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Polymorphic retroelements—a useful tool for human genetic studies
New Biotechnology · Volume 27S · April 2010
POSTERS ABSTRACTS 1
[P1.23] Application of NMR and UV spectroscopy in the study of interactions between anticancer drugs: 5-FU, LCV and their phospholipide carriers D. Pentak University of Silesia, Poland
The aim of this work was to encapsulate two drugs: 5fluorouracil (5-FU) with the hydrophobic properties and 5-formyl5,6,7,8-tetrahydropteroyl-L-glutamic acid calcium salt (LCV) with the hydrophilic properties into liposomes prepared by the modified reverse-phase evaporation (mREV) method from L-␣phosphatidylcholine dipalmitoyl (DPPC). The drugs are used in the anticancer multidrug therapy FLv (5-fluorouracil, leucovorin) and ELF (etoposide, leucovorin, 5-fluorouracil). We studied the competition for their encapsulation in liposomes by the use of two spectroscopies: 1 H NMR and UV on the basis of the analysis of the signals of each drug in the liposome – drug system. Liposomes are highly versatile structures for research, therapeutic, and analytical applications. We concluded that the liposomes obtained by the mREV method may transport more than one drug simultaneously. doi:10.1016/j.nbt.2010.01.030
[P1.24] Polymorphic retroelements—–a useful tool for human genetic studies I.Z. Mamedov ∗ , S.V. Ustyugova, D.A. Shagin, Y.B. Lebedev Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Russian Federation
Retroelements (REs) comprising about 50% of the human genome are known to influence not only neighboring genes but alter the hole genome functioning. Human genome contains several thousands RE integrations polymorphic in different populations. Recently we designed and successfully applied a new approach for experimental identification of new polymorphic RE insertions. As a result we discovered about one hundred of new polymorphic RE insertions and created a new internet resource on polymorphic REs in human genome. Our database contains information about position of RE in human genome, location of a neighboring known and predictive gene(s) and other data. The internet resource allows to perform a search on database using multiple search conditions and available online at http://labcfg.ibch.ru/home.html. Using our database we design the new set of RE based molecular genetic markers for human genetic identification. This set includes 32 polymorphic insertions of Alu randomly distributed in human genome. The discrimination power of the set was evaluated in population studies as well as in paternity identification experiments. Our 32 Alu set is the first example of application of RE based molecular genetic markers for human genome fingerprinting and for forensic studies. Our result indicates that RE insertion polymorphism becomes a useful source of new molecular genetic
S30
www.elsevier.com/locate/nbt
markers widely applied for population and medical genetics studies. doi:10.1016/j.nbt.2010.01.031
[P1.25] Molecular characterization of carboxypeptidase B (CPB) of An. stephensi with respect to TBV strategy in states under WHO/EMRO A. Raz ∗ , N. Dinparast Djadid, S. Zakeri Pasteur Institute of Iran, Islamic Republic of Iran
Malaria ranks up along with HIV and tuberculosis as one of three greatest infectious diseases of humanity. It is a complex disease that varies widely in epidemiology and clinical manifestation in different parts of the world, especially Africa and Asia. Transmission blocking vaccine (TBV) is one of the strategies that is used to prevent malaria transmission. Most of the works on TBV development have focused on parasite antigens that are expressed in the mosquito midgut. Whereas, blocking the malaria transmission could also be achieved by targeting mosquito components that are required for the successful development of the parasite in the vector. Bourgouin et al. (2007) determined that anti-CPBAg1 antibodies can block P. falciparum development in the mosquito midgut. However, the main vector in Iran and neighbour countries is An. stephensi. Therefore, the aim of current study was to characterize CPB in An. stephensi and investigate the effect of its deduced antibody on P. falciparum development. Initially cpb mRNA sequence was determined in An. stephensi by 3- and 5 -RACE (rapid amplification of cDNA ends) technique, followed by its expression in E. coli (BL21), enabling us to evaluate anti-CPB antibody effect on P. falciparum development in An. stephensi. To achieve this goal, continues in vitro culture of P. falciparum has been carried out in order to produce the gametocyte, which should be transmitted to An. stephensi local colony of Pasteur Institute of Iran through artificial membrane feeding (Tchuinkam et al., 1993). The out coming results will be discussed in detail with respect to its feasibility and applied features as a TBV. References 1 Lavazec, C. et al. (2007) Carboxypeptidases B of Anopheles gambiae as targets for a Plasmodium falciparum transmission-blocking vaccine. Infect. Immun. 75, 1635–42 2 Tchuinkam, T. et al. (1993) Experimental infections of Anopheles gambiae with Plasmodium falciparum of naturally infected gametocvte carriers in Cameroon: factors influencing the hfectiv& to mosquitoes. Trop. Med. Parasitol. 44, 271–276 doi:10.1016/j.nbt.2010.01.032
