Proinflammatory Cytokine Gene Expression by Murine Macrophages in Response to Brugia malayi Wolbachia Surface Protein

Wolbachia, an endosymbiotic bacterium found in most species of filarial parasites, is thought to play a significant role in inducing innate inflammatory responses in lymphatic filariasis patients. However, the Wolbachia-derived molecules that are recognized by the innate immune system have not yet been identified. In this study, we exposed the murine macrophage cell line RAW 264.7 to a recombinant form of the major Wolbachia surface protein (rWSP) to determine if WSP is capable of innately inducing cytokine transcription. Interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF) mRNAs were all upregulated by the rWSP stimulation in a dose-dependant manner. TNF transcription peaked at 3 hours, whereas IL-1β and IL-6 transcription peaked at 6 hours post-rWSP exposure. The levels of innate cytokine expression induced by a high-dose (9.0 μg/mL) rWSP in the RAW 264.7 cells were comparable to the levels induced by 0.1 μg/mL E. coli-derived lipopolysaccharides. Pretreatment of the rWSP with proteinase-K drastically reduced IL-1β, IL-6, and TNF transcription. However, the proinflammatory response was not inhibited by polymyxin B treatment. These results strongly suggest that the major Wolbachia surface protein molecule WSP is an important inducer of innate immune responses during filarial infections

The Genetic Polymorphisms of 24 Base Pair Duplication and Point G102S of Human Chitotriosidase to Bancroftian Filariasis at the Thai–Myanmar Border

Lymphatic filariasis, caused by lymphatic filarial parasites, Wuchereria bancrofti, and Brugia malayi, causes significant morbidity and disability to 120 million people in the tropics and subtropics. Chitin has an important role for embryogenesis in adult worms and is a component of microfilaria sheath. Human chitotriosidase (CHIT1) is a chitin-degrading enzyme which provides a protective role against chitin-containing pathogens. Here, we determined the association of CHIT1 polymorphisms with susceptibility to bancroftian filariasis (BF) in 88 individuals at the Thai–Myanmar border. Two common polymorphisms of CHIT1, contributing inactive CHIT protein, including 24 base pair (24 bp) duplication in exon 10, and p. G102S in exon 4 were genotyped by allele-specific Polymerase Chain Reaction (PCR) and PCR sequencing, respectively. Unexpectedly, genotype frequencies of 24 bp duplication insertion homozygous (INS/INS) were significantly higher in endemic normal (EN) (40.0%) than BF patients (31.4%). In contrast, genotype frequencies of p. G102S homozygous (A/A) in BF patients (21.6%) was higher than in EN (19.0%) without statistical difference. Mutant allele frequencies of 24 bp duplication were 0.6125 (98/160) and p. G102S were 0.392 (69/176). Genotype and allele frequencies of CHIT1, 24 bp duplication, and p. G102S, showed no association with BF patients

Larvicidal Activity of Synthesized Silver Nanoparticles from Curcuma zedoaria Essential Oil against Culex quinquefasciatus

Culex quinquefasciatus is the major vector of the bancroftian filarial parasite which causes human lymphatic filariasis and St. Louis encephalitis. The simple way to stop the transmission is to control the vector by using synthetic chemicals. However, herbal essential oils have biological properties, such as a larvicidal effect and are ecofriendly to use. In this study, we investigated the larvicidal activity of Curcuma zedoaria essential oil (ZEO) and biosynthesized silver nanoparticles using this essential oil (ZEO-AgNPs). The larvicidal activity against both insecticide-susceptible and -resistant strains of Cx. quinquefasciatus larvae of ZEO were investigated and compared with ZEO-AgNPs. The ZEO-AgNPs showed the utmost toxicity against both strains of Cx. quinquefasciatus. After 24 h of exposure, LC50 and LC99 of ZEO against susceptible strain were 36.32 and 85.11 ppm, respectively. While LC50 and LC99 of ZEO against the resistant strain were 37.29 and 76.79 ppm, respectively. Whereas ZEO-AgNPs offered complete larval mortality within 24 h of exposure, LC50 and LC99 of ZEO-AgNPs against the susceptible strain, were 0.57 and 8.54 ppm, respectively. For the resistant strain, LC50 and LC99 values were 0.64 and 8.88 ppm, respectively. The potency in killing Cx. quinquefasciatus and stability of ZEO-AgNPs have made this product a good candidate for the development of novel natural larvicides

The Experimental Infections of the Human Isolate of <i>Strongyloides Stercoralis</i> in a Rodent Model (The Mongolian Gerbil, <i>Meriones Unguiculatus</i>)

