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S Jongwutiwes

Publications and source records attributed to S Jongwutiwes.

At least 19 recordsLinked to original sources
PubMed2002

Molecular analysis of Cryptosporidium species isolated from HIV-infected patients in Thailand.

Cryptosporidium isolates from diarrhoeal stools of human immunodeficiency virus (HIV)-infected patients in Thailand were genetically analysed by sequencing the variable region in the 18S rRNA gene. Twenty-nine isolates from four children and 25 adults attending King Chulalongkorn Memorial Hospital in Bangkok during 1996 and 2000 were analysed. All patients suffered from chronic watery diarrhoea and had low CD4+ lymphocytes (mean +/- SD=105.5 +/- 133.2 cells/microl). Four Cryptosporidium species were identified, i.e. C. parvum (genotype 1), C. meleagridis, C. muris and C. felis occurring in 24, 3, 1 and 1 isolates, respectively. Oocysts of C. muris were significantly larger than oocysts of other species; C. felis was the smallest in these populations (P < 0.01). Sequences of the ITS1, 5.8S rRNA and ITS2 regions of C. muris and C. meleagridis identified in this study displayed unique sequences from those of other known species. Based on a limited number of isolates analysed, only C. meleagridis and C. muris were found in HIV-infected children, whereas the genotype 1 of C. parvum predominated in HIV-infected adults.

AIDS-Related Opportunistic Infections↗
PubMed2001

Diversity in the thrombospondin-related adhesive protein gene (TRAP) of Plasmodium vivax.

We analyzed 22 clinical isolates of Plasmodium vivax from Thailand and 17 from Brazil to investigate the extent of sequence variation in the thrombospondin-related adhesive protein of Plasmodium vivax (PvTRAP), a homologue of P. falciparum TRAP (PfTRAP) which has been considered to be a promising vaccine candidate. In total 54 haplotypes were identified from 73 distinct gene clones. Coexistence of different PvTRAP in circulation occurred in 10 and 13 isolates from Thailand and Brazil, respectively. Forty out of 48 substituted nucleotides are non-synonymous changes. Most of the substituted residues reside in the von Willebrand factor type A-domain (region II), a sulfated glycosaminoglycan-binding domain (region III) and a proline-rich region (region IV). All nucleotide substitutions are dimorphic. Two haplotypes from Thailand contain an inserted sequence encoding aspartic acid-serine-proline in the proline-rich region. Sequence analysis has revealed that nucleotide diversity in PvTRAP is low although Brazilian isolates display a higher degree of variation than those from Thailand. Phylogenetic construction using the neighbor joining method has shown that most of the Thai and the Brazilian isolates appear to be mainly clustered into distinct groups. Significantly greater than expected values of the mean number of non-synonymous (d(n)) than synonymous (d(s)) nucleotide substitutions per site were observed in regions II and III of PvTRAP. Analysis of the published PfTRAP sequences has shown a similar finding in regions II and IV suggesting that positive selection operates on the regions. Hence, different regions in PvTRAP and PfTRAP could be under different pressures in terms of immune selection, structural and/or functional constraints.

Amino Acid Sequence↗
PubMed2000

Intragenic recombination in the 3' portion of the merozoite surface protein 1 gene of Plasmodium vivax.

To date, little has been known about the extent of sequence variation in the C-terminal part of the Plasmodium vivax merozoite surface protein 1 (PvMSP1) which has been considered to be a potential vaccine candidate. Here, we examined the variation in the region encompassing interspecies conserved blocks (ICBs) 8 and 10 of PvMSP1 by DNA sequencing of 14 Thai isolates and three Brazilian isolates. Eighteen different alleles were detected. Three new sequence types had been identified in polymorphic region between ICB8 and CB9: one was possibly a result of intragenic recombination between the Belem and Salvador I alleles and the others displayed unique repeats. A striking variation was observed in a stretch of 38 codons in polymorphic block between conserved block CB9 and ICB10, resulting in eight different sequence types, probably generated by interallelic recombination at a single or multiple sites. There is no apparent linkage between these two polymorphic sites. On the other hand, a single or stretches of nucleotide substitutions are dimorphic like in Plasmodium falciparum MSP1 (PfMSP1) in the remaining parts, creating microheterogeneity of sequences. The C-terminal 19 kDa-encoding region was extremely conserved with a single dimorphic exchange at a known position. Thus, this study provides evidence of intragenic recombination occurring in the 3' portion of PvMSP1 and suggests that the 3' portion of PvMSP1 is more diverse than that in PfMSP1.

