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Erschienen in: Journal of Parasitic Diseases 2/2017

04.06.2016 | Original Article

Molecular identification of an old clinical isolate of Indian Kala-azar

verfasst von: Madhumita Manna, Supriya Khanra

Erschienen in: Journal of Parasitic Diseases | Ausgabe 2/2017

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Abstract

Molecular characterization is an important task for species identification of the isolates belonging to different Leishmania species. Clinical symptoms, tissue tropism, vector preference, reservoir and geographical distribution may act as distinguishing parameters but not always decisive. On the other hand, modern taxonomic tools employed to divulge characteristics of the genome or protein molecules of the parasite would be convincing and for Leishmania sp., they include nuclear and kDNA buoyant density, multi locus enzyme electrophoresis (MLEE), RAPD, RFLP or use of monoclonal antibodies etc. In the present study, we intend to establish the identification of an old clinical isolate of Indian Kala-azar, familiarly known as ‘UR6’ by MLEE, RAPD, RFLP and species specific monoclonal antibodies. UR6 has been isolated from a confirmed Kala-azar patient admitted in Calcutta School of Tropical Medicine, Kolkata in 1978. From then it is being regularly used for various scientific studies by the Leishmania Research Group of India and abroad. The isozyme profile of UR6 showed similar electrophoretic mobility that of WHO reference strain for Leishmania tropica, K27. Similar findings were obtained in the RAPD and RFLP assays. Screening with species specific monoclonal antibodies showed its strong reactivity towards L. tropica. The Jaccard’s Similarity Indices were calculated.
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Literatur
Zurück zum Zitat Abranches P, Santos-Gomes G, Rachamim N, Campino L, Schnur LF, Jaffe CL (1991) An experimental model for canine visceral leishmaniasis. Parasite Immunol 13(5):537–550CrossRefPubMed Abranches P, Santos-Gomes G, Rachamim N, Campino L, Schnur LF, Jaffe CL (1991) An experimental model for canine visceral leishmaniasis. Parasite Immunol 13(5):537–550CrossRefPubMed
Zurück zum Zitat Arora SK, Kapoor GS, Sehgal S (1998) Heterogeneity in heat shock protein genes in Leishmania isolates. Immunol Cell Biol 76(2):186–189CrossRefPubMed Arora SK, Kapoor GS, Sehgal S (1998) Heterogeneity in heat shock protein genes in Leishmania isolates. Immunol Cell Biol 76(2):186–189CrossRefPubMed
Zurück zum Zitat Bañuls AL, Dujardin JC, Guerrini F, De Doncker S, Jacquet D, Arevalo J et al (2000) Is Leishmania (Viannia) peruviana a distinct species? A MLEE/RAPD evolutionary genetics answer. J Eukaryot Microbiol 47:197–207CrossRefPubMed Bañuls AL, Dujardin JC, Guerrini F, De Doncker S, Jacquet D, Arevalo J et al (2000) Is Leishmania (Viannia) peruviana a distinct species? A MLEE/RAPD evolutionary genetics answer. J Eukaryot Microbiol 47:197–207CrossRefPubMed
Zurück zum Zitat Basu R, Bhaumik S, Basu JM, Naskar K, De T et al (2005) Kinetoplastid membrane protein-11 DNA vaccination induces complete protection against both pentavalent antimonial-sensitive and -resistant strains of Leishmania donovani that correlates with inducible nitric oxide synthase activity and IL-4 generation: evidence for mixed Th1- and Th2-like responses in visceral leishmaniasis. J Immunol 174:7160–7171CrossRefPubMed Basu R, Bhaumik S, Basu JM, Naskar K, De T et al (2005) Kinetoplastid membrane protein-11 DNA vaccination induces complete protection against both pentavalent antimonial-sensitive and -resistant strains of Leishmania donovani that correlates with inducible nitric oxide synthase activity and IL-4 generation: evidence for mixed Th1- and Th2-like responses in visceral leishmaniasis. J Immunol 174:7160–7171CrossRefPubMed
