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Amebicidal activity of plant extracts from Southeast Asia on Acanthamoeba spp.

The effect of 100 polar and 100 nonpolar plant extract materials obtained from Southeast Asia were evaluated for amebicidal activity in vitro against three species of Acanthamoeba. A. culbertsoni, A. castellanii, and A. polyphaga, the causative agents of granulomatous amebic encephalitis and amebic keratitis, were studied in vitro to determine whether the plant extracts exhibited amebicidal activity or induced encystment of the amebae. Of the 200 plant extracts tested, extracts obtained from three plants (Ipomoea sp., Kaempferia galanga, and Cananga odorata) were amebicidal for all three species of Acanthamoeba and a fourth extract prepared from Gastrochilus panduratum was lytic for A. polyphaga and growth-inhibitory for A. castellanii and A. culbertsoni. Three plant extracts induced encystment of all three species of Acanthamoeba. Select plant extracts were tested as well for tumoricidal activity against B103 neuroblastoma cells. Some plant extracts that exhibited tumoricidal activity for B103 cells were not amebicidal for Acanthamoeba spp. Additionally, the polar and nonpolar extracts that exhibited amebicidal activity were also tested for activity against primary murine peritoneal macrophage cultures. Plant extracts that demonstrated tumoricidal or amebicidal activity were not lytic for normal macrophage cultures.

Acanthamoeba↗

Development of amebicidal cell-mediated immunity in gerbils (Meriones unguiculatus) immunized with the galactose-inhibitable adherence lectin of Entamoeba histolytica.

The galactose-inhibitable adherence lectin of Entamoeba histolytica is a protective antigen in the gerbil model of amebic liver abscess. To further understand the mechanisms of vaccine efficacy, we studied the cell-mediated immune response to the lectin in gerbils. Splenocytes harvested from immunized gerbils demonstrated in vitro proliferation and production of interleukin-2 and gamma-interferon in response to purified adherence lectin (P < 0.05 for each compared to control splenocytes). Splenocytes from immunized gerbils developed direct amebicidal activity (P = 0.014) following in vitro stimulation with the adherence lectin. Splenocytes harvested from immunized gerbils following intrahepatic challenge with viable E. histolytica adherence lectin. Splenocytes harvested from immunized gerbils following intrahepatic challenge with viable E. histolytica trophozoites demonstrated proliferative and amebicidal responses (P < 0.05). In conclusion, immunization with the E. histolytica galactose-inhibitable adherence lectin elicits an amebicidal cell-mediated response that is enhanced by parasite challenge.

Animals↗

Amebicidal activity of wild animal serum.

The sera of 16 species of wild animals representing 5 classes of vertebrates were assayed for amebicidal activity against species of Naegleria. The greatest activity was observed for sera of bullfrogs, muskrats, and raccoons, all of which are animals associated with water. In contrast, the sera from animals such as toads, box turtles, sparrows, and squirrels exhibited minimal or no amebicidal activity. In general, pathogenic Naegleria tended to be less susceptible than nonpathogenic Naegleria to the lytic effect of raccoon serum. Heat-inactivated serum was not amebicidal, suggesting that perhaps complement may be involved in the serum-mediated lysis of amebas.

Amebiasis↗

Human T-lymphocyte proliferation, lymphokine production, and amebicidal activity elicited by the galactose-inhibitable adherence protein of Entamoeba histolytica.

We studied human T-lymphocyte responses to the purified Entamoeba histolytica galactose-inhibitable adherence protein. Individuals having serum anti-adherence protein antibodies possess peripheral blood lymphocytes which demonstrate antigen-specific responses to the purified adherence protein (10 micrograms/ml) and whole soluble amebic antigen (100 micrograms/ml). This was determined by incorporation of [3H]thymidine (53,080 and 73,114 dpm, respectively) and by increased production of interleukin-2 and gamma interferon (42.0 and 67.5 U/ml, respectively) (P less than 0.05 for each in comparison with values for control lymphocyte responses). Lymphocytes from antiamebic antibody-positive subjects develop in vitro amebicidal activity only when incubated for 5 days with the purified adherence protein (P = 0.02). In conclusion, the E. histolytica galactose-inhibitable adherence protein elicits an in vitro amebicidal cell-mediated immune response, further supporting the potential for the use of this protein in a subunit amebiasis vaccine.

Animals↗

The tumor necrosis factor alpha-stimulating region of galactose-inhibitable lectin of Entamoeba histolytica activates gamma interferon-primed macrophages for amebicidal activity mediated by nitric oxide.

