Cytokine gene expression in human hepatocytes infected with dengue virus serotype 3 (strain-16562)

Liver is a site of viral replication and liver dysfunction is a characteristic of severe dengue infection. To understand these mechanisms, we analyzed the response of a hepatic cell linage, HepG2 to infection with dengue 3 virus (strain 16562). Steady state levels of mRNA accumulation were assessed for 14 genes involved in modulation of the host immune responses, at 6, 24 and 48 hpi, by quantitative reverse transcription real-time PCR (qRT-PCR). Fourteen genes showed altered expression upon infection with D3V including;cytokines/chemokines (IL-1β, IL-6, IL-8, RANTES, MCP-2, IL-2Rα and TGF-βIIIR), type I interferon (IFN-α and IFN-β), and pattern-recognition receptors (TLR3, TLR8, RIG-1, MDA5 and MyD88). Although these genes are associated with mechanism of innate immune response and anti-viral activity, their altered expression does not inhibit D3V (strain 16562) growth kinetics and virus yield in HepG2 cells. Gene expression in liver may explain pathological changes associated with dengue virus infection.

Activity of chlorpromazine on nfa1 and Mp2CL5 genes of Naegleria fowleri trophozoites

Amoeba treatment of patients suffering from pri-mary amoebic meningoencephalitis caused by Naegleria fowleri has not been successful. Dam-aged morphology and effect on genes of N. fowleri as the result of its initial interaction with drug may provide clue to the success of treatment. In this study, we investigated the activity of chlorpromazine compared with amphotericin B and voriconazole against N.fowleri Khon Kaen strain using cell based assay and molecular techniques. Scanning electron and light micro-graph showed the drug interaction of treated amoebae with 0.098 ug/ml chlorpromazine was faster than 0.002 ug/ml amphotericin B and 12.5 ug/ml of voriconazole. The morphological cha-racteristics of treated amoebae with Gomori’s trichrome stain correlated to the scanning elec-tron microscope study. The effect of drugs to nfa1 and Mp2CL5 genes of treated amoebae found that at 120 min post exposure, chlorpromazine, voriconazole inhibited both genes except amphotericin B. Most of drug inhibited nfa1 except fluconazole. The results evaluated that chlorpromazine was higher potency and rapidly activity than amphotericin B and voriconazole against N. fowleri trophozoites.

ITS and pB2.5 gene expression of Naegleria fowleri in drug resistance

Naegleria fowleri was causative agent of primary amoebic meningoencephalitis (PAM). Accroding to the failure of treatment, several researches reported the activity of chemotherapeutic drugs against N.fowleri but we did not know the drug resistance of the amoebae. The purpose of this study was to examine the effects of drugs (amphotericin B, artesunate, azithromycin, voriconazole, chlorpromazine, fluconazole and gentamicin sulphate) on ITS and pB2.3 genes of Naegleria fowleri trophozoites. Our study demonstrated gene expression of treated N.fowleri by RT-PCR. The results reviewed that ITS gene of N. fowleri showed up regulate to amphotericin B, azithromicin and gentamicin sulphate, while pB2.3 gene showed up regulate to artesunate. These results compared with beta actin (house keeping gene) expression at time intervals 15 - 120 min. The change of gene expression of treated N.fowleri was possibly to cause of drug resistance. The mechanism of drug resistance genes ITS and pB2.3 of N.fowleri should be clarified in further study.