Zurück zum Zitat Basu JM, Mookerjee A, Sen P, Bhaumik S, Sen P, Banerjee S, Naskar K, Soumitra K, Choudhuri SK, Saha B, Raha S, Roy S (2006) Sodium antimony gluconate induces generation of reactive oxygen species and nitric oxide via phosphoinositide 3-kinase and mitogen-activated protein kinase activation in Leishmania donovani-infected macrophages. Antimicrob agent chemother 50:1788–1797CrossRef Basu JM, Mookerjee A, Sen P, Bhaumik S, Sen P, Banerjee S, Naskar K, Soumitra K, Choudhuri SK, Saha B, Raha S, Roy S (2006) Sodium antimony gluconate induces generation of reactive oxygen species and nitric oxide via phosphoinositide 3-kinase and mitogen-activated protein kinase activation in Leishmania donovani-infected macrophages. Antimicrob agent chemother 50:1788–1797CrossRef
Zurück zum Zitat Bhattacharyya R, Singh R, Hazra TK, Majumder HK (1993) Application of polymerase chain reaction with specific and arbitrary primers to identification and differentiation of Leishmania parasites. FEMS Microbiol Lett 114:99–104CrossRefPubMed Bhattacharyya R, Singh R, Hazra TK, Majumder HK (1993) Application of polymerase chain reaction with specific and arbitrary primers to identification and differentiation of Leishmania parasites. FEMS Microbiol Lett 114:99–104CrossRefPubMed
Zurück zum Zitat Bhattacharyya R, Das K, Sen S, Roy S, Majumder HK (1996) Development of a genus specific primer set for detection of Leishmania parasites by polymerase chain reaction. FEMS Microbiol Lett 135(2–3):195–200CrossRefPubMed Bhattacharyya R, Das K, Sen S, Roy S, Majumder HK (1996) Development of a genus specific primer set for detection of Leishmania parasites by polymerase chain reaction. FEMS Microbiol Lett 135(2–3):195–200CrossRefPubMed
Zurück zum Zitat Chatterjee M, Manna M, Bhaduri AN, Sarkar D (1995) Recent Kala-azar cases in India: isozyme profiles of Leishmania parasites. Ind J Med Res 102:165–172 Chatterjee M, Manna M, Bhaduri AN, Sarkar D (1995) Recent Kala-azar cases in India: isozyme profiles of Leishmania parasites. Ind J Med Res 102:165–172
Zurück zum Zitat Cupolillo E, Grimaldi JG, Momen H, Beverley SM (1995) Intergenic region typing (IRT): a rapid molecular approach to the characterization and evolution of Leishmania. Mol Biochem Parasitol 73:145–155CrossRefPubMed Cupolillo E, Grimaldi JG, Momen H, Beverley SM (1995) Intergenic region typing (IRT): a rapid molecular approach to the characterization and evolution of Leishmania. Mol Biochem Parasitol 73:145–155CrossRefPubMed
Zurück zum Zitat Das S, Adhya S (1990) Organization and chromosomal localization of ß-tubulin genes in Leishmania donovani. J Biosci 15:239–248CrossRef Das S, Adhya S (1990) Organization and chromosomal localization of ß-tubulin genes in Leishmania donovani. J Biosci 15:239–248CrossRef
Zurück zum Zitat Desjeux P (2004) Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27:305–318CrossRefPubMed Desjeux P (2004) Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27:305–318CrossRefPubMed
Zurück zum Zitat El Tai NO, Osman OF, el Fari M, Presber W, Schonian G (2000) Genetic heterogeneity of ribosomal internal transcribed spacer in clinical samples of Leishmania donovani spotted on filter paper as revealed by single-strand conformation polymorphisms and sequencing. Trans R Soc Trop Med Hyg 94:575–579CrossRefPubMed El Tai NO, Osman OF, el Fari M, Presber W, Schonian G (2000) Genetic heterogeneity of ribosomal internal transcribed spacer in clinical samples of Leishmania donovani spotted on filter paper as revealed by single-strand conformation polymorphisms and sequencing. Trans R Soc Trop Med Hyg 94:575–579CrossRefPubMed
