Gene discovery for the carcinogenic human liver fluke, Opisthorchis viverrini

Background: Cholangiocarcinoma (CCA) - cancer of the bile ducts - is associated with chronic infection with the liver fluke, Opisthorchis viverrini. Despite being the only eukaryote that is designated as a 'class I carcinogen' by the International Agency for Research on Cancer, little is known about its genome.\ud \ud Results: Approximately 5,000 randomly selected cDNAs from the adult stage of O. viverrini were characterized and accounted for 1,932 contigs, representing ~14% of the entire transcriptome, and, presently, the largest sequence dataset for any species of liver fluke. Twenty percent of contigs were assigned GO classifications. Abundantly represented protein families included those involved in physiological functions that are essential to parasitism, such as anaerobic respiration, reproduction, detoxification, surface maintenance and feeding. GO assignments were well conserved in relation to other parasitic flukes, however, some categories were over-represented in O. viverrini, such as structural and motor proteins. An assessment of evolutionary relationships showed that O. viverrini was more similar to other parasitic (Clonorchis sinensis and Schistosoma japonicum) than to free-living (Schmidtea mediterranea) flatworms, and 105 sequences had close homologues in both parasitic species but not in S. mediterranea. A total of 164 O. viverrini contigs contained ORFs with signal sequences, many of which were platyhelminth-specific. Examples of convergent evolution between host and parasite secreted/membrane proteins were identified as were homologues of vaccine antigens from other helminths. Finally, ORFs representing secreted proteins with known roles in tumorigenesis were identified, and these might play roles in the pathogenesis of O. viverrini-induced CCA.\ud \ud Conclusion: This gene discovery effort for O. viverrini should expedite molecular studies of cholangiocarcinogenesis and accelerate research focused on developing new interventions, drugs and vaccines, to control O. viverrini and related flukes

A combination of monosodium glutamate and high-fat and high-fructose diets increases the risk of kidney injury, gut dysbiosis and host-microbial co-metabolism.

Consumption of either monosodium glutamate (MSG) or high-fat and high-fructose (HFF) diets changes the gut microbiome and hence contributes to development of several diseases. In this study, with an emphasis on kidney injury, hamsters were divided into 4 groups as follows: (1) hamsters fed with standard diet (control); (2) hamsters fed with standard diet and MSG in drinking water (MSG); (3) hamsters fed with high-fat and high-fructose diets (HFF), and (4) animals fed MSG+HFF. After 8 months, the animals were used for the study. Despite showing normal kidney function, hamsters fed with MSG+HFF exhibited signs of kidney damage as demonstrated by the highest expression levels of high-mobility group box-1 and kidney injury molecule-1 in kidney tissues, while slight changes of histopathological features in H&E-stained sections and normal levels of creatinine were observed, indicating possible early stages of kidney injury. Sequencing of the microbial 16S rRNA gene revealed that animals fed with the MSG+HFF diet had a higher ratio of gut Firmicutes/Bacteroidetes along with marked changes in abundance and diversity of gut microbiome compared to hamsters fed with MSG or HFF alone. In addition, 1H Nuclear magnetic resonance spectroscopy showed an elevation of urine p-cresol sulfate levels in the MSG+HFF group. These results indicate that consumption of both MSG and HFF increases the risk of kidney injury, induces gut dysbiosis and an increase in the amount of p-cresol sulfate in hamsters

Effect of a health education program on reduction of pediculosis in school girls at Amphoe Muang, Khon Kaen Province, Thailand

<div><p>Background</p><p>Pediculosis caused by head lice (<i>Pediculus humanus capitis</i>) infestation is still an important health problem in schoolchildren, especially girls, worldwide, including in Thailand. Although pediculicidal agents effectively kill head lice, the re-infestation rate is still high. Thus, prevention is an important strategy for any sustainable control program. We aimed to develop and evaluate the efficacy of a health education program for increasing knowledge, changing attitudes and promoting preventive practices to reduce prevalence of pediculosis among school girls in Amphoe Muang, Khon Kaen, northeastern Thailand.</p><p>Methodology</p><p>Six schools were selected using multistage simple randomization and were allocated into intervention or control groups. A total of 267 girls was enrolled from these schools. A “knowledge, attitude and practice” (KAP) questionnaire, consent forms and health education materials were constructed and tested by experts and in one pilot school before the main investigation. Baseline prevalence of adult lice and nits was determined. The health education package was given only to the intervention group. The KAP questionnaire was re-evaluated at two months after intervention.</p><p>Results</p><p>At baseline, the prevalence and intensity of head lice infestation, and the KAP scores did not differ significantly between the two groups. After re-evaluation at two months, the KAP score was significantly greater in the intervention group. A significant decrease of the infestation rate from 59% to 44% was observed in the intervention group, whereas infestation increased in the control group (from 56% to 65%). The incidence of new cases in the intervention group (6.14%) was lower than in the control group (12.62%).</p><p>Conclusion</p><p>These findings indicated that the newly-established health education package is an effective tool for increasing KAP and reducing head lice infestation in school girls. Efforts to combat pediculosis in schoolchildren elsewhere may consider including this, or a similar, health education package in their programs.</p></div

