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Sample records for asparaginase

  1. Asparaginase Erwinia chrysanthemi

    Science.gov (United States)

    Asparaginase Erwinia chrysanthemi is used with other chemotherapy medications to treat acute lymphocytic leukemia (ALL; a type of cancer ... of allergic reactions to medications similar to asparaginase Erwinia chrysanthemi such as (asparaginase [Elspar] or pegaspargase [Oncaspar]). ...

  2. L-asparaginase treatment in acute lymphoblastic leukemia: a focus on Erwinia asparaginase

    Science.gov (United States)

    Pieters, Rob; Hunger, Stephen P; Boos, Joachim; Rizzari, Carmelo; Silverman, Lewis; Baruchel, Andre; Goekbuget, Nicola; Schrappe, Martin; Pui, Ching-Hon

    2010-01-01

    Asparaginases are a cornerstone of treatment protocols for acute lymphoblastic leukemia (ALL) and are used for remission induction and intensification treatment in all pediatric regimens and in the majority of adult protocols. Extensive clinical data have shown that intensive asparaginase treatment improves clinical outcomes in childhood ALL. Three asparaginase preparations are available; the native asparaginase derived from Escherichia coli (E. coli-asparaginase), a pegylated form of this enzyme (PEG-asparaginase) and a product isolated from Erwinia chrysanthemi, i.e. Erwinia asparaginase. Clinical hypersensitivity reactions and silent inactivation due to antibodies against E.coli-asparaginase, lead to inactivation of E-Coli asparaginase in up to 60% of cases. Current treatment protocols include E. coli-asparaginase or PEG-asparaginase for first-line treatment of ALL. Typically, patients exhibiting sensitivity to one formulation of asparaginase are switched to another product to ensure they receive the most efficacious treatment regimen possible. Erwinia asparaginase is used as a second- or third-line treatment in European and US protocols. Despite the universal inclusion of asparaginase in such treatment protocols, there is much debate regarding the optimal formulation and dosage of these agents. This manuscript provides an overview of available evidence to make recommendations for optimal use of Erwinia asparaginase in the treatment of ALL. PMID:20824725

  3. L-asparaginase treatment in acute lymphoblastic leukemia: a focus on Erwinia asparaginase.

    Science.gov (United States)

    Pieters, Rob; Hunger, Stephen P; Boos, Joachim; Rizzari, Carmelo; Silverman, Lewis; Baruchel, Andre; Goekbuget, Nicola; Schrappe, Martin; Pui, Ching-Hon

    2011-01-15

    Asparaginases are a cornerstone of treatment protocols for acute lymphoblastic leukemia (ALL) and are used for remission induction and intensification treatment in all pediatric regimens and in the majority of adult treatment protocols. Extensive clinical data have shown that intensive asparaginase treatment improves clinical outcomes in childhood ALL. Three asparaginase preparations are available: the native asparaginase derived from Escherichia coli (E. coli asparaginase), a pegylated form of this enzyme (PEG-asparaginase), and a product isolated from Erwinia chrysanthemi, ie, Erwinia asparaginase. Clinical hypersensitivity reactions and silent inactivation due to antibodies against E. coli asparaginase, lead to inactivation of E. coli asparaginase in up to 60% of cases. Current treatment protocols include E. coli asparaginase or PEG-asparaginase for first-line treatment of ALL. Typically, patients exhibiting sensitivity to one formulation of asparaginase are switched to another to ensure they receive the most efficacious treatment regimen possible. Erwinia asparaginase is used as a second- or third-line treatment in European and US protocols. Despite the universal inclusion of asparaginase in such treatment protocols, debate on the optimal formulation and dosage of these agents continues. This article provides an overview of available evidence for optimal use of Erwinia asparaginase in the treatment of ALL.

  4. Asparaginase-associated pancreatitis in children

    DEFF Research Database (Denmark)

    Raja, Raheel Altaf; Schmiegelow, Kjeld; Frandsen, Thomas Leth

    2012-01-01

    l-asparaginase has been an element in the treatment for acute lymphoblastic leukaemia (ALL) and non-Hodgkin lymphoma since the late 1960s and remains an essential component of their combination chemotherapy. Among the major toxicities associated with l-asparaginase therapy are pancreatitis......, allergic reactions, thrombotic events, hepatotoxicity and hyperlipidaemia. Acute pancreatitis is one of the most common reasons for stopping treatment with l-asparaginase. Short-term complications of asparaginase-associated pancreatitis include development of pseudocysts and pancreatic necrosis. Long......-term complications include chronic pancreatitis and diabetes. The pathophysiology of asparaginase-associated pancreatitis remains to be uncovered. Individual clinical and genetic risk factors have been identified, but they are only weak predictors of pancreatitis. This review explores the definition, possible risk...

  5. Asparaginase-Induced Hypertriglyceridemia Presenting as Pseudohyponatremia during Leukemia Treatment

    OpenAIRE

    Ashley Hinson; Dorothee Newbern; Linardic, Corinne M.

    2014-01-01

    Asparaginase is a chemotherapeutic agent used to induce disease remission in children with acute lymphoblastic leukemia (ALL). We describe the cases of two females with ALL who developed pseudohyponatremia as a presentation of hypertriglyceridemia following asparaginase treatment. Nine similar published cases of asparaginase-induced hypertriglyceridemia and its complications are also discussed. Possible mechanisms of action include inhibition of lipoprotein lipase, decreased hepatic synthesis...

  6. Asparaginase antibody and asparaginase activity in children with higher-risk acute lymphoblastic leukemia: Children's Cancer Group Study CCG-1961.

    Science.gov (United States)

    Panosyan, Eduard H; Seibel, Nita L; Martin-Aragon, Sagrario; Gaynon, Paul S; Avramis, Ioannis A; Sather, Harland; Franklin, Janet; Nachman, James; Ettinger, Lawrence J; La, Mei; Steinherz, Peter; Cohen, Lewis J; Siegel, Stuart E; Avramis, Vassilios I

    2004-04-01

    We investigated the anti-asparaginase antibody (Ab) and asparaginase enzymatic activity in the sera of 1,001 patients (CCG-1961) with high-risk acute lymphoblastic leukemia (HR-ALL). Patients received nine doses of native Escherichia coli asparaginase during induction. Half of rapid early responders (RER) were randomly assigned to standard intensity arms and continued to receive native asparaginase. The other RER patients and all slow early responders received 6 or 10 doses of PEG-asparaginase. Serum samples (n = 3,193) were assayed for determination of asparaginase Ab titers and enzymatic activity. Three hundred ninety of 1,001 patients (39%) had no elevation of Ab among multiple evaluations-that is, were Ab-negative (1.1). Among these 611 patients, 447 had no measurable asparaginase activity during therapy. Patients who were Ab-positive but had no clinical allergies continued to receive E. coli asparaginase, the activity of which declined precipitately. No detectable asparaginase activity was found in 81 of 88 Ab-positive patients shortly after asparaginase injections (94% neutralizing Ab). The Ab-positive patients with clinical allergies subsequently were given Erwinase and achieved substantial activity (0.1-0.4 IU/ml). An interim analysis of 280 patients who were followed for 30 months from induction demonstrated that the Ab-positive titers during interim maintenance-1 and in delayed intensification-1 were associated with an increased rate of events. The CCG-1961 treatment schedule was very immunogenic, plausibly due to initially administrated native asparaginase. Anti-asparaginase Ab was associated with undetectable asparaginase activity and may be correlated with adverse outcomes in HR ALL. PMID:15087948

  7. A prospective study on drug monitoring of PEGasparaginase and Erwinia asparaginase and asparaginase antibodies in pediatric acute lymphoblastic leukemia.

    Science.gov (United States)

    Tong, Wing H; Pieters, Rob; Kaspers, Gertjan J L; te Loo, D Maroeska W M; Bierings, Marc B; van den Bos, Cor; Kollen, Wouter J W; Hop, Wim C J; Lanvers-Kaminsky, Claudia; Relling, Mary V; Tissing, Wim J E; van der Sluis, Inge M

    2014-03-27

    This study prospectively analyzed the efficacy of very prolonged courses of pegylated Escherichia coli asparaginase (PEGasparaginase) and Erwinia asparaginase in pediatric acute lymphoblastic leukemia (ALL) patients. Patients received 15 PEGasparaginase infusions (2500 IU/m(2) every 2 weeks) in intensification after receiving native E coli asparaginase in induction. In case of allergy to or silent inactivation of PEGasparaginase, Erwinia asparaginase (20 000 IU/m(2) 2-3 times weekly) was given. Eighty-nine patients were enrolled in the PEGasparaginase study. Twenty (22%) of the PEGasparaginase-treated patients developed an allergy; 7 (8%) showed silent inactivation. The PEGasparaginase level was 0 in all allergic patients (grade 1-4). Patients without hypersensitivity to PEGasparaginase had serum mean trough levels of 899 U/L. Fifty-nine patients were included in the Erwinia asparaginase study; 2 (3%) developed an allergy and none silent inactivation. Ninety-six percent had at least 1 trough level ≥100 U/L. The serum asparagine level was not always completely depleted with Erwinia asparaginase in contrast to PEGasparaginase. The presence of asparaginase antibodies was related to allergies and silent inactivation, but with low specificity (64%). Use of native E coli asparaginase in induction leads to high hypersensitivity rates to PEGasparaginase in intensification. Therefore, PEGasparaginase should be used upfront in induction, and we suggest that the dose could be lowered. Switching to Erwinia asparaginase leads to effective asparaginase levels in most patients. Therapeutic drug monitoring has been added to our ALL-11 protocol to individualize asparaginase therapy.

  8. Production of L-Asparaginase by the marine luminous bacteria

    Digital Repository Service at National Institute of Oceanography (India)

    Ramaiah, N.; Chandramohan, D.

    Fortythree strains of luminous bacteria, belonging to 4 species, (Vibrio harveyi, V. fischeri, Photobacterium leiognathi and P. phosphoreum) isolated from different marine samples, were examined for the production of L-asparaginase. Presence...

  9. Pharmacokinetic/pharmacodynamic relationships of asparaginase formulations: the past, the present and recommendations for the future.

    Science.gov (United States)

    Avramis, Vassilios I; Panosyan, Eduard H

    2005-01-01

    The discovery of the tumour-inhibitory properties of asparaginase began 50 years ago with the observation that guinea-pig serum-treated lymphoma-bearing mice underwent rapid and often complete regression. Soon afterwards, the asparaginase of bacterial origin was isolated. The asparaginases of bacterial origin induce anti-asparaginase neutralising antibodies in a large proportion of patients (44-60%), thus negating the specific enzymatic activity and resulting in failure of the target amino acid deamination in serum. There is immunological cross-reaction between the antibodies against various formulations of native Escherichia coli-asparaginase and polyethylene glycol (PEG)-asparaginases, but not to Erwinia asparaginase, as suggested by laboratory preclinical findings. This evidence was strongly inferred from the interim analyses in the Children's Cancer Group (CCG)-1961 study. Thus, anti-E. coli or PEG-asparaginase antibodies seropositive patients may benefit from the Erwinia asparaginase. The inter-relationships between asparaginase activity, asparagine (ASN) and glutamine deamination remain largely unexplored in patients. Studies have shown that ASN depletion is insufficient to induce apoptosis in T lymphoblasts in vitro and that the inhibitory concentration of CEM T-cell line is correlated with the asparaginase concentration responsible for 50% glutamine deamination. The optimal catalysis of ASN and glutamine deamination in serum by asparaginase induces apoptosis of leukaemic lymphoblasts. The percentage of ASN and glutamine deamination was predicted by asparaginase activity. Asparaginase activity of 0.1 IU/mL provided insufficient depletion of both amino acids in high-risk acute lymphoblastic leukaemia (ALL) patients. With increasing glutamine deamination, mean asparaginase activities and percentages of post-treatment samples with effective ASN depletion (/=90% deamination of glutamine must occur before optimal ASN deamination takes place, due to the de novo

  10. Expression, purification and crystallization of Helicobacter pyloril-asparaginase

    Energy Technology Data Exchange (ETDEWEB)

    Dhavala, Prathusha [Turku Centre for Biotechnology, University of Turku and Åbo Akademi, Turku 20521 (Finland); Krasotkina, Julya [Institute for Biomedical Sciences, Russian Academy of Medical Sciences, Moscow (Russian Federation); Dubreuil, Christine; Papageorgiou, Anastassios C., E-mail: tassos.papageorgiou@btk.fi [Turku Centre for Biotechnology, University of Turku and Åbo Akademi, Turku 20521 (Finland)

    2008-08-01

    l-Asparaginase from H. pylori was overexpressed in E. coli, purified and crystallized. The crystals belonged to space group I222, with unit-cell parameters a = 63.6, b = 94.9, c = 100.2 Å and one molecule in the asymmetric unit. A complete data set to 1.6 Å resolution was collected using synchrotron radiation. The l-asparaginases from Escherichia coli and Erwinia chrysanthemi are effective drugs that have been used in the treatment of acute childhood lymphoblastic leukaemia for over 30 years. However, despite their therapeutic potential, they can cause serious side effects as a consequence of their intrinsic glutaminase activity, which leads to l-glutamine depletion in the blood. Consequently, new asparaginases with low glutaminase activity, fewer side effects and high activity towards l-asparagine are highly desirable as better alternatives in cancer therapy. l-Asparaginase from Helicobacter pylori was overexpressed in E. coli and purified for structural studies. The enzyme was crystallized at pH 7.0 in the presence of 16–19%(w/v) PEG 4000 and 0.1 M magnesium formate. Data were collected to 1.6 Å resolution at 100 K from a single crystal at a synchrotron-radiation source. The crystals belong to space group I222, with unit-cell parameters a = 63.6, b = 94.9, c = 100.2 Å and one molecule of l-asparaginase in the asymmetric unit. Elucidation of the crystal structure will provide insight into the active site of the enzyme and a better understanding of the structure–activity relationship in l-asparaginases.

  11. L-asparaginase treatment in acute lymphoblastic leukemia

    NARCIS (Netherlands)

    R. Pieters (Rob); S.P. Hunger (Stephen); J. Boos (Joachim); C. Rizzari (Carmelo); L.B. Silverman (Lewis); A. Baruchel (André); N. Goekbuget (Nicola); M. Schrappe (Martin); C.H. Pui (Ching-Hon)

    2011-01-01

    textabstractAsparaginases are a cornerstone of treatment protocols for acute lymphoblastic leukemia (ALL) and are used for remission induction and intensification treatment in all pediatric regimens and in the majority of adult treatment protocols. Extensive clinical data have shown that intensive a

  12. Cell-cycle inhibition by Helicobacter pylori L-asparaginase.

    Directory of Open Access Journals (Sweden)

    Claudia Scotti

    Full Text Available Helicobacter pylori (H. pylori is a major human pathogen causing chronic gastritis, peptic ulcer, gastric cancer, and mucosa-associated lymphoid tissue lymphoma. One of the mechanisms whereby it induces damage depends on its interference with proliferation of host tissues. We here describe the discovery of a novel bacterial factor able to inhibit the cell-cycle of exposed cells, both of gastric and non-gastric origin. An integrated approach was adopted to isolate and characterise the molecule from the bacterial culture filtrate produced in a protein-free medium: size-exclusion chromatography, non-reducing gel electrophoresis, mass spectrometry, mutant analysis, recombinant protein expression and enzymatic assays. L-asparaginase was identified as the factor responsible for cell-cycle inhibition of fibroblasts and gastric cell lines. Its effect on cell-cycle was confirmed by inhibitors, a knockout strain and the action of recombinant L-asparaginase on cell lines. Interference with cell-cycle in vitro depended on cell genotype and was related to the expression levels of the concurrent enzyme asparagine synthetase. Bacterial subcellular distribution of L-asparaginase was also analysed along with its immunogenicity. H. pylori L-asparaginase is a novel antigen that functions as a cell-cycle inhibitor of fibroblasts and gastric cell lines. We give evidence supporting a role in the pathogenesis of H. pylori-related diseases and discuss its potential diagnostic application.

  13. Acrylamide diminishing in potato chips by using commercial Asparaginase

    DEFF Research Database (Denmark)

    Pedreschi, Franco; Mariotti, Salomé; Granby, Kit;

    2011-01-01

    In April 2002, Swedish researchers shocked the food safety world when they presented preliminary findings of acrylamide in some fried and baked foods, most notably potato chips and French fries. Asparagine is an aminoacid precursor of acrylamide formation through Maillard reaction. Asparaginase e...

  14. Role of L-asparaginase in acute lymphoblastic leukemia: focus on adult patients

    Directory of Open Access Journals (Sweden)

    Rytting ME

    2012-06-01

    Full Text Available Michael E RyttingDepartment of Pediatrics and Leukemia, University of Texas MD Anderson Cancer Center, Houston, TX, USAAbstract: Asparaginase preparations deplete asparagine in acute lymphoblastic leukemia (ALL blasts. Asparaginase in its various forms is an important component of treatment regimens for pediatric ALL. Recently, interest and use of asparaginase in adult patients with ALL has increased, particularly in young adults. There is much less information on asparaginase use and toxicity in adult compared with pediatric populations. This review surveys prior published studies of the three most commonly used asparagine preparations as used in adult patients, and discusses important toxicities encountered in adult patients who receive asparaginase preparations.Keywords: asparaginase, leukemia, adults, children

  15. A Study on L-Asparaginase of Nocardia levis MK-VL_113

    Directory of Open Access Journals (Sweden)

    Alapati Kavitha

    2012-01-01

    Full Text Available An enzyme-based drug, L-asparaginase, was produced by Nocardia levis MK-VL_113 isolated from laterite soils of Guntur region. Cultural parameters affecting the production of L-asparaginase by the strain were optimized. Maximal yields of L-asparaginase were recorded from 3-day-old culture grown in modified asparagine-glycerol salts broth with initial pH 7.0 at temperature 30∘C. Glycerol (2% and yeast extract (1.5% served as good carbon and nitrogen sources for L-asparaginase production, respectively. Cell-disrupting agents like EDTA slightly enhanced the productivity of L-asparaginase. Ours is the first paper on the production of L-asparaginase by N. levis.

  16. Neurosurgical management of L-asparaginase induced haemorrhagic stroke.

    LENUS (Irish Health Repository)

    Ogbodo, Elisha

    2012-01-01

    The authors describe a case of L-asparaginase induced intracranial thrombosis and subsequent haemorrhage in a newly diagnosed 30-year-old man with acute lymphoblastic leukaemia who was successfully managed by surgical intervention. At presentation, he had a Glasgow Coma Score of 7\\/15, was aphasic and had dense right hemiplegia. Neuroimaging revealed an acute anterior left middle cerebral artery infarct with parenchymal haemorrhagic conversion, mass effect and subfalcine herniation. He subsequently underwent left frontal craniotomy and evacuation of large frontal haematoma and decompressive craniectomy for cerebral oedema. Six months postoperatively he underwent titanium cranioplasty. He had made good clinical recovery and is currently mobilising independently with mild occasional episodes of expressive dysphasia, difficulty with fine motor movement on the right side, and has remained seizure free. This is the first documented case of L-asparaginase induced haemorrhagic stroke managed by neurosurgical intervention. The authors emphasise the possible role of surgery in managing chemotherapy induced intracranial complications.

  17. Cerebrospinal fluid asparagine depletion during pegylated asparaginase therapy in children with acute lymphoblastic leukaemia

    DEFF Research Database (Denmark)

    Henriksen, Louise T; Nersting, Jacob; Raja, Raheel A;

    2014-01-01

    L-asparaginase is an important drug in the treatment of childhood acute lymphoblastic leukaemia (ALL). Cerebrospinal fluid (CSF) asparagine depletion is considered a marker of asparaginase effect in the central nervous system (CNS) and may play a role in CNS-directed anti-leukaemia therapy. The...

  18. PEG-asparaginase allergy in children with acute lymphoblastic leukemia in the NOPHO ALL2008 protocol

    DEFF Research Database (Denmark)

    Henriksen, Louise Tram; Harila-Saari, Arja; Ruud, Ellen;

    2015-01-01

    -asparaginase allergy in children treated according to the Nordic Society of Paediatric Haematology and Oncology (NOPHO) ALL2008 protocol. PROCEDURE: Children (1-17 years) enrolled in the NOPHO ALL2008 protocol between July 2008 and August 2011, who developed PEG-asparaginase allergy were identified through the NOPHO...

  19. Sagittal sinus thrombosis due to L-asparaginase

    Directory of Open Access Journals (Sweden)

    Nisar A Wani

    2010-01-01

    Full Text Available Cerebral Sinovenous Thrombosis (CSVT is a serious complication of L-asparaginase chemotherapy for leukemia in children. Clinical features of headache, altered consciousness, focal neurological deficit, and seizures developing during or immediately after treatment with L-asparaginase should alert the treating physician to the possibility of CSVT. Immediate imaging of the brain should be done using CT and MRI and the veins should be visualized noninvasively by CT and MR venography. We report two children on induction therapy for acute leukemia who presented with seizures, headache, and altered consciousness. Venous infarcts with and without hemorrhage were seen on CT in one patient and the empty delta sign was seen after contrast injection; however, the early changes were missed by CT. MRI detected dural sinus thrombosis relatively earlier in another patient, while the CT findings were equivocal; in this patient, contrast-enhanced MRI showed the empty delta sign and MR venography confirmed absent flow in the superior sagittal sinus, which was diagnostic of sinus thrombosis. Rapid anticoagulation was started with heparin and maintained with warfarin. The child with a unilateral small nonhemorrhagic infarct made a complete recovery while the other, with bilateral hemorrhagic infarcts, did not survive. We stress the importance of early diagnosis of CSVT using CT and MRI in children with leukemia being treated with L-asparaginase; this will permit timely treatment.

  20. Tackling Critical Catalytic Residues in Helicobacter pylori L-Asparaginase

    Directory of Open Access Journals (Sweden)

    Maristella Maggi

    2015-03-01

    Full Text Available Bacterial asparaginases (amidohydrolases, EC 3.5.1.1 are important enzymes in cancer therapy, especially for Acute Lymphoblastic Leukemia. They are tetrameric enzymes able to catalyze the deamination of L-ASN and, to a variable extent, of L-GLN, on which leukemia cells are dependent for survival. In contrast to other known L-asparaginases, Helicobacter pylori CCUG 17874 type II enzyme (HpASNase is cooperative and has a low affinity towards L-GLN. In this study, some critical amino acids forming the active site of HpASNase (T16, T95 and E289 have been tackled by rational engineering in the attempt to better define their role in catalysis and to achieve a deeper understanding of the peculiar cooperative behavior of this enzyme. Mutations T16E, T95D and T95H led to a complete loss of enzymatic activity. Mutation E289A dramatically reduced the catalytic activity of the enzyme, but increased its thermostability. Interestingly, E289 belongs to a loop that is very variable in L-asparaginases from the structure, sequence and length point of view, and which could be a main determinant of their different catalytic features.

  1. Withania somnifera (Ashwagandha): a Novel Source of L-asparaginase

    Institute of Scientific and Technical Information of China (English)

    Vishal P. Oza; Shraddha D.Trivedi; Pritesh P.Parmar; R.B.Subramanian

    2009-01-01

    Different parts of plant species belonging to Solanaceae and Fabaceae families were screened for L-asparaginase enzyme (E.C.3.5.1.1.). Among 34 plant species screened for L-asparaginase enzyme, Withania somnifera L. Was identified as a potential source of the enzyme on the basis of high specific activity of the enzyme. The enzyme was purified and characterized from W. Somnifera, a popular medicinal plant in South East Asia and Southern Europe. Purification was carried out by a combination of protein precipitation with ammonium sulfate as well as Sephadex-gel filtration. The purified enzyme is a homodimer, with a molecular mass of 72±0.5 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresisand size exclusion chromatography. The enzyme has a pH optimum of 8.5 and an optimum temperature of 37℃. The Km value for the enzyme is 6.1×10-2 mmol/L. This is the first report for L-asparaginase from W. Somnifera, a traditionally used Indian medicinal plant.

  2. Asparaginase Erwinia chrysanthemi (Erwinaze®): a guide to its use in acute lymphoblastic leukemia in the USA.

    Science.gov (United States)

    Keating, Gillian M

    2013-08-01

    Asparaginase Erwinia chrysanthemi (Erwinaze®) is approved in the USA for use in patients with acute lymphoblastic leukemia (ALL) who have developed hypersensitivity to Escherichia coli-derived asparaginase. The approved regimen of intramuscular Erwinaze® was associated with sustained, clinically meaningful asparaginase activity in patients with ALL who had to discontinue treatment with pegaspargase (a pegylated formulation of E. coli asparaginase) because of hypersensitivity. Another study revealed that development of E. coli-derived asparaginase allergy and a switch to Erwinaze® maintained event-free survival in pediatric patients with newly diagnosed ALL. In a multicenter, compassionate-use trial, Erwinaze® was generally well tolerated, with the most commonly occurring adverse events including hypersensitivity, pancreatitis, fever, hyperglycemia, and increased transaminase levels. Subclinical hypersensitivity may result in the inactivation of asparaginase and affect treatment outcome; monitoring of serum asparaginase levels may be used to identify subclinical hypersensitivity.

  3. Recombinant L-Asparaginase from Zymomonas mobilis: A Potential New Antileukemic Agent Produced in Escherichia coli

    Science.gov (United States)

    Pereira, Juliana Christina Castanheira Vicente; Costa-Amaral, Isabele Campos; da Costa, Elaine Sobral; Ribeiro, Maria Cecília Menks; Land, Marcelo Gerardin Poirot; Alves, Tito Lívio Moitinho; Larentis, Ariane Leites; Almeida, Rodrigo Volcan

    2016-01-01

    L-asparaginase is an enzyme used as a chemotherapeutic agent, mainly for treating acute lymphoblastic leukemia. In this study, the gene of L-asparaginase from Zymomonas mobilis was cloned in pET vectors, fused to a histidine tag, and had its codons optimized. The L-asparaginase was expressed extracellularly and intracellularly (cytoplasmically) in Escherichia coli in far larger quantities than obtained from the microorganism of origin, and sufficient for initial cytotoxicity tests on leukemic cells. The in silico analysis of the protein from Z. mobilis indicated the presence of a signal peptide in the sequence, as well as high identity to other sequences of L-asparaginases with antileukemic activity. The protein was expressed in a bioreactor with a complex culture medium, yielding 0.13 IU/mL extracellular L-asparaginase and 3.6 IU/mL intracellular L-asparaginase after 4 h of induction with IPTG. The cytotoxicity results suggest that recombinant L-asparaginase from Z. mobilis expressed extracellularly in E.coli has a cytotoxic and cytostatic effect on leukemic cells. PMID:27253887

  4. Recombinant L-Asparaginase from Zymomonas mobilis: A Potential New Antileukemic Agent Produced in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Karen Einsfeldt

    Full Text Available L-asparaginase is an enzyme used as a chemotherapeutic agent, mainly for treating acute lymphoblastic leukemia. In this study, the gene of L-asparaginase from Zymomonas mobilis was cloned in pET vectors, fused to a histidine tag, and had its codons optimized. The L-asparaginase was expressed extracellularly and intracellularly (cytoplasmically in Escherichia coli in far larger quantities than obtained from the microorganism of origin, and sufficient for initial cytotoxicity tests on leukemic cells. The in silico analysis of the protein from Z. mobilis indicated the presence of a signal peptide in the sequence, as well as high identity to other sequences of L-asparaginases with antileukemic activity. The protein was expressed in a bioreactor with a complex culture medium, yielding 0.13 IU/mL extracellular L-asparaginase and 3.6 IU/mL intracellular L-asparaginase after 4 h of induction with IPTG. The cytotoxicity results suggest that recombinant L-asparaginase from Z. mobilis expressed extracellularly in E.coli has a cytotoxic and cytostatic effect on leukemic cells.

  5. A critical review on properties and applications of microbial l-asparaginases.

    Science.gov (United States)

    Krishnapura, Prajna Rao; Belur, Prasanna D; Subramanya, Sandeep

    2016-09-01

    l-Asparaginase is one of the main drugs used in the treatment of acute lymphoblastic leukemia (ALL), a commonly diagnosed pediatric cancer. Although several microorganisms are found to produce l-asparaginase, only the purified enzymes from E. coli and Erwinia chrysanthemi are employed in the clinical and therapeutic applications in humans. However, their therapeutic response seldom occurs without some evidence of hypersensitivity and other toxic side effects. l-Asparaginase is also of prospective use in food industry to reduce the formation of acrylamide in fried, roasted or baked food products. This review is an attempt to compile information on the properties of l-asparaginases obtained from different microorganisms. The complications involved with the therapeutic use of the currently available l-asparaginases, and the enzyme's potential application as a food processing aid to mitigate acrylamide formation have also been reviewed. Further, avenues for searching alternate sources of l-asparaginase have been discussed, highlighting the prospects of endophytic microorganisms as a possible source of l-asparaginases with varied biochemical and pharmacological properties.

  6. Asparaginase-associated pancreatitis in children with acute lymphoblastic leukaemia in the NOPHO ALL2008 protocol

    DEFF Research Database (Denmark)

    Raja, Raheel A; Schmiegelow, Kjeld; Albertsen, BK;

    2014-01-01

    L-asparaginase is an important drug in the treatment of childhood acute lymphoblastic leukaemia (ALL). Treatment is associated with several toxicities, including acute pancreatitis. Clinical course, presentation, re-exposure to L-asparginase after pancreatitis and risk of recurrent pancreatitis...... within an asparaginase-intensive protocol has been poorly reported. Children (1-17 years) on the ongoing Nordic Society of Paediatric Haematology and Oncology (NOPHO) ALL2008 protocol with asparaginase-associated pancreatitis (AAP) diagnosed between 2008 and 2012 were identified through the online NOPHO...... with AAP with a cumulative risk of AAP of 5·9%. AAP occurred after a median of five doses (range 1-13), and 11 d (median) from the latest administration of PEG-Asparaginase. Thirteen patients developed pseudocysts (30%) and 11 patients developed necrosis (25%). One patient died from pancreatitis. Twelve...

  7. The use of asparaginase to reduce acrylamide levels in cooked food.

    Science.gov (United States)

    Xu, Fei; Oruna-Concha, Maria-Jose; Elmore, J Stephen

    2016-11-01

    Strategies proposed for reducing the formation of the suspected carcinogen acrylamide in cooked foods often rely on a reduction in the extent of the Maillard reaction, in which acrylamide is formed from the reaction between asparagine and reducing sugars. However, the Maillard reaction also provides desirable sensory attributes of cooked foods. Mitigation procedures that modify the Maillard reaction may negatively affect flavour and colour. The use of asparaginase to convert asparagine to aspartic acid may provide a means to reduce acrylamide formation, while maintaining sensory quality. This review collates research on the use of enzymes, asparaginase in particular, to mitigate acrylamide formation. Asparaginase is a powerful tool for the food industry and it is likely that its use will increase. However, the potential adverse effects of asparaginase treatment on sensory properties of cooked foods and the need to achieve sufficient enzyme-substrate contact remain areas for future research. PMID:27211635

  8. Acrylamide reduction in potato chips by using commercial asparaginase in combination with conventional blanching

    DEFF Research Database (Denmark)

    Pedreschi, Franco; Mariotti, Salomé; Granby, Kit;

    2011-01-01

    In this research acrylamide reduction in potato chips was investigated in relation to blanching and asparaginase immersion treatments before final frying. Potatoes slices (Verdi variety, diameter: 40 mm, thickness: 2.0 mm) were fried at 170 °C for 5 min (final moisture content of ∼2.0 g/100 g...... (control II). Blanching in hot water (ii) was almost as effective as asparaginase potato immersion (iii) in order to diminish acrylamide formation in potato chips (acrylamide reduction was ∼17% of the initial acrylamide concentration). When potato slices were blanched before asparaginase immersion......, the acrylamide content of the resultant potato chips was reduced considerably by almost 90%. We have demonstrated that blanching of potato slices plus asparaginase treatment is an effective combination for acrylamide mitigation during frying. It seems to be that blanching provokes changes in the microstructure...

  9. Gene sequencing, cloning, and expression of the recombinant L- Asparaginase of Pseudomonas aeruginosa SN4 strain in Escherichia coli

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    Arastoo Badoei-dalfard

    2016-03-01

    Full Text Available Introduction: L- asparaginase is in an excessive demand in medical applications and in food treating industries, the request for this therapeutic enzyme is growing several folds every year. Materials and methods: In this study, a L- asparaginase gene from Pseudomonas aeruginosa strain SN4 was sequenced and cloned in E. coli. Primers were designed based on L- asparaginase from P. aeruginosa DSM 50071, which show high similarity to SN4 strain, according to 16S rRNA sequence. The L- asparaginase gene was exposed to restriction digestion with NdeI and XhoI enzymes and then ligated into pET21a plasmid. The ligated sample was transformed into competent E. coli (DE3 pLysS DH5a cells, according to CaCl2 method. The transformed E. coli cells were grown into LB agar plate containing 100 µg/ml ampicillin, IPTG (1 mM. Results: Recombinant L- asparaginase from E. coli BL21 induced after 9 h of incubation and showed high L- asparaginase activity about 93.4 IU/ml. Recombinant L- asparaginase sequencing and alignments showed that the presumed amino acid sequence composed of 350 amino acid residues showed high similarity with P. aeruginosa L- asparaginases about 99%. The results also indicated that SN4 L- asparaginase has the catalytic residues and conserve region similar to other L- asparaginases. Discussion and conclusion: This is the first report on cloning and expression of P. aeruginosa L- asparaginases in Escherichia coli. These results indicated a potent source of L- asparaginase for in vitro and in vivio anticancer consideration. 

  10. Purification, characterization and antiproliferative activity of l-asparaginase from Aspergillus oryzae CCT 3940 with no glutaminase activity

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    Fernanda Furlan Gonçalves Dias

    2016-09-01

    Conclusions: The sensitivity of the cells lines to purified l-asparaginase from A. oryzae CCT 3940 appeared to be concentration dependent affording a more significant decrease in cell growth than that observed for the commercial l-asparaginase from Escherichia coli. The l-asparaginase from A. oryzae CCT 3940 has a high potential for pharmaceutical exploitation in the treatment of leukemia.

