Evaluation of the LH780 hematology analyzer for detection and therapeutic monitoring of malaria: Cross-reactivity with nucleated RBCs

Yoon, J.,Yun, S.G.,Kim, J.,Jung, Y.N.,Koh, Y.E.,Nam, J.H.,Han, E.T.,Lim, C.S. Verlag für Recht und Gesellschaft ; Elsevier 2016 Acta tropica Vol.164 No.-

We evaluated the diagnostic usefulness of the LH780 Coulter blood cell counter for detection and therapeutic monitoring of malaria including cross-reactivity with nucleated RBC (NRBC) samples. A total of 405 patients (43 patients with Plasmodium vivax malaria and the control group of 120 healthy subjects, 111 patients with fever, and 131 patients with NRBCs) were analyzed with routine CBC using the LH780. We analyzed the CBC results according to three selected parameters: an abnormal peak in the WBC histogram before 35fL, the presence of red dots in the nonwhite cell zone of 2D WBC Diff Dataplot, and platelet-related flags suggesting platelet clumps or giant platelets. Of the 43 malaria samples collected at diagnosis, an abnormal peak (≥2.2mm) was present in 93.0% (95% confidential interval (CI), 80.9-98.5%). Of all samples, 97.7% (95% CI, 87.7-99.9%) exhibited red dots, and platelet-related flags were observed in 81.4% (95% CI, 66.6-91.6%). The specificity of these three selected parameters was 83.1% (95% CI, 78.9-86.9%), 77.3% (95% CI, 72.7-81.6%), and 90.1% (95% CI, 86.5-92.9%), respectively. The abnormal peak (≥2.2mm) showed moderate correlation with parasite level (r=0.79). The three selected LH780 parameters were useful for identifying malaria in healthy subjects and febrile patients, but unsatisfactory for discriminating malaria in NRBC samples. The parameters showed a substantial proportion of false positives in the NRBC group, ranging from 26.7% to 49.6%. Therefore, microscopic confirmation will be necessary for application of these parameters for malaria screening and treatment monitoring.

Genetic diversity and natural selection of Duffy binding protein of Plasmodium vivax Korean isolates

Ju, H.L.,Kang, J.M.,Moon, S.U.,Bahk, Y.Y.,Cho, P.Y.,Sohn, W.M.,Park, Y.K.,Park, J.W.,Kim, T.S.,Na, B.K. Verlag für Recht und Gesellschaft ; Elsevier 2013 Acta tropica Vol.125 No.1

Plasmodium vivax Duffy binding protein (PvDBP) is a micronemal type I membrane protein that plays an essential role in erythrocyte invasion of merozoites. PvDBP is a prime blood stage vaccine candidate antigen against P. vivax, but its polymorphic nature represents a major obstacle to the successful design of a protective vaccine against vivax malaria. In this study, we analyzed the genetic polymorphism and natural selection at the N-terminal cysteine-rich region of PvDBP (PvDBPII) among 70 P. vivax isolates collected from Korean patients during 2005-2010. Seventeen single nucleotide polymorphisms (SNP), which resulted in 14 non-synonymous and 3 synonymous mutations, were found in PvDBPII among the Korean P. vivax isolates. Sequence analyses revealed that 13 different PvDBPII haplotypes, which were clustered into 3 distinct clades, were identified in Korean P. vivax isolates. The difference between the rates of nonsynomyous and synonymous mutations suggested that the region has evolved under natural selection. High selective pressure preferentially acted on regions identified or predicted to be B- and T-cell epitopes and MHC binding regions of PvDBPII. Recombination may also contribute to genetic diversity of PvDBPII. Our results suggest that PvDBPII of Korean P. vivax isolates display a limited genetic polymorphism and are under selective pressure. These results have significant implications for understanding the nature of the P. vivax population circulating in Korea and provide useful information for development of malaria vaccines based on this antigen.

Prevalence of Haplorchis taichui among humans and fish in Luang Prabang Province, Lao PDR

Sohn, W.M.,Yong, T.S.,Eom, K.S.,Min, D.Y.,Lee, D.,Jung, B.K.,Banouvong, V.,Insisiengmay, B.,Phommasack, B.,Rim, H.J.,Chai, J.Y. Verlag für Recht und Gesellschaft ; Elsevier 2014 Acta tropica Vol.136 No.-