Identification of autoantigens reacting against plasma samples from patients with CCA using M139 cell line-derived proteins.

<p>Identification of autoantigens reacting against plasma samples from patients with CCA using M139 cell line-derived proteins.</p

The rate of head lice infestation, incidence and intensity in intervention and control groups at baseline and the two-month follow-up assessment.

<p>The rate of head lice infestation, incidence and intensity in intervention and control groups at baseline and the two-month follow-up assessment.</p

Students’ demographic information in intervention and control groups at baseline.

<p>Students’ demographic information in intervention and control groups at baseline.</p

Diagnostic performance of autoantibodies against HSP70, RNH1 and ENO1 in pairwise comparisons between CCA, cholangitis, or healthy controls.

<p>AUC  =  area under the curve, 95%CI = 95% confidence interval (lower-upper).</p

Clinical and pathological features associated with HSP70, ENO1 and RNH1 autoantibodies in patients with CCA.

<p>*When the sum of subset numbers does not match patient totals, data were missing or unavailable.</p

Proteomic identification of plasma protein tyrosine phosphatase alpha and fibronectin associated with liver fluke, Opisthorchis viverrini, infection.

Opisthorchiasis caused by Opisthorchis viverrini induces periductal fibrosis via host immune/inflammatory responses. Plasma protein alteration during host-parasite interaction-mediated inflammation may provide potential diagnostic and/or prognostic biomarkers. To search for target protein changes in O. viverrini-infected hamsters, a 1-D PAGE gel band was trypsin-digested and analyzed by a LC-MS/MS-based proteomics approach in the plasma profile of infected hamsters, and applied to humans. Sixty seven proteins were selected for further analysis based on at least two unique tryptic peptides with protein ID score >10 and increased expression at least two times across time points. These proteins have not been previously identified in O. viverrini-associated infection. Among those, proteins involved in structural (19%), immune response (13%), cell cycle (10%) and transcription (10%) were highly expressed. Western blots revealed an expression level of protein tyrosine phosphatase alpha (PTPα) which reached a peak at 1 month and subsequently tended to decrease. Fibronectin significantly increased at 1 month and tended to increase with time, supporting proteomic analysis. PTPα was expressed in the cytoplasm of inflammatory cells, while fibronectin was observed mainly in the cytoplasm of fibroblasts and the extracellular matrix at periductal fibrosis areas. In addition, these protein levels significantly increased in the plasma of O. viverrini-infected patients compared to healthy individuals, and significantly decreased at 2-months post-treatment, indicating their potential as disease markers. In conclusion, our results suggest that plasma PTPα and fibronectin may be associated with opisthorchiasis and the hamster model provides the basis for development of novel diagnostic markers in the future

Western blotting analysis of HSP70 and RNH1 autoantibodies.

<p>Panel A) Recombinant human HSP70 was incubated with plasma of healthy individuals (lane 1–5) and patients with CCA (lanes 6–12). Immunoreactive bands were detected in 5/7 patients and none of the healthy controls. Panel B) Recombinant human RNH1 was incubated with plasma of healthy individuals (lane 1–5) and patients with CCA (lanes 6–12). Immunoreactive bands were detected in 6/7 patients and was weak in controls. Recombinant HSP70 and RNH1 proteins were probed with pooled healthy plasma served as negative controls (−) and pooled plasma from patients with CCA served as positive controls (+). HSP70 and RNH1 proteins were also demonstrated by staining with coomassie brilliant blue (CBB). PH  =  healthy individuals, CCA  =  cholangiocarcinoma.</p