In Vitro and in Vivo Effect of MAPK Signal Transduction Pathway Inhibitors on Echinococcus multilocularis.

To evaluate the effect of mitogen-activated protein kinase (MAPK) signal transduction pathway inhibitors against alveolar echinococcosis in vitro and in vivo, Echinococcus multilocularis metacestode cysts and protoscolices were obtained from infected mice. Protein chip technology was utilized to screen for key highly expressed target proteins in the MAPK pathway in this parasite and their corresponding inhibitors. Four-week-old Balb/c female mice used for the in vivo experiment underwent inoculation of E. multilocularis by intraperitoneal injection, as well as intragastric administration of MAPK inhibitors for 6 wk. We included 6 groups of mice: a phosphate-buffered saline (PBS) group (negative control); an albendazole-treated group (positive group); and 4 experimental groups treated with TRx0237 mesylate, GDC-0994, pifithrin-ß hydrobromide, or Selonsertib. Echinococcus multilocularis protoscolices were collected and cultured in 1066 medium with penicillin/streptomycin and 10% fetal bovine serum. The in vitro experiment included a PBS group (negative control), a dimethyl sulfoxide-treated group (solvent group), and 4 inhibitor-treated groups as in the in vivo experiment (experimental groups). Each inhibitor group received 4 drug concentrations (5, 30, 55, and 80 µM), and the experiment was performed in triplicate per sample. Fluorescence microscopy was used to evaluate the survival rate of the protoscolices every 48 hr beginning from the first 24 hr. The same grouping was used to evaluate cytotoxicity on E. multilocularis germinal cells and L02 cells. The average weights of E. multilocularis metacestode cyst tissue from each group of the in vivo experiment were 873 mg (PBS), 335 mg (albendazole), 323 mg (TRx0237 mesylate), 420 mg (GDC-0994), 340 mg (pifithrin-ß hydrobromide), and 642 mg (Selonsertib). Results showed albendazole, TRx0237 mesylate, and pifithrin-ß hydrobromide had significant inhibitory effects on inhibition of E. multilocularis. We found a positive correlation between drug concentrations and the inhibitory effects seen in the in vitro experiment, with the differences in contrast with the control group becoming statistically significant after 72 hr of treatment ( P < 0.05). The inhibition rates of TRx0237 mesylate to germinal cells by drug concentration were 23.73, 46.59, 74.71, and 77.44%. Other drugs had no effect on germinal cells. All the inhibitors had low toxicity on L02 cells. Inhibitors of the MAPK signal transduction pathway showed significant inhibitory effects on E. multilocularis, suggesting these may be potential candidates for the treatment of alveolar echinococcosis.

[Investigation on echinococcosis in animals in Gannan Tibetan Autonomous Prefecture].

OBJECTIVE: To make an investigation on echinococcosis among animals in Gannan Tibetan Autonomous Prefecture. METHODS: 21 villages from Maqu and Luqu counties were selected for the survey in August of 2004-September of 2007. Rodents were trapped in the field. Sheep and yak livers, hearts and lungs were collected from the local slaughterhouses for pathological examination. Domestic dogs (shepherd dogs) were de-wormed by 15% arecoline to receive adult worms and stray dogs were shot for dissection. RESULTS: The prevalence of alveolar echinococcosis (AE) in Ochotona dahurica was 1.2% (1/87), and 2.3% (3/132) in Myospalax fontaniere, but no infection was found in Marmota himalayana, Ochotona tibetana and Mus musculus. 113 out of 1021 (11.1%) sheep were found infected with cystic echinococcosis (CE), and 3 (0.3%) with AE. 126 out of 634 (19.9%) yaks were infected with CE, and 2 yaks (0.3%) with AE. 17 out of 74 (23.0%) dogs were infected with Echinococcus granulosus (Eg), and 4 (5.4%) with Echinococcus multilocularis (Em). CONCLUSION: The results showed that there is a widespread endemic of Echinococcus granulosus in dogs and wild animals in Gannan Tibetan Autonomous Prefecture, with less Echinococcus multilocularis infection.

Bis[(E)-4-chloro-2-(cyclo-hexyl-imino-meth-yl)phenolato]nickel(II).

In the title mononuclear nickel(II) complex, [Ni(C(13)H(15)ClNO)(2)], the Ni(II) atom is four-coordinated in a tetra-hedral geometry by the N and O atoms of the two Schiff base ligands.