6-phosphogluconate+dehydrogenase,+decarboxylating,+putative+Leishmania+major

Leishmaniasis is a metazoonosis disease caused by the gensus //Leishmania// found from a bite of a phlebotomine female sand fly of the genera //Phlebotomus// and //Lutzomyia// [1]. When a parasitic female sand fly bites into a human host, the parasite undergoes the intracellular stage where it takes on a promastigote form. Then the parasite reproduces and multiplies within the macrophages until it lyses, and then the cycle continues, affecting even “lymph nodes, spleen, liver and the bone marrow” [1]. This disease is primarily found in the tropics and “primarily in poor and underserved populations” [2]. There are 3 majors severity levels of Leishmaniasis that are visceral (kala-azar), cutaneous, and mucocutaneous. Visceral is the most severe due to the fact that the “vector-borne infection has clinical spectrums that ranges” depending successful the immune system inhibits the proliferation of the parasite [3]. Since the parasite invades the host cell by impairing cell functions by “altering the maturation process of the phagolysosome, cytokine, and chemokine production,” it is extremely difficult to diagnosis the disease [3]. Due to the fact that Leishmaniasis is a silent killer, where symptoms do not become apparent until the disease has progressed, it is the “most deadly parasitic disease after malaria” [3]. Current treatments against leishmaniasis are expensive due to its large scope of resistance against several clinical treatments. Even though there are many medical tests, for instance the Indirect Fluorescent Antibody (IFA) test, that detects the early stages of the injection, but not after six to nine months, there is a need for new treatments against Leishmaniasis [1].
 * *Target (protein/gene name): ** 6-phosphogluconate dehydrogenase decarboxylating, putative
 * *NCBI Gene # or RefSeq#: **12980488
 * *Protein ID (NP or XP #) or Wolbachia#: ** XP_003722728 [|(link!)]
 * *Organism (including strain): **// Leishmania Major //
 * Etiologic Risk Group (see link below): ** Risk Group 2 [|(link!)]
 * */ ** ** Disease Information ** ** (sort of like the Intro to your Mini ****__ Research Write __**** up): **

It is essential the pathway of the pentose phosphate pathway (PPP) and the NADP-dependent reaction where NADPH is formed. 6PGDdp plays a crucial role of the host-parasite relationship by providing the parasite the need to generate a defense against oxidative stress [6]. Inhibiting this reaction mechanism with PPP is crucial in reducing the biosynthesis pathways and the cellular redox exchange as well as reducing the synthesis of nucleotides for nucleotide and nucleic acid production
 * Link to TDR Targets page (if present): ** [|(][|**link!)**]
 * Link to Gene Database page ([|NCBI], EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **
 * Essentiality of this protein: **

Is it a monomer or multimer as biological unit **? (make prediction at** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): Dimer
 * Complex of proteins?: ** Dimer
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): ** 0.8 (TDR Target)
 * *EC#: ** 1.1.1.44
 * Link to BRENDA EC# page: **** [|(link!)] **
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic



Figure 1: Reaction Mechanism of 6-phosphogluconate dehydrogenase (E.C. 1.1.1.44) obtained from BRENDA [4].

** Enzyme Assay ** ** information (spectrophotometric, coupled assay ?, reagents): ** [] [] [] []®ion=US
 * -- links to assay reagents (substrates) pages: **
 * --- List cost and quantity of substrate reagents, supplier, and catalog # **

-- PDB # or closest PDB entry if using homology model: Homology 1PGJ -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB Query Coverage: 97% Max % Identities: 339/465% (73%) % Positives: 388/465 (83%) Chain used for homology: Chain A Figure 2: Homology Model Pair Alignment with 1PGJ (//T. Brucei)// with Query Coverage 97%, Maximum % of Identities 73%, and Positive % 83% for Chain A.
 * Structure (PDB or Homology model) **

Figure 3: PYMOL representation of the 3 inhibitors shown as purple sticks of 6PGD used with the active site shown as purple of the homology model PDB 1PGJ surface transperancy of 0.5 with carbon colored as green. The ligans are as labeled 4a) 4-phospho-D-erythronate, 4b) 2-Deoxy-6-phosphogluconate, and 4c) 5-phospho-d-rinbonate.
 * Current Inhibitors: ** 4a) 4-phospho-D-erythronate, 4b) 2-Deoxy-6-phosphogluconate, and 4c) 5-phospho-d-rinbonate.