  11. Cloning, expression, purification and characterisation of Erwinia carotovora L-asparaginase in Escherichia coli

    OpenAIRE

    Meraj Pourhossein; Hassan Korbekandi

    2014-01-01

    Background: For the past 30 years, bacterial L-asparaginases have been used as therapeutic agents in the treatment of acute childhood lymphoblastic leukemia. It is found in a variety of organisms such as microbes, plants and mammals. Their intrinsic low-rate glutaminase activity, however, causes serious side-effects, including neurotoxicity, hepatitis, coagulopathy and other dysfunctions. Erwinia carotovora asparaginase shows decreased glutaminase activity, so it is believed to have fewer sid...

  12. Isolation and Identification of L-asparaginase producing Erwinia strains which isolated from Potato Farms

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    Arastoo Badoei-Dalfard

    2016-09-01

    Full Text Available Introduction: L-Asparaginase can be effectively used for the treatment of lymphoblastic leukemia. The rapid growth of cancer cells are needed for L-asparagine abundant storage. L-asparaginase catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. The purpose of this study was to isolate and identify the L-asparaginase producing Erwinia strains from the potato farms of Jiroft. Materials and methods: Pectolytic Erwinia species isolated from crumbling potato in M9 medium. The desired L-asparaginase producing bacteria were isolated based on the color changes. Biochemical-microbial and the plant pathogenicity tests of these strains were also investigated with potato and geranium. The L-asparaginase production and molecular detection of these Erwinia strains were also investigated. Results: In this study, L-asparaginase producing Erwinia was isolated on the CVP and M9 mediums. The inoculation of Erwinia strains on the potato and geranium plants showed that Er8 and Er11 species have the ability to cause plant pathogenicity. Results showed that the maximum pathogenicity of Er8 and Er11 was observed after 48 and 15 h of inoculation in potato and geranium plants, respectively. 16S rDNA sequencing and phylogenetic analyses exhibited that Er8 and Er11 strains were similar to Erwinia chrysanthemi with 98% homology. Discussion and conclusion: Because of several applications of the Erwinia L-asparaginase in various fields, isolated Erwinia and their L-asparaginase can be suitable for applied utilization.

  13. Cloning and molecular analysis of L-asparaginase II gene (ansB

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    ZEINAT K. MOHAMED

    2015-12-01

    Full Text Available The deamination of L-asparagine to L-aspartic acid and ammonia is catalyzed by L-asparaginases (L-asparagine amino hydrolase. The enzyme L-asparaginase is widely distributed in nature from different living organisms, starting from bacteria till mammals and plants. It has been recently thought to be a therapeutic agent in treatment of various lymphoblastic leukemia diseases. There have been many attempts to isolate microorganisms that produce L-asparaginase. L-ASNase producing bacteria, Escherichia coli MG27, was previously isolated from the River Nile and identified. In this study, ansB gene, encoding L-ASNase II from E. coli MG27, was amplified by PCR, cloned and characterized by DNA sequencing. The DNA sequence was then analyzed using bioinformatics analysis and translated into amino acid sequence. Identification of highly conserved amino acid sequence motifs was conducted by comparison against the InterPro database. Analysis revealed that the protein sequence had a catalytic domain of L-asparaginase type II (IPR004550 that belong to asparaginase/glutaminase family (IPR006034 and has asparaginase/glutaminase conserved site (IPR020827. According to results predicted using PSIpred tool, ansB consists of eight α-helices and 13 β-strands.

  14. Cloning, expression, purification and characterisation of Erwinia carotovora L-asparaginase in Escherichia coli

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    Meraj Pourhossein

    2014-01-01

    Full Text Available Background: For the past 30 years, bacterial L-asparaginases have been used as therapeutic agents in the treatment of acute childhood lymphoblastic leukemia. It is found in a variety of organisms such as microbes, plants and mammals. Their intrinsic low-rate glutaminase activity, however, causes serious side-effects, including neurotoxicity, hepatitis, coagulopathy and other dysfunctions. Erwinia carotovora asparaginase shows decreased glutaminase activity, so it is believed to have fewer side-effects in leukemia therapy. Our aim was to clone, express, purify and characterize E. carotovora asparaginase. Materials and Methods: L-asparaginase from E. carotovora NCYC 1526 (ErA was cloned and expressed in Escherichia coli strain BL21 (DE3. The enzyme was purified to homogeneity by affinity chromatography. Various conditions were tested to maximize the production of recombinant asparaginase in E. coli. Results: A new L. asparaginase from E. carotovora NCYC 1526 (ErA was successfully cloned, expressed and purified in E. coli BL21 (DE3. The specific activity of the enzyme was 430 IU/mg. Conclusion: The results of the present work form the basis for a new engineered form of ErA for future therapeutic use, which could be extended with crystallographic studies.

  15. Production and optimization of L-asparaginase by an actinobacterium isolated from Nizampatnam mangrove ecosystem.

    Science.gov (United States)

    Kiranmayi, M Usha; Poda, Sudhakar; Vijayalakshmi, M

    2014-09-01

    The aim of the present study was to isolate and screen actinomycetes from the mangrove sediments of Nizampatnam that are potent to produce L-asparaginase, an enzyme that catalyses the hydrolysis of asparagine. A total of 31 actinomycetes strains were isolated, of which 6 strains were positive for L-asparaginase. Several physico-chemical parameters were optimized for maximizing L-asparaginase production by the potent strain identified as Pseudonocardia endophytica VUK-10. Production of L-asparaginase by the strain was high in modified Asparagine glucose salts broth (FM-4)(3.96 IU/ml) as compared to other tested media. Maltose(6.99 IU ml(-1)) and L-asparagine (7.42 IU ml(-1)) were found to be the most suitable carbon and nitrogen sources for optimum enzyme production. Maximum production of L-asparaginase was found in the culture medium with pH 8 and temperature 30 degrees C incubated for four days. This is the first report on the production of L-asparaginase by Pseudonocardia endophytica VUK-10 from Nizampatnam mangrove sediments.

  16. L-asparaginase production in the pseudomonas pseudoalcaligenes strain JHS-71 isolated from Jooshan Hot-spring

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    Arastoo Badoei-Dalfard

    2016-03-01

    Full Text Available L-asparaginase has lots of medical and industrial applications. Ever since L-asparaginase anti-tumor activity was first demonstrated, its production using microbial systems has attracted considerable attention owing to their cost-effective and eco-friendly nature. The aim of this study is to obtain L-asparaginase producing bacteria and determining the enzyme activity. Samples were picked up from Jooshan hot springs located in the Sirch, Kerman. The L-asparaginase producing bacteria were screened on the agar medium supplied with L-asparagine and phenol red indicator dye (pH-7.0. L-asparaginase activity was detected on the basis of pink color around the colony. Enzyme production was also performed based on ammonia detection by Nessler method. Among 24 strains, there were 7 strains which could produce L-asparaginase.Sequencing of 16S rRNA showed that, the best isolates producing L-asparaginase belongs to the Pseudomonas genus. Enzyme activity after 24 and 48 h of incubation showed that Pseudomonas pseudoalcaligenes strain JHS-71was the best strain that produced L-Asparaginase about 240 (U/ml after 48h of incubation. Results showed that, L-Asparaginase activity enhanced about 27% in the presence of Co+2. L-asparaginase JHS-71 retained more than 50% of its initial activity in the presence of Cu+2, Mn+2, Zn+2, Mg+2 and Fe+2. Because of various applications of L-asparaginase in biotechnology, P. pseudoalcaligenes strain JHS-71 can be used as a suitable candidate in these fields.

  17. Kinetic properties and inhibition of Acinetobacter glutaminase-asparaginase.

    Science.gov (United States)

    Steckel, J; Roberts, J; Philips, F S; Chou, T C

    1983-03-15

    Kinetic parameters, substrate specificity and exclusivity of ligands at binding sites of L-glutaminase-L-asparaginase purified from Acinetobacter glutaminasificans were studied in order to gain knowledge about the dual activities of this enzyme and its inhibition by structural analogs. Both L-glutamine and L-asparagine, which showed similar Km (4 approximately 7 X 10(-5) M) and Vmax (molecular activity 1.0 min-1) values, were competitive with each other for the substrate binding site. The products, L-glutamic acid and L-aspartic acid, showed competitive inhibition with respect to either L-glutamine or L-asparagine as substrates. Multiple inhibition of the glutaminase activity by L-glutamic acid and L-aspartic acid indicated that these ligands are mutually exclusive at the product-releasing site. The initial rates of both of the enzyme's activities were competitively inhibited by the following inhibitors (in rates of both of the enzyme's activities were competitively inhibited by the following inhibitors (in decreasing order of activity): 6-diazo-5-oxo-L-norleucine (DON), L-methionine sulfoximine, azaserine, and Acivicin. DON and azaserine inhibited both the asparaginase and glutaminase activities in a time-dependent and irreversible manner. The kinetic data suggest an ordered mechanism with glutamine or asparagine as the first substrate and glutamic acid or aspartic acid, respectively, as the last product. These results also suggest that a single mechanism and a single set of binding sites are responsible for catalyzing both of the enzyme's activities. The data also showed that succinylated enzyme, which has a 10-fold increase of plasma half-life in animals and humans and, thus, has benefit as a cancer chemotherapeutic agent, retained its catalytic activity and maintained Km and Vmax values similar to the native enzyme.

  18. Evaluating the potential for enzymatic acrylamide mitigation in a range of food products using an asparaginase from Aspergillus oryzae.

    Science.gov (United States)

    Hendriksen, Hanne V; Kornbrust, Beate A; Østergaard, Peter R; Stringer, Mary A

    2009-05-27

    Asparaginase, an enzyme that hydrolyzes asparagine to aspartic acid, presents a potentially very effective means for reducing acrylamide formation in foods via removal of the precursor, asparagine, from the primary ingredients. An extracellular asparaginase amenable to industrial production was cloned and expressed in Aspergillus oryzae . This asparaginase was tested in a range of food products, including semisweet biscuits, ginger biscuits, crisp bread, French fries, and sliced potato chips. In dough-based applications, addition of asparaginase resulted in reduction of acrylamide content in the final products of 34-92%. Enzyme dose, dough resting time, and water content were identified as critical parameters. Treating French fries and sliced potato chips was more challenging as the solid nature of these whole-cut products limits enzyme-substrate contact. However, by treating potato pieces with asparaginase after blanching, the acrylamide levels in French fries could be lowered by 60-85% and that in potato chips by up to 60%.

  19. L-asparaginase production by mangrove derived Bacillus cereus MAB5:optimization by response surface methodology

    Institute of Scientific and Technical Information of China (English)

    ThenmozhiC; SankarR; KaruppiahV; SampathkumarP

    2011-01-01

    Objective:To isolate marine bacteria, statistically optimize them for maximum asparaginase production. Methods:In the present study, statistically based experimental designs were applied to maximize the production of L-asparaginase from bacterial strain of Bacillus cereus (B. cereus) MAB5 (HQ675025) isolated and identified by 16S rDNA sequencing from mangroves rhizosphere sediment. Results:Plackett-Barman design was used to identify the interactive effect of the eight variables viz. yeast extract, soyabean meal, glucose, magnesium sulphate, KH2PO4, wood chips, aspargine and sodium chloride. All the variables are denoted as numerical factors and investigated at two widely spaced intervals designated as-1 (low level) and+1 (high level). The effect of individual parameters on L-asparaginase production was calculated. Soyabean meal, aspargine, wood chips and sodium chloride were found to be the significant among eight variables. The maximum amount of L-asparaginase produced (51.54 IU/mL) from the optimized medium containing soyabean meal (6.282 8 g/L), aspargine (5.5 g/L), wood chips (1.383 8 g/L) and NaCl (4.535 4 g/L). Conclusions:The study revealed that, it is useful to produce the maximum amount of L-asparaginase from B. cereus MAB5 for the treatment of various infections and diseases.

  20. Chemical modification of L-asparaginase from Cladosporium sp. for improved activity and thermal stability.

    Science.gov (United States)

    Mohan Kumar, N S; Kishore, Vijay; Manonmani, H K

    2014-01-01

    L-Asparaginase (ASNase), an antileukemia enzyme, is facing problems with antigenicity in the blood. Modification of L-asparaginase from Cladosporium sp. was tried to obtain improved stability and improved functionality. In our experiment, modification of the enzyme was tried with bovine serum albumin, ovalbumin by crosslinking using glutaraldehyde, N-bromosuccinimide, and mono-methoxy polyethylene glycol. Modified enzymes were studied for activity, temperature stability, rate constants (kd), and protection to proteolytic digestion. Modification with ovalbumin resulted in improved enzyme activity that was 10-fold higher compared to native enzyme, while modification with bovine serum albumin through glutaraldehyde cross-linking resulted in high stability of L-asparaginase that was 8.5- and 7.62-fold more compared to native enzyme at 28°C and 37°C by the end of 24 hr. These effects were dependent on the quantity of conjugate formed. Modification also markedly prolonged L-asparaginase half-life and serum stability. N-Bromosuccinimide-modified ASNase presented greater stability with prolonged in vitro half-life of 144 hr to proteolytic digestion relative to unmodified enzyme (93 h). The present work could be seen as producing a modified L-asparaginase with improved activity and stability and can be a potential source for developing therapeutic agents for cancer treatment.

  1. Purification and properties of asparaginase from the testa of immature seeds of pea (Pisum sativum L.

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    Chagas Eliana P.

    2001-01-01

    Full Text Available A K+-dependent asparaginase (E.C. 3.5.1.1. was purified 1328-fold from the testas of immature pea seeds (Pisum sativum L., var. Bolero and characterized. Antibodies raised against purified asparaginase cross-reacted with the putative asparaginase band in Western blot analyses of semi-purified extracts. However, for crude extracts of pea testas, a cross-reaction was obtained with at least four protein bands, one of which was asparaginase protein. Affinity-purified antibodies to the four strongest bands of crude extracts were fairly specific for the bands from which they were purified, suggesting a mixture of specific antibodies. The Mr of asparaginase was 69,000 by Sephacryl S200 chromatography and also by mobility on native PAGE relative to BSA. There was no evidence for dissociation into subunits on SDS-PAGE, suggesting a monomeric protein of Mr 69,000. Other properties include an apparent Km of 2.4 mM, pI between 4.5 and 5, and competitive inhibition by aspartate and glycine.

  2. Serial Ultrasound Monitoring for Early Recognition of Asparaginase Associated Pancreatitis in Children With Acute Lymphoblastic Leukemia

    DEFF Research Database (Denmark)

    Raja, Raheel Altaf; Schmiegelow, K.; Henriksen, Birthe Merete;

    2015-01-01

    BACKGROUND: Acute lymphoblastic leukemia (ALL) is the most common cancer in children and L-asparaginase is an essential component of the treatment. Cessation of L-asparaginase decreases event free survival. Acute pancreatitis is the toxicity that most commonly results in cessation of L-asparagina......BACKGROUND: Acute lymphoblastic leukemia (ALL) is the most common cancer in children and L-asparaginase is an essential component of the treatment. Cessation of L-asparaginase decreases event free survival. Acute pancreatitis is the toxicity that most commonly results in cessation of L...... protocol, with PEG-asparaginase of 2 or 6 week intervals, for 30 weeks had their pancreas monitored using serial ultrasound in order to detect early signs of inflammation. RESULTS: Nineteen of 31 eligible patients were included. Three of the included patients developed AAP. None of the patients, including...... the three patients that developed AAP, had signs of inflammatory edema or pancreas enzymes above three times the upper normal limit prior to AAP. CONCLUSION: We found no signs of inflammatory edema within the pancreas on ultrasound during treatment with PEG-asparginase in our cohort prior to development...

  3. CHARACTERIZATION OF l-ASPARAGINASE PRODUCING BACTERIA FROM WATER, FARM AND SALINE SOIL

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    K D Kamble

    2012-01-01

    Full Text Available The habitat chosen for screening the bacteria were farm soil, saline soil and water. The activity was detected on a medium containing 1% peptone, 0.6% beef extract, 0.33% KH3PO4, 0.1% L-asparagine and phenol red. L-asparaginase activity was detected on the basis of formation of red colour around the colony. Likewise efficient L- asparaginase producing bacteria were screened. These were then studied for routine microbiological and biochemical characterization. The microorganisms we characterized belonged to the genera E.coli, Serratia spp., Pseudomonas aeruginosa, Bacillus spp., Aeromonas species and Proteus spp. L-asparaginase from halophilic bacteria is expected to be non-allergic and hence halophilic bacteria from saline soil can contribute to therapeutic value of this enzyme.

  4. PRODUCTION OPTIMIZATION OF EXTRACELLULAR L-ASPARAGINASE THROUGH SOLID- STATE FERMENTATION BY ISOLATED BACILLUS SUBTILIS.

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    Susmita Shukla

    2013-02-01

    Full Text Available L-asparaginase has been used as anti-tumor agent for the treatment of acute lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. Extracellular Lasparaginase production was optimized through solid state fermentation using ground nut cake by isolated Bacillus subtilis. which was not reported in literature.Optimum production of L-asparaginase enzyme (18.4U/ml was obtained after 48h of incubation at 370C moisture content of 70% and at pH 7.

  5. E. coli-Derived L-Asparaginase Retains Enzymatic and Cytotoxic Activity In Vitro for Canine and Feline Lymphoma after Cold Storage

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    Jackie M. Wypij

    2013-01-01

    Full Text Available Background. L-asparaginase is effective in treating canine and feline lymphoma, however chemotherapy poses a significant financial cost to veterinary clients, limiting therapy for many pets. Single dose vials result in significant drug wastage, and drug shortages limit consistent availability for pets. Hypothesis. E. coli-derived asparaginase retains enzymatic and antineoplastic activity in canine and feline lymphoma cells after cold storage. Methods. E. coli-derived asparaginase was cold-stored: refrigeration (7–14 days and freezing (14 days–six months, one to three freeze/thaw cycles. Enzymatic activity of asparaginase was measured via a modified asparagine assay. Effects of cold-stored asparaginase on cell proliferation and cytotoxicity were measured in feline (MYA-1, F1B and canine (17–71, OSW lymphoma cells. Results. Cold-stored E. coli-derived asparaginase retains antineoplastic activity in all four cell lines tested. Cold-stored E. coli-derived L-asparaginase depletes asparagine and retains enzymatic activity. Duration of refrigeration, duration of freezing, and number of freeze-thaw cycles have minimal effect on asparaginase enzyme activity. Conclusions and Clinical Importance. This study establishes a scientific basis for long-term cold storage of reconstituted E. coli-derived asparaginase that may result in better utilization of limited drug resources and improve financial feasibility of E. coli-derived asparaginase as a therapeutic option for pets with lymphoma.

  6. Bio-grout based on microbially induced sand solidification by means of asparaginase activity

    Science.gov (United States)

    Li, Mengmeng; Fu, Qing-Long; Zhang, Qiuzhuo; Achal, Varenyam; Kawasaki, Satoru

    2015-11-01

    Bio-grout, a new ground improvement method, has been recently developed to improve the mechanical properties, decrease the permeability of porous materials, reinforce or repair cementitious materials and modify the properties of soil or sand. Bio-grout production depends on microbially induced calcite precipitation (MICP), which is driven mainly by an enzyme, urease. However, urease-based MICP process produces excessive ammonia, in addition to secondary pollution generated by urea that is used as substrate in it. In the present study, we reported asparaginase-based MICP process for sand bio-grout development using Bacillus megaterium, and results were also compared with urease-based bio-grouts. The asparaginase activity led to significantly less ammonia production compared to urease without compromising with desired properties of a novel grout. The UCS of bio-grout was obtained at 980 kPa, while the permeability was decreased substantially. The mineralogical composition of precipitated substance was identified as calcite using XRD and the crystal morphology was observed under SEM. The mass percentage of calcite in bio-grout was calculated by thermogravimetric analysis and XCT verified calcite precipitation in it. The results confirmed that biocalcification by means of bacterial asparaginase is a potential solution for geotechnical problems. The asparaginase-based MICP process could be of wider acceptance in future.

  7. Purification, characterization and kinetic properties of extracellular L-asparaginase produced by Cladosporium sp.

    Science.gov (United States)

    Mohan Kumar, N S; Manonmani, H K

    2013-04-01

    L-asparaginase from Cladosporium sp. grown on wheat bran by SSF was purified. Enzyme appeared to be a trimer with homodimer of 37 kDa and another 47 kDa amounting to total mass of 121 kDa as estimated by SDS-PAGE and 120 kDa on gel filtration column. The optimum temperature and pH of the enzyme were 30 °C and 6.3, respectively with Vmax of 4.44 μmol/mL/min and Km of 0.1 M. Substrate specificity studies indicated that, L-asparaginase has greater affinity towards L-asparagine with substrate hydrolysis efficiency (Vmax/Km ratio) eightfold higher than that of L-glutamine. L-asparaginase activity in presence of thiols studied showed decrease in Vmax and increase in Km, indicating nonessential mode of inactivation. Among the thiols tested, β-mercaptomethanol, exerted inhibitory effect, suggesting a critical role of disulphide linkages in maintaining a suitable conformation of the enzyme. Metal ions such as Ca(2+), Co(2+), Cu(2+), Mg(2+), Na(+), K(+) and Zn(2+) significantly affected enzyme activity whereas presence of Fe(3+), Pb(2+) and KI stimulated the activity. Detergents studied also enhanced L-asparaginase activity. In-vitro half-life of purified L-asparaginase in mammalian blood serum was 93.69 h. The enzyme inhibited acrylamide formation in potato chips by 96 % making it a potential candidate for food industry to reduce acrylamide content in starchy fried food commodities.

  8. Crystallization and preliminary crystallographic analysis of l-asparaginase from Erwinia carotovora

    Energy Technology Data Exchange (ETDEWEB)

    Wikman, Linnea E. K. [Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, Turku 20521 (Finland); Krasotkina, Julya; Kuchumova, Anastasia; Sokolov, Nikolay N. [Institute for Biomedical Chemistry, Russian Academy of Medical Sciences, 559-B, 10 Pogodinskay St, Moscow 119121 (Russian Federation); Papageorgiou, Anastassios C., E-mail: tassos.papageorgiou@btk.fi [Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, Turku 20521 (Finland)

    2005-04-01

    Er. carotovoral-asparaginase, a potential antileukaemic agent, has been crystallized. Crystals diffract to 2.6 Å using a rotating-anode source and belong to space group P2{sub 1}, with unit-cell parameters a = 78.0, b = 112.3, c = 78.7 Å, β = 101.9° and a homotetramer in the crystallographic asymmetric unit. Bacterial l-asparaginases have been used as therapeutic agents in the treatment of acute childhood lymphoblastic leukaemia for over 30 y. However, their use is limited owing to the glutaminase activity of the administered enzymes, which results in serious side effects. In contrast, l-asparaginase from Erwinia carotovora exhibits low glutaminase activity at physiological concentrations of l-asparagine and l-glutamine in the blood. Recombinant Er. carotovoral-asparaginase was crystallized in the presence of l-glutamate by the hanging-drop vapour-diffusion method using 10 mg ml{sup −1} purified enzyme, 16–18%(w/v) PEG 3350 and 0.2 M NaF. X-ray diffraction data were collected to 2.6 Å at 293 K using an in-house rotating-anode generator. The crystals belong to the monoclinic P2{sub 1} space group, with unit-cell parameters a = 78.0, b = 112.3, c = 78.7 Å, β = 101.9° and a homotetramer in the crystallographic asymmetric unit. A molecular-replacement solution has been found and refinement is currently in progress. The crystal structure may provide leads towards protein-engineering efforts aimed at safer asparaginase administration in leukaemia treatment.

  9. Erwinia asparaginase achieves therapeutic activity after pegaspargase allergy: a report from the Children's Oncology Group.

    Science.gov (United States)

    Salzer, Wanda L; Asselin, Barbara; Supko, Jeffrey G; Devidas, Meenakshi; Kaiser, Nicole A; Plourde, Paul; Winick, Naomi J; Reaman, Gregory H; Raetz, Elizabeth; Carroll, William L; Hunger, Stephen P

    2013-07-25

    AALL07P2 evaluated whether substitution of Erwinia asparaginase 25000 IU/m(2) for 6 doses given intramuscularly Monday/Wednesday/Friday (M/W/F) to children and young adults with acute lymphoblastic leukemia and clinical allergy to pegaspargase would provide a 48-hour nadir serum asparaginase activity (NSAA) ≥ 0.10 IU/mL. AALL07P2 enrolled 55 eligible/evaluable patients. NSAA ≥ 0.1 IU/mL was achieved in 38 of 41 patients (92.7%) with acceptable samples 48 hours and in 38 of 43 patients (88.4%) 72 hours after dosing during course 1. Among samples obtained during all courses, 95.8% (252 of 263) of 48-hour samples and 84.5% (125 of 148) of 72-hour samples had NSAA ≥ 0.10-IU/mL. Pharmacokinetic parameters were estimated by fitting the serum asparaginase activity-time course for all 6 doses given during course 1 to a 1-compartment open model with first order absorption. Erwinia asparaginase administered with this schedule achieved therapeutic NSAA at both 48 and 72 hours and was well tolerated with no reports of hemorrhage, thrombosis, or death, and few cases of grade 2 to 3 allergic reaction (n = 6), grade 1 to 3 hyperglycemia (n = 6), or grade 1 pancreatitis (n = 1). Following allergy to pegaspargase, Erwinia asparaginase 25000 IU/m(2) × 6 intramuscularly M/W/F can be substituted for a single dose of pegaspargase.

  10. The K+-dependent asparaginase, NSE1, is crucial for plant growth and seed production in Lotus japonicus.

    Science.gov (United States)

    Credali, Alfredo; García-Calderón, Margarita; Dam, Svend; Perry, Jillian; Díaz-Quintana, Antonio; Parniske, Martin; Wang, Trevor L; Stougaard, Jens; Vega, José M; Márquez, Antonio J

    2013-01-01

    The physiological role of K(+)-dependent and K(+)-independent asparaginases in plants remains unclear, and the contribution from individual isoforms during development is poorly understood. We have used reverse genetics to assess the phenotypes produced by the deficiency of K(+)-dependent NSE1 asparaginase in the model legume Lotus japonicus. For this purpose, four different mutants were identified by TILLING and characterized, two of which affected the structure and function of the asparaginase molecule and caused asparagine accumulation. Plant growth and total seed weight of mature mutant seeds as well as the level of both legumin and convicilin seed storage proteins were affected in the mutants. The mutants isolated in the present work are the first of their type in legumes and have enabled us to demonstrate the importance of asparagine and K(+)-dependent NSE1 asparaginase for nitrogen remobilization and seed production in L. japonicus plants.

  11. Molecular expression of l-asparaginase gene from Nocardiopsis alba NIOT-VKMA08 in Escherichia coli: A prospective recombinant enzyme for leukaemia chemotherapy.

    Science.gov (United States)

    Meena, Balakrishnan; Anburajan, Lawrance; Vinithkumar, Nambali Valsalan; Shridhar, Divya; Raghavan, Rangamaran Vijaya; Dharani, Gopal; Kirubagaran, Ramalingam

    2016-09-30

    l-Asparaginase is an antineoplastic agent that selectively reduces the level of l-asparagine in blood and diminishes the proliferation of cancerous cells. Studies were carried out on the cloning and heterologous expression of l-asparaginase biosynthesis gene (ansA) from Nocardiopsis alba NIOT-VKMA08 to achieve the stable inducible system that overproduces the glutaminase-free recombinant l-asparaginase. Overexpression of recombinant l-asparaginase was achieved with an optimized final concentration of 1.5mM of isopropyl-β-d-thiogalactoside (IPTG) and the enzyme was expressed as a soluble protein. The recombinant enzyme was purified using nickel-nitrilotriacetic acid (Ni-NTA) chromatography and the purified enzyme disclosed an elevated level of asparaginase activity (158.1IU/mL). Optimum pH and temperature of the purified l-asparaginase for the hydrolysis of l-asparagine were 8.0 and 37°C and it was very specific for its natural substrate, l-asparagine. Detailed studies were carried out on the kinetics of enzyme reaction, catalytic activity, temperature and ionic strength and the thermostability of the l-asparaginase enzyme. The functional characterisation of the recombinant l-asparaginase was studied through Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), in silico sequence analysis and protein structural modelling. Glutaminase activity was not detected in the recombinant l-asparaginase, which could reduce the probable side effects during leukaemia therapy.

  12. Molecular expression of l-asparaginase gene from Nocardiopsis alba NIOT-VKMA08 in Escherichia coli: A prospective recombinant enzyme for leukaemia chemotherapy.

    Science.gov (United States)

    Meena, Balakrishnan; Anburajan, Lawrance; Vinithkumar, Nambali Valsalan; Shridhar, Divya; Raghavan, Rangamaran Vijaya; Dharani, Gopal; Kirubagaran, Ramalingam

    2016-09-30

    l-Asparaginase is an antineoplastic agent that selectively reduces the level of l-asparagine in blood and diminishes the proliferation of cancerous cells. Studies were carried out on the cloning and heterologous expression of l-asparaginase biosynthesis gene (ansA) from Nocardiopsis alba NIOT-VKMA08 to achieve the stable inducible system that overproduces the glutaminase-free recombinant l-asparaginase. Overexpression of recombinant l-asparaginase was achieved with an optimized final concentration of 1.5mM of isopropyl-β-d-thiogalactoside (IPTG) and the enzyme was expressed as a soluble protein. The recombinant enzyme was purified using nickel-nitrilotriacetic acid (Ni-NTA) chromatography and the purified enzyme disclosed an elevated level of asparaginase activity (158.1IU/mL). Optimum pH and temperature of the purified l-asparaginase for the hydrolysis of l-asparagine were 8.0 and 37°C and it was very specific for its natural substrate, l-asparagine. Detailed studies were carried out on the kinetics of enzyme reaction, catalytic activity, temperature and ionic strength and the thermostability of the l-asparaginase enzyme. The functional characterisation of the recombinant l-asparaginase was studied through Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), in silico sequence analysis and protein structural modelling. Glutaminase activity was not detected in the recombinant l-asparaginase, which could reduce the probable side effects during leukaemia therapy. PMID:27155523

  13. Application of repeated aspartate tags to improving extracellular production of Escherichia coli L-asparaginase isozyme II.

    Science.gov (United States)

    Kim, Sun-Ki; Min, Won-Ki; Park, Yong-Cheol; Seo, Jin-Ho

    2015-11-01

    Asparaginase isozyme II from Escherichia coli is a popular enzyme that has been used as a therapeutic agent against acute lymphoblastic leukemia. Here, fusion tag systems consisting of the pelB signal sequence and various lengths of repeated aspartate tags were devised to highly express and to release active asparaginase isozyme II extracellularly in E. coli. Among several constructs, recombinant asparaginase isozyme II fused with the pelB signal sequence and five aspartate tag was secreted efficiently into culture medium at 34.6 U/mg cell of specific activity. By batch fermentation, recombinant E. coli produced 40.8 U/ml asparaginase isozyme II in the medium. In addition, deletion of the gspDE gene reduced extracellular production of asparaginase isozyme II, indicating that secretion of recombinant asparaginase isozyme II was partially ascribed to the recognition by the general secretion machinery. This tag system composed of the pelB signal peptide, and repeated aspartates can be applied to extracellular production of other recombinant proteins.

  14. Acute Pancreatitis and Diabetic Ketoacidosis following L-Asparaginase/Prednisone Therapy in Acute Lymphoblastic Leukemia

    Directory of Open Access Journals (Sweden)

    Dania Lizet Quintanilla-Flores

    2014-01-01

    Full Text Available Acute pancreatitis and diabetic ketoacidosis are unusual adverse events following chemotherapy based on L-asparaginase and prednisone as support treatment for acute lymphoblastic leukemia. We present the case of a 16-year-old Hispanic male patient, in remission induction therapy for acute lymphoblastic leukemia on treatment with mitoxantrone, vincristine, prednisone, and L-asparaginase. He was hospitalized complaining of abdominal pain, nausea, and vomiting. Hyperglycemia, acidosis, ketonuria, low bicarbonate levels, hyperamylasemia, and hyperlipasemia were documented, and the diagnosis of diabetic ketoacidosis was made. Because of uncertainty of the additional diagnosis of acute pancreatitis as the cause of abdominal pain, a contrast-enhanced computed tomography was performed resulting in a Balthazar C pancreatitis classification.

  15. Hypoglycemia associated with L-asparaginase in acute lymphoblastic leukemia treatment: a case report

    OpenAIRE

    Tanaka Ryuma; Osumi Tomoo; Miharu Masashi; Ishii Tomohiro; Hasegawa Tomonobu; Takahashi Takao; Shimada Hiroyuki

    2012-01-01

    Abstract A patient with acute lymphoblastic leukemia repeatedly developed hypoglycemia during chemotherapy. Comparison of serum glucose trends between chemotherapy with and without L-asparaginase (L-Asp) demonstrated a strong association between L-Asp and hypoglycemia. Critical blood sampling during hypoglycemia indicated hyperinsulinism, suggesting that L-Asp induced hypoglycemia in the patient through inappropriate insulin secretion. Identification of hypoglycemia as an adverse effect will ...