3' Untranslated Regions↗
PubMed2000

Heterogeneity in cyst morphology within isolates of Acanthamoeba from keratitis patients in Thailand.

We isolated Acanthamoebae from the first two keratitis patients identified in Thailand in 1988 and 1990. The patients developed decreased vision, severe photophobia, severe eye pain and foreign body sensation after minor corneal trauma. The lesions included generalized superficial punctate keratitis, stromal corneal ulcer with keratic precipitate and uveitis in one case, and corneal ulcer with abscess in the other. Both cases were diagnosed by isolation of characteristic trophozoites and cysts of Acanthamoeba from corneal tissue by non-nutrient agar culture method. Based on cyst morphology, A. castellanii and A. polyphaga were detected in one case, and A. castellanii and A. triangularis in the other. Restriction fragment length polymorphism analysis of mitochondrial DNA (mtDNA-RFLP) revealed that each patient harboured a single parasite population. One shared mtDNA-RFLP with an authentic strain of A. castellanii, and the other gave a new unique pattern. Thus species identification of Acanthamoeba based on cyst morphology per se can be arbitrary, and mtDNA-RFLP may be more appropriate for accurate species/strain differentiation amongst morphologically heterogeneous populations of Acanthamoebae.

Acanthamoeba↗
PubMed1999

Allelic recombination and linkage disequilibrium within Msp-1 of Plasmodium falciparum, the malignant human malaria parasite.

The C-terminal, cysteine-rich 19kDa domain of merozoite surface protein-1 (MSP-1) of Plasmodium falciparum is a target of the host's humoral immunity and thus a malaria vaccine candidate. Although variation in the 19kDa domain is limited among parasite isolates, tertiary structure-dependent intramolecular associations between the 19kDa domain and other parts of MSP-1 are suggested to be involved in immune evasion by allowing competitive binding of protective and non-protective antibodies directed to their epitopes, which are conformationally in close proximity but separated at the primary structure. Since allelic recombination can account for the major variability of the Msp-1 gene, we examined whether linkage disequilibrium occurs between polymorphic loci in the 5'- and the 3'-region, the latter encoding the 19kDa domain. From 184 Thai field isolates, we selected 69 isolates with a single allelic type in six variable blocks of Msp-1 as determined by PCR-based allelic typing. All the isolates showed no evidence of recombination in blocks 6 to 16, whereas recombination was apparent in blocks 2 to 6. Sequencing of the 3'-region revealed two potential recombination sites in block 17. Strong linkage disequilibrium was seen between polymorphic loci in the 5'- and 3'-regions. The strength of this disequilibrium did not correlate with distance between loci. We discuss the possible role of epistatic selection on particular association types (haplotypes) of Msp-1.

Alleles↗
PubMed1998

First outbreak of human trichinellosis caused by Trichinella pseudospiralis.

This is the first report of an epidemic of human infection with Trichinella pseudospiralis. An outbreak of trichinellosis affecting 59 individuals, of whom one died, occurred in southern Thailand during 1994-1995. The source of this epidemic was raw pork from a wild pig that was distributed to villagers by a local hunter. The most striking clinical features among 50 individuals who could be followed were muscular swelling, myalgia, and asthenia persisting for > 4 months. These were associated with significant elevations of creatine phosphokinase and lactate dehydrogenase levels. All patients had Trichinella-specific IgG antibodies in an enzyme-linked immunosorbent assay. Muscle biopsies, performed in six cases, showed nonencapsulated, actively migrating Trichinella larvae. Experimental infection of mice with larvae from human biopsies revealed nonencapsulated muscle larvae consistent with T. pseudospiralis. The identification of muscle larvae from a human specimen by random amplified polymorphic DNA analysis confirmed the causative agent to be T. pseudospiralis. Patients seemed to respond best to treatment with albendazole.