This study confirmed the prevalence of the intestinal fluke Haplorchis taichui (Trematoda: Heterophyidae) among people and fish in Luang Prabang Province, Lao PDR. Fecal specimens were collected from 559 riparian people (229 males and 330 females), residing in 4 Districts (Luang Prabang, Xieng Ngeun, Pak Ou, and Nam Bak) and were examined by the Kato-Katz fecal smear technique. The overall helminth egg positive rate was 64.9%. The positive rate for small trematode eggs (STE), which may include H. taichui and other heterophyids, Opisthorchis viverrini, and lecithodendriids, was 15.2%. For recovery of adult helminths, 10 STE-positive people were treated with 40mg/kg praziquantel and 15mg/kg pyrantel pamoate, and then purged. Mixed infections with 3 Haplorchis species (H. taichui, H. pumilio, and H. yokogawai), a species of cestode (Taenia saginata), and several species of nematodes including Enterobius vermicularis and hookworms were found. The worm load for trematodes was exclusively high for H. taichui with an average of 7691 specimens per infected person, followed by H. yokogawai (8.3 specimens) and H. pumilio (4.1 specimens). Out of 207 freshwater fish (17 species) purchased in a market in Luang Prabang District, 138 (67%) harboured H. taichui metacercariae (metacercarial burden per fish; 520). Lower prevalence of fish and lower metacercarial density were observed for H. yokogawai (52% and 50 per fish, respectively) and H. pumilio (18% and 3 per fish, respectively). STE found in the surveyed population of Luang Prabang Province were verified to be those of intestinal fukes, particularly H. taichui.

Prevalence of Opisthorchis viverrini infection in humans and fish in Kratie Province, Cambodia

Sohn, W.M.,Yong, T.S.,Eom, K.S.,Pyo, K.H.,Lee, M.Y.,Lim, H.,Choe, S.,Jeong, H.G.,Sinuon, M.,Socheat, D.,Chai, J.Y. Verlag für Recht und Gesellschaft ; Elsevier 2012 Acta tropica Vol.124 No.3

Opisthorchis viverrini is a medically important foodborne parasite in the Indochina Peninsula. In Cambodia, the prevalence of this trematode has been reported in Takeo Province, but not in other areas. In this study, we investigated the prevalence of O. viverrini infection among people in seven riparian villages along the Mekong River, Kratie Province. We also examined the status of metacercarial infection in fish hosts. Fecal specimens were collected from 2101 residents and schoolchildren, and were examined by the Kato-Katz technique. The average O. viverrini egg positive rate was 4.6%, with the highest prevalence found in Roka Kandal A village (10.4%) followed by Talous village (5.9%). In these villages, adult residents showed higher prevalences (19.4% and 9.0%, respectively) than schoolchildren (6.4% and 1.4%, respectively). O. viverrini adult worms were recovered from 2 egg-positive cases (18 and 4 specimens) after praziquantel treatment and purgation. In addition, three of seven freshwater fish species caught near the villages were positive for O. viverrini metacercariae. A total of 367 metacercariae were harvested from 19 infected fish (metacercarial density; 19 per fish). The species of the metacercariae was confirmed through adult worm recovery by experimental infection to hamsters. The results provide evidence that the surveyed areas of Kratie Province, Cambodia, are endemic for O. viverrini infection.

Determination of multiple-clone infection at allelic dimorphism site of Plasmodium vivax merozoite surface protein-1 in the Republic of Korea by pyrosequencing assay

Dinzouna-Boutamba, S.D.,Lee, S.,Son, U.h.,Yun, H.S.,Joo, S.Y.,Jeong, S.,Rhee, M.H.,Kwak, D.,Xuan, X.,Hong, Y.,Chung, D.I.,Goo, Y.K. Verlag für Recht und Gesellschaft ; Elsevier 2017 Acta tropica Vol.176 No.-

<P>Allelic diversity leading to multiple gene polymorphisms of vivax malaria parasites has been shown to greatly contribute to antigenic variation and drug resistance, increasing the potential for multiple-clone infections within the host. Therefore, to identify multiple-clone infections and the predominant haplotype of Plasmodium vivax in a South Korean population, P. vivax merozoite surface protein-1 (PvMSP-1) was analyzed by pyrosequencing. Pyrosequencing of 156 vivax malaria-infected samples yielded 97 (62.18%) output pyrograms showing two main types of peak patterns of the dimorphic allele for threonine and alanine (T1476A). Most of the samples evaluated (88.66%) carried multiple-clone infections (wild- and mutant-types), whereas 11.34% of the same population carried only the mutant-type (1476A). In addition, each allele showed a high frequency of guanine (G) base substitution at both the first and third positions (86.07% and 81.13%, respectively) of the nucleotide combinations. Pyrosequencing of the PvMSP-1 42-kDa fragment revealed a heterogeneous parasite population, with the mutant-type dominant compared to the wild-type. Understanding the genetic diversity and multiple-clone infection rates may lead to improvements in vivax malaria prevention and strategic control plans. Further studies are needed to improve the efficacy of the pyrosequencing assay with large sample sizes and additional nucleotide positions.</P>