  16. Sequential Administration of Methotrexate and Asparaginase in Relapsed or Refractory Pediatric Acute Myeloid Leukemia

    Science.gov (United States)

    Buaboonnam, Jassada; Cao, Xueyuan; Pauley, Jennifer L.; Pui, Ching-Hon; Ribeiro, Raul C.; Rubnitz, Jeffrey E.; Inaba, Hiroto

    2014-01-01

    Background The efficacy of combination chemotherapy with methotrexate (MTX) and asparaginase is not well known in relapsed and refractory acute leukemia after contemporary therapy. Procedure A retrospective study of pediatric patients with relapsed or refractory acute myeloid leukemia (AML) who received MTX and asparaginase as a salvage therapy at St. Jude Children Research Hospital was performed. MTX was given intravenously followed by a dose of asparaginase intramuscularly or intravenously 24 hours later. The chemotherapy cycle was repeated every 7-10 days. Response, survival, and toxicities were evaluated. Results Fifteen patients, median age 10.5 years (range, 1.1-18.5 years), were treated. Median number of previous therapeutic regimens was 3 (range, 1-4). Six patients responded to treatment (3 had morphologic complete remission with incomplete blood count recovery, 2 had partial remission, and 1 had stable disease for 16 months), and 4 are still alive. Three of 6 responders had monoblastic leukemia, and also developed tumor lysis syndrome. The 1- and 2-year overall survival rates are 35.6% and 17.8%, respectively. The most common adverse event was transient elevation of transaminases (9 patients). Two patients developed pancreatitis. Episodes of febrile neutropenia were rare (2 patients), and most courses (75 out of 93 total courses) were given in an outpatient setting. Conclusions Combination chemotherapy with MTX and asparaginase appears to be an effective salvage therapy and well tolerated in patients with relapsed or refractory childhood AML, even in those heavily pretreated with contemporary frontline or salvage therapy. PMID:23335430

  17. Direct long-term effects of L-asparaginase on rat and human pancreatic islets

    DEFF Research Database (Denmark)

    Clausen, Niels; Nielsen, Jens Høiriis

    1989-01-01

    L-Asparaginase, an effective agent in the treatment of acute lymphoblastic leukemia, may induce a diabetic state. The pathogenesis of the diabetogenic effect was studied in cultured pancreatic islets. Mean serum concentrations in three children with acute lymphoblastic leukemia were 2.4 U/mL (range...... 1.4-4.5) before and 31.5 U/mL (range 18.6-51.8) immediately after an intravenous injection of 1000 U/kg L-asparaginase. Glucose-induced insulin release from pancreatic islets of rat and man was measured after 3 and 7 days of culture in media with or without clinically relevant concentrations...... of Escherichia coli L-asparaginase (0.01-100 U/mL). After culture, the remaining insulin, glucagon, and DNA in the islets were determined. After 7 days of culture of adult rat or human islets, both the accumulation of insulin in the medium and the content of insulin and glucagon in the islets were significantly...

  18. Optimization, purification, and characterization of L-asparaginase from Actinomycetales bacterium BkSoiiA.

    Science.gov (United States)

    Dash, Chitrangada; Mohapatra, Sukanti Bala; Maiti, Prasanta Kumar

    2016-01-01

    Actinobacteria are promising source of a wide range of important enzymes, some of which are produced in industrial scale, with others yet to be harnessed. L-Asparaginase is used as an antineoplastic agent. The present work deals with the production and optimization of L-asparaginase from Actinomycetales bacterium BkSoiiA using submerged fermentation in M9 medium. Production optimization resulted in a modified M9 medium with yeast extract and fructose as carbon and nitrogen sources, respectively, at pH 8.0, incubated for 120 hr at 30 ± 2 °C. The crude enzyme was purified to near homogeneity by ammonium sulfate precipitation following dialysis, ion-exchange column chromatography, and finally gel filtration. The sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) revealed an apparent molecular weight of 57 kD. The enzyme was purified 95.06-fold and showed a final specific activity of 204.37 U/mg with 3.49% yield. The purified enzyme showed maximum activity at a pH 10.0 and was stable at pH 7.0 to 9.0. The enzyme was activated by Mn(2+) and strongly inhibited by Ba(2+). All these preliminary characterization suggests that the L-asparaginase from the source may be a tool useful to pharmaceutical industries after further research.

  19. PURIFICATION AND PROPERTIES OF A FUNGAL L-ASPARAGINASE FROM TRICHODERMA VIRIDE PERS: SF GREY

    Directory of Open Access Journals (Sweden)

    Lynette Lincoln

    2015-02-01

    Full Text Available A potent L-asparaginase-producing Trichoderma viride Pers: SF Grey was screened from the marine soil with the objective of studying the enzyme properties. The maximum enzyme production occurred on the third day at pH 6.5 and 37 °C when 0.5% L-asparagine supplemented with 0.5% peptone and 0.6% maltose. The enzyme was purified to homogeneity with a specific activity of 78.2 U.mg-1 and a molecular weight of 99 ± 1 kDa. It exhibited maximum activity at pH 7.0 and 37 °C. It was inhibited by Fe2+, Fe3+, Co2+ and Mn2+ but induced by Mg2+ and Na+. N-ethylemaleimide and phenylmethylsulphonylfluoride did not alter the enzyme activity, but strongly inhibited by ethylenediaminetetraacetate. L-asparaginase showed high affinity for L-asparagine with a Km of 2.56 μM. Thin layer chromatography confirmed the hydrolysis of L-asparagine. As the purified and characterized L-asparaginase of Trichoderma viride showed a good scavenging activity and reduced acrylamide level in potato products, it can further serve as an antileukemic protein and an acrylamide mitigation agent in heat-treated food stuffs rich in carbohydrates, respectively.

  20. L-Asparaginase Therapy with Concomitant Cranial Venous Thrombosis: Can Mri Help Avoiding Stroke

    International Nuclear Information System (INIS)

    To prospectively use MRI in the early detection of intracranial sino-venous thrombosis during the L-asparaginase induction therapy of acute leukemia thus preventing the evolution of brain venous infarct. Materials and Methods: The study population consisted of seventy patients receiving L-asparaginase induction therapy for acute leukemia in the National Cancer Institute of Cairo University presenting with clinical neurological signs suggestive of aseptic intracranial venous thrombosis. All the patients were studied by 1.5 Tesla magnet using conventional MRI pulse sequences and MR veno graphic studies. The imaging findings were processed as regards the detection of venous thrombosis, its signal criteria and the evaluation of any companion brain parenchymal ischemic insults. Results: Eleven patients were diagnosed with d ural venous sinus thrombosis with subsequent specific signal pattern of the thrombus that could be linked to the duration of thrombosis. The MR veno graphic studies detected the thrombosis in nine cases out of eleven. Ten cases scored brain parenchymal signal abnormality that could indicate infarction, eight of them were hemorrhagic in nature. Conclusion: L-asparaginase therapy is accompanied by high risk of venous thrombosis that could involve the intra-cranial sino-venous structures. MRI could be used effectively in the early diagnosis of such serious, curable complication using a combination of conventional spin echo pulse sequences and MR veno graphic studies. Hemorrhagic venous infarcts should draw the attention to underlying established venous thrombosis.

  1. Calcium and adenosine triphosphate control of cellular pathology: asparaginase-induced pancreatitis elicited via protease-activated receptor 2.

    Science.gov (United States)

    Peng, Shuang; Gerasimenko, Julia V; Tsugorka, Tatiana; Gryshchenko, Oleksiy; Samarasinghe, Sujith; Petersen, Ole H; Gerasimenko, Oleg V

    2016-08-01

    Exocytotic secretion of digestive enzymes from pancreatic acinar cells is elicited by physiological cytosolic Ca(2+) signals, occurring as repetitive short-lasting spikes largely confined to the secretory granule region, that stimulate mitochondrial adenosine triphosphate (ATP) production. By contrast, sustained global cytosolic Ca(2+) elevations decrease ATP levels and cause necrosis, leading to the disease acute pancreatitis (AP). Toxic Ca(2+) signals can be evoked by products of alcohol and fatty acids as well as bile acids. Here, we have investigated the mechanism by which l-asparaginase evokes AP. Asparaginase is an essential element in the successful treatment of acute lymphoblastic leukaemia, the most common type of cancer affecting children, but AP is a side-effect occurring in about 5-10% of cases. Like other pancreatitis-inducing agents, asparaginase evoked intracellular Ca(2+) release followed by Ca(2+) entry and also substantially reduced Ca(2+) extrusion because of decreased intracellular ATP levels. The toxic Ca(2+) signals caused extensive necrosis. The asparaginase-induced pathology depended on protease-activated receptor 2 and its inhibition prevented the toxic Ca(2+) signals and necrosis. We tested the effects of inhibiting the Ca(2+) release-activated Ca(2+) entry by the Ca(2+) channel inhibitor GSK-7975A. This markedly reduced asparaginase-induced Ca(2+) entry and also protected effectively against the development of necrosis.This article is part of the themed issue 'Evolution brings Ca(2+) and ATP together to control life and death'. PMID:27377732

  2. Calcium and adenosine triphosphate control of cellular pathology: asparaginase-induced pancreatitis elicited via protease-activated receptor 2

    Science.gov (United States)

    Peng, Shuang; Gerasimenko, Julia V.; Tsugorka, Tatiana; Gryshchenko, Oleksiy; Samarasinghe, Sujith; Gerasimenko, Oleg V.

    2016-01-01

    Exocytotic secretion of digestive enzymes from pancreatic acinar cells is elicited by physiological cytosolic Ca2+ signals, occurring as repetitive short-lasting spikes largely confined to the secretory granule region, that stimulate mitochondrial adenosine triphosphate (ATP) production. By contrast, sustained global cytosolic Ca2+ elevations decrease ATP levels and cause necrosis, leading to the disease acute pancreatitis (AP). Toxic Ca2+ signals can be evoked by products of alcohol and fatty acids as well as bile acids. Here, we have investigated the mechanism by which l-asparaginase evokes AP. Asparaginase is an essential element in the successful treatment of acute lymphoblastic leukaemia, the most common type of cancer affecting children, but AP is a side-effect occurring in about 5–10% of cases. Like other pancreatitis-inducing agents, asparaginase evoked intracellular Ca2+ release followed by Ca2+ entry and also substantially reduced Ca2+ extrusion because of decreased intracellular ATP levels. The toxic Ca2+ signals caused extensive necrosis. The asparaginase-induced pathology depended on protease-activated receptor 2 and its inhibition prevented the toxic Ca2+ signals and necrosis. We tested the effects of inhibiting the Ca2+ release-activated Ca2+ entry by the Ca2+ channel inhibitor GSK-7975A. This markedly reduced asparaginase-induced Ca2+ entry and also protected effectively against the development of necrosis. This article is part of the themed issue ‘Evolution brings Ca2+ and ATP together to control life and death’. PMID:27377732

  3. [The feasibility of Erwinia asparaginase for pediatric patients who developed an allergic reaction to E.coli asparaginase during treatment of acute lymphoblastic leukemia].

    Science.gov (United States)

    Takahashi, Hiroyoshi; Koh, Katsuyoshi; Kato, Motohiro; Isobe, Kiyotaka; Yasui, Naoko; Mori, Makiko; Akiyama, Kosuke; Kikuchi, Akira; Hanada, Ryoji

    2013-04-01

    Asparaginase (ASNase) is one of the most important key drugs in the treatment of acute lymphoblastic leukemia (ALL). However, clinical hypersensitivity reactions often occur and lead to the discontinuation of ASNase treatment. Here, we report a retrospective study of 68 Erwinia ASNase (Erw-ASNase) administrations in 11 patients with childhood ALL who developed allergic reactions to E.coli-ASNase in our hospital between 2006 and 2012. The median age of the patients was 6 (range, 0 to 14). Erw-ASNase purchased overseas by the patients' guardians had already been administered when we obtained informed consent from the guardians. In all patients, fibrinogen and/or anti-thrombin III levels were decreased, but thrombosis did not develop. There was only one mild adverse event (grade 2 urticaria) in one patient, in whom Erw-ASNase could be continued after increasing the doses of premedication with antihistamine and prednisolone. Erw-ASNase could be safely administered to all patients.

  4. The Human Asparaginase-Like Protein 1 hASRGL1is an Ntn Hydrolase with β-aspartyl Peptidase Activity

    OpenAIRE

    Cantor, Jason R.; Stone, Everett M.; Chantranupong, Lynne; Georgiou, George

    2009-01-01

    Herein we report the bacterial expression, purification, and enzymatic characterization of the human asparaginase-like protein 1 (hASRGL1). We present evidence that hASRGL1 exhibits β-aspartyl peptidase activity consistent with enzymes designated as plant-type asparaginases, which had thus far only been found in plants and bacteria. Similar to non-mammalian plant-type asparaginases, hASRGL1 is shown to be an Ntn hydrolase for which Thr168 serves as the essential N-terminal nucleophile for int...

  5. Reduction of acrylamide level through blanching with treatment by an extremely thermostable L-asparaginase during French fries processing.

    Science.gov (United States)

    Zuo, Shaohua; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

    2015-07-01

    Bacterial L-asparaginase catalyzes the hydrolysis of L-asparagine to L-aspartic acid. It is normally used as an antineoplastic drug applied in lymphoblastic leukemia chemotherapy and as a food processing aid in baked or fried food industry to inhibit the formation of acrylamide. The present study demonstrates cloning, expression, and characterization of a thermostable L-asparaginase from Thermococcus zilligii AN1 TziAN1_1 and also evaluates the potential for enzymatic acrylamide mitigation in French fries using this enzyme. The recombinant L-asparaginase was purified to homogeneity by nickel-affinity chromatography. The purified enzyme displayed the maximum activity at pH 8.5 and 90 °C, and the optimum temperature was the highest ever reported. The K m, k cat, and k cat/K m values toward L-asparagine were measured to be 6.08 mM, 3267 s(-1), and 537.3 mM(-1) s(-1), respectively. The enzyme retained 70 % of its original activity after 2 h of incubation at 85 °C. When potato samples were treated with 10 U/mL of L-asparaginase at 80 °C for only 4 min, the acrylamide content in final French fries was reduced by 80.5 % compared with the untreated control. Results of this study revealed that the enzyme was highly active at elevated temperatures, reflecting the potential of the T. zilligii L-asparaginase in the food processing industry. PMID:26077968

  6. Reduction of acrylamide level through blanching with treatment by an extremely thermostable L-asparaginase during French fries processing.

    Science.gov (United States)

    Zuo, Shaohua; Zhang, Tao; Jiang, Bo; Mu, Wanmeng

    2015-07-01

    Bacterial L-asparaginase catalyzes the hydrolysis of L-asparagine to L-aspartic acid. It is normally used as an antineoplastic drug applied in lymphoblastic leukemia chemotherapy and as a food processing aid in baked or fried food industry to inhibit the formation of acrylamide. The present study demonstrates cloning, expression, and characterization of a thermostable L-asparaginase from Thermococcus zilligii AN1 TziAN1_1 and also evaluates the potential for enzymatic acrylamide mitigation in French fries using this enzyme. The recombinant L-asparaginase was purified to homogeneity by nickel-affinity chromatography. The purified enzyme displayed the maximum activity at pH 8.5 and 90 °C, and the optimum temperature was the highest ever reported. The K m, k cat, and k cat/K m values toward L-asparagine were measured to be 6.08 mM, 3267 s(-1), and 537.3 mM(-1) s(-1), respectively. The enzyme retained 70 % of its original activity after 2 h of incubation at 85 °C. When potato samples were treated with 10 U/mL of L-asparaginase at 80 °C for only 4 min, the acrylamide content in final French fries was reduced by 80.5 % compared with the untreated control. Results of this study revealed that the enzyme was highly active at elevated temperatures, reflecting the potential of the T. zilligii L-asparaginase in the food processing industry.

  7. The Effects of L-Asparaginase on Blood Triglycerides, Glucose, and Albumin Levels and Coagulation State in ALL Patients in Pediatric Ward

    Directory of Open Access Journals (Sweden)

    Heydarian Farhad

    2009-10-01

    Full Text Available Asparaginase is one of the most important agents in the treatment of ALL. However, it has some side effects including dislipidemia, hyperglycemia, coagulopathy and hepatotoxicity. We studied its side ef-fects on our patients. The effects of L-asparaginase were assessed on 25 new ALL patients (case group and 25 patients with known ALL who had completed their treatment before. Sixty two percent of cases were male and remainder was female. The mean age of patients was 7.2 ± 3.8 years. In our patients, there was a rise in triglycerides (TG was seen following L- asparaginase administration (P = 0.02. Also, PT prolongation (P = 0.02 and hypoalbominemia (P = 0.002 were detected which could PT prolongation and hypoalbuminemia may be seen with L-asparaginase therapy that can be prevented with transfusion of FFP. Hypertriglyceridemia is often asymptomatic with no need for therapy.

  8. Screening and characterization of extracelluar L-asparaginase producing Bacillus subtilis strain hswx88, isolated from Taptapani hotspring of Odisha, India

    Directory of Open Access Journals (Sweden)

    Biswaprakash Pradhan

    2013-12-01

    Conclusion: The isolate is eco-friendly and useful to produce bulk quantity of extracellular, thermophilic L-asparaginase for the treatment of various tumor cases and for preparation of acrylamide free fry food preparation.

  9. Purification, characterization, cytotoxicity and anticancer activities of L-asparaginase, anti-colon cancer protein, from the newly isolated alkaliphilic Streptomyces fradiae NEAE-82

    Science.gov (United States)

    El-Naggar, Noura El-Ahmady; Deraz, Sahar F.; Soliman, Hoda M.; El-Deeb, Nehal M.; El-Ewasy, Sara M.

    2016-01-01

    L-asparaginase is an important enzyme as therapeutic agents used in combination with other drugs in the treatment of acute lymphoblastic leukemia. A newly isolated actinomycetes strain, Streptomyces sp. NEAE-82, was potentially producing extracellular L-asparaginase, it was identified as Streptomyces fradiae NEAE-82, sequencing product was deposited in the GenBank database under accession number KJ467538. L-asparaginase was purified from the crude enzyme using ammonium sulfate precipitation, dialysis and ion exchange chromatography using DEAE Sepharose CL-6B. Further the kinetic studies of purified enzyme were carried out. The optimum pH, temperature and incubation time for maximum L-asparaginase activity were found to be 8.5, 40 °C and 30 min, respectively. The optimum substrate concentration was found to be 0.06 M. The Km and Vmax of the enzyme were 0.01007 M and 95.08 Uml−1min−1, respectively. The half-life time (T1/2) was 184.91 min at 50 °С, while being 179.53 min at 60 °С. The molecular weight of the subunits of L-asparaginase was found to be approximately 53 kDa by SDS–PAGE analysis. The purified L-asparaginase showed a final specific activity of 30.636 U/mg protein and was purified 3.338-fold. The present work for the first time reported more information in the production, purification and characterization of L-asparaginase produced by newly isolated actinomycetes Streptomyces fradiae NEAE-82. PMID:27605431

  10. L-Asparaginase Activity of Fungal Endophytes from Tabernaemontana heyneana Wall. (Apocynaceae), Endemic to the Western Ghats (India)

    OpenAIRE

    Manasa, Chandramouli; Nalini, Monnanda Somaiah

    2014-01-01

    “Endophytes,” the microbes residing within the plant tissues, are important sources of secondary metabolites. Tabernaemontana heyneana Wall., a medicinal tree, endemic to the Western Ghats with rich ethnobotanical history and unique chemical diversity, was selected to study fungal endophytes and evaluate them for L-asparaginase activity. Healthy plant parts were selected for the isolation of endophytes following standard isolation protocols. A total of 727 isolates belonging to 20 taxa were o...

  11. Effective treatment for suppression of acrylamide formation in fried potato chips using L-asparaginase from Bacillus subtilis

    OpenAIRE

    Onishi, Yohei; Prihanto, Asep A.; Yano, Shigekazu; Takagi, Kazuyoshi; Umekawa, Midori; Wakayama, Mamoru

    2015-01-01

    It has been reported that acrylamide, a potential carcinogen, is formed from the reaction of L-asparagine (L-Asn) and reducing sugars contained in foods during heating processes and free asparagine is a limiting factor for acrylamide formation. It has been reported that potato products such as potato chips, which are made through heating processes, contain high levels of acrylamide. To decrease the amount of L-Asn in potatoes using L-asparaginase, effective treatment conditions of sliced pota...

  12. L-Asparaginase Activity of Fungal Endophytes from Tabernaemontana heyneana Wall. (Apocynaceae), Endemic to the Western Ghats (India)

    Science.gov (United States)

    Manasa, Chandramouli; Nalini, Monnanda Somaiah

    2014-01-01

    “Endophytes,” the microbes residing within the plant tissues, are important sources of secondary metabolites. Tabernaemontana heyneana Wall., a medicinal tree, endemic to the Western Ghats with rich ethnobotanical history and unique chemical diversity, was selected to study fungal endophytes and evaluate them for L-asparaginase activity. Healthy plant parts were selected for the isolation of endophytes following standard isolation protocols. A total of 727 isolates belonging to 20 taxa were obtained. The isolates comprised of bark (11%), twig (22%), leaf (43%), fruit (12.0%), and seeds (12%). Endophytes such as Colletotrichum, Curvularia, Fusarium, Phomopsis, Verticillium, and Volutella colonized T. heyneana plant parts. Fusarium sp., Phomopsis spp., isolate Thlf01, and Fusarium solani were the dominant genera of bark, twig, leaf, fruits, and seed samples, respectively. The endophytes were screened for their ability to utilize L-asparagine by plate assay method. Fusarium spp. exhibited a high level of activity among the nine endophytes tested positive for L-asparaginase activity. Studies underline the potentials of endophyte-derived fungal L-asparaginases as sources of chemotherapeutic agents.

  13. Subgroups of Paediatric Acute Lymphoblastic Leukaemia Might Differ Significantly in Genetic Predisposition to Asparaginase Hypersensitivity.

    Directory of Open Access Journals (Sweden)

    Nóra Kutszegi

    Full Text Available L-asparaginase (ASP is a key element in the treatment of paediatric acute lymphoblastic leukaemia (ALL. However, hypersensitivity reactions (HSRs to ASP are major challenges in paediatric patients. Our aim was to investigate genetic variants that may influence the risk to Escherichia coli-derived ASP hypersensitivity. Sample and clinical data collection was carried out from 576 paediatric ALL patients who were treated according to protocols from the Berlin-Frankfurt-Münster Study Group. A total of 20 single nucleotide polymorphisms (SNPs in GRIA1 and GALNT10 genes were genotyped. Patients with GRIA1 rs4958351 AA/AG genotype showed significantly reduced risk to ASP hypersensitivity compared to patients with GG genotype in the T-cell ALL subgroup (OR = 0.05 (0.01-0.26; p = 4.70E-04, while no such association was found in pre-B-cell ALL. In the medium risk group two SNPs of GRIA1 (rs2055083 and rs707176 were associated significantly with the occurrence of ASP hypersensitivity (OR = 0.21 (0.09-0.53; p = 8.48E-04 and OR = 3.02 (1.36-6.73; p = 6.76E-03, respectively. Evaluating the genders separately, however, the association of rs707176 with ASP HSRs was confined only to females. Our results suggest that genetic variants of GRIA1 might influence the risk to ASP hypersensitivity, but subgroups of patients can differ significantly in this respect.

  14. Expanding targets for a metabolic therapy of cancer: L-asparaginase.

    Science.gov (United States)

    Covini, Daniele; Tardito, Saverio; Bussolati, Ovidio; Chiarelli, Laurent R; Pasquetto, Maria V; Digilio, Rita; Valentini, Giovanna; Scotti, Claudia

    2012-01-01

    The antitumour enzyme L-asparaginase (L-asparagine amidohydrolase, EC 3.5.1.1, ASNase), which catalyses the deamidation of L-asparagine (Asn) to L-aspartic acid and ammonia, has been used for many years in the treatment of acute lymphoblastic leukaemia. Also NK tumours, subtypes of myeloid leukaemias and T-cell lymphomas respond to ASNase, and ovarian carcinomas and other solid tumours have been proposed as additional targets for ASNase, with a potential role for its glutaminase activity. The increasing attention devoted to the antitumour activity of ASNase prompted us to analyse recent patents specifically concerning this enzyme. Here, we first give an overview of metabolic pathways affected by Asn and Gln depletion and, hence, potential targets of ASNase. We then discuss recent published patents concerning ASNases. In particular, we pay attention to novel ASNases, such as the recently characterised ASNase produced by Helicobacter pylori, and those presenting amino acid substitutions aimed at improving enzymatic activity of the classical Escherichia coli enzyme. We detail modifications, such as natural glycosylation or synthetic conjugation with other molecules, for therapeutic purposes. Finally, we analyse patents concerning biotechnological protocols and strategies applied to production of ASNase as well as to its administration and delivery in organisms. PMID:21854356

  15. Screening of Streptomycetes for L-Asparaginase, Therapeutic Agent of Lymphocytic Leukemia from Western Ghats of Karnataka, India

    Directory of Open Access Journals (Sweden)

    Mamatha B Salimath

    2016-03-01

    Full Text Available Actinomycetes are the most economically and biotechnologically valuable prokaryotes and are responsible for the production of about half of the discovered bioactive secondary metabolites, antibiotics, anticancer agents and enzymes. The present study was successful in characterizing 25 Streptomycetes isolates inhabiting Agumbe province, of Western Ghats soil of Karnataka, India, for potential source of L-asparaginase enzyme. L- Asparaginase (L-asparagine amido hydrolase E.C.3.5.1.1 is an extracellular enzyme has anti-carcinogenic potential, received increasing awareness in the current years because of its use as an effective agent against Acute Lymphocytic Leukemia (ALL. L-asparagine is a source of essential amino acid necessary for the growth of leukemic cells in higher amounts. Depletion of L-asparagine from the circulatory blood leads to death of malignant cells. Streptomyces isolates have been collected from soil samples by serial dilution and plating method employing Starch Casein Nitrate agar and ISP media. They were identified based on cultural, morphological and biochemical characteristics. When primarily screened for L-asparaginase production by Rapid plate assay using Modified M9 medium containing L-asparagine and Phenol red as indicator, 23 isolates were found to be positive by showing change in color of medium from yellow to pink. Strain V17 isolate proved to be potent producer of the enzyme with higher amount of enzyme up to 22.45 IU/ml. About 92% of the tested isolates were positive for anti-cancerous potential, indicating the Western Ghats soils as potential sources for anti-cancerous agents. Further studies on purification and characterization of enzyme are under progress.

  16. L-Asparaginase as potent anti-leukemic agent and its significance of having reduced glutaminase side activity for better treatment of acute lymphoblastic leukaemia.

    Science.gov (United States)

    Ramya, L N; Doble, Mukesh; Rekha, V P B; Pulicherla, K K

    2012-08-01

    Acute lymphoblastic leukaemia (ALL) is one of the leading types of malignant disorder seen in children. Viral infections, genetic factors and exposure to chemical carcinogens are some of the factors responsible for causing ALL. Treatment strategies followed for curing ALL include chemotherapy or radiation therapy, wherein, chemotherapy involves the use of the enzymatic drug L-Asparaginase. The enzyme can be produced from various plants, animals, bacterial and fungal sources but, among them, bacterial sources are widely used for production of this enzyme. The enzyme is non-human in origin having certain bottle necks with L-Asparaginase therapy in the form of side effects such as pancreatitis, thrombosis which are mainly due to glutaminase side activity. Hence, present-day research is mainly focussed on minimizing or completely eliminating the glutaminase activity of the enzyme L-Asparaginase. This review is focussed on the complications associated with glutaminase side activity and use of glutaminase free enzymatic drug L-Asparaginase in treating ALL and the other developments related to the modification of the drug for quality treatment. PMID:22684410

  17. The K+ dependent asparaginase, NSE1, is crucial for plant growth and seed production in Lotus japonicus

    DEFF Research Database (Denmark)

    Credali, Alfredo; Garcia-Calderón, Margarita; Dam, Svend Secher;

    2013-01-01

    The physiological role of K+-dependent and K+-independent asparaginases in plants remains unclear, and the contribution from individual isoforms during development is poorly understood. We have used reverse genetics to assess the phenotypes produced by the deficiency of K+-dependent NSE1 asparagi...

  18. Screening and characterization of extracelluar L-asparaginase producing Bacillus subtilis strain hswx88, isolated from Taptapani hotspring of Odisha, India

    Institute of Scientific and Technical Information of China (English)

    Biswaprakash Pradhan; Sashi K Dash; Sabuj Sahoo

    2013-01-01

    Objective: To screen and isolate an eco-friendly, a thermophilic and potent L-asparaginase producing bacterium, with novel immunological properties that may obviates hypersensitivity reactions. Methods: In the present study bacterial strain isolated for extracellular L-asparaginase production from hotspring, identified by morphological, biochemical and physiological tests followed by 16S rDNA technology and the L-asparaginase production ability was tested by both semi quantitative and quantitative enzymatic assay. Result:The bacterial strain was identified as Bacillus subtilis strain hswx88 (GenBank Accession Number: JQ237656.1). The extracellular enzyme yielding capacity isolate Bacillus subtilis strain hswx88 (23.8 IU/mL) was found to be 1.7 and 14.5 times higher than the reference organism Pectobacterium carotovorum MTCC 1428 (14.2 IU/mL) and Bacillus sp. BCCS 034 (1.64 IU/mL). Conclusion: The isolate is eco-friendly and useful to produce bulk quantity of extracellular, thermophilic L-asparaginase for the treatment of various tumor cases and for preparation of acrylamide free fry food preparation.

  19. Proteins from Erwinia asparaginase Erwinase ® and E. coli asparaginase 2 MEDAC ® for treatment of human leukemia, show a multitude of modifications for which the consequences are completely unclear.

    Science.gov (United States)

    Bae, Narkhyun; Pollak, Arnold; Lubec, Gert

    2011-07-01

    L-Asparaginase from Erwinia chrysanthemi (ASPG_ERWCH; UniProtKB accession number P06608 (Erwinase(®))) and L-asparaginase 2 from Escherichia coli (ASPG2_ECOLI; UniProtKB accession number P00805 (Medac(®))), both L-asparagine amidohydrolases, are widely used for the treatment of acute lymphoblastic leukemia. A series of serious side effects have been reported and this warrants studies into the protein chemistry of the medical products sold. Mass spectrometry (MS) data on ASPG_ERWCH and ASPG2_ECOLI have not been published so far and herein a gel-based proteomics study was performed to provide information about sequence and modifications of the commercially available medical products. ASPG_ERWCH and ASPG2_ECOLI were applied onto two-dimensional gel electrophoresis, spots were in-gel digested with several proteases and resulting peptides and protein modifications were analysed by nano-ESI-LC-MS/MS. Four spots were observed for ASPG_ERWCH, six spots were observed for ASPG2_ECOLI and the identified proteins showed high sequence coverage without sequence conflicts. Several protein modifications including technical and posttranslational modifications were demonstrated. Protein modifications are known to change physicochemical, immunochemical, biological and pharmacological properties and results from this work may challenge re-designing of the product including possible removal of the modifications by the manufacturer because it is not known whether they are contributing to the serious adverse effects of the protein drug.

  20. Purification and Characterization of Glutaminase Free Asparaginase from Enterobacter cloacae: In-Vitro Evaluation of Cytotoxic Potential against Human Myeloid Leukemia HL-60 Cells.

    Directory of Open Access Journals (Sweden)

    Islam Husain

    Full Text Available Asparaginase is an important antileukemic agent extensively used worldwide but the intrinsic glutaminase activity of this enzymatic drug is responsible for serious life threatening side effects. Hence, glutaminase free asparaginase is much needed for upgradation of therapeutic index of asparaginase therapy. In the present study, glutaminase free asparaginase produced from Enterobacter cloacae was purified to apparent homogeneity. The purified enzyme was found to be homodimer of approximately 106 kDa with monomeric size of approximately 52 kDa and pI 4.5. Purified enzyme showed optimum activity between pH 7-8 and temperature 35-40°C, which is close to the internal environment of human body. Monovalent cations such as Na+ and K+ enhanced asparaginase activity whereas divalent and trivalent cations, Ca2+, Mg2+, Zn2+, Mn2+, and Fe3+ inhibited the enzyme activity. Kinetic parameters Km, Vmax and Kcat of purified enzyme were found to be 1.58×10-3 M, 2.22 IU μg-1 and 5.3 × 104 S-1, respectively. Purified enzyme showed prolonged in vitro serum (T1/2 = ~ 39 h and trypsin (T1/2 = ~ 32 min half life, which is therapeutically remarkable feature. The cytotoxic activity of enzyme was examined against a panel of human cancer cell lines, HL-60, MOLT-4, MDA-MB-231 and T47D, and highest cytotoxicity observed against HL-60 cells (IC50 ~ 3.1 IU ml-1, which was comparable to commercial asparaginase. Cell and nuclear morphological studies of HL-60 cells showed that on treatment with purified asparaginase symptoms of apoptosis were increased in dose dependent manner. Cell cycle progression analysis indicates that enzyme induces apoptosis by cell cycle arrest in G0/G1 phase. Mitochondrial membrane potential loss showed that enzyme also triggers the mitochondrial pathway of apoptosis. Furthermore, the enzyme was found to be nontoxic for human noncancerous cells FR-2 and nonhemolytic for human erythrocytes.

  1. A clinically attainable dose of L-asparaginase targets glutamine addiction in lymphoid cell lines.

    Science.gov (United States)

    Sugimoto, Koichi; Suzuki, Hiroshi I; Fujimura, Tsutomu; Ono, Asami; Kaga, Naoko; Isobe, Yasushi; Sasaki, Makoto; Taka, Hikari; Miyazono, Kohei; Komatsu, Norio

    2015-11-01

    L-asparaginase (L-ASNase) is an important branch of chemotherapy for acute lymphoblastic leukemia (ALL) and some types of non-Hodgkin's lymphoma, including natural killer (NK)-cell lymphoma. Although it mediates hydrolysis of asparagine (Asn) and glutamine (Gln), which are variably required for cancer cell survival, the relative contribution of Asn and Gln depletion to the anti-tumor activity in therapeutic doses is unclear in ALL and malignant lymphoma. Here we demonstrate that L-ASNase exerts cytotoxicity through targeting the Gln addiction phenotype in lymphoid cell lines. A clinically attainable intermediate dose of L-ASNase induced massive apoptosis in ALL Jurkat and mantle cell lymphoma Jeko cell lines, while a low dose of L-ASNase effectively killed NK-cell lymphoma cells. In the lymphoid cell lines Jurkat and Jeco, deprivation of Gln but not Asn specifically suppressed cell growth and survival, and phenocopied the action of L-ASNase. L-ASNase treatment and Gln deprivation dramatically disrupted the refilling of the tricarboxylic acid (TCA) cycle by intracellular glutamate (Glu) and disturbed the mitochondrial integrity, which were alleviated by various anaplerotic TCA cycle intermediates, suggesting a direct contribution of glutaminase activity of L-ASNase. The action of L-ASNase differs between Jurkat cells and NK-cell lymphoma cells, according to their dependence on Gln and Asn. Furthermore, we observed that high expression of glutaminase GLS1 is associated with increased sensivity to L-ASNase in pediatric B lineage ALL. Our results redefine L-ASNase as a therapeutic agent targeting Gln addiction in certain lymphoid cells and offer an additional basis for predicting L-ASNase sensitivity and engineering selective L-ASNase derivatives for leukemia and lymphoma.

