- Email: [email protected]
Promising experimental Calcium silicate-methacrylate hybrid composites able of Calcium and Fluoride releasing for clinical application in dentistry
S462
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
[P-M.95] Promising experimental Calcium silicate-methacrylate hybrid composites able of Calcium and Fluoride releasing for clinical application in dentistry Cesar Henriqhe Zanchi 2 , Carlo Prati 1 , Francesco Siboni 1,∗ , Fabricio Aulo Ogliari 2 , Evandro Piva 2 , Maria Giovanna Gandolfi 1 1
Laboratory of Biomaterials, Department of Odontostomatological Sciences, University of Bologna, Bologna, Italy 2 School of Dentistry, Federal University of Pelotas, Pelotas, RS, Brazil
Fig. 1. Overview of the Magnetic Capture Hybridization Technology for the isolation and analysis of methylation status of cdh1 promoter sequence.
Results: The method showed a high specificity and sensitivity, with a detection level of 20-50 cdh1 copies. Captured fragments were rinsed out of background, denatured and preliminary experiments to analyze methylation status of cdh1 promoter by Pyrosequencing were also carried out. Results indicated that isolated DNA maintains its methylation allowing bisulfite conversion and therefore methylation can be investigated. Conclusions: The MCH technology can be proposed as a valid and versatile tool to isolate potential biomarkers from whole plasma samples after the selection of a specific probe sequence that can be conjugated to paramagnetic nanoparticles. Particularly, the technology could be useful in the screening of a large number of samples, since it is rapid, sensitive and well suited for automation.
Introduction: Dual-curing calcium-silicate-filled methacrylate composites were developed to obtain “smart” materials able of calcium and fluoride releasing for biomimetic remineralization. The composites were prepared incorporating UDMA/HEMA/TEGDMA resin in calcium-silicate powders to obtain fast setting materials for dental applications such as enamel sealant and restorative materials for posterior teeth. Methods: Three calcium-silicate powders were used: grey MTA free from radiopacifing agent (Angelus, Brazil), F10 TC (GandolfiPrati, University of Bologna, Italy) and PC (white Portland cement as control powder). The dual-curing resin contained UDMA, HEMA, TEGDMA and PEG. Each calcium-silicate powders (60% wt) was mixed with the dual-curing UDMA/HEMA/TEGDMA resin (40% wt) obtaining MTA-Rc, F10 TC-Rc and PC-Rc. A resin-reinforced glassionomer cement (Vitrebond, 3 M ESPE, Germany) was tested as control. Ten samples of each composite (8 mm diameter and 1.6 mm thick) were soaked in 10 mL of deionized water and maintained in a cabinet at 37 ◦ C. Calcium and fluorine leached in soaking water and pH of soaking water were measured after 3, 24 hours and 3,7,14,28 days using ion-selective electrodes connected to a meter. ESEM/EDX was performed after soaking in simulated body fluid at 37 ◦ C. Results: All the experimental materials are able to basify the soaking water from pH 6.8 to 8-10 (Table 1). All the experimenTable 1 pH Of Soaking Water (mean ± SD, n = 10) Materials
3 hours
1 day
3 days
7 days
14 days
28 days
MTA-Rc FIOTC-Rc PC-Re Vitrebond Water
10.01 ± 0.18 9.93 ± 0.14 9.99 ± 0.26 6.64 ± 0.14 6.87 ± 0.04
8.72 ± 0.24 9.08 ± 0.52 8.55 ± 0.8 7.53 ± 0.12 6.87 ± 0.26
3.78 ± 0.22 9.00 ± 0.09 8.22 ± 0.5 7.40 ± 0.16 7.02 ± 0.18
8.22 ± 0.06 8.36 ± 0.30 8.36 ± 0.23 8.36 ± 0.12 6.93 ± 0.8
8.06 ± 0.02 8.37 ± 0.10 8.26 ± 0.5 5.90 ± 0.23 7.06 ± 0.48
8.24 ± 0.06 8.11 ± 0.18 8.50 ± 0.8 7.77 ± 0.03 6.98 ± 0.16