Strongyloidiasis is life-threatening disease which is mainly caused by Strongyloides stercoralis infection. Autoinfection of the parasite results in long-lasting infection and fatal conditions, hyperinfection and dissemination (primarily in immunosuppressed hosts). However, mechanisms of autoinfection and biology remain largely unknown. Rodent models including mice and rats are not susceptible to the human isolate of S. stercoralis. Variations in susceptibility of the human isolate of S. stercoralis are found in dogs. S. ratti and S. venezuelensis infections in rats are an alternative model without the ability to cause autoinfection. The absence of appropriate model for the human isolate of strongyloidiasis hampers a better understanding of human strongyloidiasis. We demonstrated the maintenance of the human isolate of the S. stercoralis life cycle in the Mongolian gerbil (Meriones unguiculatus). The human isolate of S. stercoralis caused a patent infection in immunosuppressed gerbils, more than 18 months. The mean number of recovery adult parasitic worms were 120 &#177; 23 (1.2% of the initial dose) and L1s were 12,500 &#177; 7500 after day 28 post-inoculation (p.i.). The prepatent period was 9&#8315;14 days. Mild diarrhoea was found in gerbils carrying a high number of adult parasitic worms. Our findings provided a promising model for studying biology and searching new alternative drugs against the parasites. Further studies about the hyperinfection and dissemination would be performed

Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of -2

<p><b>Copyright information:</b></p><p>Taken from "Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of "</p><p>http://www.filariajournal.com/content/6/1/6</p><p>Filaria Journal 2007;6():6-6.</p><p>Published online 30 Jul 2007</p><p>PMCID:PMC1976415.</p><p></p>tructed by PAUP software. Bootstrap values are shown above the branches where greater than 50% obtained after 1000 replicates. Thai 1-Thai 6: genetic populations of the Thai strain of ; Myanmar 7-Myanmar12: genetic populations of the Myanmar strain of

Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of -0

<p><b>Copyright information:</b></p><p>Taken from "Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of "</p><p>http://www.filariajournal.com/content/6/1/6</p><p>Filaria Journal 2007;6():6-6.</p><p>Published online 30 Jul 2007</p><p>PMCID:PMC1976415.</p><p></p>rnally periodic (Myanmar strain) in four Myanmar patients based on the average counts of microfilariae in triplicated of 20 μl blood samples. Values shown are means ± SD

Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of -1

<p><b>Copyright information:</b></p><p>Taken from "Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of "</p><p>http://www.filariajournal.com/content/6/1/6</p><p>Filaria Journal 2007;6():6-6.</p><p>Published online 30 Jul 2007</p><p>PMCID:PMC1976415.</p><p></p>00 bp DNA marker; Lane 1–6: the Thai strain of ; Lane 7–12: the Myanmar strain of ; Lane C: Negative Control (uninfected human blood sample). The 300 bp and 795 bp fragments (arrows) are specific to the Myanmar strain of . The 645, 705, 1290, and 1400 bp fragments were common in both the Thai and Myanmar strains of

A Simple Genotyping Method for Rapid Differentiation of <i>Blastocystis</i> Subtypes and Subtype Distribution of <i>Blastocystis</i> spp. in Thailand

Blastocystis spp. is one of the most common protozoa of humans and animals worldwide. The genetic diversity of Blastocystis spp. might be associated with a wide range of symptoms. However, the prevalence of each subtype is different in each country. Until now, there is no standard method for subtyping of Blastocystis spp. We developed a sequential restriction fragment length polymorphism (RFLP) analysis for the rapid differentiation of human Blastocystis subtypes. A large-scale study was also conducted to determine the subtype distribution of Blastocystis spp. in Thailand. Stool samples were collected from 1025 school-age students in four regions of Thailand. Blastocystis infections were identified by direct smear, formalin ethyl-acetate concentration technique (FECT), Boeck and Drbohlav&#8217;s Locke-Egg-Serum (LES) medium culture, and polymerase chain reaction (PCR) of small-subunit ribosomal DNA (SSU rDNA). Subtypes of Blastocystis spp. were determined by RFLP. Phylogenetic tree of partial SSU rDNA sequences of Blastocystis spp. was constructed using the Maximum Likelihood (ML) method. Out of 1025 students, 416 (40.6%) were positive for Blastocystis spp. Using two steps of RFLP reactions, we could determine subtype one&#8211;three among these students. Subtype 3 was the most common subtype (58.72%) in Thai students, followed by subtype 1 (31.2%), and subtype 2 (10.1%). Blastocystis subtype 3 was the most prevalent in all regions of Thailand. The subtype distribution of Blastocystis spp. in Thailand was different from other countries

Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of -3

<p><b>Copyright information:</b></p><p>Taken from "Random Amplified Polymorphic DNA (RAPD) for differentiation between Thai and Myanmar strains of "</p><p>http://www.filariajournal.com/content/6/1/6</p><p>Filaria Journal 2007;6():6-6.</p><p>Published online 30 Jul 2007</p><p>PMCID:PMC1976415.</p><p></p>rnally periodic (Myanmar strain) in four Myanmar patients based on the average counts of microfilariae in triplicated of 20 μl blood samples. Values shown are means ± SD