  2. PRODUCTION PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR ANTI-LEUKAEMIC L-ASPARAGINASE FROM ISOLATED BACILLUS SUBTILIS USING SOLID STATE FERMENTATION.

    Directory of Open Access Journals (Sweden)

    Susmita Shukla

    2013-08-01

    Full Text Available Bacterial L-asparaginase has been widely used as therapeutic agent in treatment of various lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. The present work deals with production and purification of extracellular L-asparaginase from soil isolates using solid state fermentation. The isolate was characterized by big chemical test and identified as Bacillus subtilis. The enzyme production was carried out by solid state fermentation comparing the results with submerged fermentation. The enzyme was purified to near homogeneity by ammonium sulphate precipitation, dialysis, followed by gel filtration on Sephadex G-100 column, CM Sephadex C-50 and SDS-PAGE. The enzyme was purified at 110.2 folds and showed a final specific activity of 1785.7 IU/mg with 26.5% yield. SDS-PAGE of the purified enzyme revealed an apparent molecular weight of 109 kDa. The purified enzyme showed maximum activity at pH 9 when it was incubated at 50°C for 35 min. The enzyme was activated by Mg+2 and strongly inhibited by EDTA.

  3. Minimally-Myelosuppressive Asparaginase-Containing Induction Regimen for Treatment of a Jehovah’s Witness with mutant IDH1/NPM1/NRAS Acute Myeloid Leukemia

    Directory of Open Access Journals (Sweden)

    Ashkan Emadi

    2016-03-01

    Full Text Available Treatment of patients with acute myeloid leukemia (AML who do not wish to accept blood product transfusion, including Jehovah’s Witnesses, is extremely challenging. The use of conventional chemotherapy for induction of complete remission (CR results in profound anemia and thrombocytopenia requiring frequent transfusions of blood products, without which such treatment will be life-threatening. Finding a well tolerable, minimally myelosuppressive induction regimen for such patients with AML is a clear example of area of unmet medical need. Here, we report a successful treatment of a 52-year-old Jehovah’s Witness with newly diagnosed AML with peg-asparaginase, vincristine and methylprednisolone. The AML was characterized with normal karyotype, and mutations in isocitrate dehydrogenase 1 (IDH1-Arg132Ser, nucleophosmin 1 (NPM1-Trp289Cysfs*12 and neuroblastoma RAS viral oncogene homolog (NRAS-G1y12Va1. After one 28-day cycle of treatment, the patient achieved complete remission with incomplete count recovery (CRi and after the second cycle, he achieved CR with full blood count recovery. The patient has never received any blood products. Notwithstanding that myeloperoxidase-induced oxidative degradation of vincristine results in its lack of activity as monotherapy in AML, its combination with corticosteroid and asparaginase has resulted in a robust remission in this patient. Diminished steroid clearance by asparaginase activity as well as reduction in serum glutamine level induced by glutaminase enzymatic activity of asparaginase may have contributed to effective killing of the myeloblasts that carry IDH1/NPM1/NRAS mutations. In conclusion, asparaginase-containing regimens, which are approved for treatment of acute lymphoblastic leukemia (ALL but not AML, can be used to treat patients with AML who do not accept blood transfusion.

  4. Allergic complications of L-asparaginase therapy in children with acute lymphoblastic leukaemia

    Directory of Open Access Journals (Sweden)

    Konstantinidis Georgios

    2011-01-01

    Full Text Available Introduction. L-asparaginase (L-ASP is one of the most effective medications for the treatment of acute lymphoblastic leukaemia (ALL in children, and allergic reactions to the therapy are considered the most significant side effects. Objective. The aim of this study was to determine the prevalence and type of allergic reactions, as well as to identify potential risk factors for the development of allergic reactions during L-ASP therapy in children with ALL. Methods. The study encompassed 70 patients under 18 years of age, who were treated at the Institute for Child and Youth Healthcare of Vojvodina, Novi Sad in the period January 2000 - June 2009. We analyzed the frequency and type of allergic reactions during the administration of L-ASP, the onset of allergic reaction in relation to the phase of therapy of underlying disease, as well as the prevalence of allergic reactions in relation to drug administration method. Results. Allergic reaction manifested in 17 patients (24%. In 14 patients (82% allergic reaction to L-ASP manifested as urticaria, bronchospasm or anaphylaxis, whereas a mild local reaction was observed in only three patients (18%. In a group treated, according to the high-risk protocol, the prevalence of allergic reactions was statistically significantly higher in the intermediate-risk group of patients (p<0.01, i.e. statistically significantly more frequent, as compared to the standard-risk group of patients (p<0.05. The majority of patients (11; 65% developed allergic reactions to the 9th dose of L-ASP, i.e. the first dose during the reinduction phase. The time interval between the last L-ASP dose in the induction phase and the 1st dose in the reinduction phase was at least four weeks. With respect to administration method, the majority of patients (16; 94% developed allergic reaction after intravenous application of L-ASP. Conclusion. Potential risk factors for the development of allergic reaction to L-ASP are a high-risk therapy

  5. Minimal contribution of severe hypertriglyceridemia in L-asparaginase-associated pancreatitis developed in a child with acute lymphocytic leukemia.

    Science.gov (United States)

    Goto, Yoshinori; Nishimura, Ryosei; Nohara, Atsushi; Mase, Shintaro; Fujiki, Toshihiro; Irabu, Hitoshi; Kuroda, Rie; Araki, Raita; Ikawa, Yasuhiro; Maeba, Hideaki; Yachie, Akihiro

    2016-08-01

    A 10-year-old girl developed L-asparaginase (ASP)-associated pancreatitis during chemotherapy for acute lymphocytic leukemia. Her symptoms showed alleviation with continuous regional arterial infusion of protease inhibitor and systemic somatostatin analog therapy. She had intermittent and marked hypertriglyceridemia, an initial trigger for pancreatitis, probably as a side effect of ASP and steroids. However, we considered the pancreatitis to have developed mainly because of factors other than hypertriglyceridemia as lipoprotein analysis confirmed chylomicron levels to be nearly undetectable. Extremely large chylomicrons contribute directly to the onset of pancreatitis by causing blockage of small vessels. Although it is necessary to examine patients for dyslipidemia developing as a side effect of ASP, therapeutic intervention for hypertriglyceridemia is not considered to prevent the onset of ASP-associated pancreatitis. PMID:27599414

  6. Identification and Structural Analysis of an l-Asparaginase Enzyme from Guinea Pig with Putative Tumor Cell Killing Properties*

    Science.gov (United States)

    Schalk, Amanda M.; Nguyen, Hien-Anh; Rigouin, Coraline; Lavie, Arnon

    2014-01-01

    The initial observation that guinea pig serum kills lymphoma cells marks the serendipitous discovery of a new class of anti-cancer agents. The serum cell killing factor was shown to be an enzyme with l-asparaginase (ASNase) activity. As a direct result of this observation, several bacterial l-asparaginases were developed and are currently approved by the Food and Drug Administration for the treatment of the subset of hematological malignancies that are dependent on the extracellular pool of the amino acid asparagine. As drugs, these enzymes act to hydrolyze asparagine to aspartate, thereby starving the cancer cells of this amino acid. Prior to the work presented here, the precise identity of this guinea pig enzyme has not been reported in the peer-reviewed literature. We discovered that the guinea pig enzyme annotated as H0W0T5_CAVPO, which we refer to as gpASNase1, has the required low Km property consistent with that possessed by the cell-killing guinea pig serum enzyme. Elucidation of the ligand-free and aspartate complex gpASNase1 crystal structures allows a direct comparison with the bacterial enzymes and serves to explain the lack of l-glutaminase activity in the guinea pig enzyme. The structures were also used to generate a homology model for the human homolog hASNase1 and to help explain its vastly different kinetic properties compared with gpASNase1, despite a 70% sequence identity. Given that the bacterial enzymes frequently present immunogenic and other toxic side effects, this work suggests that gpASNase1 could be a promising alternative to these bacterial enzymes. PMID:25320094

  7. Effects of Prednisolone, L-Asparaginase, Gemfibrozil, and Combinations of These Elements on Mice Lipid Profile, Liver, and Pancreas.

    Science.gov (United States)

    Kose, Dogan; Tarakci, Nuriye; Celik, Zeliha Esin; Vatansev, Husamettin; Cimbek, Emine Ayca; Ugras, Serdar; Sen, Yasar; Caliskan, Umran; Koksal, Yavuz

    2016-01-01

    The aim of this study is to determine the effects of L-asparaginase (L-ASP), corticosteroids (CSs), and antilipidemics, separately and in combination, on the lipid profiles and the liver and pancreas histology in mice. This study included 8 groups of 7 mice each. Before any drug administration, serum samples were taken from all of the mice. Then, normal saline was applied to the control group, and a medication or combination of medications was applied to the other groups. Levels of triglycerides, cholesterol (COL), and high-density lipoprotein (HDL) and low-density lipoprotein (LDL) were determined, and the livers and pancreases were evaluated histologically at the end of the study. Triglycerides increased significantly in the CS-only and the L-ASP-only groups, COL increased significantly in the CS-only group, and HDL increased significantly in the CS-only and the antilipidemic-only groups. LDL was significantly lower in the CS-only and the L-ASP-only groups. CSs and L-ASP were significantly effective in liver necrosis, L-ASP was significantly effective in liver balloon degeneration, and CS were significantly effective in pancreas vacuolization. Triglyceride measurement is recommended before/during CS and/or L-ASP treatment. Starting with an antilipidemic agent can be considered to avoid possible complications in patients with significantly high rates. Indicators of a possible liver or pancreas injury should also be considered. PMID:26599986

  8. 门冬酰胺酶致急淋白血病患儿两次脑血栓形成1例%Asparaginase Induced Cerebral Thrombosis For Twice In One Childhood Acute Lymphoblastic Leukemia Case

    Institute of Scientific and Technical Information of China (English)

    王成军; 汪俭; 李艳; 许喆; 陈天平

    2015-01-01

    Asparaginase depletion can specific affect the synthesis of asparagine protein in tumor cell, it is one of the core drugs for treating childhood acute lymphoblastic leukemia, it can improve the cure rate. Effect of asparaginase on coagulation is great influence, and a two-way risk of both thrombosis and bleeding exist. We report that asparaginase induced cerebral thrombosis for twice in one childhood ALL patient and our clinical treatment course, which should provide reference for clinical treatment in these patients treated with asparaginase for future.%门冬酰胺酶能特异性消耗门冬酰胺影响肿瘤细胞蛋白质的合成,是儿童急性淋巴细胞白血病治疗的核心药物之一,对提高儿童急淋治愈率的贡献很大.门冬酰胺酶对机体凝血功能的影响也很大,同时有血栓形成及出血的双向风险.该文报道了1例门冬酰胺酶致急性淋巴细胞白血病患儿两次脑血栓形成及临床干预经过,为以后此类患儿的临床治疗提供参考.

  9. Preparation and nanoencapsulation of l-asparaginase II in chitosan-tripolyphosphate nanoparticles and in vitro release study

    Science.gov (United States)

    Bahreini, Elham; Aghaiypour, Khosrow; Abbasalipourkabir, Roghayeh; Mokarram, Ali Rezaei; Goodarzi, Mohammad Taghi; Saidijam, Massoud

    2014-07-01

    This paper describes the production, purification, and immobilization of l-asparaginase II (ASNase II) in chitosan nanoparticles (CSNPs). ASNase II is an effective antineoplastic agent, used in the acute lymphoblastic leukemia chemotherapy. Cloned ASNase II gene ( ansB) in pAED4 plasmid was transformed into Escherichia coli BL21pLysS (DE3) competent cells and expressed under optimal conditions. The lyophilized enzyme was loaded into CSNPs by ionotropic gelation method. In order to get optimal entrapment efficiency, CSNP preparation, chitosan/tripolyphosphate (CS/TPP) ratio, and protein loading were investigated. ASNase II loading into CSNPs was confirmed by Fourier transform infrared (FTIR) spectroscopy, and morphological observation was carried out by transmission electron microscopy. Three absolute CS/TPP ratios were studied. Entrapment efficiency and loading capacity increased with increasing CS and TPP concentration. The best ratio was applied for obtaining optimal ASNase II-loaded CSNPs with the highest entrapment efficiency. Size, zeta potential, entrapment efficiency, and loading capacity of the optimal ASNase II-CSNPs were 340 ± 12 nm, 21.2 ± 3 mV, 76.2% and 47.6%, respectively. The immobilized enzyme showed an increased in vitro half-life in comparison with the free enzyme. The pH and thermostability of the immobilized enzyme was comparable with the free enzyme. This study leads to a better understanding of how to prepare CSNPs, how to achieve high encapsulation efficiency for a high molecular weight protein, and how to prolong the release of protein from CSNPs. A conceptual understanding of biological responses to ASNase II-loaded CSNPs is needed for the development of novel methods of drug delivery.

  10. 门冬酰胺酶相关儿童急性胰腺炎诊治研究进展%Progress of diagnosis and treatment of asparaginase associated pancreatitis in children

    Institute of Scientific and Technical Information of China (English)

    石苇

    2016-01-01

    门冬酰胺酶(ASP)是儿童急性淋巴细胞白血病和非霍奇金淋巴瘤联合化疗方案中的关键药物之一.ASP相关急性胰腺炎(AAP)是ASP的主要严重不良反应.现通过复习有关儿童AAP的近年国内外文献和我国《培门冬酶治疗急性淋巴细胞白血病和恶性淋巴瘤的专家共识》,以及对网络收集的历年来我国儿童AAP报道资料进行归纳与统计分析,参照国际AAP诊断标准,提出儿童AAP流行病学、临床表现、早期诊断和有效治疗要点,以及左旋门冬酰胺酶和培门冬酶的AAP临床对比.全文资料数据详尽,分析归纳依据充分,相关经验的临床可操作性强,对临床有效诊治儿童AAP具有较大的参考价值.%Asparaginase(ASP) is an important drug in the treatment of childhood acute lymphoblastic leukemia and non-Hodgkin lymphoma.Asparaginase associated pancreatitis (AAP) is the main treatment-adverse events of asparaginase.After reviewing the recent foreign literatures about AAP and the Chinese expert about polyethylene glycol conjugated asparaginase (PEG-ASP) in the treatment of acute lymphoblastic leukemia and malignant lymphoma with asparaginase,conclude and analysis the data about childhood AAP and show the epidemiology,clinical features,early diagnosis and effective treatment of children with AAP.Make clinical compare of L-asparaginase and PEG-ASP.Based on the full grasp of the relevant data,analyzing,introducing and integrating,this may be helpful to the diagnosis and treatment of childhood AAP.

  11. Clinical diagnosis and treatment of asparaginase associated pancreatitis in adults%成人门冬酶相关胰腺炎的临床诊治

    Institute of Scientific and Technical Information of China (English)

    李梦洁; 何牧卿; 沈益民

    2015-01-01

    目的 探讨成人门冬酶相关胰腺炎(AAP)的临床特点及诊治过程,旨在提高AAP诊治水平.方法 回顾性分析2009年1月至2015年6月间浙江医院血液内科及温州医科大学附属第二医院血液科收治的384例急性淋巴细胞白血病(ALL)患者的临床资料,所有患者均给予含培门冬酶或左旋门冬酰胺酶(L-asp)的多药联合化疗,分析AAP的发生情况、临床表现、诊断、治疗及转归.结果 384例ALL患者中18例发生AAP,发生率为4.7%,其中轻症AAP (MAAP) 13例,重症AAP(SAAP)5例.16例AAP发生在诱导化疗期,2例发生在维持强化期.腹痛为主要临床表现,血淀粉酶及脂肪酶活性均升高.治疗后,MAAP患者的腹痛症状缓解,血淀粉酶及脂肪酶活性明显下降,继续使用培门冬酶或L-asp化疗,未见胰腺炎复发.5例SAAP患者血淀粉酶及脂肪酶反复升高,其中1例因合并重症感染及囊肿破裂出血而病死.结论 成人ALL患者在门冬酶化疗期间出现腹痛症状应考虑AAP可能,加强血淀粉酶及脂肪酶检测,辅以B超和CT检查有助于早期诊治AAP,并改善患者预后.%Objective To investigate the clinical characteristics and the course of diagnosis and therapy of asparaginase associated pancreatitis (AAP) in adults, in order to improve the ability of diagnosis and treatment.Methods Data of 384 cases of acute lymphoblastic leukemia (ALL) who received treatment in Department of Hematology, Zhejiang Hospital, and Department of Hematology, Second Hospital Affiliated to Wenzhou Medical University from January 2009 to June 2015 was retrospectively analyzed.All patients were given multi-drug chemotherapy including PEG-asparaginase or L-asparaginase, the incidence of AAP, clinical manifestations, diagnosis, treatment and prognosis were analyzed.Results Among the 384 cases, 18 patients developed AAP, and the incidence of AAP was 4.7%, including 13 cases of mild AAP (MAAP), 5 cases of severe AAP (SAAP).Sixteen cases of

  12. Clinical Utility of Ammonia Concentration as a Diagnostic Test in Monitoring of the Treatment with L-Asparaginase in Children with Acute Lymphoblastic Leukemia

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    Małgorzata Czogała

    2014-01-01

    Full Text Available L-asparaginase (ASP is an enzyme used as one of the basic regimens in the acute lymphoblastic leukemia (ALL therapy. Because of the possibility of the enzyme inactivation by antibodies, monitoring of ASP activity is essential. The aim of the study was to examine if plasma concentration of ammonia, a direct product of the reaction catalyzed by ASP, can be used in the assessment of ASP activity. A group of 87 patients with acute lymphoblastic leukemia treated in the Department of Pediatric Oncology and Hematology in Krakow was enrolled to the study. ASP activity and ammonia concentration were measured after ASP administrations during induction. A positive correlation was found between the ammonia concentration and ASP activity (R=0.44; P<0.0001 and between the medium values of ammonia concentration and ASP activity (R=0.56; P<0.0001. The analysis of ROC curves revealed the moderate accuracy of the ammonia concentration values in the ASP activity assessment. It was also found that the medium value of ammonia concentrations can be useful in identification of the patients with low (<100 IU/L and undetectable (<30 IU/L ASP activity. The plasma ammonia concentration may reflect ASP activity and can be useful when a direct measurement of the activity is unavailable.

  13. L-asparaginase-induced abnormality in plasma glucose level in patients of acute lymphoblastic leukemia admitted to a tertiary care hospital of Odisha

    Directory of Open Access Journals (Sweden)

    Mousumee Panigrahi

    2016-01-01

    Full Text Available Objectives: The objective of this study was to evaluate any abnormal change in plasma glucose levels in patients treated with L-asparaginase (L-Asp-based chemotherapy regimen in patients of acute lymphoblastic leukemia (ALL. Materials and Methods: This retrospective, hospital-based study was conducted in patients of ALL, admitted to the Clinical Haematology Department of a tertiary care hospital of Odisha from August 2014 to July 2015. Indoor records of 146 patients on multi-centered protocol-841 were evaluated for any alteration in plasma glucose level, time of onset of hypo/hyperglycemia, and persistence of plasma glucose alteration. Results: Twenty-one percent of patients showed abnormal plasma glucose level. Most of these patients developed hypoglycemia and were of lower age group. Most of these patients developed hypoglycemia and were of lower age group, whereas a majority of higher age group patients developed hyperglycemia. In majority of the cases, abnormal glucose developed after three doses of L-Asp. Hypoglycemia subsided whereas hyperglycemia persisted till the end of our observation period. Conclusions: L-Asp produces more incidences of hypoglycemia than hyperglycemia in a good number of ALL patients towards which clinicians should be more vigilant. However, hyperglycemia persists for a longer duration than hypoglycemia.

  14. Microchip CE-LIF method for the hydrolysis of L-glutamine by using L-asparaginase enzyme reactor based on gold nanoparticle.

    Science.gov (United States)

    Qiao, Juan; Qi, Li; Yan, Huijuan; Li, Yaping; Mu, Xiaoyu

    2013-02-01

    L-Asparaginase (L-Asnase) can suppress the growth of malignant cells by rapid depletion of two essential amino acids, L-glutamine (L-Gln) and L-asparagine (L-Asn). To study the cytotoxic effect and the secondary complications of L-Asnase in the treatment of acute lymphoblastic leukemia, the development of a novel enzyme reactor of L-Asnase for the hydrolysis of L-Gln, employing the enzyme-gold nanoparticle conjugates in capillary, was reported in this work. First, a microchip CE (MCE)-LIF was established for the separation of L-amino acids (L-Gln and L-glutamic acid) and studying the hydrolysis of L-Gln by using L-Asnase enzyme reactor. Then, using L-Gln as target analyte, the enzyme kinetics of L-Asnase in free solution, enzyme-gold nanoparticle conjugates (E-GNP), and the enzyme-gold nanoparticle conjugates immobilized in capillary (E-GNP-C) were investigated in detail with the proposed MCE-LIF method. Moreover, for optimizing the enzymatic reaction efficiency, three important parameters, including the length of capillary, the enzyme concentration reacted with gold nanoparticle and the amount of L-Asnase immobilized on the gold nanoparticle, have been studied. Owing to the high specific activity, the E-GNP-C enzyme reactor exhibited the best performance for the hydrolysis of L-Gln.

  15. Side effects of L-asparaginase with different sources and administrations%不同菌种来源左旋门冬酰胺酶及不同用法不良反应的观察

    Institute of Scientific and Technical Information of China (English)

    王华; 耿其荣; 吕跃

    2011-01-01

    Objective To investigate the side effects of L - asparagninase ( L - ASP ) from 2 different sources, Erwinia and Escherichia coli, with consecutive or every - other -day usage. Methods Liver function, blood clotting, ( PT, APTT, TT ), fibrinogen concentration, albumin, blood glucose and plasma infusion were recorded for analysis. Results Significantly more fresh plasma infusion was required in use of Escherichia coli - asparaginase than that of Erwiniaasparaginase. There was no significant difference in side effects, between consecutive day and every other day administrations. Conclusion Erwinia -asparaginase and Escherichia coli- asparaginase have the same safety in consecutive or every other day use in treating acute lymphoid leukemia and lymphoblastic lymphoma.%目的 比较欧文菌株与埃希大肠杆菌株来源、连用与隔日使用左旋门冬酰胺酶(L-asparaginase,L-ASP)的不良反应发生情况.方法 采用病例分析方法,比较欧文菌株与埃希大肠杆菌株来源L-ASP以及L-ASP连用与隔日使用肝功能、凝血三项(PT、APTT、TT)、纤维蛋白原以及血浆白蛋白、血糖的变化,所需输注的血浆量.结果 欧文菌株与埃希大肠杆菌株来源L-ASP相比,后者所需输注新鲜血浆量高于前者,不良反应发生率差异无统计学,连用与隔日使用差异无统计学意义.结论 欧文菌株与埃希大肠杆菌株来源L-ASP具有相似的安全性.连用与隔日使用L-ASP治疗急性淋巴细胞白血病和淋巴母细胞淋巴瘤同样安全.

  16. Biochemical characterization and immobilization of Erwinia carotovoral-asparaginase in a microplate for high-throughput biosensing of l-asparagine.

    Science.gov (United States)

    Labrou, Nikolaos E; Muharram, Magdy Mohamed

    2016-10-01

    l-Asparaginases (l-ASNase, E.C. 3.5.1.1) catalyze the conversion of l-asparagine to l-aspartic acid and ammonia. In the present work, a new form of l-ASNase from a strain of Erwinia carotovora (EcaL-ASNase) was cloned, expressed in Escherichia coli as a soluble protein and characterized. The enzyme was purified to homogeneity by a single-step procedure comprising ion-exchange chromatography. The properties of the recombinant enzyme were investigated employing kinetic analysis and molecular modelling and the kinetic parameters (Km, kcat) were determined for a number of substrates. The enzyme was used to assemble a microplate-based biosensor that was used for the development of a simple assay for the determination of l-asparagine in biological samples. In this sensor, the enzyme was immobilized by crosslinking with glutaraldehyde and deposited into the well of a microplate in 96-well format. The sensing scheme was based on the colorimetric measurement of ammonia formation using the Nessler's reagent. This format is ideal for micro-volume applications and allows the use of the proposed biosensor in high-throughput applications for monitoring l-asparagine levels in serum and foods samples. Calibration curve was obtained for l-asparagine, with useful concentration range 10-200μΜ. The biosensor had a detection limit of 10μM for l-asparagine. The method's reproducibility was in the order of ±3-6% and l-asparagine mean recoveries were 101.5%.

  17. Allergic shock induced by L-asparaginase enzyme injection%培门冬酶注射液致过敏性休克

    Institute of Scientific and Technical Information of China (English)

    李静; 曹翠明; 魏芳芳

    2012-01-01

    One 63-year-old male patient was diagnosed as non Hodgkin lymphoma, ALK negative anaplastic large cell type I B for 38 days. The patient has no drug and food allergy history. During the treatment, the chest, back, buttocks and upper limbs wheal were itchy when L-asparaginase enzyme injection 3750 IU was injected on three parts of uniform muscles in the third time after 90 minutes. After 60 minutes, when the patient went to toilet, he was suffered from severe headache, chest pain, sore throat, tongue numbness, blurred vision, decreased blood pressure, without dyspnea. Immediately, he was given allergy, expand blood volume therapy, after 30 minutes, the patient had his remission from the symptoms. Even though, there was still patchy rubella skin. One day later, all the symptoms disappeared completely. All medicine were kept using and there was no longer similar reaction.%1例63岁男性患者,确诊“非霍奇金淋巴瘤、ALK阴性间变大细胞型、Ⅰ期B”38 d,既往无药物和食物过敏史.治疗期间在第三次使用培门冬酶注射液3750IU分三个部位匀速肌肉注射90 min后,患者出现前胸、后背部、臀部及双上肢风团,诉奇痒无比,60 min后患者入厕时诉剧烈头痛、胸痛、咽喉痛,舌尖麻木,视物模糊,血压降低,无呼吸困难.立即给予抗过敏、扩充血容量治疗,30 min后症状缓解,仍有皮肤斑片状风疹.1d后症状完全消失,其余药物继续使用,未再发生类似反应.

  18. Experience in diagnosis and treatment of asparaginase-associated pancreatitis in children%门冬酰胺酶相关胰腺炎的诊治体会

    Institute of Scientific and Technical Information of China (English)

    陈再生; 李健

    2014-01-01

    Objective To analyze the clinical characteristics and the course of diagnosis and therapy of PEG-asparaginase associated pancreatitis (AAP)in childhood,and to reveal the pathophysiology of AAP,enhance the ability of diagnosis and treament.Method Data of 13 cases with AAP in childhood seen from March 2011 to March 2014 were analyzed with regard to clinical manifestations,laboratory findings,imaging feature and treatment.Result AAP was found in 12 of acute lymphoblastic leukemia (ALL) and 1 of non-Hodgkin's lymphoma (NHL),8 were boys and 5 were girls,with a mean age 6 years.In 12 cases AAP occurred during the induction-remission treatment,in 1 case during the maintenance-intensification phase.AAP occurred after a median of two doses,and 9 d (median) from the latest administration of PEG-asparaginase.The major manifestations of AAP was abdominal pain (11/13).At the time of AAP diagnosis during the first 48 hours the median peak serum amylase and serum lipase levels were 559 U/L (range 118 -1 585,upper normal limit:125) and 934 U/L (range 221-1 673,upper normal limit:300).Three cases with serum amylase and serum lipase levels above 3 times upper normal limit were repeatedly complicated with pancreatic pseudocyst; 11 patients had abnormal CT imaging,8 cases revealed effusion around the pancreas,and 4 cases had pseudocyst.Therapy with ulinastatin,octreotide acetate,glucocorticoid could relieve abdominal pain significantly.Three cases underwent abdominal puncture drainage and 5 cases fulfilled nasojejunal nutrition therapy.Nine of them were cured,4 developed pseudocyst,in 2 AAP vanished gradually and 2 died with pseudocyst.Conclusion The major manifestations of AAP were abdominal pain,but sometimes apparent and sometimes latent.Condition of acute pancreatitis may exacerbate rapidly and easily.Early identification had significance.Pancreatic pseudocyst suggested a poor prognosis.%目的 分析门冬酰胺酶相关胰腺炎(AAP)的临床特征和诊治经过.方法 收集

  19. Pharmacokinetics of Recombinant E.coli L-asparaginase in Rats%重组E.coli L-天冬酰胺酶在大鼠中的药物代谢动力学

    Institute of Scientific and Technical Information of China (English)

    陈建华; 吴梧桐; 平野和行

    2002-01-01

    应用放射性核素示踪技术研究125I重组E.coli L-天冬酰胺酶在大鼠体内的药物代谢动力学.125I重组L-天冬酰胺酶静脉注射后24h内,在尿,粪,胆汁中的排泄量分别占注射剂量的68.95%,4.44%和5.36%.测定血浆中125I重组E.coli L-天冬酰胺酶浓度,应用聚丙烯酰胺凝胶电泳和生物成像分析系统结合方法评价原药水平,由房室模型评价药物动力学参数,静注后,浓度时间曲线符合二房室模型.初期和末端的t1/2分别为0.52~0.63 h和2.39~2.76h.AUC与剂量成正比.重组E.coli L-天冬酰胺酶的分解代谢产物主要随尿液排泄,重组E.coli L-天冬酰胺酶在大鼠中的药物动力学参数为临床试验提供了有用依据.%The distribution of 125[ recombinant E. coli L-asparaginase in tissues or organs and the excretion in urine,feces and bile were studied with in vivo radioactive tracer technique. The amount of radioactivity excreted in urine, feces and bile within 24 h after intravenous administration of125I recombinant E.col L- asparaginase to rats was 68.95%,4.44%and 5.36% of the dose respectively. 125I recombinant E. coli L- asparaginase in plasma samples was determined. The levels of structural intact molecule in plasma samples were evaluated by SDS-PAGE and bio-imnging analyzer system.Pharmacokinetic parameters were assessed with a model-dependent method. The concentration-time curves of recombinant with the two-compartment model. The first and terminal elimination t1/2 were 0.52 ~ 0.63 h and 2.39 ~ 2.76 h respectively.AUC was linearly related to the doses. The results of distribution in tissues or organs and excretion in urine suggested that the metabolites of the enzyme were cleared by mechanisms of urinary excretion. Pharmacokinetics parameters of recombinant E . coli L- asparaginase in rats are warranted for the design of future clinical trials.

  20. Comparison of gemcitabine, oxaliplatin and L-asparaginase and etoposide, vincristine, doxorubicin, cyclophosphamide and prednisone as first-line chemotherapy in patients with stage IE to IIE extranodal natural killer/T-cell lymphoma: a multicenter retrospective study.

    Science.gov (United States)

    Wang, Hua; Wuxiao, Zhi-Jun; Zhu, Jiayu; Wang, Zhihui; Wang, Ke-Feng; Li, Su; Chen, Xiaoqin; Lu, Yue; Xia, Zhong-Jun

    2015-04-01

    Optimal chemotherapy regimen for Extranodal natural killer/T-cell lymphoma (ENKTL) has not yet been defined. We retrospectively compared the outcome of 93 patients newly diagnosed with stage IE to IIE ENKTL who received gemcitabine, oxaliplatin and L-asparaginase (GELOX) (n = 40) or etoposide, vincristine, doxorubicin, cyclophosphamide and prednisone (EPOCH) (n = 53) as induction chemotherapy. After induction chemotherapy, the complete response (CR) rate and overall response rate (ORR) for the GELOX group were higher than those for the EPOCH group (70.0% vs. 41.5%, p = 0.007 for CR; 87.5% vs. 67.9%, p = 0.047 for ORR). The GELOX regimen resulted in significantly superior 5-year progression-free survival (PFS) (79.0% vs. 46.5%, p = 0.005) and overall survival (OS) rates (78.9% vs. 50.4%, p = 0.003). Toxicity of both regimens was acceptable. The GELOX regimen produces a better long outcome with less toxicity than the EPOCH regimen for patients with early stage ENKTL. PMID:24991715

  1. 左旋门冬酰胺酶诱发儿童急性胰腺炎临床分析%Clinical Analysis of Acute Pancreatitis Induced by L-asparaginase in Children

    Institute of Scientific and Technical Information of China (English)

    高海丽; 李彦格; 刘炜

    2014-01-01

    目的:分析左旋门冬酰胺酶(L-ASP)诱发儿童急性胰腺炎的临床特点,以降低L-ASP后急性胰腺炎发病率,提高临床预后。方法:对本院应用L-ASP后7例急性胰腺炎的诱因、临床表现、实验室检查、及治疗结果进行总结分析。结果:351例次应用L-ASP化疗的患儿中,发生急性胰腺炎7例,发生率2%,其中重症胰腺炎6例,发病前有暴饮暴食者3例,临床表现中腹痛6例、腹胀5例、发热6例、休克5例,所有病例均有血淀粉酶及脂肪酶增高,胰腺彩超均异常。7例胰腺炎中存活3例,中位年龄11岁(7~14岁),死亡4例,中位年龄4岁(2.6~6岁),均死于休克。结论:暴饮暴食、年龄>7岁为胰腺炎发生的重要危险因素,儿童急性胰腺炎症状不典型,重症胰腺炎死亡率高,早期诊断是提高预后的关键。%Objective:To analyze clinical features of children with acute pancreatitis induced by L-asparaginase (L-ASP),so as to reduce incidence of pancreatitis caused by L-ASP and improve clinical outcomes.Method:The incentives,clinical manifestations,laboratory tests,and outcome of 7 acute pancreatitis caused by L-ASP in our hospital were analyzed.Result:Of 351 cases with L-ASP application,7 cases suffered acute pancreatitis,the incidence was 2%, 6 cases were severe pancreatitis,3 cases were before the onset of overeating,6 cases had abdominal pain,5 cases had stomachache,6 cases had fever,and 5 cases suffered shock.All patients had elevated serum amylase and lipase,and all pancreatic ultrasound showed abnormalities.3 cases survived with a median age of 11 years(7-14 years old),4 cases died with a median age of 4 years(2.6-6 years old),all died of shock.Conclusion:Overeating,age more than 7 years old are important risk factors for pancreatitis.Symptoms of pancreatitis in children are atypical,severe pancreatitis have high mortality rate,early diagnosis is key to improve prognosis.

