Potent and broad anticancer activities of leaf extracts from Melia azedarach L. of the subtropical Okinawa islands

Plant extracts have been traditionally used for various therapeutic applications. By conducting an initial screening of several subtropical plants, in this study, we evaluated the anticancer activities of Melia azedarach L. The extract from Melia azedarach L. leaves (MLE) show high cytotoxic effects on cancer cells and in vivo mouse and dog tumor models. During the initial screening, MLE showed strong antiproliferative activity against HT-29 colon, A549 lung, and MKN1 gastric cancer cells. In subsequent tests, using 39 human tumor cell lines, we confirmed the potent anticancer activities of MLE. The anticancer activity of MLE was also confirmed in vivo. MLE markedly inhibited the growth of transplanted gastric MKN1 cancer xenografts in mice. To elucidate the mechanism underlying the anticancer effects of MLE, MLE-treated MKN1 cells were observed using an electron microscope; MLE treatment induced autophagy. Furthermore, western blot analysis of proteins in lysates of MLE-treated cells revealed induction of light chain 3 (LC3)-II autophagosomal proteins. Thus, MLE appeared to suppress MKN1 cell proliferation by inducing autophagy. In addition, in the mouse macrophage cell line J774A.1, MLE treatment induced TNF-alpha production, which might play a role in tumor growth suppression in vivo. We also performed a preclinical evaluation of MLE treatment on dogs with various cancers in veterinary hospitals. Dogs with various types of cancers showed a mean recovery of 76% when treated with MLE. Finally, we tried to identify the active substances present in MLE. All the active fractions obtained by reverse-phase chromatography contained azedarachin B-related moieties, such as 3-deacetyl-12-hydroxy-amoorastatin, 12-hydroxy-amoorastatin, and 12-hydroxyamoorastaton. In conclusion, MLE contains substances with promising anticancer effects, suggesting their future use as safe and effective anticancer agents

LYSOGENY IN CORYNEBACTERIUM RENALE

In order to isolate temperate phages, ultraviolet irradiation was applied to the total 87 strains of C. renale. These strains included those of 3 types of C. renale, and were isolated mainly in Hokkaido except for 4,of which were received from Scotland. Phages were isolated from 29 of the 46 (63%) strains of type I. No phages were obtained from 31 strains of type II or 10 strains of type III. Of the 29 phages isolated from type I strains, 28 were from the strains isolated in Hokkaido and 1 was from a strain isolated in Scotland. The former phage group, all had a similar host range and lysed strains of C. renale isolated in Hokkaido while the latter only lysed another strain isolated in Scotland. No common host range was found between the two phage groups. This suggests that phage-typing is applicable for C. renale and that different phage-types exist in Japan and Scotland. It was found on a farm, in Hokkaido, that the urine and urinary tracts of many apparently healthy cattle were parasitized by C. renale of the same phage-type. Pyelonephritis occurred only in a small percentage of these cattle. Thus, based on the knowledge obtained from the phage-typing of C. renale, we concluded that the factors influencing the establishment of the disease, were largely predisposing factors of the host animal and were not the type of C. renale

Functional growth inhibition of influenza A and B viruses by liquidand powder components of leaves from the subtropical plant Meliaazedarach L.

AbstractWe evaluated the anti-influenza-virus effects of Melia components and discuss the utility of these components. The effects ofleaf components of Melia azedarach L. on viruses were examined, and plaque inhibition tests were performed. The in vivoefficacy of M. azedarach L. was tested in a mouse model. Leaf components of Melia azedarach L. markedly inhibited thegrowth of various influenza viruses. In an initial screening, multiplication and haemagglutination (HA) activities of H1N1,H3N2, H5, and B influenza viruses were inactivated by the liquid extract of leaves of M. azedarach L. (MLE). Furthermore,plaque inhibition titres of H1N1, H3N2, and B influenza viruses treated with MLE ranged from 103.7to 104.2.MLE possessedhigh plaque-inhibitory activity against pandemic avian H5N1, H7N9, and H9N2 vaccine candidate strains, with aplaque inhibition titre of more than 104.2.Notably, the buoyant density decreased from 1.175 to 1.137 g/cm3, and spikelessparticles appeared. We identified four anti-influenza virus substances: pheophorbide b, pheophorbide a, pyropheophorbidea, and pheophytin a. Photomorphogenesis inside the envelope may lead to removal of HA and neuraminidase spikes fromviruses. Thus, MLE could efficiently remove floating influenza virus in the air space without toxicity. Consistent with thisfinding, intranasal administration of MLE in mice significantly decreased the occurrence of pneumonia. Additionally, leafpowder of Melia (MLP) inactivated influenza viruses and viruses in the intestines of chickens. MLE and MLP may haveapplications as novel, safe biological disinfectants for use in humans and poultry

Advancing usability of an influenza hemagglutinin virus-like particle vaccine expressing a chimeric cytokine

Abstract Vaccine efficacy of conventional influenza vaccines depend on the antigenic similarity between the selected vaccine strain and annual epidemic strain. Since the influenza virus evolves yearly, a vaccine which is independent from viral antigenic mutation is desired. We have developed chimeric cytokine (CC) and hemagglutinin (HA) incorporated virus-like particle (CCHA-VLP) as a universal influenza vaccine candidate. Using mouse models, it was shown that the vaccine provided broad-based protective activity against several types of human and avian influenza A viruses. In this report, nasal immunization and mixture form (CC- and HA-VLP) were tested to improve usability of this vaccine. Immunogenicity was evaluated by induction of IgG, IgA, and IFN-γ secreting cells. Protective activity was measured as mouse survival rate against lethal challenge with H1N1 and H5N1 viruses and against H3N2 virus by lung viral titer. Nasal immunization showed low immunogenicity and low protective efficacy, but the addition of a sesame oil adjuvant improved vaccine efficacy. Mixture form of CC- and HA-VLP showed comparable or higher vaccine efficacy when compared to the incorporated form, CCHA-VLP. These results contribute to improved usability, such as needle-less administration and easy HA subtypes alteration