Adolescent↗
PubMed1997

Interallelic recombination in the merozoite surface protein 1 (MSP-1) gene of Plasmodium vivax from Thai isolates.

The merozoite of Plasmodium vivax possesses a high molecular mass surface protein called Pv-merozoite surface protein 1, PvMSP-1, which exhibits antigenic diversity among isolates. In this study, the extent of sequence variation in the polymorphic region and the flanking interspecies conserved blocks (ICBs) 5 and 6 of the PvMSP-1 gene was analyzed using the polymerase chain reaction to amplify the DNA fragment encompassing these regions, followed by sequencing. Twenty different alleles were obtained from 15 Thai isolates. Results revealed five distinct sequence types of the polymorphic region, two of which were newly identified in this study: one probably generated by intragenic recombination at a site different from that previously reported and the other by duplication of a 30 nucleotide (nt) sequence at the 3' end of the region. On the other hand, almost all nucleotide substitutions in the flanking regions, ICB5 and ICB6, were dimorphic, creating microheterogeneity in the region. Furthermore, stretches of nucleotide substitutions were found to be linked in ICB6, suggesting the potential recombination sites between these stretches. It is also noted that extensive sequence variation in the PvMSP-1 gene and coinfection with different PvMSP-1 alleles occurred among the P. vivax population in the endemic areas of Thailand.

Animals↗
PubMed1996

The merozoite surface protein 2 (MSP2) gene of Plasmodium falciparum from a Thai isolate.

The merozoite surface protein 2 (MSP2) of Plasmodium falciparum was a malaria vaccine candidate. The gene encoding MSP2 of a Thai isolate was amplified by polymerase chain reaction followed by subcloning into a phagemid vector and sequencing. Sequence alignment with other previously published sequences revealed that the MSP2 allele in this isolate belonged to FC27 allelic family. The central variable sequence of the MSP2 allele in this study was related to an allele from Indonesia. The flanking sequences of the variable region were highly conserved.

Amino Acid Sequence↗
PubMed1994

Allelic variation in the circumsporozoite protein of Plasmodium falciparum from Thai field isolates.

Allelic variation in the Plasmodium falciparum circumsporozoite (CS) protein gene has been examined by sequencing the entire gene in 15 isolates from an endemic area of Thailand. The isolates contain a total of six new allelic forms of the tetrapeptide repeats and eight variants of the T cell epitope (TCE) region of the CS gene. All nucleotide substitutions in the TCE are nonsynonymous. There is no apparent association between the sequence patterns in the repeats and in the TCE. Comparison of the TCE with published sequences has shown that most variants of our isolates are not identical to those found in different geographic areas, suggesting geographic variation in genetic diversity of the CS protein. In a phylogenetic tree, the new Thai alleles did not cluster together, suggesting a considerable heterogeneity within some geographic areas. Furthermore, analyses of tetrapeptide repeats from a number of isolates and strains showed evidence of three genetic mechanisms for the generation of variation in the repeats of the CS gene: point mutation, duplication of one or more repeat units, and intragenic recombination.

Alleles↗
PubMed1993

Sequence conservation in the C-terminal part of the precursor to the major merozoite surface proteins (MSP1) of Plasmodium falciparum from field isolates.

The C-terminal part of the precursor to the major merozoite surface proteins (MSP1) of Plasmodium falciparum contains potential protective epitopes and two cleavage sites for processing which take place prior to erythrocyte invasion by the merozoite. Since sequences available to date are limited and derived from cultured parasites, we have examined the extent of variations of this important part of the MSP1 gene from natural populations. Our sequence analyses of 1.6-1.7 kb from blocks 13-17 of the gene obtained from 19 Thai wild isolates have identified a deletion of a codon and 18 nucleotide substitutions, all of which are dimorphic substitutions and all but one create amino acid exchanges. However, residues at two cleavage sites for the C-terminus 42 kDa polypeptide and the 19-kDa polypeptide, a subfragment of the former, are conserved. Furthermore, all 12 cysteine residues at the C-terminal 19-kDa polypeptide are perfectly conserved, allowing the formation of 2 epidermal growth factor-like structures. These results indicate that in contrast to extensive variations at the N-terminal part of MSP1, limited variations occur at the C-terminal part.