Entamoeba histolytica adheres via galactose-lectin (Gal-lectin) to human colonic mucins and intestinal epithelial cells as a prerequisite to amebic invasion. Native Gal-lectin is a protective antigen in the gerbil model of amebiasis. Amino acids 596 to 1082 of Gal-lectin mediate E. histolytica adherence to target cells and stimulate tumor necrosis factor alpha (TNF-alpha) production by naive murine bone marrow macrophages (BMM). Resistance to amebiasis requires an effective cell-mediated immune response against E. histolytica trophozoites mediated by nitric oxide (NO) released from activated macrophages. Herein, we determine whether the TNF-alpha-stimulating region of Gal-lectin can activate gamma interferon (IFN-gamma)-primed BMM for NO production and amebicidal activity. Native Gal-lectin (100 to 500 ng/ml) stimulated TNF-alpha and inducible nitric oxide synthase (iNOS) mRNA expression in IFN-gamma-primed BMM as did lipopolysaccharide (100 ng/ml). Primed BMM produced TNF-alpha and NO in response to Gal-lectin in a dose-dependent manner. Antilectin monoclonal antibody IG7, which recognizes a domain (amino acids 596 to 818) of the TNF-alpha mRNA-stimulating region of Gal-lectin, specifically inhibited TNF-alpha and iNOS mRNA induction and TNF-alpha and NO production by primed BMM in response to Gal-lectin (100 ng/ml). Simultaneous treatment of BMM with IFN-gamma and Gal-lectin (100 ng/ml) activated the cells to kill E. histolytica trophozoites, whereas IFN-gamma treatment alone had no effect. In the presence of monoclonal antibody 1G7 or aminoguanidine (an iNOS inhibitor), NO production and amebicidal activity were inhibited >80%. These results suggest that the TNF-alpha-stimulating region of native Gal-lectin is a potent stimulus of IFN-gamma-primed BMM for NO production, which is essential for host defense against amebiasis.

Animals↗

Genotoxicity of amebicide and anthelmintic drugs in Escherichia coli pol A+/pol A-.

The amebicides dehydroemetine and chloroquine diphosphate and the anthelmintic pyrvinium pamoate, previously reported to be mutagenic in Salmonella typhimurium (Cortinas de Nava et al., 1983), were clearly shown to be genotoxic in the Escherichia coli pol A+/pol A- assay. Two other antiparasitic drugs, diiodohydroxyquin and 4-hexylresorcinol, were also found to be genotoxic in E. coli, while iodochlorhydroxyquin preferentially inhibited the pol A+ strain. From the 3 alternative testing methods employed, the liquid suspension succeeded in detecting 5 antiparasitic drugs as genotoxic; the microsuspension identified 2, and the disc diffusion method only 1. However, the metabolic activation system could only be coupled successfully and in a reproducible way to the microsuspension assay.

Amebicides↗

[Evaluation of the tolerance and efficiency of quinfamide, a new intraluminal amebicide, in man (one day treatment). Double blind study].

A new intraluminal amebicide (Quinfamide) was tested to assess its effectivity and tolerance for treatment of non-dysenteric intestinal amebiasis. The drug was administered to three groups of ten patients each, whom received 300, 600 and 1 200 mg. on a 24 hours schedule. Another group of ten patients received Teclozan as control drug. Diagnosis and results were judged by rectosigmoidoscopy before, 15 and 30 days after treatment. In addition, microscopic investigation of ameba was performed in freshly passed stools, before, and after 8, 15 and 30 days of treatment. Success after treatment with the three doses of Quinfamide was obtained in 89.2% of the cases. Side reactions were clinically non-significant. More experience is needed before the effectivity of the drug is stablished.

Acetamides↗

[New amebicides].

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Amebicides↗

In vitro evaluation of the amebicidal activity of Buddleia cordata (Loganiaceae, H.B.K.) on several strains of Acanthamoeba.

Infectious diseases produced by free-living amoebae from the genus Acanthamoeba have been recently recognized. The need for antiamebic compounds is urgent as the occurrence of these diseases is being registered more frequently since the late sixties. We screened the aqueous and methanolic extract of a plant used by folk medicine (Buddleia cordata) against eye and skin inflammation for antiamebic activity. We tested the extracts on 29 strains of free-living amoebae, with the result that they were amebostatic for 14 and 15 strains, respectively. We obtained linarin and vanillic acid from the extracts, but only linarin was amebostatic to all the strains and vanillic acid had no activity. However, acetyl vanillic acid had similar effects on amoebae to linarin. Threshold values of these two active compounds ranged from 31.25 microg/ml to 4 mg/ml and from 31.25 microg/ml to 8 mg/ml for linarin and acetyl vanillic acid, respectively. These differences in threshold values were observed even on several strains belonging to the same species (as in the case of A. castellanii and A. polyphaga) indicating the need of susceptibility testing for each clinical isolate of free-living amoebae.

Acanthamoeba↗

An integrated concept of amebicidal action: electron transfer and oxy radicals.

Cyclic voltammetry data were obtained for most of the main categories of antiamebic agents, specifically, quinones, heterocyclic nitro compounds, metal derivatives and chelators, and iminium-type ions. The reductions (our data and literature values) were for the most part reversible, with potentials usually in the favorable range of +0.10 to -0.56 V. The drug effect is believed to result generally from the catalytic production of oxidative stress usually arising from the formation of superoxide via electron transfer. In addition, relevant literature data are provided.

Amebicides↗