Zurück zum Zitat Evans DA (1989) Handbook on isolation characterization and cryopreservation of Leishmania. World Health Organ, Geneva, pp 15–40 Evans DA (1989) Handbook on isolation characterization and cryopreservation of Leishmania. World Health Organ, Geneva, pp 15–40
Zurück zum Zitat Evans DA, Lanham SM, Baldwin CI, Peters W (1984) The isolation and isoenzyme characterization of Leishmania braziliensis subsp. from patients with cutaneous leishmaniasis acquired in Belize. Trans R Soc Trop Med Hyg 78:35–42CrossRefPubMed Evans DA, Lanham SM, Baldwin CI, Peters W (1984) The isolation and isoenzyme characterization of Leishmania braziliensis subsp. from patients with cutaneous leishmaniasis acquired in Belize. Trans R Soc Trop Med Hyg 78:35–42CrossRefPubMed
Zurück zum Zitat Fraga J, Montalvo AM, De Doncker S, Dujardin JC, Van der Auwera G (2010) Phylogeny of Leishmania species based on the heat-shock protein 70 gene. Infect Genet Evol 10:238–245CrossRefPubMed Fraga J, Montalvo AM, De Doncker S, Dujardin JC, Van der Auwera G (2010) Phylogeny of Leishmania species based on the heat-shock protein 70 gene. Infect Genet Evol 10:238–245CrossRefPubMed
Zurück zum Zitat Garcia L, Kindt A, Bermudez H, Llanos-Cuentas A, De Doncker S, Arevalo J et al (2004) Culture-independent species typing of neotropical Leishmania for clinical validation of a PCR-based assay targeting heat shock protein 70 genes. J Clin Microbiol 42:2294–2297CrossRefPubMedPubMedCentral Garcia L, Kindt A, Bermudez H, Llanos-Cuentas A, De Doncker S, Arevalo J et al (2004) Culture-independent species typing of neotropical Leishmania for clinical validation of a PCR-based assay targeting heat shock protein 70 genes. J Clin Microbiol 42:2294–2297CrossRefPubMedPubMedCentral
Zurück zum Zitat Guha R, Gupta D, Rastogi R, Vikram R, Krishnamurthy G, Bimal S, Roy S, Mukhopadhyay A (2013) Vaccination with Leishmania hemoglobin receptor-encoding DNA protects against visceral leishmaniasis. Sci Transl Med 5:202ra121CrossRefPubMed Guha R, Gupta D, Rastogi R, Vikram R, Krishnamurthy G, Bimal S, Roy S, Mukhopadhyay A (2013) Vaccination with Leishmania hemoglobin receptor-encoding DNA protects against visceral leishmaniasis. Sci Transl Med 5:202ra121CrossRefPubMed
Zurück zum Zitat Guha R, Das S, Ghosh J, Sundar S, Dujardin JC, Roy S (2014) Antimony resistant Leishmania donovani but not sensitive ones drives greater frequency of potent TR regulatory cells upon interaction with human PBMCs: role of IL-10 and TGF-b in early immune response. Plos Negl Trop Dis 8:e2995CrossRefPubMedPubMedCentral Guha R, Das S, Ghosh J, Sundar S, Dujardin JC, Roy S (2014) Antimony resistant Leishmania donovani but not sensitive ones drives greater frequency of potent TR regulatory cells upon interaction with human PBMCs: role of IL-10 and TGF-b in early immune response. Plos Negl Trop Dis 8:e2995CrossRefPubMedPubMedCentral
Zurück zum Zitat Haldar AK, Yadav V, Singhal E, Bisht KK, Singh A, Bhaumik S, Basu R, Roy S, Sen P (2010) Leishmania donovani isolates with antimony-resistant but not group b phenotype inhibit sodium antimony gluconate-induced dendritic cell activation. PLoS Pathog 6:e1000907CrossRefPubMedPubMedCentral Haldar AK, Yadav V, Singhal E, Bisht KK, Singh A, Bhaumik S, Basu R, Roy S, Sen P (2010) Leishmania donovani isolates with antimony-resistant but not group b phenotype inhibit sodium antimony gluconate-induced dendritic cell activation. PLoS Pathog 6:e1000907CrossRefPubMedPubMedCentral