Amino Acid Sequence↗
PubMed1992

Sequence variation in the tripeptide repeats and T cell epitopes in P190 (MSA-1) of Plasmodium falciparum from field isolates.

The N-terminal part of p190, the precursor to the major merozoite surface antigens of Plasmodium falciparum, contains the T and B cell epitopes and tripeptide repeats. The p190 gene exhibits allelic dimorphism, but the tripeptide repeat-encoding region is the only exception to the dimorphic variations of the gene. To date, sequences available to document variations in the epitopes are very limited. Thus, in this study, the extent of the variation in these regions was analyzed using the polymerase chain reaction to amplify the DNA fragment encompassing these regions, followed by sequencing. Twenty-five gene clones were obtained from 19 isolates from the Mae Sod district in Thailand and their sequences were compared with those reported elsewhere. Results reveal 3 sequence types in the tripeptide-encoding region, and each contains a novel repetitive consensus nucleotide sequence. On the other hand, almost all nucleotide substitutions in block III are dimorphic. Identification of linkages of the dimorphic substitutions has led us to postulate 6 potential crossover sites, where intragenic recombinations of p190 alleles could occur. Sequences of T and B cell epitopes are highly conserved among these wild isolates and those from geographically diverse culture-adapted parasites.

Amino Acid Sequence↗
PubMed1991

Coexistence of GP195 alleles of Plasmodium falciparum in a small endemic area.

Dimorphic variations in the genotype of the precursor to Plasmodium falciparum major merozoite surface antigens or gp195, among wild isolates in a small malaria parasite population were examined using Southern blot hybridization techniques. Hybridization, with DNA fragment probes and oligonucleotide probes derived from variable blocks of known gp195 alleles against 18 wild isolates from Mae Sod district in Thailand, revealed the existence of seven gp195 alleles, two of which were newly identified in this study. In four out of 17 patients, two different alleles coexisted in the circulation. It was furthermore noted that the seven alleles did not occur at the same frequency, but rather several alleles predominated in the population of P. falciparum in this small malaria field.

Alleles↗
PubMed1990

Cryptosporidiosis among orphanage children in Thailand: a one year prospective study.

The prevalence of cryptosporidiosis and other parasitic infections was studied among the orphanage children in Nonthaburi province, Thailand, during May 1986--April 1987. Cryptosporidium oocysts were detected in fecal specimens of 22 out of 303 diarrheal cases (7.3%) by using a modified cold Kinyoun acid-fast technique. None were found among 513 children without diarrhea. Most of the cryptosporidiosis cases were those under 3 years old. The duration of diarrhea in 20 cases varied from 1 to 25 days (mean = 6.6 days). All cases recovered uneventfully without repeated infection. No sexual preponderance was found. Most of the infections were documented during March to June when both temperature and rainfall were high. It was also noted that these children had high rates of other parasitic infections, 38.1% of the total population. Thus, cryptosporidiosis was not uncommon among children with diarrhea and could be detected almost throughout the year.

Adolescent↗
PubMed1990

Cryptosporidiosis: report of a case with life-threatening diarrhea.

A case of life-threatening chronic diarrhea, caused by Cryptosporidium spp., was reported. The patient was a seven month old boy from an orphanage suffering from voluminous watery diarrhea, fever and vomiting three days before admission. He presented with severe dehydration and severe hypokalemia. Numerous cryptosporidial oocysts were detected in the stools without other enteropathogens. Although co-trimoxazole and gentamicin were administered in the first week of illness, the diarrheal symptom was still persistent. At the end of the third week, spontaneous remission was observed. The overall diarrheal period lasted twenty-five days. In this case, cryptosporidiosis manifested as severe chronic diarrhea in the immunocompetent and supportive management was very important because an effective drug is not available at present.

Cryptosporidiosis↗