  2. 门冬酰胺酶治疗血液肿瘤致高甘油三酯血症的临床研究%Asparaginase for therapy of transient hypertriglyceridemia caused by hematologic neoplasms

    Institute of Scientific and Technical Information of China (English)

    于亚平; 史平; 刘海宁; 翟勇平; 宋萍; 李峰; 周晓刚; 安志明; 唐玉梅

    2011-01-01

    为了研究门冬酰胺酶(ASP)治疗血液肿瘤时致高甘油三酯血症(HTG)的发生率、临床特点和治疗,以引起临床对该副反应的重视,对24例儿童和成人急性淋巴细胞白血病(ALL)、淋巴母细胞淋巴瘤和NK/T细胞淋巴瘤患者接受了含ASP的联合化疗方案治疗,测定治疗前后的血胆固醇和甘油三酯(TG)水平,确定血脂异常的发生率和特点.24例患者共接受36个周期含ASP的联合方案化疗,2例次出现HTG,发生率为5.6%.2例均为成人ALL,均在初次缓解后4和5个月,再次使用ASP进行巩固治疗时出现HTG,峰值TG分别为32.57和15.77 mmol/L.1例同时伴有下肢剧烈疼痛.停用ASP并加用胰岛素静脉滴入治疗,9~11 d后血脂均恢复正常.初步研究结果提示,接受ASP治疗的血液肿瘤患者有发生HTG的危险,尽管此种HTG在停药多后多可恢复,但可继发胰腺炎、高黏滞综合征或血栓.因而ASP治疗时应常规监测血TG水平.%The objective of this study was to investigate the incidence, clinical characteristics and treatment of asparaginase (ASP)-associated hypertriglyceridemia in adult and children with hematologic neoplasms, and draw attention to this side effects. Twenty-four newly diagnosed children and adult with acute lymphoblastic leukemia(ALL),lymphoblastic lymphoma and NK/T lymphoma were retrospectively evaluated for lipid abnormalities. They were treated with a regimen containing ASP between 2006 and 2010. Total cholesterol and triglycerides (TG) levels were measured in all patients before and after use of ASP. All 24 patients received 36 cycles chemotherapy including ASP.ASP associated hypertriglyceridemia occured in 2 cycles of 2 adult patients with ALL(5.6 %). Transient hypertriglyceridemia (32.57 mmol/L and 15. 77 mmol/L) was observed following administration of ASP as intensification treatment after 4 and 5 months of complete remission. One patient reported pain in both lower extremities associated with

  3. ALL患儿L-Asp治疗相关性血浆氨基酸水平变化及其意义%Contents of L-Asparaginase-Related Amino Acids in the Plasma of Children with ALL

    Institute of Scientific and Technical Information of China (English)

    江华; 顾龙君; 陈静; 潘慈; 薛惠良

    2003-01-01

    Objective To detect the contents of L-asparaginase(L-Asp)-related amino acids in ALL children receiving L-Asp treatment; and to study the relationship between changes in L-Asp contents and clinical efficacy on ALL. Methods Plasma concentrations of asparagine (Asn), aspartic acid (Aspa), glutamine (Gln) and glutamic acid (Glu) in different stages of L-Asp treatment were measured by the HPLC-FLD method in 20 children with ALL (17 cases of B-ALL and 3 of T-ALL). Results The plasma Asn level after the 1st administration of L-Asp decreased significantly. With the administration of L-Asp according to the induction remission formula of All-XH99, the Asn level remained low, even to nil level. This status remained for 7 days after cessation of L-Asp, and even 10 days in 15 cases of B-ALL, but the Asn level in all the cases of T-ALL and only 2 cases of B-ALL increased and even returned to nomal 7 days after L-Asp treatment ended. The concentration of Glu after the second and the last administrations of L-Asp increased significantly and it returned to normal on the 7th day after cessation of L-Asp, while the concentration of Aspa increased and failed to return to normal on day 10 after cessation of of L-Asp. The concentration of Gln slightly decreased during the course of treatment with L-Asp, but the difference was not significant compared with that before treatment. Conclusions The Asn level of children with T-ALL after cessation of L-Asp recovers earier than that of children with B-ALL, indicating that it may be helpful for individualization of L-Asp administration in the treatment of ALL on the basis of the immunophenotype of children. The glutaminase activity of L-Asp does not exert effects on the treatment.%目的检测ALL患儿血浆左旋门冬酰胺酶(L-Asp)治疗过程中相关氨基酸水平变化,探讨这种变化与临床疗效的相关性,为L-Asp的个体化用药提供依据.方法通过HPLC-FLD技术检测20例初治ALL患儿(17例为B-ALL,3例为T-ALL)在L

  4. Meta-analysis and meta-regression analysis to compare the outcomes of chemotherapy for T- and B-lineage acute lymphoblastic leukemia (ALL): the use of dexamethasone, L-asparaginase, and/or methotrexate may improve the outcome of T-lineage ALL.

    Science.gov (United States)

    Kako, Shinichi; Akahoshi, Yu; Harada, Naonori; Nakano, Hirofumi; Kameda, Kazuaki; Ugai, Tomotaka; Wada, Hidenori; Yamasaki, Ryoko; Ishihara, Yuko; Kawamura, Koji; Sakamoto, Kana; Sato, Miki; Ashizawa, Masahiro; Terasako-Saito, Kiriko; Kimura, Shun-ichi; Kikuchi, Misato; Nakasone, Hideki; Yamazaki, Rie; Kanda, Junya; Nishida, Junji; Kanda, Yoshinobu

    2016-01-01

    The effects of intensive regimens and the roles of drugs used might differ between T- and B-lineage acute lymphoblastic leukemia (ALL). We performed a literature search for clinical studies published from January 1998 to March 2013. Studies were eligible for inclusion in the analyses if they included more than 80 patients with adult ALL who were treated with a uniform regimen and compared T- and B-lineage ALL. Studies that included only adolescent or elderly patients were excluded. We identified 11 clinical studies, which included a total of 381 and 1366 patients with T- and B-lineage ALL, respectively, and performed meta-analyses using the selected studies. Nine studies included patients with Philadelphia chromosome-positive (Ph+) ALL. A meta-analysis using the random-effect model demonstrated superior survival in patients with T-lineage ALL compared to those with B-lineage ALL (hazard ratio 1.78, 95 % confidence interval 1.50-2.11), though the inclusion of patients with Ph+ ALL in B-lineage ALL must have influenced this result strongly. We performed meta-regression analyses, adjusted according to whether or not patients with Ph+ ALL were included in each study. Use of dexamethasone (Dex), higher dose of methotrexate (MTX), and higher dose of L-asparaginase (L-asp) were associated with a significant trend toward a better outcome in T-lineage ALL. A meta-regression analysis including Dex and the dose of L-asp or MTX together as covariates showed that these factors were independently significant. In conclusion, the use of Dex and high-dose L-asp or MTX may improve the outcome of T-lineage ALL. This hypothesis should be tested in a prospective study including only patients with Ph-negative ALL.

  5. 重组L-天门冬酰胺酶II的液相色谱/电喷雾离子化质谱法分析%HIGH PERFORMANCE LIQUID CHROMATOGRAPHY / ELECTROSPRAY IONIZATON MASS SPECTROMETRIC CHARACTERIZATION OF RECOMBINANT L-ASPARAGINASE II

    Institute of Scientific and Technical Information of China (English)

    韩俊; 盛龙生; 杨仲元; 相秉仁; 安登魁

    2001-01-01

    AIM To characterize the primary structure of recombinant L-asparaginase II product. METHODS The molecular weight of the protein was measured by pneumatically-assisted electrospray ionization mass spectrometry with flow injection mode. Subsequently, tryptic peptide mapping was performed by high performance liquid chromatography on a C8 column with tandem UV and MS detection. An easy-to-use and simple denaturation process with trichloroacetic acid was conducted prior to tryptic digest so as to release the digest resistance from the protein structure. The amino acid sequences of the tryptic peptides were elucidated based on their in-source collision-induced dissociation spectra. RESULTS The measured molecular mass was different from the theoretical value. Three amino acid variations were unambiguously detected along the peptide backbone derived from the gene-encoding sequence. CONCLUSION This paper revealed that LC/ESI/MS had provided a promising and robust technique in primary structure analysis and quality control of DNA-derived recombinant protein pharmaceuticals.%目的 对基因重组L-天门冬酰胺酶II产品进行一级结构分析。方法 采用流动注射方式,电喷雾离子化质谱法测定分子量;三氯乙酸变性,胰蛋白酶水解后,HPLC测定肽图谱;结合质谱源内碰撞诱导解离技术,解析酶解肽段的氨基酸顺序。结果 分子量测定值与理论值不符,质谱解析表明样品中存在3个氨基酸变异现象,由此计算的分子量与测定值吻合。结论 液相色谱/电喷雾质谱法为基因重组蛋白质药物的一级结构分析和质量控制提供了新途径。

  6. 达沙替尼联合甲氨蝶呤和左旋门冬酰胺酶挽救性治疗费城染色体阳性急性淋巴细胞白血病致重度药物性肝损伤一例并文献复习%Dasatinib combined with methotrexate and L-asparaginase in the treatment of patients with Philadelphia chromosome positive acute lymphoblastic leukemia cause severe drug-induced liver injury: one case report and literature review

    Institute of Scientific and Technical Information of China (English)

    王璐; 魏旭东; 尹青松; 汪萍; 米瑞华; 艾昊

    2015-01-01

    目的 提高对达沙替尼联合大剂量甲氨蝶呤(HD-MTX)和左旋门冬酰胺酶(L-Asp)方案挽救性治疗费城染色体阳性急性淋巴细胞白血病(Ph+ ALL)致重度药物性肝损伤的认识.方法 报道达沙替尼联合HD-MTX及L-Asp方案挽救性治疗Ph+ ALL致重度药物性肝损伤1例并文献复习.结果 此例Ph+ ALL患者在该方案化疗后第7天暴发重度药物性肝损伤,肝功能:总胆红素(TBIL)221.7 μmol/L,直接胆红素(DBIL) 156.1 μmol/L,间接胆红素(IBIL) 65.6μmol/L,丙氨酸氨基转移酶(ALT)111U/L,天冬氨酸氨基转移酶(AST)131U/L,碱性磷酸酶(ALP) 354 U/L,谷氨酰转肽酶(GGT) 256U/L,总胆汁酸(TBA) 199.2 μmol/L.经积极保肝治疗2周后,胆红素及氨基转移酶降至正常,且疾病达完全缓解.结论 达沙替尼联合HD-MTX及L-Asp方案挽救性治疗Ph+ ALL疗效显著,但三药均有明显肝脏毒性,联合使用时应注意暴发性药物性肝损伤的发生.%Objective To improve the cognition of sever liver injury of treating Philadelphia positive acute lymphoblastic leukemia (Ph+ ALL) with salvage chemotherapy of dasatinib combined with high-dose methotrexate (HD-MTX) and L-asparaginase (L-Asp).Methods Severe drug-induced liver injury caused by dasatinib with HD-MTX and L-Asp in one patient with Ph+ ALL was reported.Results Severe drug-induced liver injury happened on the seventh day after treatment,TBIL 221.7 μmol/L,DBIL 156.1 μmol/L,IBIL 65.6 μmol/L,ALT 111 U/L,AST 131 U/L,ALP 354 U/L,GGT 256 U/L,TBA 199.2 μmol/L.Through proper treatment,the patient recovered quite good,and the patient achieved complete remission after this chemotherapy.Conclusion Salvage chemotherapy which contains dasatinib,MTX and L-Asp can be effectively used in Ph+ ALL,but they are all of the hepatotoxicity,so drug-induced Liver injury may happen while they are used together.

  7. Etoposide, Prednisone, Vincristine Sulfate, Cyclophosphamide, and Doxorubicin Hydrochloride With Asparaginase in Treating Patients With Acute Lymphoblastic Leukemia or Lymphoblastic Lymphoma

    Science.gov (United States)

    2016-04-26

    B Acute Lymphoblastic Leukemia; B Lymphoblastic Lymphoma; Recurrent Adult Acute Lymphoblastic Leukemia; Recurrent B Lymphoblastic Lymphoma; Recurrent T Lymphoblastic Leukemia/Lymphoma; Refractory B Lymphoblastic Lymphoma; Refractory T Lymphoblastic Lymphoma; T Acute Lymphoblastic Leukemia; T Lymphoblastic Lymphoma

  8. Antimicrobial and L-asparaginase activities of endophytic fungi isolated from Datura innoxia and Hyoscyamus muticus medicinal plants

    OpenAIRE

    El-Said, Ahmed H. M.; YASSMIN M. SHEBANY; Hussein, Mohamed A.; El-Dawy, Eman G. A.

    2016-01-01

    Thirty-six species and two varieties belonging to 16 genera of fungal endophytes were isolated from the leaves of Datura innoxia and Hyoscyamus muticus plants. The most prevailing fungi were: Aspergillus fumigatus, A. niger, A. terreus var. africanus, Cladosporium cucumerinum, C. oxysporum, Penicillium aurantiogriseum and P. chrysogenum. Endophytic fungi from D. innoxia and H. muticus plants were tested for antibacterial and antifungal activities from which 68.98 and  78.26% respectively...

  9. EST Table: AV404106 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available aa pdb|2ZAK|A Chain A, Orthorhombic Crystal Structure Of Precursor E. Coli Isoaspartyl PeptidaseL-Asparaginase (Ecaiii) With Active...r E. Coli Isoaspartyl PeptidaseL-Asparaginase (Ecaiii) With Active- Site T179a Mu...tation pdb|3C17|A Chain A, Hexagonal Crystal Structure Of Precursor E. Coli Isoaspartyl PeptidaseL-Asparaginase (Ecaiii) With Active.... Coli Isoaspartyl PeptidaseL-Asparaginase (Ecaiii) With Active- Site T179a Mutat

  10. Sequential DICE combined with l-asparaginase chemotherapy followed by involved field radiation in newly diagnosed, stage IE to IIE, nasal and extranodal NK/T-cell lymphoma.

    Science.gov (United States)

    Dong, Li-Hua; Zhang, Li-Juan; Wang, Wen-Jia; Lei, Wen; Sun, Xing; Du, Jian-Wei; Gao, Xue; Li, Gang-Ping; Li, Yu-Fu

    2016-07-01

    Extranodal natural killer (NK)/T-cell lymphoma is an aggressive lymphoid tumor. Optimal treatment strategies have not yet been fully defined. To explore a more effective treatment, we conducted sequential chemoradiotherapy (SCRT) and evaluated the safety and efficacy. Seventy-eight patients (51 males, 27 females) were analyzed. The complete response (CR) rate was higher for patients in the SCRT group (90.9%) than in the radiotherapy group (77.8%; p = 0.124). The relapse rate (RR) and death rate (DR) were lower in the SCRT group (RR: 6.7% vs 33.3%, p < 0.001; DR: 15.2% vs. 55.6%, p < 0.001). Progression-free survival (PFS) and overall survival (OS) rates of 5 years after diagnosis were significantly higher for patients in the SCRT group (PFS: 89%; OS: 82%) than in the radiotherapy group (PFS: 49%, p < 0.001; OS: 49%, p < 0.001). Treatment-related adverse events were more common in the SCRT group. However, the adverse events were controlled. PMID:26726970

  11. A Pilot Study of Intensified PEG-Asparaginase in High-risk Acute Lymphoblastic Leukemia: Children's Oncology Group Study AALL08P1.

    Science.gov (United States)

    Rodriguez, Vilmarie; Kairalla, John; Salzer, Wanda L; Raetz, Elizabeth A; Loh, Mignon Lc; Carroll, Andrew J; Heerema, Nyla A; Wood, Brent L; Borowitz, Michael J; Burke, Michael J; Asselin, Barbara L; Devidas, Meenakshi; Winick, Naomi J; Carroll, William L; Hunger, Stephen P; Dreyer, ZoAnn E

    2016-08-01

    AALL08P1 was designed to determine whether biweekly intensified pegaspargase (I-PEG) was feasible and safe in pediatric patients with newly diagnosed high-risk B-precursor lymphoblastic leukemia when given with Children's Oncology Group hemiaugmented BFM therapy. High-risk average (HR-Avg) patients received standard pegaspargase dosing (6 doses), whereas high-risk high (HR-High) patients received I-PEG biweekly from the start of Consolidation until day 1 of Maintenance. Feasibility and safety were defined in advance as ≥65% of patients tolerating at least 8 doses of I-PEG and 90% requiring ≤49 weeks from day 1 of Consolidation to the initiation of Maintenance. Targeted toxicities included allergic reactions, anaphylaxis, pancreatitis, thrombosis, bleeding, central nervous system events, and sepsis. AALL08P1 enrolled 104 patients; 54 were classified as HR-Avg and 30 as HR-High after completion of induction therapy. Only 53% (16/30) of the HR-High patients received ≥8 total doses of I-PEG and 50% (15/30) took ≤49 weeks from start of Consolidation to the initiation of Maintenance. I-PEG did not significantly increase grade 2 to 5 targeted toxicities. I-PEG was not feasible or safe as defined in AALL08P1. Complete assessment of this regimen was limited due to removal of patients from I-PEG regimen and early closure of the study. PMID:27299599

  12. Lymphatic system and lymphoma

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    2008108 Comparison of polyethylene glycol conjugated asparaginase and L-asparaginase for treatment of childhood acute lymphoblastic leukemia. ZHU Xiaofan(竺晓凡), et al. Instit Hemat & Blood Dis Hosp, CAMS & PUMC, Tianjin 300020. Chin J Hematol 2008;29(1):29-33. Objective To evaluate efficacy and side-effect of polyethylene glycol conjugated asparaginase (PEG-Asp) containing VDPAP regimen in newly diagnosed childhood acute lymphoblastic leukemia (ALL).

  13. Efficacy of pegaspargase in extra nodal natural killer/T-cell lymphoma nasal type: A case report from China

    Directory of Open Access Journals (Sweden)

    Xingan Xiong

    2015-01-01

    Full Text Available Extranodal natural killer (NK/T-cell lymphoma, nasal type, is a rare and highly aggressive disease with a grim prognosis. There is no known satisfactory treatment. The author herein to report one case of L-asparaginase extranodal NK/T-cell lymphoma primary treated with L-asparaginase methotrexate and dexamethasone.

  14. Effects of Acrylamide Inhibition by Asparaginase and Sugar Substitution on Cookie Dough Rheology and Baking Attributes%丙烯酞胺抑制剂对曲奇面团流变学和烘焙特性的影响

    Institute of Scientific and Technical Information of China (English)

    Mohamed ABDEL-SHAFI ABDEL-SAMIE; 黄卫宁; 李珍妮; Okkyung Kim CHUNG

    2011-01-01

    通过引入天冬酞胺酶和/或甜菊苷到曲奇配方中替代部分糖以抑制其生产过程中丙烯酞胺的生成,分析单独或同时添加这两种配料时曲奇面团的动态流变学特性、硬度和曲奇饼干的烘焙感官特性.结果表明:当单独添加天冬酸胺酶(1000 ASNU)时可降低曲奇样品中天冬酞胺含量(0.045mg/g)的67%,从而抑制95%丙烯酞胺的生成,且不会影响曲奇产品的烘焙特性.而天冬酞胺酶和甜菊苷同时添加时可抑制样品中96%丙烯酞胺的生成.动态流变学结果表明:引入天冬酞胺酶不会影响曲奇面团的流变学性质,而甜菊苷的加入会增加面团的弹性模量G,和黏性模量G",这是因为甜菊苷替代面团中糖成分后促进曲奇面团面筋网络的形成.但是,部分糖被取代后会改变曲奇的一些烘焙特性,如:水分含量增加,曲奇饼干颜色变淡,延展率和破碎力降低.感官分析结果表明:感官评定者不能接受45%和60%糖取代量的曲奇,但可以接受15%或30%糖取代量的曲奇.

  15. Drugs Approved for Leukemia

    Science.gov (United States)

    ... Ask about Your Treatment Research Drugs Approved for Leukemia This page lists cancer drugs approved by the ... not listed here. Drugs Approved for Acute Lymphoblastic Leukemia (ALL) Abitrexate (Methotrexate) Arranon (Nelarabine) Asparaginase Erwinia chrysanthemi ...

  16. L-Asparaginaz Allerjisi Sürekli Adrenalin ?nfüzyonu E?li?inde Yapılan Desensitizasyonla A??labilir miş

    OpenAIRE

    2, Semra KARA; 1, Nurullah ÇEL?K; 2, Ömer CEV?T; 2, Hayri B. TOKSOY 2 Dilara ?ÇA?ASIO?LU

    2010-01-01

    ABSTRACT May L-asparaginase allergies be removed by desen-sitization with continous adrenaline infusion? The allergic reactions related to chemotherapeutic drugs represent a very significiant problem in clinical practice as all these antineoplastic drugs must be used in a phase associated schedule. L-asparaginase is an anti-neoplastic drug in which hypersensitivity reactions can be seen commonly. It is estimated that hyper-sensitivity reactions may occur in 5-35% and life threatening a...

  17. Role of γ-glutamyltranspeptidase in the pathogenesis of Helicobacter pylori infection.

    Science.gov (United States)

    Rimbara, Emiko; Mori, Shigetarou; Kim, Hyun; Shibayama, Keigo

    2013-10-01

    γ-Glutamyltranspeptidase and asparaginase have been shown to play important roles in Helicobacter pylori colonization and cell death induced by H. pylori infection. In this study, the association of γ-glutamyltranspeptidase and asparaginase was elucidated by comparing activities of both deamidases in H. pylori strains from patients with chronic gastritis, gastric and duodenal ulcers, and gastric cancer. γ-Glutamyltranspeptidase activities in H. pylori strains from patients with gastric cancer were significantly higher than in those from patients with chronic gastritis or gastric ulcers. There was a wide range of asparaginase activities in H. pylori strains from patients with gastric cancer and these were not significantly than those from patients with other diseases. To identify the contributions of γ-glutamyltranspeptidase and asparaginase to gastric cell inflammation, human gastric epithelial cells (AGS line) were infected with H. pylori wild-type and knockout strains and inflammatory responses evaluated by induction of interleukin-8 (IL-8). IL-8 response was significantly decreased by knockout of the γ-glutamyltranspeptidase-encoding gene but not by knockout of the asparaginase-encoding gene. Additionally, IL-8 induction by infection with the H. pylori wild-type strain was significantly decreased by adding glutamine during infection. These findings indicate that IL-8 induction caused by γ-glutamyltranspeptidase activity in H. pylori is mainly attributable to depletion of glutamine. These data suggest that γ-glutamyltranspeptidase plays a significant role in the chronic inflammation caused by H. pylori infection.

  18. 抗体夹心酶联免疫吸附法测定重组E.coli L-天冬酰胺酶及药代动力学研究%Antibody sandwich enzyme-linked immunoadsorbent assay for determination of recombinant E.coli L-asparaginase in rat plasma and its pharmacokinetics

    Institute of Scientific and Technical Information of China (English)

    陈建华; 吴梧桐; 平野和行

    2003-01-01

    目的建立大鼠血浆中重组E.coli L-天冬酰胺酶的抗体夹心酶联免疫吸附测定法,并进行药代动力学研究.方法应用重组E.coli L-天冬酰胺酶免疫家兔,分离IgG,用DEAE-纤维素柱色谱纯化,辣根过氧化物酶标记抗体,建立抗体夹心酶联免疫吸附法,测定大鼠血浆中重组E.coli L-天冬酰胺酶浓度.结果方法的线性范围为1~64 U*L-1,血药浓度与时间的关系符合二房室模型,初期和末端的T1/2分别为0.50~0.57 h和2.45~3.02 h,AUC与剂量成正比.结论建立的抗体夹心酶联免疫吸附法在灵敏度、特异性、线性范围、精密度和回收率等方面,满足药代动力学研究要求.实验方法和重组E.coli L-天冬酰胺酶在大鼠中的药代动力学参数为临床研究提供了手段和依据.

  19. Effect of Recombinant -L-Asparaginase Proliposome on the Acute Lymphatic Leukaemia in Mice%重组L-门冬酰胺酶前体脂质体对急性淋巴白血病小鼠的治疗效果

    Institute of Scientific and Technical Information of China (English)

    王弘; 吴梧桐; 顾学裘; 郭青龙

    2000-01-01

    考察L-门冬酰胺酶前体脂质体(L-ASNasePL)与L-ASNase对移植性小鼠急性淋巴细胞白血病的作用.采用DBA/2小鼠两种L1210、P388腹水瘤模型,脂质体组和L-ASNase组小鼠每天ip一次(按L-门冬酰胺酶计7400,3700,1850KU/kg), 连续10d.设生理盐水组(0.9%NaCl,0.4ml/20g)及阿霉素注射剂(2mg/kg)组为阴性和阳性对照组.结果L-ASNase PL及L-门冬酰胺酶(L-ASNase)皆能显著延长移植性淋巴细胞白血病的小鼠存活天数(P<0.01), L-ASNase PL对 L1210、P388移植小鼠,生命延长率分别最高达184.7%和183.3%.L-ASNase组生命延长率最高分别为132.9%和132.1%.L-ASNasePL与相同剂量的L-ASNase相比,其高、中剂量组(7400,3700kU/kg)皆显著延长L1210、P388小鼠的存活天数(P<0.05).

  20. Comparison of the Anti-Leukemia Effect and Mechanism of L-Asparaginase between Two Different Strains%两种菌种来源的左旋门冬酰胺酶抗白血病作用与机制比较

    Institute of Scientific and Technical Information of China (English)

    王开美; 徐宏贵; 郭海霞; 金少文; 罗湘琴; 方建培

    2014-01-01

    比较大肠杆菌来源的左旋门冬酰胺酶(E coli-L-Asp)与欧文菌来源的左旋门冬酰胺酶(Erwinia-L-Asp)体外抗白血病作用,并探讨其作用机制.以两种来源的L-Asp分别处理急性淋巴细胞白血病(ALL)细胞株(REH、Jurkat),采用CCK-8试剂盒检测细胞增殖变化(IC50),Annexin V/PI双染流式细胞术检测细胞凋亡,高压液相色谱(HPLC)检测用药后培养液中4种氨基酸[门冬酰胺(Asn)、门冬氨酸(Aspa)、谷氨酰胺(G1n)、谷氨酸(G1u)]浓度的变化.结果表明,REH、Jurkat细胞株对两种L-Asp的抑制作用均敏感,且在一定时间内存在明显剂量效应关系,即在较低的药物浓度下,细胞增殖抑制率处于较低水平,而药物浓度增加后,细胞的增殖抑制率明显升高,其中作用后24 h,Erwinia-L-Asp组细胞抑制率、细胞凋亡率均明显高于E coli-L-Asp组(P<0.05),但至48 h两组比较差异无显著性(P>0.05).两种药物在低浓度时均能耗竭培养液中的Asn,且作用相当(P>0.05),高浓度时耗竭Gin作用才凸显,而Erwinia-L-Asp的作用明显强于E.coli-L-Asp(P<0.05).结论:两种菌种来源的L-Asp抗白血病作用与剂量存在量效关系,除耗竭外环境中的Asn和Gin外,诱导细胞凋亡也是其抗白血病作用之一.Erwinia-L-Asp具有起效快、Gin耗竭作用较强的特点,可作为临床儿童ALL治疗的一线药物选择.

  1. Kinetics of inclusion body formation and its correlation with the characteristics of protein aggregates in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Arun K Upadhyay

    Full Text Available The objective of the research was to understand the structural determinants governing protein aggregation into inclusion bodies during expression of recombinant proteins in Escherichia coli. Recombinant human growth hormone (hGH and asparaginase were expressed as inclusion bodies in E.coli and the kinetics of aggregate formation was analyzed in details. Asparaginase inclusion bodies were of smaller size (200 nm and the size of the aggregates did not increase with induction time. In contrast, the seeding and growth behavior of hGH inclusion bodies were found to be sequential, kinetically stable and the aggregate size increased from 200 to 800 nm with induction time. Human growth hormone inclusion bodies showed higher resistance to denaturants and proteinase K degradation in comparison to those of asparaginase inclusion bodies. Asparaginase inclusion bodies were completely solubilized at 2-3 M urea concentration and could be refolded into active protein, whereas 7 M urea was required for complete solubilization of hGH inclusion bodies. Both hGH and asparaginase inclusion bodies showed binding with amyloid specific dyes. In spite of its low β-sheet content, binding with dyes was more prominent in case of hGH inclusion bodies than that of asparaginase. Arrangements of protein molecules present in the surface as well as in the core of inclusion bodies were similar. Hydrophobic interactions between partially folded amphiphillic and hydrophobic alpha-helices were found to be one of the main determinants of hGH inclusion body formation. Aggregation behavior of the protein molecules decides the nature and properties of inclusion bodies.

  2. Conservative Management of Pancreatic Pseudocysts in Children with Acute Lymphoblastic Leukemia

    OpenAIRE

    Spraker, Holly L.; Spyridis, Georgios P.; Pui, Ching-Hon; Howard, Scott C.

    2009-01-01

    Treatment with asparaginase for acute lymphoblastic leukemia (ALL) can cause acute pancreatitis. Complication of pancreatitis by pancreatic pseudocyst formation can prolong the hospital stay, delay chemotherapy, and necessitate long-term parenteral nutrition. We report five children with ALL who developed acute pancreatitis complicated by pancreatic pseudocysts. They required modifications to their chemotherapy regimen and prolonged parenteral nutrition but no surgical intervention. All five ...

  3. Endoscopic Cystogastrostomy: Minimally Invasive Approach for Pancreatic Pseudocyst

    Directory of Open Access Journals (Sweden)

    Gull-Zareen Khan Sial

    2014-12-01

    Full Text Available Pancreatic pseudocysts in children are not uncommon. Non-resolving pseudocysts often require surgical intervention. Endoscopic cystogastrostomy is a minimally invasive procedure which is recommended for this condition. We report a large pancreatic pseudocyst in a 4-year old child, which developed following therapy with PEG-Asparaginase for acute lymphoblastic leukemia. It was managed with minimally invasive procedure.

  4. Cell-based laboratory evaluation of coagulation activation by antineoplastic drugs for the treatment of lymphoid tumors

    Science.gov (United States)

    Tsunaka, Misae; Arai, Reina; Ohashi, Ayaka; Koyama, Takatoshi

    2016-01-01

    Objectives: Combining vorinostat, L-asparaginase, and doxorubicin (Dox) led to improved response rates in the treatment of lymphoid tumors. However, deep-vein thrombosis has been noted as one of the most serious side effects with these drugs, and how these regimens cause deep-vein thrombosis is unclear. Methods: We investigated the procoagulant effects of vorinostat, L-asparaginase, and doxorubicin in lymphoid tumors, focusing on tissue factor, phosphatidylserine, and antithrombin. The human vascular endothelial cell line EAhy926 as well as the lymphoid neoplastic cell lines HUT78 (cutaneous T-cell lymphoma), Molt4 (acute T-lymphoblastic leukemia), and Ramos (Burkitt lymphoma) were employed to investigate these procoagulant effects. Results: Vorinostat, L-asparaginase, and doxorubicin induced exposure of phosphatidylserine and procoagulant activity on the surface of lymphoid tumor cells. Vorinostat and doxorubicin also induced phosphatidylserine exposure and increased procoagulant activity on EAhy926 cells. Expression of tissue factor antigen was induced by doxorubicin on the surface of each type of cells, whereas expression of tissue factor mRNA was unchanged. Secretion of antithrombin from HepG2 cells was reduced only by L-asparaginase. Conclusion: These data suggest that vorinostat and doxorubicin may induce procoagulant activity in vessels through apoptosis of tumor cells and through phosphatidylserine exposure and/or tissue factor expression on vascular endothelial cells. L-asparaginase may induce a thrombophilic state by reducing the secretion of anticoagulant proteins such as antithrombin. The laboratory methods described here could be useful to evaluate the procoagulant effects of antineoplastic drugs. PMID:27504186

  5. ATTEMPT TO REDUCE ACRYLAMIDE CONTENT IN ROASTED CHICORY

    Directory of Open Access Journals (Sweden)

    Gabriela Zięć

    2015-02-01

    Full Text Available The aim of this study was to reduce the formation of acrylamide during roasting of chicory roots by soaking the fresh roots in a solution of calcium chloride, by the use of different temperature and time of roasting of dried roots, as well as by the addition of the enzyme (asparaginase during roasting of dried roots. It was shown, that with increasing roasting temperature of chicory roots from 100 - 175 ° C the acrylamide content also increased, while at a temperature of 210 ° C the growth was inhibited. Increasing roasting time from 10 - 25 minutes resulted in an increased acrylamide content. Soaking the roots in the CaCl2 solution for 20 minutes reduced the formation of acrylamide during the roasting approximately by 40%, similarly as the application of asparaginase to the dried roots during the roasting process.