Visceral leishmaniasis: Revisiting current treatments and approaches for future discoveries

Verlag für Recht und Gesellschaft ; Elsevier 2016 Acta tropica Vol.155 No.-

<P>The current treatments for visceral leishmaniasis are old and toxic with limited routes of administration. The emergence of drug-resistant Leishmania threatens the efficacy of the existing reservoir of antileishmanials, leading to an urgent need to develop new treatments. It is particularly important to review and understand how the current treatments act against Leishmania in order to identify valid drug targets or essential pathways for next-generation antileishmanials. It is equally important to adapt newly emerging biotechnologies to facilitate the current research on the development of novel antileishmanials in an efficient fashion. This review covers the basic background of the current visceral leishmaniasis treatments with an emphasis on the modes of action. It briefly discusses the role of the immune system in aiding the chemotherapy of leishmaniasis, describes potential new antileishmanial drug targets and pathways, and introduces recent progress on the utilization of high-throughput phenotypic screening assays to identify novel antileishmanial compounds. (C) 2016 Elsevier B.V. All rights reserved.</P>

Identification and characterization of a serine protease inhibitor of Clonorchis sinensis

Kang, J.M.,Sohn, W.M.,Ju, J.W.,Kim, T.S.,Na, B.K. Verlag für Recht und Gesellschaft ; Elsevier 2010 Acta tropica Vol.116 No.2

A gene encoding a serine protease inhibitor of Clonorchis sinensis (CsSERPIN) was identified and characterized. CsSERPIN contained an open reading frame of 1158bp that encoded 385 amino acid residues. Sequence analysis of the primary structure of CsSERPIN revealed that it had essential structural motifs including a reactive central loop (RCL), which well conserved in the serine protease inhibitor (serpin) superfamily. CsSERPIN was classified as a member of the ovalbumin-type serpin family on the basis of phylogenetic analysis and the absence of a classical N-terminal signal peptide. Recombinant CsSERPIN showed an inhibitory effect on chymotrypsin in a dose-dependent manner, but did not effectively inhibit trypsin, thrombin, elastases or cathepsin G. Optimal pH values of CsSERPIN were between 7.0 and 9.0, as evidenced by the rapid loss of inhibitory activity under acidic conditions. CsSERPIN was expressed at various developmental stages of the parasite, from eggs to adult worms, but its expression level was higher in eggs and adult worms than in metacercariae and juvenile worms. CsSERPIN was identified in the soluble extract of the parasite, but not in the excretory and secretory products (ESP) or insoluble extract of the parasite. Immunolocalization analysis of CsSERPIN showed that it mainly localized to the eggs and vitelline glands of the adult worm. These results suggest that intracellular CsSERPIN may be possibly involved in maintaining the physiology of eggs as well as in egg production of C. sinensis by regulating endogenous serine proteases.

Genetic polymorphism in pvmdr1 and pvcrt-o genes in relation to in vitro drug susceptibility of Plasmodium vivax isolates from malaria-endemic countries

Lu, F.,Lim, C.S.,Nam, D.H.,Kim, K.,Lin, K.,Kim, T.S.,Lee, H.W.,Chen, J.H.,Wang, Y.,Sattabongkot, J.,Han, E.T. Verlag für Recht und Gesellschaft ; Elsevier 2011 Acta Tropica Vol.117 No.2

Treatment failure of chloroquine for Plasmodium vivax infection has increased in endemic countries. However, the molecular mechanisms for resistance and in vitro susceptibility of P. vivax to chloroquine remain elusive. We investigated the prevalence of mutations in the pvmdr1 and pvcrt-o genes, and the copy number of the pvmdr1 gene in isolates from the Republic of Korea (ROK), Thailand, the Union of Myanmar (Myanmar), and Papua New Guinea (PNG). We also measured in vitro susceptibility of Korean isolates to antimalarial drugs. The pvmdr1 analysis showed that mutations at amino acid position Y976F of pvmdr1 were found in isolates from Thailand (17.9%), Myanmar (13.3%), and PNG (100%), but none from the ROK, and mutation at position F1076L was present in isolates from the ROK (100%), Thailand (60.7%), and Myanmar (46.7%). One copy of the pvmdr1 gene was observed in most isolates and double copy numbers of the gene were observed in two Thai isolates. In the exons of the pvcrt-o gene that were sequenced, a K10 insertion was present in isolates from Thailand (56.0%) and Myanmar (46.2%), and the wild type was found in all Korean isolates. The results suggest that gene polymorphisms and copy number variation was observed in isolates of P. vivax from Southeast Asian countries. In Korean isolates polymorphism as limited to the F1076L variant, and no isolates with high level of resistance were found by in vitro susceptibility determinations. Moreover, our results provide a baseline for future prospective drug studies in malaria-endemic areas.