new hemophilia patients are born every year .On a conservative estimate 50% of them have severe disease and with modern treatment, will require additional USD 36,000/year/patient i.e a 36 million dollar incremental additional health care burden each year to manage these 1000 severe hemophilia patient.Which is added to our existing pool of haemophiliacs, estimated at 1,00,000-1,20,000.One of the ways to manage this challenge is to establish prenatal diagnostic centres, for hemophilia, spread widely for easy accessibility.These diagnostic centers can eventually develop into prenatal diagnosis centres for other diseases like common hemoglobinopathies and they should be equipped with modest molecular diagnostic facilities.Presently there are a series of techniques of different levels of difficulties and complexities, available for foetal tissue sampling and laboratory detection of prenatal diagnosis of hemophilia, allowing a centre to chose the technology suitable for their expertise and financial capability.
Beta-thalassemia is one of the most prevalent autosomal disorders in the world.Mutations in the HBB gene underlie deficiencies in hemoglobin production, which can interfere with oxygen delivery resulting in wide range of disease severity.Although >535 mutations have been characterized in the HBB gene, beta-thalassemia is broadly classified into three groups, based on clinical severity: beta-thalassemia major, beta-thalassemia intermedia and beta-thalassemia minor.In this article we review: 1) the molecular and biochemical basis of beta-thalassemia; 2) clinical features; 3) the range of common molecular variants of beta-thalassemia in a subset of geographic regions within the Indian Subcontinent and the Middle East; 4) potential molecular diagnostics; and 5) current and future treatments.We suggest that efforts to more completely characterize the HBB mutation distribution in high-risk areas, such as the Indian Subcontinent and the Middle East, may lead to improved diagnosis with earlier and more effective intervention strategies.
Oral anticoagulant therapy (OAT) calls for continuous control by prothrombin time (PT) test, as the therapeutic range in INR units is very narrow.Warfarin (or coumarin) inhibits coagulation factor synthesis in the liver, but at the same time inactive coagulation factors are formed.The aim here was to measure "active coagulation factors" and inhibition in calibrator kits and patient plasmas by a new method for Quick and Owren PT.Four calibration kits and 200 plasma samples obtained from OAT patients were assessed using Quick and Owren PT for INR Tot (active coagulation factors + inhibition) and INR Acf (only active coagulation factors).Conspicuous variation in inhibition was noted between the four calibration kits.The new-generation PT method develops anticoagulation therapy based on active coagulation factors in vivo and improves INR result harmonization for Quick and Owren PT reagents.This new approach improves Quick PT reliability.
Non-myeloablative allogeneic stem cell transplantation (NST) has been one of the most exciting developments in the treatment of hematologic malignancies in the last years.Since 1999, we have chosen to employ in México a regimen to conduct NST, introducing some changes with the main goal of decreasing the cost of the procedure and in turn, making it available to a larger number of patients in developing countries.Using this method we have done over 400 allografts in Latin American patients with different both malignant and non-malignant diseases: Chronic myelogenous leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, myelodysplasia, thalassemia major, relapsed Hodg-kin´s disease, Blackfan-Diamond syndrome, adrenoleukodystrophy, Hunter´s syndrome, aplastic anemia and several solid tumors.In the whole group, the median granulocyte recovery time to 0.5 x 10 9 /L was 13 days, whereas the median platelet recovery time to 20 x 10 9 /L was 12 days.Around one third of the patients did not need red blood cell transfusions and also one third did not need platelet transfusions.In more than 70% of cases the procedure could be completed totally on an outpatient basis.The follow up time of the patients ranges between 30 and 2000 days.Approximately 50% of the allografted individuals have developed acute graft versus host disease (GVHD), whereas around 30% developed chronic GVHD.The median post-allograft overall survival (SV) has not been reached and the 2000 day overall SV is 54%, the 100-day mortality being 16%.In the whole group of patients, the median cost of each NST was 18 000 USD, a figure which contrasts with that informed from developing countries.More than 95% of the patients who were allografted in México and Latin America using this method could not have afforded the cost of a conventional or more expensive stem cell transplant; accordingly, this procedure has enabled doctors in México and Latin America to offer this therapeutic approach to a larger number of individuals.
Benzene, an aromatic hydrocarbon that is a natural component of crude oil and natural gas, is toxic to the blood and blood-forming organs.Epidemiological studies have established an association between benzene exposure and acute myeloid leukemia, and increasing evidence also indicates a possible association between benzene and multiple myeloma.A specific benzene-associated myelodysplastic syndrome has also been suggested.Chronic hematotoxic effects of benzene exposure, including reduced lymphocyte, neutrophil and platelet counts in peripheral blood, have been detected at occupational exposure below a level that had previously been considered not to cause any health effects.Whether these abnormalities represent bone marrow damage and/or initial events in the development of a true neoplastic disease is not known.Together with a reported nonlinear relationship between benzene exposure and the level of various metabolites, favoring production of biologically reactive quinones at exposure below 1 part per million, these observations suggest that benzene even at low exposure levels may contribute to the risk of acute myeloid leukemia or myelodysplastic syndrome, especially among genetically susceptible individuals.
The Eph receptor tyrosine kinases form the largest sub-family of the Receptor tyrosine kinases.Eph receptors interact with membrane bound ligands, termed ephrins.Eph-ephrin interactions have significant roles during mammalian development where they mediate such processes as cell adhesion and migration.Although Eph/ephrin expression is low in adult tissues evidence is accumulating which indicates that Eph and ephrins continue to play significant roles in postembryonic tissues.In this review we describe the current understanding of the function of Eph-ephrin signaling in human and mouse hematopoiesis.Ephs and ephrins have been shown to regulate the differentiation of hematopoietic cells, to direct lymphoid development as well as immune function.Whilst much still remains to be learned about the role of Ephephrin signaling in hematopoieis it is already clear that this system exerts important regulatory effects on differentiation, cell migration and effector function.The possibility that these molecules may be therapeutic targets is also discussed.