  6. Enzyme-based flow injection analysis system for glutamine and glutamate in mammalian cell culture media.

    Science.gov (United States)

    Mayer, C; Frauer, A; Schalkhammer, T; Pittner, F

    1999-03-01

    We present the setup of a flow injection analysis system designed for on-line monitoring of glutamate and glutamine. These amino acids represent a major energy source in mammalian cell culture. A cycling assay consisting of glutamate dehydrogenase and aspartate aminotransferase produces NADH proportional to the glutamate concentration in the sample. NADH is then measured spectrophotometrically. Glutamine is determined by conversion to glutamate which is fed into the cycling assay. The conversion of glutamine to glutamate is catalyzed by asparaginase. Asparaginase was used in place of glutaminase due to its relatively high reactivity with glutamine and a pH optimum similar to that of glutamate dehydrogenase. The enzymes were immobilized covalently to activated controlled pore glass beads and integrated into the flow injection analysis system. The application of the immobilized enzymes and the technical setup are presented in this paper.

  7. Asparagine Depletion Potentiates the Cytotoxic Effect of Chemotherapy Against Brain Tumors

    OpenAIRE

    Panosyan, Eduard H.; Wang, Yuntao; Xia, Peng; Lee, Wai-Nang Paul; Pak, Youngju; Laks, Dan R.; Lin, Henry J.; Moore, Theodore B.; Cloughesy, Timothy F.; Kornblum, Harley I.; Lasky, Joseph L.

    2014-01-01

    Targeting amino acid metabolism has therapeutic implications for aggressive brain tumors. Asparagine is an amino acid that is synthesized by normal cells. However, some cancer cells lack asparagine synthetase (ASNS), the key enzyme for asparagine synthesis. Asparaginase (ASNase) contributes to eradication of acute leukemia by decreasing asparagine levels in serum and cerebrospinal fluid. However, leukemic cells may become ASNase-resistant by up-regulating ASNS. High expression of ASNS has als...

  8. GenBank blastx search result: AK112031 [KOME

    Lifescience Database Archive (English)

    Full Text Available inkage group 13 Contains part of two novel genes, a novel gene for a protein similar to asparaginases, a novel... gene similar to RAB1B (RAS oncogene family member 1B), a novel gene, two novel genes similar to SLC1A4 an...d SLC1A1 (solute carrier family 1 members 4 and 1) and ORF1 and ORF2 of a novel a

  9. GenBank blastx search result: AK111986 [KOME

    Lifescience Database Archive (English)

    Full Text Available inkage group 13 Contains part of two novel genes, a novel gene for a protein similar to asparaginases, a novel... gene similar to RAB1B (RAS oncogene family member 1B), a novel gene, two novel genes similar to SLC1A4 an...d SLC1A1 (solute carrier family 1 members 4 and 1) and ORF1 and ORF2 of a novel a

  10. GenBank blastx search result: AK243329 [KOME

    Lifescience Database Archive (English)

    Full Text Available N13 in linkage group 13 Contains part of two novel genes, a novel gene for a protein similar to asparaginases, a novel... gene similar to RAB1B (RAS oncogene family member 1B), a novel gene, two novel genes similar to S...LC1A4 and SLC1A1 (solute carrier family 1 members 4 and 1) and ORF1 and ORF2 of a novel

  11. GenBank blastx search result: AK242961 [KOME

    Lifescience Database Archive (English)

    Full Text Available N13 in linkage group 13 Contains part of two novel genes, a novel gene for a protein similar to asparaginases, a novel... gene similar to RAB1B (RAS oncogene family member 1B), a novel gene, two novel genes similar to S...LC1A4 and SLC1A1 (solute carrier family 1 members 4 and 1) and ORF1 and ORF2 of a novel

  12. GenBank blastx search result: AK061969 [KOME

    Lifescience Database Archive (English)

    Full Text Available inkage group 13 Contains part of two novel genes, a novel gene for a protein similar to asparaginases, a novel... gene similar to RAB1B (RAS oncogene family member 1B), a novel gene, two novel genes similar to SLC1A4 an...d SLC1A1 (solute carrier family 1 members 4 and 1) and ORF1 and ORF2 of a novel a

  13. Extra Nodal NK/T-Cell Lymphoma Nasal Type: Efficacy of Pegaspargase Report of Two Patients from the United Sates and Review of Literature

    OpenAIRE

    Reyes, Vincent Edgar; Al-Saleem, Tahseen; Robu, Valentin G.; Smith, Mitchell R.

    2009-01-01

    Extranodal NK/T cell lymphoma nasal type is an EBV driven non-Hodgkin lymphoma, rare in the United States, with no known satisfactory treatment. Two patients with this entity refractory to CHOP chemotherapy responded to single agent pegaspargase (pegylated L-asparaginase). A 64-year-old Caucasian woman presented with extranodal NK/T lymphoma nasal type on her left buttocks. After initial treatment with loco-regional irradiation and CHOP, she developed extensive lower extremity lesions, and su...

  14. Management of Ontario children with acute lymphoblastic leukemia by the Dana-Farber Cancer Institute protocols.

    OpenAIRE

    Desai, S J; Barr, R D; Andrew, M.; deVeber, L L; Pai, M K

    1989-01-01

    There is ample evidence of the value of intensive therapeutic strategies in the management of acute lymphoblastic leukemia (ALL), the commonest form of malignant disease in children. Such a program, devised at the Dana-Farber Cancer Institute (DFCI), Boston, and incorporating high-dose L-asparaginase, was adopted in 1984 by the Children's Hospital at Chedoke-McMaster, Hamilton, Ont., and the Children's Hospital of Western Ontario, London. We describe the experience of these institutions in th...

  15. Occurrence of Streptomyces aurantiacus in Mangroves of Bhitarkanika

    Directory of Open Access Journals (Sweden)

    Gupta, N.

    2007-01-01

    Full Text Available Thirteen strains of Streptomyces were isolated from phyllosphere of nine mangrove tree species found in Bhitarkanika mangrove ecosystem of Orissa. According to physiological, biochemical data, all 13 of the isolates were taxonomically identified to the genus Streptomyces as aurantiacus species. All strains are grayish, spirals and forming amorphous colony. Almost all utilized araginose, produced H2S, resistant towards rifampicin and penicillin, urea except few strains. However, they exhibited different extracellular activity like phosphate solubilization, lipase and L asparaginase production. This is a unique report from this mangrove ecosystem as far as Streptomyces occurrence is concerned.

  16. Uncoupling Intramolecular Processing and Substrate Hydrolysis in the N-terminal Nucleophile Hydrolase hASRGL1 by Circular Permutation

    OpenAIRE

    Li, Wenzong; Cantor, Jason R.; Yogesha, S.D.; Yang, Shirley; Chantranupong, Lynne; Liu, June Qingxia; Agnello, Giulia; Georgiou, George; Stone, Everett M.; Yan ZHANG

    2012-01-01

    The human asparaginase-like protein 1 (hASRGL1) catalyzes the hydrolysis of l-asparagine and isoaspartyl-dipeptides. As an N-terminal nucleophile (Ntn) hydrolase superfamily member, the active form of hASRGL1 is generated by an intramolecular cleavage step with Thr168 as the catalytic residue. However, in vitro, autoprocessing is incomplete (~50 %), fettering the biophysical characterization of hASRGL1. We circumvented this obstacle by constructing a circularly permuted hASRGL1 that uncoupled...

  17. Thrombotic complications in children with non-Hodgkin lymphoma

    OpenAIRE

    N. V. Lipay; A. S. Fedorova; Dmitriev, V. V.

    2014-01-01

    Our study was aimed at identifying of risk factors of venous thrombosis (VT) in children with non-Hodgkin lymphomas. VT episodes were registered in 13 of 174 children treated (7.5 %). Possible impact of morphological type, initial mediastinal involvement, gender, age and use of L-asparaginase as a risk factor of thrombosis development were analyzed. Using multivariate analysis primary mediastinal tumor (OR = 4.73 [CI: 1.42–17.10]) and patient age older than 13 years (OR = 4.3 [CI: 1.19–20.28)...

  18. RALLE pilot: response-guided therapy for marrow relapse in acute lymphoblastic leukemia in children.

    Science.gov (United States)

    Saarinen-Pihkala, Ulla M; Parto, Katriina; Riikonen, Pekka; Lähteenmäki, Päivi M; Békàssy, Albert N; Glomstein, Anders; Möttönen, Merja

    2012-05-01

    Despite improved treatment results of childhood acute lymphoblastic leukemia (ALL), 20% to 30% have a relapse, and then the outcome is very poor. We studied 40 children with ALL marrow relapse piloting an ALL relapse protocol with well-known drugs and drug combinations by using a concept of response-guided design. We also measured response in logarithmic fashion. Our primary end points were achievement of M1 marrow status, minimal residual disease status below 10, and second remission. The remission induction rate was 90% with 10% induction mortality. After the A blocks (dexamethasone, vincristine, idarubicin and pegylated L-asparaginase), 85% had M1 status, 39% had minimal residual disease ≤1×10, and 66% had 2 to 3 log response. After B1 block (cyclo, VP-16) the figures were 92%, 58%, and 83%, respectively. Twenty-five of 40 patients received allogeneic stem cell transplantation. Three-year event-free survival of the whole cohort was 37%, and the relapse rate was 38%. Three-year event-free survival by risk group was 53% for late, 34% for early, and 21% for very early relapses. An ALL marrow relapse nonresponsive to steroids, vincristine, asparaginase, anthracyclines, and alkylating agents is uncommon, and these classic drugs can still be advocated for induction of ALL relapse. The problems lie in creating a consolidation capable of preventing particularly posttransplant relapses. PMID:22246158

  19. Analysis of the efficacy and safety of a combined gemcitabine, oxaliplatin and pegaspargase regimen for NK/T-cell lymphoma

    Science.gov (United States)

    Xia, Zhong-jun; Huang, Hui-qiang; Jiang, Wen-qi; Lu, Yue

    2016-01-01

    Extranodal natural killer/T-cell lymphoma (ENKTL) is an aggressive neoplasm with a poor outcome. Novel L-asparaginase-based treatment regimens, such as GELOX (gemcitabine, oxaliplatin, and L-asparaginase) and P-gemox (gemcitabine, oxaliplatin, and pegaspargase), have shown promising results against stage IE/IIE ENKTL. To define the general applicability of P-gemox, in a retrospective analysis we examined the efficacy and safety of P-gemox in a cohort of 117 patients with newly diagnosed or relapsed/refractory ENKTL. Treatment included 2 to 8 cycles of P-gemox: intravenous gemcitabine (1250 mg/m2) and oxaliplatin (85 mg/m2) and intramuscular pegaspargase (2500 IU/m2) on day 1 and repeated every 2 weeks, or intravenous gemcitabine (1000 mg/m2) on days 1 and 8 and intravenous oxaliplatin (130 mg/m2) and intramuscular pegaspargase (2500 IU/m2) on day 1 and repeated every 3 weeks. Upon completion of treatment, the overall response rate was 88.8%, and responses were similar for newly diagnosed and relapsed/refractory patients. After a median follow-up of 17 months, the 3-year overall and progression-free survival rates were 72.7% and 57.8%, respectively. Multivariate analysis showed that CR after treatment was the most significant factor affecting survival. P-gemox thus appears to be an effective and well-tolerated treatment for patients with ENKTL. PMID:27072578

  20. Resveratrol induces cell cycle arrest and apoptosis in malignant NK cells via JAK2/STAT3 pathway inhibition.

    Directory of Open Access Journals (Sweden)

    Ly Quoc Trung

    Full Text Available Natural killer (NK cell malignancies, particularly aggressive NK cell leukaemias and lymphomas, have poor prognoses. Although recent regimens with L-asparaginase substantially improved outcomes, novel therapeutic approaches are still needed to enhance clinical response. Resveratrol, a naturally occurring polyphenol, has been extensively studied for its anti-inflammatory, cardioprotective and anti-cancer activities. In this study, we investigated the potential anti-tumour activities of resveratrol against the NK cell lines KHYG-1, NKL, NK-92 and NK-YS. Resveratrol induced robust G0/G1 cell cycle arrest, significantly suppressed cell proliferation and induced apoptosis in a dose- and time-dependent manner for all four cell lines. In addition, resveratrol suppressed constitutively active STAT3 in all the cell lines and inhibited JAK2 phosphorylation but had no effect on other upstream mediators of STAT3 activation, such as PTEN, TYK2, and JAK1. Resveratrol also induced downregulation of the anti-apoptotic proteins MCL1 and survivin, two downstream effectors of the STAT3 pathway. Finally, resveratrol induced synergistic effect on the apoptotic and antiproliferative activities of L-asparaginase against KHYG-1, NKL and NK-92 cells. These results suggest that resveratrol may have therapeutic potential against NK cell malignancies. Furthermore, our finding that resveratrol is a bonafide JAK2 inhibitor extends its potential benefits to other diseases with dysregulated JAK2 signaling.

  1. Hypersensitivity reactions to anticancer agents: Data mining of the public version of the FDA adverse event reporting system, AERS

    Directory of Open Access Journals (Sweden)

    Sakaeda Toshiyuki

    2011-10-01

    Full Text Available Abstract Background Previously, adverse event reports (AERs submitted to the US Food and Drug Administration (FDA database were reviewed to confirm platinum agent-associated hypersensitivity reactions. The present study was performed to confirm whether the database could suggest the hypersensitivity reactions caused by anticancer agents, paclitaxel, docetaxel, procarbazine, asparaginase, teniposide, and etoposide. Methods After a revision of arbitrary drug names and the deletion of duplicated submissions, AERs involving candidate agents were analyzed. The National Cancer Institute Common Terminology Criteria for Adverse Events version 4.0 was applied to evaluate the susceptibility to hypersensitivity reactions, and standardized official pharmacovigilance tools were used for quantitative detection of signals, i.e., drug-associated adverse events, including the proportional reporting ratio, the reporting odds ratio, the information component given by a Bayesian confidence propagation neural network, and the empirical Bayes geometric mean. Results Based on 1,644,220 AERs from 2004 to 2009, the signals were detected for paclitaxel-associated mild, severe, and lethal hypersensitivity reactions, and docetaxel-associated lethal reactions. However, the total number of adverse events occurring with procarbazine, asparaginase, teniposide, or etoposide was not large enough to detect signals. Conclusions The FDA's adverse event reporting system, AERS, and the data mining methods used herein are useful for confirming drug-associated adverse events, but the number of co-occurrences is an important factor in signal detection.

  2. Radiation sensitivity of fungal microflora isolated from some pharmaceutical ingredients

    International Nuclear Information System (INIS)

    The total number of fungal microflora of D-glucose, NaCl, KCl and their solutions was determined. The fungal isolates were identified as Aspergillus fumigatus. Aspergillus niger; Spicaria divaricate and Spicaria silvatica and their response to γ-radiation was determined, the most predominant isolate Asp. fumigatus was also the most irradiation resistant. The Dio and the lethal dose were determined for each isolate in a pure spore suspension as well as in the presence of the other isolates. The higher lethal dose values obtained for pure spore suspension as compared to that obtained for the natural fungal flora a D-glucose are discussed in terms of spore clumping. The activity of amylase, protease and L-asparaginase of Asp. fumigatus was examined prior to and after exposure to different doses of γ-radiation. Though all were inhibited at high doses, the effect was not as drastic as it was on cell viability

  3. Primary NK/T cell lymphoma nasal type of the colon

    Directory of Open Access Journals (Sweden)

    Ana María Chirife

    2013-02-01

    Full Text Available Since nasal NK/T-cell lymphoma and NK/T-cell lymphoma nasal type are rare diseases, colonic involvement has seldom been seen. We report a case of a patient with a primary NK/T-cell lymphoma nasal type of the colon. The patient had no history of malignant diseases and was diagnosed after exhaustive study in the context of fever of unknown origin. The first therapeutic approach followed the DAEPOCH-protocol: etoposide, prednisone, doxor-rubicin, vincristine and cyclophosphamide. The persistence of constitutional symptoms after the first treatment course motivated the switch to a second line following the SMILE-protocol: dexamethasone, metotrexate, ifosfamide, E.coli L-asparaginase, and etoposide. Despite intensive chemotherapy, the patient died 2 months after the diagnose of an extranodal NK/T-cell lymphoma of the colon and 4 months after the first symptomatic appearance of disease.

  4. Computational approach for enzymes present in Capsicum annuum: A review

    Directory of Open Access Journals (Sweden)

    Shivendu Ranjan

    2013-12-01

    Full Text Available Capasicum annuumor sweet bell pepper is one of the more economical and agriculturally viable vegetable grown all over the world owing to its antioxidant and other medicinal properties. This review highlights the essential enzymes present and its mode of action using bioinformatics online tools viz.uniprot, swissprot and Brenda enzyme db and ExPAsy protein databases. The enzymes viz. peroxidase, polyphenol oxidase, tyrosinase, catecholase, Pectin esterase, Catalase, 9-lipoxygenase, L-asparaginase, Polygalactouronase, Capsanthin and Ribulose-Phosphate 3-Epimerase contribute to its properties by various molecular mechanisms. Understanding of these mechanisms will be helpful for application of these enzymes in food processing and in the production of food ingredients. The increasing sophistication of processing industries creates a demand for abroad variety of enzymes with characteristics compatible with food processing conditions for e.g. shelf life of fruits and vegetables canbe increased by decreasing the levels of peroxidase and polyphenoloxidase

  5. PRODUCTION OF EXTRACELLULAR ENZYMES BY HALOPHILIC BACTERIA ISOLATED FROM SOLAR SALTERNS

    Directory of Open Access Journals (Sweden)

    Jhuma Biswas

    2013-12-01

    Full Text Available During the course of survey of halophilic microorganisms, a total of sixteen bacterial isolates were obtained from coastal solar salterns of Orissa and West Bengal, India. Morphological, physiological and biochemical characteristics of these isolates indicate that majority of them belong to the genus Halomonas, however, members belonging to Cobetia and Halococcus were not uncommon. These isolates were screened for the production of extracellular enzymes such as amylase, glutaminase, asparaginase, xylanase, cellulase, gelatinase, inulinase, caseinase, pectinase, urease and lipase. Among these hydrolytic enzymes, glutamine and asparagine hydrolytic activities were predominant, although lipid and casein degrading activities were not inferior. However, amylase and gelatinase production were rare. None of these halophiles was able to degrade cellulose, inulin, pectin and xylan and only one isolate was capable of hydrolyzing urea

  6. Asparagine Metabolic Pathways in Arabidopsis.

    Science.gov (United States)

    Gaufichon, Laure; Rothstein, Steven J; Suzuki, Akira

    2016-04-01

    Inorganic nitrogen in the form of ammonium is assimilated into asparagine via multiple steps involving glutamine synthetase (GS), glutamate synthase (GOGAT), aspartate aminotransferase (AspAT) and asparagine synthetase (AS) in Arabidopsis. The asparagine amide group is liberated by the reaction catalyzed by asparaginase (ASPG) and also the amino group of asparagine is released by asparagine aminotransferase (AsnAT) for use in the biosynthesis of amino acids. Asparagine plays a primary role in nitrogen recycling, storage and transport in developing and germinating seeds, as well as in vegetative and senescence organs. A small multigene family encodes isoenzymes of each step of asparagine metabolism in Arabidopsis, except for asparagine aminotransferase encoded by a single gene. The aim of this study is to highlight the structure of the genes and encoded enzyme proteins involved in asparagine metabolic pathways; the regulation and role of different isogenes; and kinetic and physiological properties of encoded enzymes in different tissues and developmental stages. PMID:26628609

  7. Consensus definitions of 14 severe acute toxic effects for childhood lymphoblastic leukaemia treatment

    DEFF Research Database (Denmark)

    Schmiegelow, Kjeld; Attarbaschi, Andishe; Barzilai, Shlomit;

    2016-01-01

    toxic effects, that no two protocols shared identical definitions of all toxic effects, and that no toxic effect definition was shared by all protocols. Using the Delphi method over three face-to-face plenary meetings, consensus definitions were obtained for all 14 toxic effects. In the overall......Although there are high survival rates for children with acute lymphoblastic leukaemia, their outcome is often counterbalanced by the burden of toxic effects. This is because reported frequencies vary widely across studies, partly because of diverse definitions of toxic effects. Using the Delphi...... method, 15 international childhood acute lymphoblastic leukaemia study groups assessed acute lymphoblastic leukaemia protocols to address toxic effects that were to be considered by the Ponte di Legno working group. 14 acute toxic effects (hypersensitivity to asparaginase, hyperlipidaemia, osteonecrosis...

  8. Thrombotic complications in children with non-Hodgkin lymphoma

    Directory of Open Access Journals (Sweden)

    N. V. Lipay

    2014-07-01

    Full Text Available Our study was aimed at identifying of risk factors of venous thrombosis (VT in children with non-Hodgkin lymphomas. VT episodes were registered in 13 of 174 children treated (7.5 %. Possible impact of morphological type, initial mediastinal involvement, gender, age and use of L-asparaginase as a risk factor of thrombosis development were analyzed. Using multivariate analysis primary mediastinal tumor (OR = 4.73 [CI: 1.42–17.10] and patient age older than 13 years (OR = 4.3 [CI: 1.19–20.28 were identified as prognostic factors of thrombosis development (р < 0,05.

  9. Thrombotic complications in children with non-Hodgkin lymphoma

    Directory of Open Access Journals (Sweden)

    N. V. Lipay

    2013-01-01

    Full Text Available Our study was aimed at identifying of risk factors of venous thrombosis (VT in children with non-Hodgkin lymphomas. VT episodes were registered in 13 of 174 children treated (7.5 %. Possible impact of morphological type, initial mediastinal involvement, gender, age and use of L-asparaginase as a risk factor of thrombosis development were analyzed. Using multivariate analysis primary mediastinal tumor (OR = 4.73 [CI: 1.42–17.10] and patient age older than 13 years (OR = 4.3 [CI: 1.19–20.28 were identified as prognostic factors of thrombosis development (р < 0,05.

  10. Current issues in dietary acrylamide:formation,mitigation and risk assessment

    DEFF Research Database (Denmark)

    Pedreschi, F.; Salome Mariotti, M.; Granby, Kit

    2014-01-01

    the investigation of AA precursors, mechanisms of AA formation and AA mitigation technologies in potato, cereal and coffee products. Additionally, most relevant issues of AA risk assessment are discussed. New technologies tested from laboratory to industrial scale face, as a major challenge, the reduction of AA...... content of browned food, while still maintaining its attractive organoleptic properties. Reducing sugars such as glucose and fructose are the major contributors to AA in potato-based products. On the other hand, the limiting substrate of AA formation in cereals and coffee is the free amino acid asparagine....... For some products the addition of glycine or asparaginase reduces AA formation during baking. Since, for potatoes, the limiting substrate is reducing sugars, increases in sugar content in potatoes during storage then introduce some difficulties and potentially quite large variations in the AA content...

  11. The complex clinical picture of presumably allergic side effects to cytostatic drugs: symptoms, pathomechanism, reexposure, and desensitization.

    Science.gov (United States)

    Pagani, Mauro

    2010-07-01

    The number of drugs used for the treatment of different types of cancers is constantly increasing and actually exceeds 100 distinct chemical formulations. The use of most cytotoxic agents is associated with potential hypersensitivity reactions, and the constant increase of their administration has caused an increase in incidence of these adverse effects, thus becoming a relevant problem for clinicians. Hypersensitivity reactions are common with platinum compounds, L-asparaginase, taxanes, procarbazine, and epipodophyllotoxins, whereas they are unusual, but always possible, with the other chemotherapeutic drugs. Reactions associated with individual drugs are discussed in detail. The mechanism underlying these hypersensitivity reactions involves IgE-mediated hypersensitivity reactions, nonallergic hypersensitivity reactions, and a few pathogenetically unclear reactions. More studies are needed to better understand, diagnose, treat, and prevent these reactions. To achieve this goal, a multidisciplinary approach to treat patients with cancer who have potential allergies is needed.

  12. Reversible posterior leukoencephalopathy syndrome in childhood: report of nine cases and review of the literature.

    Science.gov (United States)

    Gümüş, Hakan; Per, Hüseyin; Kumandaş, Sefer; Yikilmaz, Ali

    2010-04-01

    Reversible posterior leukoencephalopathy syndrome (RPLS) is recently described disorder with typical radiological findings in the posterior regions of the cerebral hemisphere and cerebellum. Its clinical symptoms include headache, decreased alertness, mental abnormalities, such as confusion, diminished spontaneity of speech, and changed behavior ranging from drowsiness to stupor, seizures, vomiting and abnormalities of visual perception like cortical blindness. RPLS is caused by various heterogeneous factors, the commonest being hypertension, followed by non-hypertensive causes such as eclampsia, renal diseases and immunosuppressive therapy. We presented nine patients with RPLS who had primary diagnoses such as acute post-streptococcal glomerulonephritis, idiopathic hypertension, the performing of intravenous immunoglobulin for infection with crescentic glomerulonephritis, erythrocyte transfusion for severe iron deficiency, L: -asparaginase treatment for acute lymphoblastic leukemia and performing of granulocyte-colony stimulating factor for ulcerative colitis due to neutropenia. Early recognition of RPLS as complication during different diseases and therapy in childhood may facilitate precise diagnosis and appropriate treatment. PMID:19809787

  13. DIVERSITY OF EXTREMOPHILIC MAGENSITE BACTERIA AND IT’S ENZYMATIC POTENTIAL

    Directory of Open Access Journals (Sweden)

    Ramya Suseenthar

    2012-11-01

    Full Text Available The present study focuses on the diversity of extremophilic bacteria from magnesite soil which are less explored and screened for the multitude of commercially important enzymes produced. Five different soil samples varying in color were collected from magnesite deposit Salem, Tamil Nadu. The soil samples had pH of 8-9, high nitrogen content, low phosphorous, potassium and trace elements. A remarkable reduction in total heterotrophic bacterial diversity of the ranging between 2.0x 107 CFU/g and 5.2x105 CFU/g. Generic composition of 25 isolates comprised of 28% Bacillus spp., followed by 20% Staphylococcus spp., 16% Enterobacter spp., 12% Pseudomonas spp., 12% Escherichia coli, 8% Micrococcus spp. and 4% Serratia spp. Maximum of 72% of isolates showed lipase activity followed by 68% L-asparaginase, 60% protease, 52% cellulase, 44% L-glutaminase, 20% amylase and 4% tyrosinase activity were observed. The dialysate of culture filtrate exhibited 27000 U/ml of lipase, 5393 U/ml and 5923 U/ml of L-asparaginase and 4074 U/ml of -amylase activity. The 16s rDNA sequence analysis of S5B2 showed 100% similarity with Bacillus pumilus (X8, S1B4 with 99% of Bacillus pumilus (X22 and 99% of S2B2 with Bacillus licheniformis (2J-1. This is the first report on the bacterial diversity from Magnesite soil with the potential to produce industrially robust enzymes for various biotechnological applications.

  14. Treatment of feline lymphoma using a 12-week, maintenance-free combination chemotherapy protocol in 26 cats.

    Science.gov (United States)

    Limmer, S; Eberle, N; Nerschbach, V; Nolte, I; Betz, D

    2016-08-01

    The aim of this prospective clinical trial was to investigate the efficacy and toxicity of a short-term, maintenance-free chemotherapy protocol in feline lymphoma. Twenty-six cats with confirmed diagnosis of high-/intermediate-grade lymphoma were treated with a 12-week protocol consisting of cyclic administration of l-asparaginase, vincristine, cyclophosphamide, doxorubicin and prednisolone. Complete (CR) and partial remission (PR) rates were 46 and 27%, respectively. Median duration of first CR was 394 days compared with a median PR duration of 41 days. No factor was identified to significantly influence the likelihood to reach CR. Overall survival amounted to 78 days (range: 9-2230 days). Median survival in CR cats was 454 days and in PR cats was 82 days. Toxicosis was mainly low grade with anorexia seen most frequently. In cats achieving CR, maintenance-free chemotherapy may be sufficient to attain long-term remission and survival. Factors aiding in prognosticating the likelihood for CR, strategies enhancing response and targeting chemotherapy-induced anorexia need to be identified in future. PMID:24548273

  15. Infections During Induction Therapy of Protocol CCLG-2008 in Childhood Acute Lymphoblastic Leukemia: A Single-center Experience with 256 Cases in China

    Directory of Open Access Journals (Sweden)

    Si-Dan Li

    2015-01-01

    Full Text Available Background: Infections remain a major cause of therapy-associated morbidity and mortality in children with acute lymphoblastic leukemia (ALL. Methods: We retrospectively analyzed the medical charts of 256 children treated for ALL under the CCLG-2008 protocol in Beijing Children′s Hospital. Results: There were 65 infectious complications in 50 patients during vincristine, daunorubicin, L-asparaginase and dexamethasone induction therapy, including microbiologically documented infections (n = 12; 18.5%, clinically documented infections (n = 23; 35.3% and fever of unknown origin (n = 30; 46.2%. Neutropenia was present in 83.1% of the infectious episodes. In all, most infections occurred around the 15 th day of induction treatment (n = 28, and no patients died of infection-associated complications. Conclusions: The infections in this study was independent of treatment response, minimal residual diseases at the end of induction therapy, gender, immunophenotype, infection at first visit, risk stratification at diagnosis, unfavorable karyotypes at diagnosis and morphologic type. The infection rate of CCLG-2008 induction therapy is low, and the outcome of patients is favorable.

  16. Final results of a single institution experience with a pediatric-based regimen, the augmented Berlin-Frankfurt-Münster, in adolescents and young adults with acute lymphoblastic leukemia, and comparison to the hyper-CVAD regimen.

    Science.gov (United States)

    Rytting, Michael E; Jabbour, Elias J; Jorgensen, Jeffrey L; Ravandi, Farhad; Franklin, Anna R; Kadia, Tapan M; Pemmaraju, Naveen; Daver, Naval G; Ferrajoli, Alessandra; Garcia-Manero, Guillermo; Konopleva, Marina Y; Borthakur, Gautam; Garris, Rebecca; Wang, Sa; Pierce, Sherry; Schroeder, Kurt; Kornblau, Steven M; Thomas, Deborah A; Cortes, Jorge E; O'Brien, Susan M; Kantarjian, Hagop M

    2016-08-01

    Several studies reported improved outcomes of adolescents and young adults (AYA) with acute lymphoblastic leukemia (ALL) treated with pediatric-based ALL regimens. This prompted the prospective investigation of a pediatric Augmented Berlin-Frankfurt-Münster (ABFM) regimen, and its comparison with hyper-fractionated cyclophosphamide, vincristine, Adriamycin, and dexamethasone (hyper-CVAD) in AYA patients. One hundred and six AYA patients (median age 22 years) with Philadelphia chromosome- (Ph) negative ALL received ABFM from October 2006 through March 2014. Their outcome was compared to 102 AYA patients (median age 27 years), treated with hyper-CVAD at our institution. The complete remission (CR) rate was 93% with ABFM and 98% with hyper-CVAD. The 5-year complete remission duration (CRD) were 53 and 55%, respectively (P = 0.98). The 5-year overall survival (OS) rates were 60 and 60%, respectively. The MRD status on Day 29 and Day 84 of therapy was predictive of long-term outcomes on both ABFM and hyper-CVAD. Severe regimen toxicities with ABFM included hepatotoxicity in 41%, pancreatitis in 11%, osteonecrosis in 9%, and thrombosis in 19%. Myelosuppression-associated complications were most significant with hyper-CVAD. In summary, ABFM and hyper-CVAD resulted in similar efficacy outcomes, but were associated with different toxicity profiles, asparaginase-related with ABFM and myelosuppression-related with hyper-CVAD. Am. J. Hematol. 91:819-823, 2016. © 2016 Wiley Periodicals, Inc. PMID:27178680

  17. Elevated temperature altered photosynthetic products in wheat seedlings and organic compounds and biological activity in rhizopshere soil under cadmium stress

    Science.gov (United States)

    Jia, Xia; Zhao, Yonghua; Wang, Wenke; He, Yunhua

    2015-09-01

    The objective of this study was to investigate the effects of slightly elevated atmospheric temperature in the spring on photosynthetic products in wheat seedlings and on organic compounds and biological activity in rhizosphere soil under cadmium (Cd) stress. Elevated temperature was associated with increased soluble sugars, reducing sugars, starch, and total sugars, and with decreased amino acids in wheat seedlings under Cd stress. Elevated temperature improved total soluble sugars, free amino acids, soluble phenolic acids, and organic acids in rhizosphere soil under Cd stress. The activity of amylase, phenol oxidase, invertase, β-glucosidase, and L-asparaginase in rhizosphere soil was significantly improved by elevated temperature under Cd stress; while cellulase, neutral phosphatase, and urease activity significantly decreased. Elevated temperature significantly improved bacteria, fungi, actinomycetes, and total microorganisms abundance and fluorescein diacetate activity under Cd stress. In conclusion, slightly elevated atmospheric temperature in the spring improved the carbohydrate levels in wheat seedlings and organic compounds and biological activity in rhizosphere soil under Cd stress in the short term. In addition, elevated atmospheric temperature in the spring stimulated available Cd by affecting pH, DOC, phenolic acids, and organic acids in rhizosphere soil, which resulted in the improvement of the Cd uptake by wheat seedlings.

  18. Infections During Induction Therapy of Protocol CCLG-2008 in Childhood Acute Lymphoblastic Leukemia: A Single-center Experience with 256 Cases in China

    Institute of Scientific and Technical Information of China (English)

    Si-Dan Li; Yong-Bing Chen; Zhi-Gang Li; Run-Hui Wu; Mao-Quan Qin; Xuan Zhou; Jin Jiang

    2015-01-01

    Background:Infections remain a major cause of therapy-associated morbidity and mortality in children with acute lymphoblastic leukemia (ALL).Methods:We retrospectively analyzed the medical charts of 256 children treated for ALL under the CCLG-2008 protocol in Beijing Children's Hospital.Results:There were 65 infectious complications in 50 patients during vincristine,daunorubicin,L-asparaginase and dexamethasone induction therapy,including microbiologically documented infections (n =12; 18.5%),clinically documented infections (n =23; 35.3%) and fever of unknown origin (n =30; 46.2%).Neutropenia was present in 83.1% of the infectious episodes.In all,most infections occurred around the 15t1h day of induction treatment (n =28),and no patients died of infection-associated complications.Conclusions:The infections in this study was independent of treatment response,minimal residual diseases at the end of induction therapy,gender,immunophenotype,infection at first visit,risk stratification at diagnosis,unfavorable karyotypes at diagnosis and morphologic type.The infection rate of CCLG-2008 induction therapy is low,and the outcome of patients is favorable.