POSTERS ABSTRACTS 1
[P1.23] Application of NMR and UV spectroscopy in the study of interactions between anticancer drugs: 5-FU, LCV and their phospholipide carriers D. Pentak University of Silesia, Poland
The aim of this work was to encapsulate two drugs: 5fluorouracil (5-FU) with the hydrophobic properties and 5-formyl5,6,7,8-tetrahydropteroyl-L-glutamic acid calcium salt (LCV) with the hydrophilic properties into liposomes prepared by the modified reverse-phase evaporation (mREV) method from L-␣phosphatidylcholine dipalmitoyl (DPPC). The drugs are used in the anticancer multidrug therapy FLv (5-fluorouracil, leucovorin) and ELF (etoposide, leucovorin, 5-fluorouracil). We studied the competition for their encapsulation in liposomes by the use of two spectroscopies: 1 H NMR and UV on the basis of the analysis of the signals of each drug in the liposome – drug system. Liposomes are highly versatile structures for research, therapeutic, and analytical applications. We concluded that the liposomes obtained by the mREV method may transport more than one drug simultaneously. doi:10.1016/j.nbt.2010.01.030
[P1.24] Polymorphic retroelements—–a useful tool for human genetic studies I.Z. Mamedov ∗ , S.V. Ustyugova, D.A. Shagin, Y.B. Lebedev Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Russian Federation
Retroelements (REs) comprising about 50% of the human genome are known to influence not only neighboring genes but alter the hole genome functioning. Human genome contains several thousands RE integrations polymorphic in different populations. Recently we designed and successfully applied a new approach for experimental identification of new polymorphic RE insertions. As a result we discovered about one hundred of new polymorphic RE insertions and created a new internet resource on polymorphic REs in human genome. Our database contains information about position of RE in human genome, location of a neighboring known and predictive gene(s) and other data. The internet resource allows to perform a search on database using multiple search conditions and available online at http://labcfg.ibch.ru/home.html. Using our database we design the new set of RE based molecular genetic markers for human genetic identification. This set includes 32 polymorphic insertions of Alu randomly distributed in human genome. The discrimination power of the set was evaluated in population studies as well as in paternity identification experiments. Our 32 Alu set is the first example of application of RE based molecular genetic markers for human genome fingerprinting and for forensic studies. Our result indicates that RE insertion polymorphism becomes a useful source of new molecular genetic
S30
www.elsevier.com/locate/nbt
markers widely applied for population and medical genetics studies. doi:10.1016/j.nbt.2010.01.031
[P1.25] Molecular characterization of carboxypeptidase B (CPB) of An. stephensi with respect to TBV strategy in states under WHO/EMRO A. Raz ∗ , N. Dinparast Djadid, S. Zakeri Pasteur Institute of Iran, Islamic Republic of Iran
Malaria ranks up along with HIV and tuberculosis as one of three greatest infectious diseases of humanity. It is a complex disease that varies widely in epidemiology and clinical manifestation in different parts of the world, especially Africa and Asia. Transmission blocking vaccine (TBV) is one of the strategies that is used to prevent malaria transmission. Most of the works on TBV development have focused on parasite antigens that are expressed in the mosquito midgut. Whereas, blocking the malaria transmission could also be achieved by targeting mosquito components that are required for the successful development of the parasite in the vector. Bourgouin et al. (2007) determined that anti-CPBAg1 antibodies can block P. falciparum development in the mosquito midgut. However, the main vector in Iran and neighbour countries is An. stephensi. Therefore, the aim of current study was to characterize CPB in An. stephensi and investigate the effect of its deduced antibody on P. falciparum development. Initially cpb mRNA sequence was determined in An. stephensi by 3- and 5 -RACE (rapid amplification of cDNA ends) technique, followed by its expression in E. coli (BL21), enabling us to evaluate anti-CPB antibody effect on P. falciparum development in An. stephensi. To achieve this goal, continues in vitro culture of P. falciparum has been carried out in order to produce the gametocyte, which should be transmitted to An. stephensi local colony of Pasteur Institute of Iran through artificial membrane feeding (Tchuinkam et al., 1993). The out coming results will be discussed in detail with respect to its feasibility and applied features as a TBV. References 1 Lavazec, C. et al. (2007) Carboxypeptidases B of Anopheles gambiae as targets for a Plasmodium falciparum transmission-blocking vaccine. Infect. Immun. 75, 1635–42 2 Tchuinkam, T. et al. (1993) Experimental infections of Anopheles gambiae with Plasmodium falciparum of naturally infected gametocvte carriers in Cameroon: factors influencing the hfectiv& to mosquitoes. Trop. Med. Parasitol. 44, 271–276 doi:10.1016/j.nbt.2010.01.032