tal materials release calcium (higher release by MTA-Rc and PC-Rc) (Table 2). Only F10 TC-Rc and Vitrebond release fluorine (Table 3). Conclusions: The study support the concept that calcium-silicatefilled methacrylate resin composites are “smart” materials able to release calcium and fluoride for remineralizing caries-like dentine
doi:10.1016/j.jbiotec.2010.09.682 Table 2 Materials
3 hours
1 day
3 days
7 days
14 days
28 days
MTA-Rc FIOTC-Rc PC-Re Vitrebond water
1478 ± 1.91 4.25 ± 1.43 13.02 ± 1.04 3.60 ± 1.17 0.60 ± 0.39
14.72 ± 4.09 7.15 ± 1.05 12.98 ± 0.46 2.60 ± 0.84 0.93 ± 0.41
10.27 ± 1.53 3.55 ± 0.45 10.19 ± 2.57 0.60 ± 0.52 1.08 ± 0.30
18.81 ± 1.67 3.20 ± 0.25 29.36 ± 1.82 0.30 ± 0.43 0.98 ± 0.44
15.61 ± 2.03 12.41 ± 3.39 12.09 ± 1.49 0.60 ± 0.52 1.05 ± 0.13
30.01 ± 1.85 6.43 ± 1.87 19.72 ± 1.77 0.40 ± 0.52 0.95 ± 0.31
Table 3 Materials
3 hours
1 day
3 days
7 days
14 days
28 days
MTA-Rc FIOTC-Rc PC-Re Vitrebond water
0.31 ± 0.23 4.11 ± 0.73 0.61 ± 0.27 9.40 ± 0.89 0.17 ± 0.05
0.16 ± 0.13 11.86 ± 1.04 0.65 ± 0.25 11.40 ± 2.51 0.37 ± 0.15
0.23 ± 0.25 15.45 ± 1.87 0.45 ± 0.24 12.40 ± 1.67 0.37 ± 0.17
0.51 ± 0.28 20.85 ± 1.29 0.56 ± 0.24 18.20 ± 4.21 0.35 ± 0.26
0.43 ± 0.21 18.54 ± 1.37 0.84 ± 0.10 14.40 ± 1.14 0.37 ± 0.25
1.05 ± 0.18 14.98 ± 1.81 1.30 ± 0.06 6.00 ± 1.00 0.42 ± 0.22
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
lesions. The incorporation of calcium-silicate particles in resin composite material may extend the range of their clinical applications. doi:10.1016/j.jbiotec.2010.09.683 [P-M.96] Therapeutic potential of plant bioactive molecules L. Bertini 1 , S. Proietti 1 , C. Caporale 1 , F. Mondello 2 , A. Cassone 2 , C. Caruso 1,∗ 1 Dipartimento di Agrobiologia e Agrochimica, Università della Tuscia, Viterbo, Italy, Italy 2 Dipartimento di Malattie Infettive Parassitarie ed Immunomediate, Istituto Superiore di Sanità, Roma, Italy, Italy Keywords: pathogenesis related proteins PR4; antifungal activity; human pathogen; plant bioactive molecules
The search for biologically active compounds from natural sources has always been of great interest to scientist looking for new sources of drugs useful to fight infectious diseases. To this aim, considerable attention has been recently focussed on bio-friendly plant-based products. The plant kingdom represents an extraordinary reservoir of novel molecules and the potential of higher plants as source for new drug is still largely unexplored. Studying the plant defence mechanisms, we focused our attention on Pathogenesis-Related proteins of class 4 (PR4). In the last years, we have isolated and sequenced four PR4 proteins from wheat kernels, named wheatwin1 to wheatwin4, that strongly inhibit both host and non-specific phytopathogenic fungi. We have also demonstrated that they have ribonuclease activity and that their antifungal activity is correlated with the enzymatic one. In this work we report on the isolation of the ortholog gene from Arabidopsis thaliana, coding for a PR4 protein named HEL. This protein was produced at high level in E. coli, which showed ribonuclease and antifungal activities stronger than those shown by the wheatwins. The growing interest around the research of new natural bioactive molecules prompted us to test the activity of this recombinant protein against human pathogens. Preliminary tests carried out on Candida albicans, the etiologic agent of opportunistic mucosal and systemic infections in humans, showed that recombinant HEL has a cytocidal effect on budding cells at a very low concentration. To the best of our knowledge, this is the first time that PR4 proteins have been tested against human pathogens. These results add significance to the exploitation of this molecule in biotechnological process.