Zurück zum Zitat Handman E, Hocking RE (1982) Stage-specific, Strain-specific and cross reactive antigens of Leishmania species identified by monoclinal antibodies. Infect Immun 37:28–33PubMedPubMedCentral Handman E, Hocking RE (1982) Stage-specific, Strain-specific and cross reactive antigens of Leishmania species identified by monoclinal antibodies. Infect Immun 37:28–33PubMedPubMedCentral
Zurück zum Zitat Jaffe CL, McMahoon Pratt D (1983) Monoclonal antibodies specific for Leishmania tropica. I. Characterization of antigens associated with stage and species-specific determinants. J Immunol 131:1987–1993PubMed Jaffe CL, McMahoon Pratt D (1983) Monoclonal antibodies specific for Leishmania tropica. I. Characterization of antigens associated with stage and species-specific determinants. J Immunol 131:1987–1993PubMed
Zurück zum Zitat Jaffe CL, Sharfstein R (1987) Species -specific antibodies to Leishmania tropica (minor) recognize somatic antigens and exometabolites. J Immunol 139:1310PubMed Jaffe CL, Sharfstein R (1987) Species -specific antibodies to Leishmania tropica (minor) recognize somatic antigens and exometabolites. J Immunol 139:1310PubMed
Zurück zum Zitat Khanra S, Bandopadhyay SK, Chakraborty P, Datta S, Mondal D, Chatterjee M, Naskar K, Roy S, Manna M (2011) Characterization of the recent clinical isolates of Indian Kala-azar patients by RAPD-PCR method. J Parasit Dis 35:116–122CrossRefPubMedPubMedCentral Khanra S, Bandopadhyay SK, Chakraborty P, Datta S, Mondal D, Chatterjee M, Naskar K, Roy S, Manna M (2011) Characterization of the recent clinical isolates of Indian Kala-azar patients by RAPD-PCR method. J Parasit Dis 35:116–122CrossRefPubMedPubMedCentral
Zurück zum Zitat Khanra S, Datta S, Mondal D, Saha P, Bandopadhyay SK, Roy S, Manna M (2012) RFLPs of ITS, ITS1 and hsp70 amplicons and sequencing of ITS1 of recent clinical isolates of Kala-azar from India and Bangladesh confirms the association of L. tropica with the disease. Acta Trop 124:229–234CrossRefPubMed Khanra S, Datta S, Mondal D, Saha P, Bandopadhyay SK, Roy S, Manna M (2012) RFLPs of ITS, ITS1 and hsp70 amplicons and sequencing of ITS1 of recent clinical isolates of Kala-azar from India and Bangladesh confirms the association of L. tropica with the disease. Acta Trop 124:229–234CrossRefPubMed
Zurück zum Zitat Kreutzer RD, Christensen HA (1980) Characterization of Leishmania spp by isozyme electrophoresis. Am J Trop Med Hyg 29:199–208CrossRefPubMed Kreutzer RD, Christensen HA (1980) Characterization of Leishmania spp by isozyme electrophoresis. Am J Trop Med Hyg 29:199–208CrossRefPubMed
Zurück zum Zitat Kumar R, Bumb RA, Ansari NA, Mehta RD, Salotra P (2007) Cutaneous leishmaniasis caused by Leishmania tropica in Bikaner, India: parasite identification and characterization using molecular and immunologic tools. Am J Trop Med Hyg 76:896–901PubMed Kumar R, Bumb RA, Ansari NA, Mehta RD, Salotra P (2007) Cutaneous leishmaniasis caused by Leishmania tropica in Bikaner, India: parasite identification and characterization using molecular and immunologic tools. Am J Trop Med Hyg 76:896–901PubMed
Zurück zum Zitat Manna M (1998) Molecular taxonomic analysis of clinical isolates of Indian Kala-azar patients. PhD Thesis, Jadavpur University, Kolkata Manna M (1998) Molecular taxonomic analysis of clinical isolates of Indian Kala-azar patients. PhD Thesis, Jadavpur University, Kolkata