** Purification Method ** ** : ** Metal Affinity Chromatography (NIckle-NTA) Figure 4: PYMOL representation of homology model of PDB 1PGJ of target protein shown as surface transperency 0.5 and overall with green carbons. Active region shown as cyan.
 * Expression Information (has it been expressed in bacterial cells): ** E.Coli pNIC-Bsa4
 * Image of protein (PyMol with features delineated and shown separately): **

** Molecular Weight ** ** of your protein in kiloDaltons using the **** [|Expasy ProtParam] **** website :** 52314.88 g/mol
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **
 * MSNDLGIIGLGVMGANLALNIAEKGFKVAVFNRTYAKTTSFLKEHESEKFAANLNGYETM ||
 * KEFAASLKKPRRAFILVQAGAATDSTIEQLKEVFENGDIIIDTGNANFKDQDKRAAQLES ||
 * QGLRFLGMGISGGEEGARKGPAFFPGGTPSVWEEVRPIVEAAAAKAEDGRPCVTFNGKGG ||
 * AGSCVKMYHNAGEYAVLQIWGEAYSALLAFGFDNDQIADVFESWKADGFLKSYMLDISIA ||
 * ACRAREATGNYLSEKVKDRIGSKGTGLWSAQEALEIGVPAPSLNMAVISRQMTMYKGERI ||
 * ANCKAFPNFPRGPSEEATDKSPNSPEAKKLYHAVSLCIIASYAQMFQCLRELDKVYGFGL ||
 * NLPATIATFRAGCILQGYLLGPMTKAFEENPNLPNLMDAFTKEIAAGLDDCRQILAKLTV ||
 * NTAVSLPVMMASLSYINAMYTETLPYGQLVSLQRDVFGRHGYERTDKDGRESFEWPALQX ||
 * *length of your protein in Amino Acids :** 430 aa
 * Molar ** ** Extinction coefficient ** ** of your protein at 280 nm wavelength: ** 51840 Lmol-1cm-1
 * TMpred graph Image ** ( @http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
 * *CDS ** ** Gene Sequence ** ** (paste as text only): **
 * ATGTCTAACGACCTGGGCATCATTGGCCTGGGTGTTATGGGTGCGAACCTGGCGCTGAAC ||
 * ATCGCAGAAAAAGGTTTCAAAGTAGCGGTTTTTAACCGCACCTACGCCAAGACCACCTCT ||
 * TTCCTGAAGGAACATGAATCTGAGAAATTCGCCGCTAATCTGAACGGTTACGAAACCATG ||
 * AAAGAATTCGCTGCTTCTCTCAAGAAGCCGCGTCGTGCTTTCATCCTCGTTCAGGCAGGT ||
 * GCAGCGACTGACTCTACTATCGAGCAACTCAAGGAAGTGTTCGAAAATGGTGACATCATT ||