S463
steps. Many improvements have been focused in large scale production using bioreactors (Carnes et al., 2006; Sousa et al., 2005). In order to use the equipment (flasks) and growth media available in a common research laboratory, this work studied size of plasmids and protocols (conditions in flasks production) in order to improve sc-pDNA yields. Methods: Escherichia coli DH5␣ was transformed with VR1012NH36 (6 kb) and pVAX-NH36 (4 kb) plasmids and growth in three culture media (LB (control), SDCAS (O’Kennedy et al., 2000) and ZMY505 (Studier, 2005)), in 250 mL flasks at 37 ◦ C 100 and 200 rpm, were gotten characteristic kinetics of dry cellular growth and plasmid DNA production for both conditions. Results: Laboratory scale cultures using LB showed that pDNA volumetric and specific yields were 11 mg/L and 10.7 mg/g dry cell mass for both plasmids, using ZMY505 pDNA yieds were 16 and 18.5 mg/L (VR1012-NH36 and pVAX-NH36 respectively). DNA volumetric yield were increased at 200 rpm giving 24 mg/L and 30 mg/L respectively, in all cases specific and volumetric pDNA yields were highest in 10 or 12 h. Additionally pDNA production was inducible for increased of temperature using different growth phases or times for induction of specific pDNA, theses increased until 2 or 3 fold (VR1012-NH36 and pVAX-NH36, respectively) giving volumetric yields between 40 and 55 mg/L. Discussion: A specialized medium for pDNA production could help for increasing biomass and specific pDNA yields with increase temperature, improving final volumetric pDNA yields. Plasmid yields were highest after the culture has entered the stationary phase because of accumulation (Danquah, 2007). This project was funded by Grant from 167077 and grant PIFUTP08-108 JOL from ICyTDF. References Carnes, A.E., Hodgson, C.P., y Williams, J.A., 2006. Inducible Escherichia coli fermentation for increased plasmid DNA production. Biotechnol. Appl. Biochem 45, 155–166. Danquah, M., y Forde, G., 2007. Growth Medium selection and its economic impact on plasmid DNA production. J. Bios. Bioeng. 104 (6), 490–497. Gamboa-León, R., Paraguai de Souza, E., Borja-Cabrera, G.P., Santos, F.N., Myashiro, L.M., Pinheiro, R.O., Dumonteil, E., Palatnik-de-Sousa, C.B., 2006. Immunotherapy against visceral leishmaniasis with the nucleoside hydrolase-DNA vaccine of Leishmania donovani. Vaccine 24, 4863–4873. O’Kennedy, R.D., Baldwin, C., y Keshavarz-Moore, E., 2000. Effects of growth medium selection on plasmid DNA production and initial processing steps. J. Biotech. 76, 175–183. Sousa, F., Tomaz, C.T., Prazeres, D.M.F., Queiroz, J.A., 2005. Separation of supercoiled and open circular plasmid DNA isoforms by chromatography with a histidine–agarose support. Analytical Biochemistry 343, 183–185. Studier, W.F., 2005. Protein production by auto-induction in high density shaking cultures. Protein Exp. Purif. 41 (1), 207–234.
doi:10.1016/j.jbiotec.2010.09.684
doi:10.1016/j.jbiotec.2010.09.685
[P-M.97]
[P-M.98]
Production of DNA vaccine against leishmaniasis: comparison of culture media and plasmid size
Submicroscopic and multiple Plasmodium falciparum infections in Sudanese pregnant women
Myriam Sanchez 1,∗ , Jaime Ortega 1 , Eric Dumonteil 2
Samia Omer 1,∗ , Eltahir Khalil 2 , Hashim Ali 1 , Sharief Abdalla 1
1
1
2
2
Introduction: Plasmid DNA (pDNA) vaccines induce a protective or therapeutic immune response against murine Leishmaniasis (Gamboa-León et al., 2006), however high amounts of pure and supercoiled plasmid DNA (sc-pDNA) is needed. The established protocols to produce and purify DNA at laboratory scale, do not satisfy the requirement of pDNA even at the first experimentation
Control of malaria in pregnancy remains a public health challenge. Malarial parasitaemia below the threshold of microscopy but detectable by PCR assays is common in endemic regions. Little information is presented on the frequency of submicroscopic malaria infection, as well as on multiplicity of Plasmodium
CINVESTAV, Mexico 2Centro de Investigaciones Regionales Hideyo Noguchi, Mexico Keywords: pDNA; ZMY505; flasks; induction
Tropical medicine Research Institute, Sudan Institute of Endemic Diseases, Sudan3 Omdurman Maternity Hospital, Omdurman, Sudan Keywords: Malaria; Multiplicity of infection; pregnancy
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
[P-M.95] Promising experimental Calcium silicate-methacrylate hybrid composites able of Calcium and Fluoride releasing for clinical application in dentistry Cesar Henriqhe Zanchi 2 , Carlo Prati 1 , Francesco Siboni 1,∗ , Fabricio Aulo Ogliari 2 , Evandro Piva 2 , Maria Giovanna Gandolfi 1 1
Laboratory of Biomaterials, Department of Odontostomatological Sciences, University of Bologna, Bologna, Italy 2 School of Dentistry, Federal University of Pelotas, Pelotas, RS, Brazil
Fig. 1. Overview of the Magnetic Capture Hybridization Technology for the isolation and analysis of methylation status of cdh1 promoter sequence.