Zurück zum Zitat Manna M, Majumder HK, Sundar S, Bhaduri AN (2005) The molecular characterization of clinical isolates from Indian Kala-azar patients by MLEE and RAPD-PCR. Med Sci Monit 11:220–227 Manna M, Majumder HK, Sundar S, Bhaduri AN (2005) The molecular characterization of clinical isolates from Indian Kala-azar patients by MLEE and RAPD-PCR. Med Sci Monit 11:220–227
Zurück zum Zitat McMahon-Pratt D, David JR (1981) Monoclonal antibodies that distinguish between New World species of Leishmania. Nature 291:581–583CrossRef McMahon-Pratt D, David JR (1981) Monoclonal antibodies that distinguish between New World species of Leishmania. Nature 291:581–583CrossRef
Zurück zum Zitat McMahon-Pratt D, Bennett E, David JR (1982) Monoclonal antibodies that distinguish subspecies of Leishmania braziliensis. J Immunol 129:926–927PubMed McMahon-Pratt D, Bennett E, David JR (1982) Monoclonal antibodies that distinguish subspecies of Leishmania braziliensis. J Immunol 129:926–927PubMed
Zurück zum Zitat Miles MA, Povoa MM, de Souza AA, Lainson R, Shaw JJ (1980) Some methods for the enzymic characterization of Latin- American Leishmania with particular reference to Leishmania mexicana amazonensis and subspecies of Leishmania hertigi. Trans R Soc Trop Med Hyg 74:243–252CrossRefPubMed Miles MA, Povoa MM, de Souza AA, Lainson R, Shaw JJ (1980) Some methods for the enzymic characterization of Latin- American Leishmania with particular reference to Leishmania mexicana amazonensis and subspecies of Leishmania hertigi. Trans R Soc Trop Med Hyg 74:243–252CrossRefPubMed
Zurück zum Zitat Montalvo AM, Fraga J, Monzote L, Montano I, De Doncker S, Dujardin JC, Van der Auwera G (2010) Heat-shock protein 70 PCR-RFLP: a universal simple too for Leishmania species discrimination in the New and Old World. Parasitology 137:1159–1168CrossRefPubMed Montalvo AM, Fraga J, Monzote L, Montano I, De Doncker S, Dujardin JC, Van der Auwera G (2010) Heat-shock protein 70 PCR-RFLP: a universal simple too for Leishmania species discrimination in the New and Old World. Parasitology 137:1159–1168CrossRefPubMed
Zurück zum Zitat Mukhopadhyay S, Bhattacharyya S, Majhi R, De T, Naskar K, Majumdar S, Roy S (2000) Use of an attenuated Leishmanial Parasite as an immunoprophylactic and immunotherapeutic agent against Murine Visceral Leishmaniasis. Clin Diagn Lab Immunol 7:233–240PubMedPubMedCentral Mukhopadhyay S, Bhattacharyya S, Majhi R, De T, Naskar K, Majumdar S, Roy S (2000) Use of an attenuated Leishmanial Parasite as an immunoprophylactic and immunotherapeutic agent against Murine Visceral Leishmaniasis. Clin Diagn Lab Immunol 7:233–240PubMedPubMedCentral
Zurück zum Zitat Mukhopadhyay R, Mukherjee S, Mukherjee B, Naskar K, Mondal D, Decuypere S, Ostyn B, Prajapati VK, Sundar S, Dujardin JC, Roy S (2011) Characterization of antimony-resistant Leishmania donovani isolates: biochemical and biophysical studies and interaction with host cells. Int J Parasitol 41:1311–1321CrossRefPubMed Mukhopadhyay R, Mukherjee S, Mukherjee B, Naskar K, Mondal D, Decuypere S, Ostyn B, Prajapati VK, Sundar S, Dujardin JC, Roy S (2011) Characterization of antimony-resistant Leishmania donovani isolates: biochemical and biophysical studies and interaction with host cells. Int J Parasitol 41:1311–1321CrossRefPubMed
Zurück zum Zitat Rioux JA, Lanotte G, Serres E, Pratlong F, Bastien P, Periéres J (1990) Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification. Ann Parasitol Hum Comp 65:111–125CrossRefPubMed Rioux JA, Lanotte G, Serres E, Pratlong F, Bastien P, Periéres J (1990) Taxonomy of Leishmania. Use of isoenzymes. Suggestions for a new classification. Ann Parasitol Hum Comp 65:111–125CrossRefPubMed