  19. Towards a metabolic therapy of cancer?

    Science.gov (United States)

    Chiu, Martina; Ottaviani, Laura; Bianchi, Massimiliano G; Franchi-Gazzola, Renata; Bussolati, Ovidio

    2012-12-01

    It is increasingly appreciated that cancer cells must be endowed with specific metabolic adaptations to support enhanced growth and to ensure survival under stressful conditions. On the other hand, many oncogenic mutations of protooncogenes and tumor suppressor genes directly cause metabolic derangements and, conversely, mutations of enzymes have been found to underlie several forms of cancer. Thus, cancer-specific metabolic alterations are now considered among the hallmarks of malignant tumors. Most commonly, cancer cells exhibit enhanced glycolysis under aerobic conditions (the Warburg effect) but alterations in the metabolism of amino acids, such as glutamine, serine and proline are increasingly described as important metabolic features of selected tumor types. In theory, all these deranged cancer-specific metabolic pathways may constitute novel therapeutic targets, although the only "metabolic" drug in clinical use is still represented by the enzyme L-asparaginase. However, the increasing amount of experimental evidence, as well as the number of trials in progress, suggests that metabolic drugs will soon complement standard anti-cancer chemotherapy and modern biological drugs. PMID:23762991

  20. IMPACT OF PRE-TREATMENTS ON THE ACRYLAMIDE FORMATION AND ORGANOLEPTIC EVOLUTION OF FRIED POTATO CHIPS

    Directory of Open Access Journals (Sweden)

    Samir Abdel-Monem Ahmed Ismial

    2013-01-01

    Full Text Available The main objective of this investigation was to study the effect of different pre-frying treatments on reduction of acrylamide formation of fried potato Moreover; the impact of different phenol compounds and leaves on acrylamide formation was evaluated. In addition, the effects of these treatments on the sensorial quality of fried potato chips were studied. Results showed that blanching process caused significant decreases in acrylamide content of fried potato. The highest decrease was observed for those samples blanched in MgCl2 (0.1 M, L-cysteine (0.05 M and 0.01 M of citric acid solutions, 97.97, 97.17 and 93.43%, respectively. Soaking of potato slices in water or different solutions significantly reduced the formation of acrylamide. The decreases in acrylamide content ranged from 61.61 to 97.47%. Soaking in crude, semi-purified asparaginase solutions, blanching in hot water plus immersing in the enzyme solutions and soaking in phenolic acid solutions caused significant reduction in the formation of acrylamide of potato chips. Addition of fresh leaves into frying oil significantly influenced acrylamide formation. Oregano, rosemary, bamboo, guava and olive leaves caused the greatest reductions. Potato slices blanched in distilled water at 65°C, NaCl, Mg Cl2 and 0.1 M glutamine had significantly the highest scores of overall acceptability.

  1. Successful therapy of convoluted T-lymphoblastic lymphoma in the adult

    International Nuclear Information System (INIS)

    Fifteen adult patients with biopsy-proven convoluted T-lymphoblastic lymphoma were treated with an aggressive regimen, modified from the LSA2-L2 protocol used for childhood lymphoma. The treatment schema consisted of induction phase, including cyclophosphamide, vincristine, prednisone, adriamycin, and 2000 rads to mediastinum, as well as intrathecal methotrexate. Consolidation phase included cytosine arabinoside, 6-thioguanine, L-asparaginase, and CCNU, along with cranial irradiation and further intrathecal methotrexate. Maintenance consisted of cyclical chemotherapy and intrathecal methotrexate, continuing for a total of 3 yr. Median age in the group was 25 yr (range 16-73). There were 8 males and 7 females. At diagnosis, 9 patients had mediastinal involvement, and 9 had bone marrow involvement. Five of these demonstrated malignant cells in the peripheral blood. Complete clinical response was attained in 11 patients. Three patients achieved partial response. Four complete responders have relapsed, 1 in the central nervous system at 6 mo. and 1 in nodal sites at 3 mo, 1 in multiple sites at 24 mo. and 1 in bone marrow at 42 mo while off all chemotherapy for 6 mos. At this time, median survival of all patients is 28.3 mo. and median relapse-free survival is 21 mo. The median survival for complete responders in excess of 71 mo. while the median relapse-free survival for this group is 41 mo

  2. Five-year analysis from phase 2 trial of "sandwich" chemoradiotherapy in newly diagnosed, stage IE to IIE, nasal type, extranodal natural killer/T-cell lymphoma.

    Science.gov (United States)

    Zhang, Li; Jiang, Ming; Xie, Li; Zhang, Hong; Jiang, Yu; Yang, Qun-pei; Liu, Wei-ping; Zhang, Wen-yan; Zhuo, Hong-yu; Li, Ping; Chen, Nian-yong; Zhao, Sha; Wang, Feng; Zou, Li-qun

    2016-01-01

    The "sandwich" protocol, was first proposed by us and comprised of l-asparaginase, vincristine, and prednisone chemotherapy with radiotherapy, results in 2-year overall survival and progression-free survival rates that surpass traditional therapies for patients with newly diagnosed, stage IE-IIE, nasal type, extranodal natural killer/T-cell lymphoma. The results had been published by cancer. These patients were followed up over a median period of 67 months, for which updates and the results of prognostic factors analyses are presented. The 5-year overall survival and progress-free survival rates were both 64%. The highest rates of death occurred during the first 6 months, and between the second and third year after enrollment. The initial therapeutic response (odds ratio = 5.83; P = 0.001) and B symptoms (odds ratio = 6.13; P = 0.043) were significant prognostic factors for overall survival. However, the international prognostic index was not significant for progress-free survival and overall survival. There were no severe long-term side effects. These results indicate that the "sandwich" protocol may benefit the long-term survival of patients with newly diagnosed stage IE-IIE, nasal type, extranodal natural killer/T-cell lymphoma. However, additional studies with larger samples are required to confirm these results. This study is registered at www.Chictr.org (ChicTR-TNC-09000394). PMID:26633585

  3. Fanconi Syndrome: A Rare Initial Presentation of Acute Lymphoblastic Leukemia.

    Science.gov (United States)

    Sahu, Kamal Kant; Law, Arjun Datt; Jain, Nidhi; Khadwal, Alka; Suri, Vikas; Malhotra, Pankaj; Varma, Subhash Chander

    2016-06-01

    A-14-year old boy, presented with a short history of excessive thirst and increased urine output. Clinical examination showed pallor, generalized lymphadenopathy and hepatosplenomegaly. For evaluation of his polyuric state he underwent routine laboratory investigations, including renal function test, acid-base studies, urine analysis. Blood tests suggested hypokalemia, hypouricemia, hypocalcemia and hyperchloremia with normal liver and kidney function tests. The arterial blood gas analysis was suggestive of normal anion gap metabolic acidosis. Urine analysis was suggestive of hyperuricosuria, hypercalciuria and glycosuria with a positive urine anion gap. His hemogram showed pancytopenia with differential count showing 88% blasts. Bone marrow examination and flowcytometry confirmed the diagnosis of B cell acute lymphoblastic leukemia. Hence this case was atypical and very interesting in the sense that the Fanconi syndrome is very rare to be an initial presenting feature of acute lymphoblastic leukemia. The patient was started on oral as well intravenous supplementation with potassium, bicarbonate, calcium and phosphorus. Simultaneously, as per the modified BFM -90 protocol (four drug based regimen-Prednisolone, vincristine, daunorubicin, cyclophosphamide along with l-asparaginase), he was started on induction protocol. By the end of 3rd week of induction therapy, his urine output started normalizing and finally settled at the end of induction therapy. At present he is in the maintenance phase of chemotherapy. PMID:27408343

  4. Lasting engraftment of histoincompatible bone marrow cells in dogs

    Energy Technology Data Exchange (ETDEWEB)

    Vriesendorp, H.M.; Klapwijk, W.M.; van Kessel, A.M.C.; Zurcher, C.; van Bekkum, D.W.

    1981-05-01

    Conditioning protocols were tested for their efficacy in increasng the incidence of engraftment of histoincompatible dog bone marrow cells. Cyclophosphamide and total body irradiation (TBI), Corynebacterium parvum and TBI, a 3- or 5-day delayed transfusion of bone marrow cells after TBI, or an increase in the number of donor bone marrow cells or lymphocytes appeared to be ineffective. These protocols were previously reported to promote recovery of splenic hemopoiesis in mice in short-term assays. The noted discrepancy between studies with mice and dogs invalidated allogeneic resistance as measured in the mouse spleen assay as a model for bone marrow allograft rejection. Intravenous treatment with silica particles or L-asparaginase did improve the engraftment rate after 7.5 Gy TBI. Low efficiency and significant extra toxicity restrict the applicability of these procedures. The most promising conditioning schedule found appeared to be two fractions of 6.0 Gy TBI separated by a 72-h interval. Prolonged survival was noted after transplantation of bone marrow cells from a one-DLA haplotype-mismatched donor. Possibilities for further improvement of this protocol are discussed.

  5. Lasting engraftment of histoincompatible bone marrow cells in dogs

    Energy Technology Data Exchange (ETDEWEB)

    Vriesendorp, H.M.; Klapwijk, W.M.; van Kessel, A.M.; Zurcher, C.; van Bekkum, D.W.

    1981-05-01

    Conditioning protocols were tested for their efficacy in increasing the incidence of engraftment of histoincompatible dog bone marrow cells. Cyclophosphamide and total body irradation (TBI), Corynebacterium parvum and TBI, a 3- or 5-day delayed transfusion of bone marrow cells after TBI, or an increase in the number of donor bone marrow cells or lymphocytes appeared to be ineffective. These protocols were previously reported to promote recovery of splenic hemopoiesis in mice in short-term assays. The noted discrepancy between studies with mice and dogs invalidated allogeneic resistance as measured in the mouse spleen assay as a model for bone marrow allograft rejection. Intravenous treatment with silica particles or L-asparaginase did improve the engraftment rate after 7.5 Gy TBI. Low efficiency and significant extra toxicity restrict the applicability of these procedures. The most promising conditioning schedule found appeared to be two fractions of 6.0 Gy TBI separated by a 72-hr interval. Prolonged survival was noted after transplantation of bone marrow cells from a one-DLA haplo-type-mismatched donor. Possibilities for further improvement of this protocol are discussed.

  6. Biotechnology development for biomedical applications.

    Energy Technology Data Exchange (ETDEWEB)

    Kuehl, Michael; Brozik, Susan Marie; Rogers, David Michael; Rempe, Susan L.; Abhyankar, Vinay V.; Hatch, Anson V.; Dirk, Shawn M.; Hedberg-Dirk, Elizabeth (University of New Mexico, Albuquerque, NM); Sukharev, Sergei (University of Maryland, College Park, MD); Anishken, Andriy (University of Maryland, College Park, MD); Cicotte, Kirsten; De Sapio, Vincent; Buerger, Stephen P.; Mai, Junyu

    2010-11-01

    Sandia's scientific and engineering expertise in the fields of computational biology, high-performance prosthetic limbs, biodetection, and bioinformatics has been applied to specific problems at the forefront of cancer research. Molecular modeling was employed to design stable mutations of the enzyme L-asparaginase with improved selectivity for asparagine over other amino acids with the potential for improved cancer chemotherapy. New electrospun polymer composites with improved electrical conductivity and mechanical compliance have been demonstrated with the promise of direct interfacing between the peripheral nervous system and the control electronics of advanced prosthetics. The capture of rare circulating tumor cells has been demonstrated on a microfluidic chip produced with a versatile fabrication processes capable of integration with existing lab-on-a-chip and biosensor technology. And software tools have been developed to increase the calculation speed of clustered heat maps for the display of relationships in large arrays of protein data. All these projects were carried out in collaboration with researchers at the University of Texas M. D. Anderson Cancer Center in Houston, TX.

  7. Posterior reversible encephalopathy syndrome in leukemic children: a sensitive issue.

    Science.gov (United States)

    Kridis, Wala Ben; Mdhaffer, Moez; Hentati, Yosr; Kammoun, Fatma; Milad, Abir; Haddar, Sondes; Mahfoudh, Khaireddine Ben; Triki, Chahinez; Elloumi, Moez

    2015-01-01

    Posterior reversible encephalopathy syndrome (PRES) is an acute central nervous system disorder characterized by reversible brain vasogenic edema. We report here a new case of a nine-year-old boy with B-cell acute lymphoblastic leukemia (B-ALL) who developed PRES secondary to induction chemotherapy including dexamethasone (dexamethasone®), vincristine (oncovin(®)), daunorubicin (adriblastine(®)) and intrathecal injection. Cerebral magnetic resonance imaging (MRI) showed high signal intensity on T2 at cortical and sub cortical region of parieto-frontal and parieto-occipital lobes. The patient was put under sodium valproate (depakine(®)) and we decided to continue dexamethasone (dexamethasone(®)) and daunorubicin (adriblastine(®)) injection. The MRI, after four weeks, was normal. So, we resumed vincristine (oncovin(®)) and we started L-asparaginase injections. Then, the outcome was favorable. The treatment of PRES is based on the withdrawal of the triggering factor to avoid the risk of irreversible lesions. But, due to the severity of leukemia the discontinuation of chemotherapy is difficult because of the risk of disease progression. PMID:24919742

  8. Clofarabine for the treatment of adult acute lymphoid leukemia: the Group for Research on Adult Acute Lymphoblastic Leukemia intergroup.

    Science.gov (United States)

    Huguet, Françoise; Leguay, Thibaut; Raffoux, Emmanuel; Rousselot, Philippe; Vey, Norbert; Pigneux, Arnaud; Ifrah, Norbert; Dombret, Hervé

    2015-04-01

    Clofarabine, a second-generation purine analog displaying potent inhibition of DNA synthesis and favorable pharmacologic profile, is approved for the treatment of acute lymphoblastic leukemia (ALL) after failure of at least two previous regimens in patients up to 21 years of age at diagnosis. Good neurologic tolerance, synergy with alkylating agents, management guidelines defined through pediatric ALL and adult acute myeloid leukemia, have also prompted its administration in more than 100 adults with Philadelphia chromosome-positive and negative B lineage and T lineage ALL, as single agent (40 mg/m(2)/ day for 5 days), or in combination. In a Group for Research on Adult Acute Lympho- blastic Leukemia (GRAALL) retrospective study of two regimens (clofarabine ± cyclophosphamide + / - etoposide (ENDEVOL) ± mitoxantrone ± asparaginase ± dexamethasone (VANDEVOL)), remission was achieved in 50% of 55 relapsed/refractory patients, and 17-35% could proceed to allogeneic stem cell. Clofarabine warrants further exploration in advanced ALL treatment and bridge-to-transplant. PMID:24996442

  9. Enzymes approved for human therapy: indications, mechanisms and adverse effects.

    Science.gov (United States)

    Baldo, Brian A

    2015-02-01

    Research and drug developments fostered under orphan drug product development programs have greatly assisted the introduction of efficient and safe enzyme-based therapies for a range of rare disorders. The introduction and regulatory approval of 20 different recombinant enzymes has enabled, often for the first time, effective enzyme-replacement therapy for some lysosomal storage disorders, including Gaucher (imiglucerase, taliglucerase, and velaglucerase), Fabry (agalsidase alfa and beta), and Pompe (alglucosidase alfa) diseases and mucopolysaccharidoses I (laronidase), II (idursulfase), IVA (elosulfase), and VI (galsulfase). Approved recombinant enzymes are also now used as therapy for myocardial infarction (alteplase, reteplase, and tenecteplase), cystic fibrosis (dornase alfa), chronic gout (pegloticase), tumor lysis syndrome (rasburicase), leukemia (L-asparaginase), some collagen-based disorders such as Dupuytren's contracture (collagenase), severe combined immunodeficiency disease (pegademase bovine), detoxification of methotrexate (glucarpidase), and vitreomacular adhesion (ocriplasmin). The development of these efficacious and safe enzyme-based therapies has occurred hand in hand with some remarkable advances in the preparation of the often specifically designed recombinant enzymes; the manufacturing expertise necessary for commercial production; our understanding of underlying mechanisms operative in the different diseases; and the mechanisms of action of the relevant recombinant enzymes. Together with information on these mechanisms, safety findings recorded so far on the various adverse events and problems of immunogenicity of the recombinant enzymes used for therapy are presented.

  10. L-type amino-acid transporter 1 (LAT1): a therapeutic target supporting growth and survival of T-cell lymphoblastic lymphoma/T-cell acute lymphoblastic leukemia.

    Science.gov (United States)

    Rosilio, C; Nebout, M; Imbert, V; Griessinger, E; Neffati, Z; Benadiba, J; Hagenbeek, T; Spits, H; Reverso, J; Ambrosetti, D; Michiels, J-F; Bailly-Maitre, B; Endou, H; Wempe, M F; Peyron, J-F

    2015-06-01

    The altered metabolism of cancer cells is a treasure trove to discover new antitumoral strategies. The gene (SLC7A5) encoding system L amino-acid transporter 1 (LAT1) is overexpressed in murine lymphoma cells generated via T-cell deletion of the pten tumor suppressor, and also in human T-cell acute lymphoblastic leukemia (T-ALL)/lymphoma (T-LL) cells. We show here that a potent and LAT1 selective inhibitor (JPH203) decreased leukemic cell viability and proliferation, and induced transient autophagy followed by apoptosis. JPH203 could also alter the in vivo growth of luciferase-expressing-tPTEN-/- cells xenografted into nude mice. In contrast, JPH203 was nontoxic to normal murine thymocytes and human peripheral blood lymphocytes. JPH203 interfered with constitutive activation of mTORC1 and Akt, decreased expression of c-myc and triggered an unfolded protein response mediated by the C/EBP homologous protein (CHOP) transcription factor associated with cell death. A JPH203-resistant tPTEN-/-clone appeared CHOP induction deficient. We also demonstrate that targeting LAT1 may be an efficient broad spectrum adjuvant approach to treat deadly T-cell malignancies as the molecule synergized with rapamycin, dexamethasone, doxorubicin, velcade and l-asparaginase to alter leukemic cell viability. PMID:25482130

  11. Purification and characterization of a glutaminase enzyme accounting for the majority of glutaminase activity in Aspergillus sojae under solid-state culture.

    Science.gov (United States)

    Ito, Kotaro; Hanya, Yoshiki; Koyama, Yasuji

    2013-10-01

    Glutaminase, an enzyme that hydrolyzes L-glutamine to L-glutamate, plays an important role in the production of fermented foods by enhancing the umami taste. In this study, we found ten glutaminase genes in the Aspergillus sojae genome by conducting a BLAST search of the characterized glutaminase sequence. We subsequently constructed glutaminase gene disruptants. The glutaminase activity of the gahB disruptant was decreased by approximately 90 % in A. sojae and Aspergillus oryzae, indicating that this enzyme (GahB) accounted for the majority of the glutaminase activity in Aspergillus species. Subsequently, GahB protein was purified from the AsgahB-overexpressing transformant and characterized. The molecular mass was estimated to be approximately 110 and 259 kDa by SDS-PAGE and gel filtration chromatography, respectively, indicating that the native form of AsGahB was a dimer. The optimal pH was 9.0, and the optimal temperature was 50 °C. Analysis of substrate specificity revealed that AsGahB had peptidoglutaminase-asparaginase activity, similar to AsGahA, but preferred free L-glutamine to free L-asparagine, C-terminal glutaminyl, and asparaginyl residues in peptides.

  12. The periplasmic enzyme, AnsB, of Shigella flexneri modulates bacterial adherence to host epithelial cells.

    Directory of Open Access Journals (Sweden)

    Divya T George

    Full Text Available S. flexneri strains, most frequently linked with endemic outbreaks of shigellosis, invade the colonic and rectal epithelium of their host and cause severe tissue damage. Here we have attempted to elucidate the contribution of the periplasmic enzyme, L-asparaginase (AnsB to the pathogenesis of S. flexneri. Using a reverse genetic approach we found that ansB mutants showed reduced adherence to epithelial cells in vitro and attenuation in two in vivo models of shigellosis, the Caenorhabditis elegans and the murine pulmonary model. To investigate how AnsB affects bacterial adherence, we compared the proteomes of the ansB mutant with its wild type parental strain using two dimensional differential in-gel electrophoresis and identified the outer membrane protein, OmpA as up-regulated in ansB mutant cells. Bacterial OmpA, is a prominent outer membrane protein whose activity has been found to be required for bacterial pathogenesis. Overexpression of OmpA in wild type S. flexneri serotype 3b resulted in decreasing the adherence of this virulent strain, suggesting that the up-regulation of OmpA in ansB mutants contributes to the reduced adherence of this mutant strain. The data presented here is the first report that links the metabolic enzyme AnsB to S. flexneri pathogenesis.

  13. 重组人源胰岛素原C肽在大肠杆菌中的克隆、表达与纯化%Cloning,expression and purification of recombinant human proinsulin C-peptide in E.coli

    Institute of Scientific and Technical Information of China (English)

    王学军; 顾凯; 曹荣月; 林洁; 吴洁; 刘景晶

    2006-01-01

    目的:构建一种新型表达载体(pEDCC),表达并纯化得到人源胰岛素原C肽.方法:将编码截短的门冬酰胺酶突变体(ansB-C),天然C肽,人IgG1铰链区(hinge),额外的酸敏感二肽(DP)以及富含碱性氨基酸的8肽(KRKRKKSR)的核苷酸序列依次分别插入pET28a载体中,构建表达载体pEDCC.在乳糖的诱导下,融合蛋白ansB-C-hinge-DPKRKRKKSRNGSGR-C-peptide以包涵体形式高效表达.融合蛋白经洗涤和乙醇分级沉淀纯化后,通过酸水解将PKRKRKKSRNGSGR-C-peptide释放出来.C肽N端14肽用胰蛋白酶切割,通过DE52柱与C-peptide分离.结果:构建的表达载体pEDCC序列正确,融合蛋白经分离纯化得到了高纯度的重组人源胰岛素原C肽.结论:以截短的门冬酰胺酶作为融合伙伴,并以富含碱性氨基酸的8肽调节等电点是一种生产重组人源胰岛素原C肽的有效方法.%Aim:To obtain recombinant human proinsulin C-peptide,a novel expression vector pEDCC was constructed to facilitate the expression and purification of C-peptide. Methods:Gene fragments encoding a truncated asparaginase fragment mutant,native C-peptide,a hinge fragment of human IgG1,an extra acid-labile dipeptide and a basic-amino-acid-riched octopeptide were introduced in turn into plasmid pET28a. The fusion protein ansB-C-hinge-DPKRKRKKSRNGSGR-C-peptide was expressed effectively as inclusion bodies after induced by lactose and partially purified by means of washing and ethanol fractionation. After being hydrolyzed,the polypeptide PKRKRKKSRNGSGR-C-peptide was liberated from the fusion partner. The N-terminal tetradecapeptide extension of C-peptide was subsequently cleaved by trypsin and removed by DE52 column. Results:The nucleotides sequence of plasmid pEDCC was confirmed to be identical with that of designed fusion protein. Recombinant human proinsulin C-peptide was obtained with high purity after purification. Conclusion:Employing truncated asparaginase as the fusion partner and basic

  14. Preparation and characterization of silk fibroin as a biomaterial with potential for drug delivery

    Directory of Open Access Journals (Sweden)

    Zhang Hao

    2012-06-01

    Full Text Available Abstract Background Degummed silk fibroin from Bombyx mori (silkworm has potential carrier capabilities for drug delivery in humans; however, the processing methods have yet to be comparatively analyzed to determine the differential effects on the silk protein properties, including crystalline structure and activity. Methods In this study, we treated degummed silk with four kinds of calcium-alcohol solutions, and performed secondary structure measurements and enzyme activity test to distinguish the differences between the regenerated fibroins and degummed silk fibroin. Results Gel electrophoresis analysis revealed that Ca(NO32-methanol, Ca(NO32-ethanol, or CaCl2-methanol treatments produced more lower molecular weights of silk fibroin than CaCl2-ethanol. X-ray diffraction and Fourier-transform infrared spectroscopy showed that CaCl2-ethanol produced a crystalline structure with more silk I (α-form, type II β-turn, while the other treatments produced more silk II (β-form, anti-parallel β-pleated sheet. Solid-State 13C cross polarization and magic angle spinning-nuclear magnetic resonance measurements suggested that regenerated fibroins from CaCl2-ethanol were nearly identical to degummed silk fibroin, while the other treatments produced fibroins with significantly different chemical shifts. Finally, enzyme activity test indicated that silk fibroins from CaCl2-ethanol had higher activity when linked to a known chemotherapeutic drug, L-asparaginase, than the fibroins from other treatments. Conclusions Collectively, these results suggest that the CaCl2-ethanol processing method produces silk fibroin with biomaterial properties that are appropriate for drug delivery.

  15. Central Nervous System Disease in Hematological Malignancies: Historical Perspective and Practical Applications

    Science.gov (United States)

    Pui, Ching-Hon; Thiel, Eckhard

    2009-01-01

    Acute lymphoblastic leukemia (ALL) 5-year survival rates are approaching 90% in children and 50% in adults who are receiving contemporary risk-directed treatment protocols. Current efforts focus not only on further improving cure rate but also on patient quality of life. Hence, all protocols decrease or limit the use of cranial irradiation as central nervous system (CNS)-directed therapy, even in patients with high-risk presenting features, such as the presence of leukemia cells in the cerebrospinal fluid (even resulting from traumatic lumbar puncture), adverse genetic features, T-cell immunophenotype, and a large leukemia-cell burden. Current strategies for CNS-directed therapy involve effective systemic chemotherapy (eg, dexamethasone, high-dose methotrexate, intensive asparaginase, ifosfamide) and early intensification and optimization of intrathecal therapy. Options under investigation for the treatment of relapsed or refractory CNS leukemia in ALL patients include thiotepa and intrathecal liposomal cytarabine. CNS involvement in non-Hodgkin’s lymphoma (NHL) is associated with young age, advanced stage, number of extranodal sites, elevated lactate dehydrogenase, and International Prognostic Index score. Refractory CNS lymphoma in patients with NHL carries a poor prognosis, with a median survival of 2 to 6 months; the most promising treatment, autologous stem cell transplant, can extend median survival from 10 to 26 months. CNS prophylaxis is required during the initial treatment of NHL subtypes that carry a high risk of CNS relapse, such as B-cell ALL, Burkitt’s lymphoma, and lymphoblastic lymphoma. The use of CNS prophylaxis in the treatment of diffuse large B-cell lymphoma is controversial because of the low risk of CNS relapse (~5%) in this population. In this article, we review current and past practice of intrathecal therapy in ALL and NHL and the risk-models that aim to identify predictors of CNS relapse in NHL. PMID:19660680

  16. Treatment of Childhood Acute Lymphoblastic Leukemia Without Prophylactic Cranial Irradiation

    Science.gov (United States)

    Pui, Ching-Hon; Campana, Dario; Pei, Deqing; Bowman, W. Paul; Sandlund, John T.; Kaste, Sue C.; Ribeiro, Raul C.; Rubnitz, Jeffrey E.; Raimondi, Susana C.; Onciu, Mihaela; Coustan-Smith, Elaine; Kun, Larry E.; Jeha, Sima; Cheng, Cheng; Howard, Scott C.; Simmons, Vickey; Bayles, Amy; Metzger, Monika L.; Boyett, James M.; Leung, Wing; Handgretinger, Rupert; Downing, James R.; Evans, William E.; Relling, Mary V.

    2009-01-01

    Background We conducted a clinical trial to test whether prophylactic cranial irradiation could be omitted in all children with newly diagnosed acute lymphoblastic leukemia. Methods A total of 498 evaluable patients were enrolled. Treatment intensity was based on presenting features and the level of minimal residual disease after remission induction treatment. Continuous complete remission was compared between the 71 patients who previously would have received prophylactic cranial irradiation and the 56 historical controls who received it. Results The 5-year event-free and overall survival probabilities (95% confidence interval) for all 498 patients were 85.6% (79.9% to 91.3%) and 93.5% (89.8% to 97.2%), respectively. The 5-year cumulative risk of isolated central-nervous-system (CNS) relapse was 2.7% (1.1% to 4.2%), and that of any CNS relapse (isolated plus combined) was 3.9% (1.9% to 5.9%). The 71 patients had significantly better continuous complete remission than the 56 historical controls (P=0.04). All 11 patients with isolated CNS relapse remain in second remission for 0.4 to 5.5 years. CNS leukemia (CNS-3 status) or a traumatic lumbar puncture with blasts at diagnosis and a high level of minimal residual disease (≥ 1%) after 6 weeks of remission induction were significantly associated with poorer event-free survival. Risk factors for CNS relapse included the presence of the t(1;19)[TCF3-PBX1], any CNS involvement at diagnosis, and T-cell immunophenotype. Common adverse effects included allergic reactions to L-asparaginase, osteonecrosis, thrombosis, and disseminated fungal infection. Conclusions With effective risk-adjusted chemotherapy, prophylactic cranial irradiation can be safely omitted in the treatment of childhood acute lymphoblastic leukemia. PMID:19553647

  17. Improved Prognosis for Older Adolescents With Acute Lymphoblastic Leukemia

    Science.gov (United States)

    Pui, Ching-Hon; Pei, Deqing; Campana, Dario; Bowman, W. Paul; Sandlund, John T.; Kaste, Sue C.; Ribeiro, Raul C.; Rubnitz, Jeffrey E.; Coustan-Smith, Elaine; Jeha, Sima; Cheng, Cheng; Metzger, Monika L.; Bhojwani, Deepa; Inaba, Hiroto; Raimondi, Susana C.; Onciu, Mihaela; Howard, Scott C.; Leung, Wing; Downing, James R.; Evans, William E.; Relling, Mary V.

    2011-01-01

    Purpose The prognosis for older adolescents and young adults with acute lymphoblastic leukemia (ALL) has been historically much worse than that for younger patients. We reviewed the outcome of older adolescents (age 15 to 18 years) treated in four consecutive Total Therapy studies to determine if recent improved treatment extended to this high-risk group. Patients and Methods Between 1991 and 2007, 963 pediatric patients, including 89 older adolescents, were enrolled on Total Therapy studies XIIIA, XIIIB, XIV, and XV. In the first three studies, treatment selection was based on presenting clinical features and leukemic cell genetics. In study XV, the level of residual disease was used to guide treatment, which featured intensive methotrexate, glucocorticoid, vincristine, and asparaginase, as well as early triple intrathecal therapy for higher-risk ALL. Results The 89 older adolescents were significantly more likely to have T-cell ALL, the t(4;11)(MLL-AF4), and detectable minimal residual disease during or at the end of remission induction; they were less likely to have the t(12;21)(ETV6-RUNX1) compared with younger patients. In the first three studies, the 44 older adolescents had significantly poorer event-free survival and overall survival than the 403 younger patients. This gap in prognosis was abolished in study XV: event-free survival rates at 5 years were 86.4% ± 5.2% (standard error) for the 45 older adolescents and 87.4% ± 1.7% for the 453 younger patients; overall survival rates were 87.9% ± 5.1% versus 94.1% ± 1.2%, respectively. Conclusion Most older adolescents with ALL can be cured with risk-adjusted intensive chemotherapy without stem-cell transplantation. PMID:21172890

  18. Acrylamide mitigation strategies: critical appraisal of the FoodDrinkEurope toolbox.

    Science.gov (United States)

    Palermo, M; Gökmen, V; De Meulenaer, B; Ciesarová, Z; Zhang, Y; Pedreschi, F; Fogliano, V

    2016-06-15

    FoodDrinkEurope Federation recently released the latest version of the Acrylamide Toolbox to support manufacturers in acrylamide reduction activities giving indication about the possible mitigation strategies. The Toolbox is intended for small and medium size enterprises with limited R&D resources, however no comments about the pro and cons of the different measures were provided to advise the potential users. Experts of the field are aware that not all the strategies proposed have equal value in terms of efficacy and cost/benefit ratio. This consideration prompted us to provide a qualitative science-based ranking of the mitigation strategies proposed in the acrylamide Toolbox, focusing on bakery and fried potato products. Five authors from different geographical areas having a publication record on acrylamide mitigation strategies worked independently ranking the efficacy of the acrylamide mitigation strategies taking into account three key parameters: (i) reduction rate; (ii) side effects; and (iii) applicability and economic impact. On the basis of their own experience and considering selected literature of the last ten years, the authors scored for each key parameter the acrylamide mitigation strategies proposed in the Toolbox. As expected, all strategies selected in the Toolbox turned out to be useful, however, not at the same level. The use of enzyme asparaginase and the selection of low sugar varieties were considered the best mitigation strategies in bakery and in potato products, respectively. According to authors' opinion most of the other mitigation strategies, although effective, either have relevant side effects on the sensory profile of the products, or they are not easy to implement in industrial production. The final outcome was a science based commented ranking which can enrich the acrylamide Toolbox supporting individual manufacturer in taking the best actions to reduce the acrylamide content in their specific production context. PMID:26666890

  19. Arginase treatment prevents the recovery of canine lymphoma and osteosarcoma cells resistant to the toxic effects of prolonged arginine deprivation.