 * ATTGACACGGGTAATGCGAACTTCAAAGACCAGGACAAGCGTGCCGCGCAACTGGAATCT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CAGGGTCTCCGTTTCCTGGGTATGGGTATCTCTGGTGGTGAAGAAGGCGCACGTAAGGGT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CCAGCGTTCTTTCCGGGTGGTACCCCGTCTGTTTGGGAAGAGGTTCGTCCGATCGTAGAA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GCGGCTGCGGCAAAAGCTGAGGACGGTCGTCCATGCGTTACCTTTAACGGCAAGGGCGGT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GCGGGTAGCTGCGTTAAAATGTACCATAACGCGGGTGAGTATGCTGTGCTCCAAATCTGG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GGTGAGGCGTACTCTGCGCTGCTGGCATTCGGCTTTGACAATGACCAGATCGCCGACGTT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TTTGAGTCTTGGAAGGCGGACGGTTTTCTCAAATCTTATATGCTCGACATCTCTATCGCT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GCGTGTCGCGCACGCGAAGCAACCGGTAACTACCTGTCTGAAAAAGTTAAAGACCGTATC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GGTTCTAAGGGTACCGGCCTGTGGAGCGCGCAGGAAGCGCTGGAAATCGGTGTTCCGGCA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CCGAGCCTCAACATGGCTGTTATCTCTCGTCAGATGACCATGTATAAGGGCGAGCGTATC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GCGAACTGCAAAGCATTCCCGAACTTTCCTCGTGGTCCGTCTGAAGAGGCAACGGACAAA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TCTCCGAACTCTCCGGAAGCAAAGAAGCTGTATCACGCGGTCTCCCTGTGCATTATCGCG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TCTTACGCGCAGATGTTCCAGTGCCTGCGTGAACTGGACAAAGTTTACGGCTTCGGTCTG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AACCTGCCAGCCACGATCGCCACTTTCCGCGCTGGCTGCATCCTGCAAGGCTATCTGCTG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GGCCCTATGACCAAGGCATTTGAGGAAAATCCGAATCTGCCAAACCTGATGGACGCGTTC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ACCAAAGAAATTGCTGCGGGCCTGGATGACTGCCGCCAGATCCTGGCGAAACTGACGGTT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AACACCGCAGTTTCTCTGCCTGTGATGATGGCCTCTCTGTCTTACATCAACGCGATGTAT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ACGGAAACGCTCCCGTATGGTCAGCTGGTGTCTCTGCAACGTGACGTATTCGGCCGTCAC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GGTTATGAACGTACCGATAAAGATGGCCGTGAATCTTTTGAATGGCCGGCTCTCCAGTAA ||