Results: The method showed a high specificity and sensitivity, with a detection level of 20-50 cdh1 copies. Captured fragments were rinsed out of background, denatured and preliminary experiments to analyze methylation status of cdh1 promoter by Pyrosequencing were also carried out. Results indicated that isolated DNA maintains its methylation allowing bisulfite conversion and therefore methylation can be investigated. Conclusions: The MCH technology can be proposed as a valid and versatile tool to isolate potential biomarkers from whole plasma samples after the selection of a specific probe sequence that can be conjugated to paramagnetic nanoparticles. Particularly, the technology could be useful in the screening of a large number of samples, since it is rapid, sensitive and well suited for automation.
Introduction: Dual-curing calcium-silicate-filled methacrylate composites were developed to obtain “smart” materials able of calcium and fluoride releasing for biomimetic remineralization. The composites were prepared incorporating UDMA/HEMA/TEGDMA resin in calcium-silicate powders to obtain fast setting materials for dental applications such as enamel sealant and restorative materials for posterior teeth. Methods: Three calcium-silicate powders were used: grey MTA free from radiopacifing agent (Angelus, Brazil), F10 TC (GandolfiPrati, University of Bologna, Italy) and PC (white Portland cement as control powder). The dual-curing resin contained UDMA, HEMA, TEGDMA and PEG. Each calcium-silicate powders (60% wt) was mixed with the dual-curing UDMA/HEMA/TEGDMA resin (40% wt) obtaining MTA-Rc, F10 TC-Rc and PC-Rc. A resin-reinforced glassionomer cement (Vitrebond, 3 M ESPE, Germany) was tested as control. Ten samples of each composite (8 mm diameter and 1.6 mm thick) were soaked in 10 mL of deionized water and maintained in a cabinet at 37 ◦ C. Calcium and fluorine leached in soaking water and pH of soaking water were measured after 3, 24 hours and 3,7,14,28 days using ion-selective electrodes connected to a meter. ESEM/EDX was performed after soaking in simulated body fluid at 37 ◦ C. Results: All the experimental materials are able to basify the soaking water from pH 6.8 to 8-10 (Table 1). All the experimenTable 1 pH Of Soaking Water (mean ± SD, n = 10) Materials
3 hours
1 day
3 days
7 days
14 days
28 days
MTA-Rc FIOTC-Rc PC-Re Vitrebond Water
10.01 ± 0.18 9.93 ± 0.14 9.99 ± 0.26 6.64 ± 0.14 6.87 ± 0.04
8.72 ± 0.24 9.08 ± 0.52 8.55 ± 0.8 7.53 ± 0.12 6.87 ± 0.26
3.78 ± 0.22 9.00 ± 0.09 8.22 ± 0.5 7.40 ± 0.16 7.02 ± 0.18
8.22 ± 0.06 8.36 ± 0.30 8.36 ± 0.23 8.36 ± 0.12 6.93 ± 0.8
8.06 ± 0.02 8.37 ± 0.10 8.26 ± 0.5 5.90 ± 0.23 7.06 ± 0.48
8.24 ± 0.06 8.11 ± 0.18 8.50 ± 0.8 7.77 ± 0.03 6.98 ± 0.16
tal materials release calcium (higher release by MTA-Rc and PC-Rc) (Table 2). Only F10 TC-Rc and Vitrebond release fluorine (Table 3). Conclusions: The study support the concept that calcium-silicatefilled methacrylate resin composites are “smart” materials able to release calcium and fluoride for remineralizing caries-like dentine
doi:10.1016/j.jbiotec.2010.09.682 Table 2 Materials
3 hours
1 day
3 days
7 days
14 days
28 days
MTA-Rc FIOTC-Rc PC-Re Vitrebond water
1478 ± 1.91 4.25 ± 1.43 13.02 ± 1.04 3.60 ± 1.17 0.60 ± 0.39