    Directory of Open Access Journals (Sweden)

    James W Wells

    Full Text Available Rapidly growing tumor cells require a nutrient-rich environment in order to thrive, therefore, restricting access to certain key amino acids, such as arginine, often results in the death of malignant cells, which frequently display defective cell cycle check-point control. Healthy cells, by contrast, become quiescent and remain viable under arginine restriction, displaying full recovery upon return to arginine-rich conditions. The use of arginase therapy to restrict available arginine for selectively targeting malignant cells is currently under investigation in human clinical trials. However, the suitability of this approach for veterinary uses is unexplored. As a prelude to in vivo studies in canine malignancies, we examined the in vitro effects of arginine-deprivation on canine lymphoid and osteosarcoma cell lines. Two lymphoid and 2 osteosarcoma cell lines were unable to recover following 6 days of arginine deprivation, but all remaining cell lines displayed full recovery upon return to arginine-rich culture conditions. These remaining cell lines all proved susceptible to cell death following the addition of arginase to the cultures. The lymphoid lines were particularly sensitive to arginase, becoming unrecoverable after just 3 days of treatment. Two of the osteosarcoma lines were also susceptible over this time-frame; however the other 3 lines required 6-8 days of arginase treatment to prevent recovery. In contrast, adult progenitor cells from the bone marrow of a healthy dog were able to recover fully following 9 days of culture in arginase. Over 3 days in culture, arginase was more effective than asparaginase in inducing the death of lymphoid lines. These results strongly suggest that short-term arginase treatment warrants further investigation as a therapy for lymphoid malignancies and osteosarcomas in dogs.

  20. Extranodal NK/T-Cell Lymphoma, Nasal Type, Presenting as a Breast Mass.

    Science.gov (United States)

    Rahal, Ahmad; Reddy, Pavan S; Alvares, Carmelita

    2015-01-01

    Extranodal natural killer/T-cell lymphoma, nasal type, is a rare type of non-Hodgkin cell lymphoma endemic to East Asia and parts of Central and South America. In most cases, it is driven by Epstein-Barr virus infections, with a broad range of morphologic appearances, frequent necrosis, and angioinvasion. It is designated as NK/T reflecting uncertainty in its cellular origins. These tumors usually arise in the nasal region, typically presenting with symptoms of nasal obstruction, epistaxis, and/or a destructive mass involving the nose, sinuses, or palate. The treatment of patients with extranodal NK/T-cell lymphoma, nasal type, is largely determined by the extent of disease. Localized disease is usually treated with radiation and chemotherapy. The disseminated disease requires combination chemotherapy. This report describes the case of a 30-year-old Caucasian female presenting with a left breast mass of two months duration. Excisional biopsy was done, and the pathological exam confirmed the diagnosis of extranodal NK/T-cell lymphoma, nasal type. Our patient received a systemic combination chemotherapy with steroid (dexamethasone), methotrexate, ifosfamide, L-asparaginase, and etoposide (SMILE) regimen, resulting in a complete clinical and radiological remission. On the basis of our review of the literature, extranodal NK/T non-Hodgkin cell lymphoma, nasal type, presenting as a breast mass is very rare and very uncommon in the United States. Awareness of this occurrence may be valuable as this case may be a forerunner of additional similar cases developing in the future. PMID:26824008

  1. Elevated atmospheric CO2 affected photosynthetic products in wheat seedlings and biological activity in rhizosphere soil under cadmium stress.

    Science.gov (United States)

    Jia, Xia; Liu, Tuo; Zhao, Yonghua; He, Yunhua; Yang, Mingyan

    2016-01-01

    The objective of this study was to investigate the effects of elevated CO2 (700 ± 23 μmol mol(-1)) on photosynthetic products in wheat seedlings and on organic compounds and biological activity in rhizosphere soil under cadmium (Cd) stress. Elevated CO2 was associated with decreased quantities of reducing sugars, starch, and soluble amino acids, and with increased quantities of soluble sugars, total sugars, and soluble proteins in wheat seedlings under Cd stress. The contents of total soluble sugars, total free amino acids, total soluble phenolic acids, and total organic acids in the rhizosphere soil under Cd stress were improved by elevated CO2. Compared to Cd stress alone, the activity of amylase, phenol oxidase, urease, L-asparaginase, β-glucosidase, neutral phosphatase, and fluorescein diacetate increased under elevated CO2 in combination with Cd stress; only cellulase activity decreased. Bacterial abundance in rhizosphere soil was stimulated by elevated CO2 at low Cd concentrations (1.31-5.31 mg Cd kg(-1) dry soil). Actinomycetes, total microbial abundance, and fungi decreased under the combined conditions at 5.31-10.31 mg Cd kg(-1) dry soil. In conclusion, increased production of soluble sugars, total sugars, and proteins in wheat seedlings under elevated CO2 + Cd stress led to greater quantities of organic compounds in the rhizosphere soil relative to seedlings grown under Cd stress only. Elevated CO2 concentrations could moderate the effects of heavy metal pollution on enzyme activity and microorganism abundance in rhizosphere soils, thus improving soil fertility and the microecological rhizosphere environment of wheat under Cd stress.

  2. Rituximab in B-Lineage Adult Acute Lymphoblastic Leukemia.

    Science.gov (United States)

    Maury, Sébastien; Chevret, Sylvie; Thomas, Xavier; Heim, Dominik; Leguay, Thibaut; Huguet, Françoise; Chevallier, Patrice; Hunault, Mathilde; Boissel, Nicolas; Escoffre-Barbe, Martine; Hess, Urs; Vey, Norbert; Pignon, Jean-Michel; Braun, Thorsten; Marolleau, Jean-Pierre; Cahn, Jean-Yves; Chalandon, Yves; Lhéritier, Véronique; Beldjord, Kheira; Béné, Marie C; Ifrah, Norbert; Dombret, Hervé

    2016-09-15

    Background Treatment with rituximab has improved the outcome for patients with non-Hodgkin's lymphoma. Patients with B-lineage acute lymphoblastic leukemia (ALL) may also have the CD20 antigen, which is targeted by rituximab. Although single-group studies suggest that adding rituximab to chemotherapy could improve the outcome in such patients, this hypothesis has not been tested in a randomized trial. Methods We randomly assigned adults (18 to 59 years of age) with CD20-positive, Philadelphia chromosome (Ph)-negative ALL to receive chemotherapy with or without rituximab, with event-free survival as the primary end point. Rituximab was given during all treatment phases, for a total of 16 to 18 infusions. Results From May 2006 through April 2014, a total of 209 patients were enrolled: 105 in the rituximab group and 104 in the control group. After a median follow-up of 30 months, event-free survival was longer in the rituximab group than in the control group (hazard ratio, 0.66; 95% confidence interval [CI], 0.45 to 0.98; P=0.04); the estimated 2-year event-free survival rates were 65% (95% CI, 56 to 75) and 52% (95% CI, 43 to 63), respectively. Treatment with rituximab remained associated with longer event-free survival in a multivariate analysis. The overall incidence rate of severe adverse events did not differ significantly between the two groups, but fewer allergic reactions to asparaginase were observed in the rituximab group. Conclusions Adding rituximab to the ALL chemotherapy protocol improved the outcome for younger adults with CD20-positive, Ph-negative ALL. (Funded by the Regional Clinical Research Office, Paris, and others; ClinicalTrials.gov number, NCT00327678 .). PMID:27626518

  3. Pharmacokinetic modeling of an induction regimen for in vivo combined testing of novel drugs against pediatric acute lymphoblastic leukemia xenografts.

    Directory of Open Access Journals (Sweden)

    Barbara Szymanska

    Full Text Available Current regimens for induction therapy of pediatric acute lymphoblastic leukemia (ALL, or for re-induction post relapse, use a combination of vincristine (VCR, a glucocorticoid, and L-asparaginase (ASP with or without an anthracycline. With cure rates now approximately 80%, robust pre-clinical models are necessary to prioritize active new drugs for clinical trials in relapsed/refractory patients, and the ability of these models to predict synergy/antagonism with established therapy is an essential attribute. In this study, we report optimization of an induction-type regimen by combining VCR, dexamethasone (DEX and ASP (VXL against ALL xenograft models established from patient biopsies in immune-deficient mice. We demonstrate that the VXL combination was synergistic in vitro against leukemia cell lines as well as in vivo against ALL xenografts. In vivo, VXL treatment caused delays in progression of individual xenografts ranging from 22 to >146 days. The median progression delay of xenografts derived from long-term surviving patients was 2-fold greater than that of xenografts derived from patients who died of their disease. Pharmacokinetic analysis revealed that systemic DEX exposure in mice increased 2-fold when administered in combination with VCR and ASP, consistent with clinical findings, which may contribute to the observed synergy between the 3 drugs. Finally, as proof-of-principle we tested the in vivo efficacy of combining VXL with either the Bcl-2/Bcl-xL/Bcl-w inhibitor, ABT-737, or arsenic trioxide to provide evidence of a robust in vivo platform to prioritize new drugs for clinical trials in children with relapsed/refractory ALL.

  4. Treatment of isolated testicular relapse in childhood acute lymphoblastic leukemia: an Italian multicenter study. Associazione Italiana Ematologia ed Oncologia Pediatrica.

    Science.gov (United States)

    Uderzo, C; Grazia Zurlo, M; Adamoli, L; Zanesco, L; Aricò, M; Calculli, G; Comelli, A; Cordero di Montezemolo, L; Di Tullio, M T; Guazzelli, C

    1990-04-01

    Between May 1980 and April 1987, 49 children with acute lymphoblastic leukemia (ALL) in isolated testicular and first leukemia relapse (ITR) were enrolled in the Associazione Italiana Ematologia ed Oncologia Pediatrica (AIEOP) multicenter study REC80-ITR. According to the Rome Workshop criteria, 77% were at standard and 23% at high initial prognostic risk. In 33% of the cases, ITR occurred during first treatment. The REC80-ITR protocol consisted of an induction phase regimen of vincristine (VCR), cytarabine (ARA-C), methotrexate (MTX), and asparaginase (L-asp), and bilateral testicular irradiation, and CNS prophylaxis with intrathecal MTX and a maintenance phase with a multidrug rotating regimen. Total treatment duration was 30 months. The median time of observation after ITR was 51 months. The Kaplan-Meier estimates of survival and disease-free survival (DFS) at 4 years were 67.7% and 41%, respectively. Patients who had an ITR on therapy or within the first off-therapy year showed the poorest outcome. The DFS at 3 years was 20%, 47.6%, and 100%, respectively, for children who had an ITR on treatment (n = 16), within the first year of treatment withdrawal (n = 22), or later (n = 10) (P = .001). Patients with an asymptomatic occult testicular infiltrate at treatment discontinuation had a very unfavorable prognosis. Eighty-one percent of second relapses involved the bone marrow. In our experience, children presenting an early ITR (ie, within 6 months of treatment withdrawal) need a very aggressive treatment because of the high probability of an underlying systemic disease. On the other hand, patients with a late ITR seem to have a truly local recurrence and can apparently be cured by standard protocols, as shown in protocol REC80-ITR.

  5. Cloning, expression, and purification of a recombinant Tat-HA-NR2B9c peptide.

    Science.gov (United States)

    Zhou, Hai-Hui; Zhang, Ai-Xia; Zhang, Yu; Zhu, Dong-Ya

    2012-10-01

    To design a peptide disrupting the interaction between N-methyl-d-aspartate receptors-2B (NR2B) and postsynaptic density protein-95 (PSD-95), a gene fragment encoding a chimeric peptide was constructed using polymerase chain reaction and ligated into a novel expression vector for recombinant expression in a T7 RNA polymerase-based expression system. The chimeric peptide contained a fragment of the cell membrane transduction domain of the human immunodeficiency virus type1 (HIV-1) Tat, a influenza virus hemagglutinin (HA) epitope-tag, and the C-terminal 9 amino acids of NR2B (NR2B9c). We named the chimeric peptide Tat-HA-NR2B9c. The expression plasmid contained a gene fragment encoding the Tat-HA-NR2B9c was ligated to the C-terminal fragment of l-asparaginase (AnsB-C) via a unique acid labile Asp-Pro linker. The recombinant fusion protein was expressed in inclusion body in Escherichia coli under isopropyl β-d-1-thiogalactopyranoside (IPTG) and purified by washing with 2M urea, solubilizing in 4M urea, and then ethanol precipitation. The target chimeric peptide Tat-HA-NR2B9c was released from the fusion partner following acid hydrolysis and purified by isoelectric point precipitation and ultrafiltration. SDS-PAGE analysis and MALDI-TOF-MS analysis showed that the purified Tat-HA-NR2B9c was highly homogeneous. Furthermore, we investigated the effects of Tat-HA-NR2B9c on ischemia-induced cerebral injury in the rats subjected to middle cerebral artery occlusion (MCAO) and reperfusion, and found that the peptide reduced infarct size and improved neurological functions. PMID:22944204

  6. A therapeutic trial of decitabine and vorinostat in combination with chemotherapy for relapsed/refractory acute lymphoblastic leukemia.

    Science.gov (United States)

    Burke, Michael J; Lamba, Jatinder K; Pounds, Stanley; Cao, Xueyuan; Ghodke-Puranik, Yogita; Lindgren, Bruce R; Weigel, Brenda J; Verneris, Michael R; Miller, Jeffrey S

    2014-09-01

    DNA hypermethylation and histone deacetylation are pathways of leukemia resistance. We investigated the tolerability and efficacy of decitabine and vorinostat plus chemotherapy in relapse/refractory acute lymphoblastic leukemia (ALL). Decitabine (15 mg/m(2) iv) and vorinostat (230 mg/m(2) PO div BID) were given days 1-4 followed by vincristine, prednisone, PEG-asparaginase, and doxorubicin. Genome wide methylation profiles were performed in 8 matched patient bone marrow (BM) samples taken at day 0 and day 5 (postdecitabine). The median age was 16 (range, 3-54) years. All patients had a prior BM relapse, with five relapsing after allogeneic transplant. The most common nonhematological toxicities possibly related to decitabine or vorinostat were infection with neutropenia (grade 3; n = 4) and fever/neutropenia (grade 3, n = 4; grade 4, n = 1). Of the 13 eligible patients, four achieved complete remission without platelet recovery (CRp), two partial response (PR), one stable disease (SD), one progressive disease (PD), two deaths on study and three patients who did not have end of therapy disease evaluations for an overall response rate of 46.2% (CRp + PR). Following decitabine, significant genome-wide hypo-methylation was observed. Comparison of clinical responders with nonresponders identified methylation profiles of clinical and biological relevance. Decitabine and vorinostat followed by re-Induction chemotherapy was tolerable and demonstrated clinical benefit in relapsed patients with ALL. Methylation differences were identified between responders and nonresponders indicating interpatient variation, which could impact clinical outcome. This study was registered at www.clinicaltrials.gov as NCT00882206.

  7. Acrylamide formation and antioxidant level in biscuits related to recipe and baking.

    Science.gov (United States)

    Haase, N U; Grothe, K-H; Matthäus, B; Vosmann, K; Lindhauer, M G

    2012-08-01

    Heated plant foods may contain compounds with adverse health effects (e.g. acrylamide). On the other hand, health-promoting compounds (e.g. antioxidants) have also been identified in such foods. Therefore, a baking experiment with biscuits was carried out to study the potential impact of both acrylamide and antioxidants in that food. Two different wheat flour types - wholemeal (WMF) and white flour type 550 (T550; 0.55% mineral content) - as well as recipe (fat and leavening agent) and thermal input (temperature × time) were changed. Furthermore, the effect of an enzymatic asparagine hydrolysation was tested. Antioxidants were determined with two independent procedures ABTS - (2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate)) and FRAP-assay (ferric reducing ability of plasma). WMF samples resulted in an unchanged acrylamide level, but in a significantly higher antioxidant concentration when compared with T550 samples (149 and 141 µg kg(-1) acrylamide and 9.1 and 5.1 mmol TE kg(-1) FW ABTS for WMF and T550, respectively). A reduced fat content yielded in an increased volume. Raising agents had no effect on acrylamide levels, but antioxidants were higher in samples with sodium bicarbonate (SBC) than with ammonium bicarbonate (ABC). Thermal input (temperature × time; 150°C × 25 min to 240°C × 9 min) indicated an exponential acrylamide increase especially at higher temperatures (from 75 to 236 µg kg(-1)), whereas antioxidant increase was linear (from 7.0 to 7.7 mmol TE kg(-1) FW, ABTS). FRAP and ABTS values were correlated on a low level, whereas acrylamide content of biscuits was correlated with FRAP and lightness (R (2 )= 0.62 and 0.47, and 0.71 and 0.85 for WMF and T550, respectively). The enzyme asparaginase reduced acrylamide formation by about 50% for both raising agents (SBC and ABC, respectively), whereas antioxidant levels were not affected. An evaluation of recipe variants with low acrylamide and high

  8. Changes in Free Amino Acid Concentration in Rye Grain in Response to Nitrogen and Sulfur Availability, and Expression Analysis of Genes Involved in Asparagine Metabolism.

    Science.gov (United States)

    Postles, Jennifer; Curtis, Tanya Y; Powers, Stephen J; Elmore, J S; Mottram, Donald S; Halford, Nigel G

    2016-01-01

    Free asparagine plays a central role in nitrogen storage and transport in many plant species due to its relatively high ratio of nitrogen to carbon. However, it is also a precursor for acrylamide, a Class 2a carcinogen that forms during high-temperature processing and cooking. The concentration of free asparagine was shown to increase by approximately 70% in rye grain in response to severe sulfur deficiency (F-test, p = 0.004), while the concentration of both free asparagine and free glutamine increased (by almost threefold and approximately 62%, respectively) in response to nitrogen application (F-test, p supply on other free amino acids: The concentration of free proline, for example, showed a significant (F-test, p = 0.019) effect of nitrogen interacting with sulfur, with the highest concentration occurring when the plants were deprived of both nitrogen and sulfur. Polymerase chain reaction products for several genes involved in asparagine metabolism and its regulation were amplified from rye grain cDNA. These genes were asparagine synthetase-1 (ScASN1), glutamine synthetase-1 (ScGS1), potassium-dependent asparaginase (ScASP), aspartate kinase (ScASK), and general control non-derepressible-2 (ScGCN2). The expression of these genes and of a previously described sucrose non-fermenting-1-related protein kinase-1 gene (ScSnRK1) was analyzed in flag leaf and developing grain in response to nitrogen and sulfur supply, revealing a significant (F-test, p supply on ScGS1 expression in the grain at 21 days post-anthesis. There was also evidence of an effect of sulfur deficiency on ScASN1 gene expression. However, although this effect was large (almost 10-fold) it was only marginally statistically significant (F-test, 0.05 < p < 0.10). The study reinforced the conclusion that nutrient availability can have a profound impact on the concentrations of different free amino acids, something that is often overlooked by plant physiologists but which has important implications for

  9. Identification of genes differentially expressed during interaction of Mexican lime tree infected with "Candidatus Phytoplasma aurantifolia"

    Directory of Open Access Journals (Sweden)

    Nekouei Mojtaba

    2011-01-01

    Full Text Available Abstract Background "Candidatus Phytoplasma aurantifolia", is the causative agent of witches' broom disease in Mexican lime trees (Citrus aurantifolia L., and is responsible for major losses of Mexican lime trees in Southern Iran and Oman. The pathogen is strictly biotrophic, and thus is completely dependent on living host cells for its survival. The molecular basis of compatibility and disease development in this system is poorly understood. Therefore, we have applied a cDNA- amplified fragment length polymorphism (AFLP approach to analyze gene expression in Mexican lime trees infected by "Ca. Phytoplasma aurantifolia". Results We carried out cDNA-AFLP analysis on grafted infected Mexican lime trees of the susceptible cultivar at the representative symptoms stage. Selective amplifications with 43 primer combinations allowed the visualisation of 55 transcript-derived fragments that were expressed differentially between infected and non-infected leaves. We sequenced 51 fragments, 36 of which were identified as lime tree transcripts after homology searching. Of the 36 genes, 70.5% were down-regulated during infection and could be classified into various functional groups. We showed that Mexican lime tree genes that were homologous to known resistance genes tended to be repressed in response to infection. These included the genes for modifier of snc1 and autophagy protein 5. Furthermore, down-regulation of genes involved in metabolism, transcription, transport and cytoskeleton was observed, which included the genes for formin, importin β 3, transducin, L-asparaginase, glycerophosphoryl diester phosphodiesterase, and RNA polymerase β. In contrast, genes that encoded a proline-rich protein, ubiquitin-protein ligase, phosphatidyl glycerol specific phospholipase C-like, and serine/threonine-protein kinase were up-regulated during the infection. Conclusion The present study identifies a number of candidate genes that might be involved in the

  10. Research progress of T cell lymphoma: reports from 2014 international conference on T cell lymphoma in clinical treatment%T细胞淋巴瘤研究新进展:2014年国际T细胞淋巴瘤临床治疗大会报道

    Institute of Scientific and Technical Information of China (English)

    马军; 朱军; 石远凯; 姜文奇; 黄慧强; 邱林

    2014-01-01

    The treatment status and progress in T cell lymphoma including epigenetic involved mutations that control DNA and histone methylation were reported and intensively discussed in 2014 international T cell lymphoma forum.According to the theory,treatment with HDAC inhibitor belinostat and romidepsin for peripheral T cell lymphoma (PTCL) can achieve 29 %-38 % overall response rate (ORR) and 13.6 months median relief time.NK/T cell lymphoma in southeast asia is a common malignant lymphoma,15 %-28 % of the NHL accounted in China and Japan for,which is significantly higher than that in the European and American countries,mainly related to EB virus widespread infection.L-asparaginase enzymes,such as SMILE and AspMetDex,can make the early cases with more than 70 % long-term survival rate,advanced cases with 40 % response rate.Some new drugs,such as pralatrexate,combined with romidepsin can be used in PTCL cases to improve the complete remission rate.%2014年国际T细胞淋巴瘤临床大会主要报告了T细胞淋巴瘤的治疗状况及进展,其中包括靶向表观遗传学在T细胞淋巴瘤中的应用.有研究发现在外周T细胞淋巴瘤(PTCL)中控制DNA和组蛋白甲基化的基因存在突变.根据这一研究结果应用组蛋白去乙酰化酶(HDAC)抑制剂belinostat和romidepsin可使PTCL获得29% ~ 38%的总反应率,中位缓解时间为13.6个月.NK/T细胞淋巴瘤是东南亚国家常见的恶性淋巴瘤,在中国和日本占非霍奇金淋巴瘤的15%~ 28%,明显高于欧美国家,主要与亚洲人群EB病毒感染率高有关.应用含左旋门冬酰胺酶的方案如SMILE和AspMetDex方案,可使早期病例长期生存率超过70%,晚期病例可达到近40%.对于PTCL可以将新药如普拉曲沙、罗咪酯肽等联合应用,从而提高完全缓解率.

  11. Evaluation of certain food additives.

    Science.gov (United States)

    2009-01-01

    This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, including flavouring agents, with a view to recommending acceptable daily intakes (ADIs) and to preparing specifications for identity and purity. The first part of the report contains a general discussion of the principles governing the toxicological evaluation and assessment of intake of food additives (in particular, flavouring agents). A summary follows of the Committee's evaluations of technical, toxicological and intake data for certain food additives (asparaginase from Aspergillus niger expressed in A. niger, calcium lignosulfonate (40-65), ethyl lauroyl arginate, paprika extract, phospholipase C expressed in Pichia pastoris, phytosterols, phytostanols and their esters, polydimethylsiloxane, steviol glycosides and sulfites [assessment of dietary exposure]) and 10 groups of related flavouring agents (aliphatic branched-chain saturated and unsaturated alcohols, aldehydes, acids and related esters; aliphatic linear alpha,beta-unsaturated aldehydes, acids and related alcohols, acetals and esters; aliphatic secondary alcohols, ketones and related esters; alkoxy-substituted allylbenzenes present in foods and essential oils and used as flavouring agents; esters of aliphatic acyclic primary alcohols with aliphatic linear saturated carboxylic acids; furan-substituted aliphatic hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids and related esters, sulfides, disulfides and ethers; miscellaneous nitrogen-containing substances; monocyclic and bicyclic secondary alcohols, ketones and related esters; hydroxy- and alkoxy-substituted benzyl derivatives; and substances structurally related to menthol). Specifications for the following food additives were revised: canthaxanthin; carob bean gum and carob bean gum (clarified); chlorophyllin copper complexes, sodium and potassium salts; Fast Green FCF; guar gum and guar gum (clarified

  12. Drug sensitivities of extranodal NK/T-cell Lymphoma cells cultured in different culture systems%结外NK/T细胞淋巴瘤敏感药物筛选

    Institute of Scientific and Technical Information of China (English)

    刘蕾; 郭宏强; 杨树军

    2015-01-01

    目的 应用无血清培养基(serum free medium,SFM)初步富集肿瘤耐药细胞,筛选出结外NK/T细胞淋巴瘤(extranodal NK/T-celllymphoma,ENKTCL)的敏感药物.方法 应用加入10%人血清和700 U/mL人白细胞介素-2(interlukin-2,IL-2)的1640培养基和加入700 U/mL人IL-2的SFM VIVO-15TM,分别培养SNK-6细胞;MTT法分别检测2种培养体系中表柔比星(adriamycin,ADM)、奥沙利铂(oxaliplatin,L-OHP)、吉西他滨(gemcitabine,GEM)、左旋门冬酰胺酶(L-Asparaginase,L-ASP)、阿糖胞苷(cytosine arabinoside,Ara-C)和甲氨蝶呤(methotrexate,MTX)等药物作用的IC50;2种培养体系中分别加入上述各种药物作用32 h后,7AAD/PE-Annexin Ⅴ染色法流式细胞术检测各处理组细胞的凋亡率,并分析比较.结果 无血清培养体系中细胞部分悬浮生长,每个悬浮球由>50个数目不等的细胞组成,悬浮球体积较有血清中明显增大.有血清和SFM中IC50含量,ADM分别为(0.12±0.018)和(0.751±0.14) μg/mL,P<0.001;Ara-C分别为(0.217±0.002)和(0.822±0.028) μg/mL,P<0.001; MTX分别为(0.023±0.001)和(0.032±0.002) μg/mL,P=0.002.药物作用后,有血清和SFM中SNK-6细胞的凋亡率,ADM分别为(45.23±2.58)%和(30.91±2.13)%,P=0.041;Ara-C分别为(56.12±2.32)%和(41.47±2.46)%,P=0.034;MTX分别为(24.42±1.92)%和(13.01±2.28)%,P=0.045;GEM分别为(15.05±2.05)%和(41.45±1.74)%,P<0.001.结论 SFM VIVO-15TM可用于ENKTCL SNK-6细胞培养;无血清培养后的SNK-6细胞对ADM、Ara-C和MTX耐药性增强,对L-OHP、GEM和L-ASP敏感.%OBJECTIVE To enrich tumor-initiating cells from serum free medium (SFM) supplemented with human interleukin-2 (IL-2).To screen the chemosensitivity of drugs for extranodal NK/T-cell lymphoma (ENKTCL) in vitro.METHODS SNK-6 cells were cultured in 10% human serum culture medium and SFM VIVO-15TM supplemented with human IL-2.The growth status of cells cultured in two kinds of system were closely observed and recorded

  13. SMILE方案治疗复发难治NK/T细胞淋巴瘤的初步临床报告%Preliminary outcomes of patients with relapsed or refractory NK/T-cell lymphoma treated by SMILE regimen

    Institute of Scientific and Technical Information of China (English)

    周颖; 蔡清清; 林旭滨; 高岩; 卜庆; 王潇潇; 黄慧强

    2009-01-01

    目的 探索SMILE方案治疗NK/T细胞淋巴瘤的疗效和不良反应.方法 2006年11月至2008年2月,5例初治和5例复发NK/T细胞淋巴瘤患者接受SMILE方案(甲氨蝶呤、异环磷酰胺、左旋门冬酰胺酶、依托泊苷等)化疗.1例患者进一步接受了自体外周血造血干细胞支持下的超大剂量化疗,2例患者进一步接受局部放疗.结果 10例患者中有8例可以评价疗效,总有效率50%(4/8),无完全缓解.其中初治和复发患者有效率均为50%.主要不良反应为骨髓抑制及氨基转移酶升高,其中Ⅲ~Ⅳ度粒细胞减少占65%,发热性粒细胞减少占25%,Ⅲ度氨基转移酶升高占10%,其他不良反应少见,无治疗相关死亡.26.1%患者由于严重的不良反应中止治疗.结论 SMILE治疗复发耐药的NK/T细胞淋巴瘤有一定疗效,但不良反应明显,目前尚不能作为复发难治NK/T细胞淋巴瘤的标准一线方案.%Objective To evaluate the efficacy and toxicity of SMILE regimen for NK/T-cell lymphoma. Methods From November 2006 to February 2008, 5 patients with relapsed and 5 with first treatment NK/T-cell lymphoma were involved in this study. These patients were treated with SMILE regimen including methotrexate, isofosfamide, L-asparaginase and etoposide.1 patient were treated with autolognus hematopoietic stem cell transplantation (AHSCT), and 2 patients received local regional radiation following SMILE. Results Among 10 patients, 8 were eligible to response evaluation. The overall response rate for whole group was 50 %(4/8) without complete remission. The overall response rate for both previously untreated and relapsed patients were 50 %(2/4). Major toxicity were bone marrow supression and transient transaminase elevation, the incidence of grade Ⅲ -Ⅳ neutroponia was 65 %, and febrile neutropenia was 25 %, Grade Ⅲ transaminase elevation was 10 %. Other toxicities were mild, no treatment-related mortality occurred. 26.1% cycles discontinued due to

  14. 成人Ph染色体阴性急性淋巴细胞白血病患者预后因素分析%Prognostic factors of adult Philadelphia chromosome negative acute lymphoblastic leukemia

    Institute of Scientific and Technical Information of China (English)

    王婧; 黄晓军; 江滨; 贾晋松; 杨申淼; 鲍立; 江浩; 路瑾; 主鸿鹄

    2015-01-01

    一个或一个以上不良预后因素的患者,采用allo-HSCT更具生存优势.%Objective To analyze the prognostic factors in adult Philadelphia chromosome negative acute lymphoblastic leukaemia (Ph ALL).Methods From December 1999 to December 2013,353 consecutive hospitalized 18-65-year-old adult Ph-ALL patients were retrospectively analyzed.Induction therapy was CODP±L-asparaginase (L-Asp) regimen,and consolidation therapy included CODP and high dose methotrexate or revised Hyper-CVAD A and B regimens for 8 cycles.178 patients (50.4%) performed allo-HSCT after three to five cycles of consolidation treatment,and 172 patients didn't receive allo-HSCT.The median follow-up was 39.9 months (2.0 to 171.0 months) for the 184 survivors.Results Three patients (0.85%) happened early death.CR rate after the first cycle of induction chemotherapy was 77.4% (271/350) among evaluated 350 patients.Overall CR rate was 92.9% (325/350).WBC ≥100.0× 109/L (P=0.010) and hepatomegaly/splenomegaly/lymphadenopathy (P=0.036) were independent adverse factors for overall CR.Among the 325 CR patients,117 patients developed relapse,cumulative incidence of relapse (CIR) at 5 years was 43.2%,disease-free survival (DFS) and overall survival (OS) rates at 5 years were 44.7% and 45.6% respectively.Multivariate analysis showed that harboring central nervous system leukaemia (CNSL) at diagnose (P=0.004,P=0.002,P< 0.001,respectively),induction regimen without L-Asp (P=0.023,P=0.009,P=0.004,respectively),time to CR more than 4 weeks (P=0.034,P=0.024,P=0.003,respectively),and non-allo-HSCT (P < 0.001,P < 0.001,P < 0.001,respectively) were adverse factors of relapse,DFS and OS.In addition,high WBC count at diagnosis (≥30.0 × 109/L for B lineage and ≥ 100.0× 109/L for T lineage) was poor factor of DFS (P=0.044).Based on the four adverse prognostic factors of DFS above mentioned (including WBC at diagnose,harboring CNSL at diagnose,induction regimen with or without L

  15. Pegasparaginase as ifrst-line treatment of children with leukemia and lymphoma%培门冬酶一线治疗儿童淋巴系统肿瘤的临床研究

    Institute of Scientific and Technical Information of China (English)

    王宏胜; 翟晓文; 陆凤娟; 李军; 苗慧; 钱晓文; 朱晓华; 吴玥

    2014-01-01

    Background and purpose: L-asparaginase (L-Asp) is an important drug in the treatment of childhood lymphoid neoplasms at present, but a lot of adverse reactions of L-Asp were observed. Pegasparaginase (PEG-Asp) is available in China in recent years. This study aimed to explore efifcacy and side-effect of PEG-Asp as ifrst-line treatment in childhood acute lymphoblastic leukemia (ALL) and lymphoblastic lymphoma (LBL). Methods:A total number of 211 ALL or LBL patients were treated with CCLG 2008 or BFM-90 protocol with PEG-Asp or L-Asp between Apr. 2008 and Mar. 2013;42 patients, among whom, were 35 ALL patients and 7 LBL patients, were treated with PEG-Asp as ifrst-line treatment;169 patients were treated with L-Asp as ifrst-line treatment (including 53 patients treated with L-Asp during induction protocol; with PEG-Asp during consolidate protocol). The clinical outcome and adverse reaction of PEG-Asp with L-Asp were observe and compared. Results: There were 35 ALL patients in PEG-Asp ifrst-line treatment group and the complete remission rate after 1 course of PEG-Asp was 97.1%,however, which was 83.3%of high risk ALL patients. The complete remission rate of 7 LBL patients of PEG-Asp ifrst-line treatment group was 57.1%. There was no signiifcant difference between 2 groups (P>0.05). Thirty-four patients relapsed including 5 patients of PEG-Asp ifrst-line treatment group, 16 patients of L-Asp ifrst-line treatment group and 13 patients treated with L-Asp during induction protocol and with PEG-Asp during consolidate protocol. Thirty-one patients died including 3, 18, 10 patients in 3 groups respectively. Twenty-two patients died of relapse, 4 died without remission, 5 died of complications. There was also no signiifcant difference between 2 groups (P>0.05). The incidence rates of adverse reactions were 47.6% and 63.3% respectively. Anaphylaxis, liver functions abnormalities, blood coagulation abnormalities, gastrointestinal reaction, hyperglycemia and pancreatitis were