 * *GC% Content for gene: **** 53.125% **
 * *CDS Gene Sequence (codon optimized) - copy from ** ** output ** ** of ** ** Primer Design ** ** Protocol (paste as text only): **
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ATGTCTAACGACCTGGGCATCA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GATGTTCAGCGCCAGGTTCGCACCCATAACACCCAGGCCAATGATGCCCAGGTCGTTAGA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AACCTGGCGCTGAACATCGCAGAAAAAGGTTTCAAAGTAGCGGTTTTTAACCGCACCTAC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TCTCAGATTCATGTTCCTTCAGGAAAGAGGTGGTCTTGGCGTAGGTGCGGTTAAAAACCG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CCTGAAGGAACATGAATCTGAGAAATTCGCCGCTAATCTGAACGGTTACGAAACCATGAA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AAAGCACGACGCGGCTTCTTGAGAGAAGCAGCGAATTCTTTCATGGTTTCGTAACCGTTC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AGCCGCGTCGTGCTTTCATCCTCGTTCAGGCAGGTGCAGCGACTGACTCTACTATCGAGC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CAATAATGATGTCACCATTTTCGAACACTTCCTTGAGTTGCTCGATAGTAGAGTCAGTCG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TTCGAAAATGGTGACATCATTATTGACACGGGTAATGCGAACTTCAAAGACCAGGACAAG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AGGAAACGGAGACCCTGAGATTCCAGTTGCGCGGCACGCTTGTCCTGGTCTTTGAAGTTC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CTCAGGGTCTCCGTTTCCTGGGTATGGGTATCTCTGGTGGTGAAGAAGGCGCACGTAAGG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CTTCCCAAACAGACGGGGTACCACCCGGAAAGAACGCTGGACCCTTACGTGCGCCTTCTT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CCCCGTCTGTTTGGGAAGAGGTTCGTCCGATCGTAGAAGCGGCTGCGGCAAAAGCTGAGG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CCCGCACCGCCCTTGCCGTTAAAGGTAACGCATGGACGACCGTCCTCAGCTTTTGCCGCA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AAGGGCGGTGCGGGTAGCTGCGTTAAAATGTACCATAACGCGGGTGAGTATGCTGTGCTC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CCGAATGCCAGCAGCGCAGAGTACGCCTCACCCCAGATTTGGAGCACAGCATACTCACCC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CGCTGCTGGCATTCGGCTTTGACAATGACCAGATCGCCGACGTTTTTGAGTCTTGGAAGG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AGAGATGTCGAGCATATAAGATTTGAGAAAACCGTCCGCCTTCCAAGACTCAAAAACGTC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CAAATCTTATATGCTCGACATCTCTATCGCTGCGTGTCGCGCACGCGAAGCAACCGGTAA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ACCCTTAGAACCGATACGGTCTTTAACTTTTTCAGACAGGTAGTTACCGGTTGCTTCGCG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ACCGTATCGGTTCTAAGGGTACCGGCCTGTGGAGCGCGCAGGAAGCGCTGGAAATCGGTG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TCTGACGAGAGATAACAGCCATGTTGAGGCTCGGTGCCGGAACACCGATTTCCAGCGCTT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TGGCTGTTATCTCTCGTCAGATGACCATGTATAAGGGCGAGCGTATCGCGAACTGCAAAG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CGTTGCCTCTTCAGACGGACCACGAGGAAAGTTCGGGAATGCTTTGCAGTTCGCGATACG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">CCGTCTGAAGAGGCAACGGACAAATCTCCGAACTCTCCGGAAGCAAAGAAGCTGTATCAC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ATCTGCGCGTAAGACGCGATAATGCACAGGGAGACCGCGTGATACAGCTTCTTTGCTTCC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GCGTCTTACGCGCAGATGTTCCAGTGCCTGCGTGAACTGGACAAAGTTTACGGCTTCGGT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">AGCCAGCGCGGAAAGTGGCGATCGTGGCTGGCAGGTTCAGACCGAAGCCGTAAACTTTGT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ACTTTCCGCGCTGGCTGCATCCTGCAAGGCTATCTGCTGGGCCCTATGACCAAGGCATTT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TGAACGCGTCCATCAGGTTTGGCAGATTCGGATTTTCCTCAAATGCCTTGGTCATAGGGC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ACCTGATGGACGCGTTCACCAAAGAAATTGCTGCGGGCCTGGATGACTGCCGCCAGATCC ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">ATCACAGGCAGAGAAACTGCGGTGTTAACCGTCAGTTTCGCCAGGATCTGGCGGCAGTCA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GCAGTTTCTCTGCCTGTGATGATGGCCTCTCTGTCTTACATCAACGCGATGTATACGGAA ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GTCACGTTGCAGAGACACCAGCTGACCATACGGGAGCGTTTCCGTATACATCGCGTTGAT ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">GGTGTCTCTGCAACGTGACGTATTCGGCCGTCACGGTTATGAACGTACCGATAAAGATGG ||
 * <span style="color: #4f81bd; font-family: 'Times New Roman',serif;">TTACTGGAGAGCCGGCCATTCAAAAGATTCACGGCCATCTTTATCGGTACGTTCATAA ||