14.72 ± 4.09 7.15 ± 1.05 12.98 ± 0.46 2.60 ± 0.84 0.93 ± 0.41
10.27 ± 1.53 3.55 ± 0.45 10.19 ± 2.57 0.60 ± 0.52 1.08 ± 0.30
18.81 ± 1.67 3.20 ± 0.25 29.36 ± 1.82 0.30 ± 0.43 0.98 ± 0.44
15.61 ± 2.03 12.41 ± 3.39 12.09 ± 1.49 0.60 ± 0.52 1.05 ± 0.13
30.01 ± 1.85 6.43 ± 1.87 19.72 ± 1.77 0.40 ± 0.52 0.95 ± 0.31
Table 3 Materials
3 hours
1 day
3 days
7 days
14 days
28 days
MTA-Rc FIOTC-Rc PC-Re Vitrebond water
0.31 ± 0.23 4.11 ± 0.73 0.61 ± 0.27 9.40 ± 0.89 0.17 ± 0.05
0.16 ± 0.13 11.86 ± 1.04 0.65 ± 0.25 11.40 ± 2.51 0.37 ± 0.15
0.23 ± 0.25 15.45 ± 1.87 0.45 ± 0.24 12.40 ± 1.67 0.37 ± 0.17
0.51 ± 0.28 20.85 ± 1.29 0.56 ± 0.24 18.20 ± 4.21 0.35 ± 0.26
0.43 ± 0.21 18.54 ± 1.37 0.84 ± 0.10 14.40 ± 1.14 0.37 ± 0.25
1.05 ± 0.18 14.98 ± 1.81 1.30 ± 0.06 6.00 ± 1.00 0.42 ± 0.22
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
lesions. The incorporation of calcium-silicate particles in resin composite material may extend the range of their clinical applications. doi:10.1016/j.jbiotec.2010.09.683 [P-M.96] Therapeutic potential of plant bioactive molecules L. Bertini 1 , S. Proietti 1 , C. Caporale 1 , F. Mondello 2 , A. Cassone 2 , C. Caruso 1,∗ 1 Dipartimento di Agrobiologia e Agrochimica, Università della Tuscia, Viterbo, Italy, Italy 2 Dipartimento di Malattie Infettive Parassitarie ed Immunomediate, Istituto Superiore di Sanità, Roma, Italy, Italy Keywords: pathogenesis related proteins PR4; antifungal activity; human pathogen; plant bioactive molecules
The search for biologically active compounds from natural sources has always been of great interest to scientist looking for new sources of drugs useful to fight infectious diseases. To this aim, considerable attention has been recently focussed on bio-friendly plant-based products. The plant kingdom represents an extraordinary reservoir of novel molecules and the potential of higher plants as source for new drug is still largely unexplored. Studying the plant defence mechanisms, we focused our attention on Pathogenesis-Related proteins of class 4 (PR4). In the last years, we have isolated and sequenced four PR4 proteins from wheat kernels, named wheatwin1 to wheatwin4, that strongly inhibit both host and non-specific phytopathogenic fungi. We have also demonstrated that they have ribonuclease activity and that their antifungal activity is correlated with the enzymatic one. In this work we report on the isolation of the ortholog gene from Arabidopsis thaliana, coding for a PR4 protein named HEL. This protein was produced at high level in E. coli, which showed ribonuclease and antifungal activities stronger than those shown by the wheatwins. The growing interest around the research of new natural bioactive molecules prompted us to test the activity of this recombinant protein against human pathogens. Preliminary tests carried out on Candida albicans, the etiologic agent of opportunistic mucosal and systemic infections in humans, showed that recombinant HEL has a cytocidal effect on budding cells at a very low concentration. To the best of our knowledge, this is the first time that PR4 proteins have been tested against human pathogens. These results add significance to the exploitation of this molecule in biotechnological process.