Zurück zum Zitat Sacks DL, Kenney RT, Neva FA, Kreutzer RD, Jaffe CL, Gupta AK et al (1995) Indian Kala-azar caused by Leishmania tropica. Lancet 345:959–961CrossRefPubMed Sacks DL, Kenney RT, Neva FA, Kreutzer RD, Jaffe CL, Gupta AK et al (1995) Indian Kala-azar caused by Leishmania tropica. Lancet 345:959–961CrossRefPubMed
Zurück zum Zitat Schnur LF, Chance ML, Ebert F, Thomas SC, Peters W (1981) The biochemical and serological taxonomy of visceralizing Leishmania. Ann Trop Med Parasitol 75:131–144CrossRef Schnur LF, Chance ML, Ebert F, Thomas SC, Peters W (1981) The biochemical and serological taxonomy of visceralizing Leishmania. Ann Trop Med Parasitol 75:131–144CrossRef
Zurück zum Zitat Schonian G, Nasereddin A, Dinse N, Schweynoch C, Schallig HD, Presber W et al (2003) PCR diagnosis and characterization of Leishmania in local and imported clinical samples. Diagn Microbiol Infect Dis 47:349–358CrossRefPubMed Schonian G, Nasereddin A, Dinse N, Schweynoch C, Schallig HD, Presber W et al (2003) PCR diagnosis and characterization of Leishmania in local and imported clinical samples. Diagn Microbiol Infect Dis 47:349–358CrossRefPubMed
Zurück zum Zitat Sneath PHA, Sokal RR (1973) Numerical taxonomy. Freeman, San Francisco, USA Sneath PHA, Sokal RR (1973) Numerical taxonomy. Freeman, San Francisco, USA
Zurück zum Zitat Tibayrenc M, Neubauer K, Barnabé C, Guerrini F, Skarecky D, Ayala FJ et al (1993) Genetic characterization of six parasitic protozoa: parity between random-primer DNA typing and multilocus enzyme electrophoresis. Proc Natl Acad Sci USA 90:1335–1339CrossRefPubMedPubMedCentral Tibayrenc M, Neubauer K, Barnabé C, Guerrini F, Skarecky D, Ayala FJ et al (1993) Genetic characterization of six parasitic protozoa: parity between random-primer DNA typing and multilocus enzyme electrophoresis. Proc Natl Acad Sci USA 90:1335–1339CrossRefPubMedPubMedCentral
Zurück zum Zitat Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl Acids Res 18:6531–6535CrossRefPubMedPubMedCentral Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl Acids Res 18:6531–6535CrossRefPubMedPubMedCentral
Zurück zum Zitat World Health Organization (1990) Control of Leishmaniases. WHO, Geneva World Health Organization (1990) Control of Leishmaniases. WHO, Geneva
Zurück zum Zitat World Health Organization (1994) Report on the consultative meeting of Leishmania/HIV co-infection. Co-sponsored by the Instituto Superiore Di Sanita and The Division of the Control of Tropical Diseases, WHO, Rome, pp 4–8 World Health Organization (1994) Report on the consultative meeting of Leishmania/HIV co-infection. Co-sponsored by the Instituto Superiore Di Sanita and The Division of the Control of Tropical Diseases, WHO, Rome, pp 4–8
Zurück zum Zitat Zemanova E, Jirku M, Mauricio IL, Miles MA, Lukes J (2004) Genetic polymorphism within the Leishmania donovani complex: correlation with geographic origin. Am J Trop Med Hyg 70:613–617PubMed Zemanova E, Jirku M, Mauricio IL, Miles MA, Lukes J (2004) Genetic polymorphism within the Leishmania donovani complex: correlation with geographic origin. Am J Trop Med Hyg 70:613–617PubMed
Metadaten
Titel
Molecular identification of an old clinical isolate of Indian Kala-azar
verfasst von
Madhumita Manna
Supriya Khanra
Publikationsdatum
04.06.2016
Verlag
Springer India
Erschienen in
Journal of Parasitic Diseases / Ausgabe 2/2017
Print ISSN: 0971-7196
Elektronische ISSN: 0975-0703
DOI
https://doi.org/10.1007/s12639-016-0799-9

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