Do Not Need this info for Spring (but still copy these lines to your Target page for now) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to. __ TACTTCCAATCC __<span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">ATGTCTAACGACCTGGGCATCATTGGCCTGGGTGTTATGGGTGCGAACCTGGCGCTGAACATCGCAGAAAAAGGTTTCAAAGTAGCGGTTTTTAACCGCACCTACGCC <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">AAGACCACCTCTTTCCTGAAGGAACATGAATCTGAGAAATTCGCCGCTAATCTGAACGGTTACGAAACCATGAAAGAATTCGCTGCTTCTCTCAAGAAGCCGCGTCGTGCTTTCATCCTC <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">GTTCAGGCAGGTGCAGCGACTGACTCTACTATCGAGCAACTCAAGGAAGTGTTCGAAAATGGTGACATCATTATTGACACGGGTAATGCGAACTTCAAAGACCAGGACAAGCGTGCCGC <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">GCAACTGGAATCTCAGGGTCTCCGTTTCCTGGGTATGGGTATCTCTGGTGGTGAAGAAGGCGCACGTAAGGGTCCAGCGTTCTTTCCGGGTGGTACCCCGTCTGTTTGGGAAGAGGTTCG <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">TCCGATCGTAGAAGCGGCTGCGGCAAAAGCTGAGGACGGTCGTCCATGCGTTACCTTTAACGGCAAGGGCGGTGCGGGTAGCTGCGTTAAAATGTACCATAACGCGGGTGAGTATGCTGT <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">GCTCCAAATCTGGGGTGAGGCGTACTCTGCGCTGCTGGCATTCGGCTTTGACAATGACCAGATCGCCGACGTTTTTGAGTCTTGGAAGGCGGACGGTTTTCTCAAATCTTATATGCTCGAC <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">ATCTCTATCGCTGCGTGTCGCGCACGCGAAGCAACCGGTAACTACCTGTCTGAAAAAGTTAAAGACCGTATCGGTTCTAAGGGTACCGGCCTGTGGAGCGCGCAGGAAGCGCTGGAAAT <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">CGGTGTTCCGGCACCGAGCCTCAACATGGCTGTTATCTCTCGTCAGATGACCATGTATAAGGGCGAGCGTATCGCGAACTGCAAAGCATTCCCGAACTTTCCTCGTGGTCCGTCTGAAGAG <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">GCAACGGACAAATCTCCGAACTCTCCGGAAGCAAAGAAGCTGTATCACGCGGTCTCCCTGTGCATTATCGCGTCTTACGCGCAGATGTTCCAGTGCCTGCGTGAACTGGACAAAGTTTAC <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">GGCTTCGGTCTGAACCTGCCAGCCACGATCGCCACTTTCCGCGCTGGCTGCATCCTGCAAGGCTATCTGCTGGGCCCTATGACCAAGGCATTTGAGGAAAATCCGAATCTGCCAAACCTG <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">ATGGACGCGTTCACCAAAGAAATTGCTGCGGGCCTGGATGACTGCCGCCAGATCCTGGCGAAACTGACGGTTAACACCGCAGTTTCTCTGCCTGTGATGATGGCCTCTCTGTCTTACATC <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">AACGCGATGTATACGGAAACGCTCCCGTATGGTCAGCTGGTGTCTCTGCAACGTGACGTATTCGGCCGTCACGGTTATGAACGTACCGATAAAGATGGCCGTGAATCTTTTGAATGGCCGG <span style="color: #5b9bd5; font-family: 'Times New Roman',serif;">CTCTCCAGTAA __ CAGTAAAGGTGGATA __
 * *GC% Content for gene (codon optimized): ** 52.688679%
 * Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **
 * ( **** link to DNA Works output ** ** text file ** ** - ** that should be saved in your Google Docs folder after you did the primer design protocol)

Forward Primer: 5’ __TACTTCCAATCCATG__TCTAACGACCTGGGCATCATTGG 3’ 38 bp <span style="color: #4f81bd; font-family: 'Times New Roman',serif; font-size: 12pt;">Reverse Primer: 5’ __TATCCACCTTTACTG__TTACTGGAGAGCCGGCCATTC 3’ 36 b
 * Primer design results for 'tail' primers (this is just 2 sequences): **

<span style="font-family: 'Times New Roman',serif; font-size: 12pt;">References: <span style="font-family: 'Times New Roman',serif; font-size: 12pt;">(1) Dawit, G. //J. Bacteriol. Parasitol.// **2013**, //4//, 2–7. <span style="font-family: 'Times New Roman',serif; font-size: 12pt;">(2) Miranda, A.; Saldaña, A.; González, K.; Paz, H.; Santamaría, G.; Samudio, F.; Calzada, J. E. //Trans. R. Soc. Trop. Med. Hyg.// **2012**, //106// (9), 544–548. <span style="font-family: 'Times New Roman',serif; font-size: 12pt;">(3) Savoia, D. //J. Infect. Dev. Ctries.// **2015**, //9// (6), 588–596.