S463
steps. Many improvements have been focused in large scale production using bioreactors (Carnes et al., 2006; Sousa et al., 2005). In order to use the equipment (flasks) and growth media available in a common research laboratory, this work studied size of plasmids and protocols (conditions in flasks production) in order to improve sc-pDNA yields. Methods: Escherichia coli DH5␣ was transformed with VR1012NH36 (6 kb) and pVAX-NH36 (4 kb) plasmids and growth in three culture media (LB (control), SDCAS (O’Kennedy et al., 2000) and ZMY505 (Studier, 2005)), in 250 mL flasks at 37 ◦ C 100 and 200 rpm, were gotten characteristic kinetics of dry cellular growth and plasmid DNA production for both conditions. Results: Laboratory scale cultures using LB showed that pDNA volumetric and specific yields were 11 mg/L and 10.7 mg/g dry cell mass for both plasmids, using ZMY505 pDNA yieds were 16 and 18.5 mg/L (VR1012-NH36 and pVAX-NH36 respectively). DNA volumetric yield were increased at 200 rpm giving 24 mg/L and 30 mg/L respectively, in all cases specific and volumetric pDNA yields were highest in 10 or 12 h. Additionally pDNA production was inducible for increased of temperature using different growth phases or times for induction of specific pDNA, theses increased until 2 or 3 fold (VR1012-NH36 and pVAX-NH36, respectively) giving volumetric yields between 40 and 55 mg/L. Discussion: A specialized medium for pDNA production could help for increasing biomass and specific pDNA yields with increase temperature, improving final volumetric pDNA yields. Plasmid yields were highest after the culture has entered the stationary phase because of accumulation (Danquah, 2007). This project was funded by Grant from 167077 and grant PIFUTP08-108 JOL from ICyTDF. References Carnes, A.E., Hodgson, C.P., y Williams, J.A., 2006. Inducible Escherichia coli fermentation for increased plasmid DNA production. Biotechnol. Appl. Biochem 45, 155–166. Danquah, M., y Forde, G., 2007. Growth Medium selection and its economic impact on plasmid DNA production. J. Bios. Bioeng. 104 (6), 490–497. Gamboa-León, R., Paraguai de Souza, E., Borja-Cabrera, G.P., Santos, F.N., Myashiro, L.M., Pinheiro, R.O., Dumonteil, E., Palatnik-de-Sousa, C.B., 2006. Immunotherapy against visceral leishmaniasis with the nucleoside hydrolase-DNA vaccine of Leishmania donovani. Vaccine 24, 4863–4873. O’Kennedy, R.D., Baldwin, C., y Keshavarz-Moore, E., 2000. Effects of growth medium selection on plasmid DNA production and initial processing steps. J. Biotech. 76, 175–183. Sousa, F., Tomaz, C.T., Prazeres, D.M.F., Queiroz, J.A., 2005. Separation of supercoiled and open circular plasmid DNA isoforms by chromatography with a histidine–agarose support. Analytical Biochemistry 343, 183–185. Studier, W.F., 2005. Protein production by auto-induction in high density shaking cultures. Protein Exp. Purif. 41 (1), 207–234.
doi:10.1016/j.jbiotec.2010.09.684
doi:10.1016/j.jbiotec.2010.09.685
[P-M.97]
[P-M.98]
Production of DNA vaccine against leishmaniasis: comparison of culture media and plasmid size
Submicroscopic and multiple Plasmodium falciparum infections in Sudanese pregnant women
Myriam Sanchez 1,∗ , Jaime Ortega 1 , Eric Dumonteil 2
Samia Omer 1,∗ , Eltahir Khalil 2 , Hashim Ali 1 , Sharief Abdalla 1
1
1
2
2
Introduction: Plasmid DNA (pDNA) vaccines induce a protective or therapeutic immune response against murine Leishmaniasis (Gamboa-León et al., 2006), however high amounts of pure and supercoiled plasmid DNA (sc-pDNA) is needed. The established protocols to produce and purify DNA at laboratory scale, do not satisfy the requirement of pDNA even at the first experimentation
Control of malaria in pregnancy remains a public health challenge. Malarial parasitaemia below the threshold of microscopy but detectable by PCR assays is common in endemic regions. Little information is presented on the frequency of submicroscopic malaria infection, as well as on multiplicity of Plasmodium
CINVESTAV, Mexico 2Centro de Investigaciones Regionales Hideyo Noguchi, Mexico Keywords: pDNA; ZMY505; flasks; induction
Tropical medicine Research Institute, Sudan Institute of Endemic Diseases, Sudan3 Omdurman Maternity Hospital, Omdurman, Sudan Keywords: Malaria; Multiplicity of infection; pregnancy