RESUMEN
The combination of cytotoxic treatment modalities, including oncolytic viral gene therapies and immunotherapy, usually yields a synergistic effect. In the current study, a bicistronic adenoviral vector, Ad-CD-GMCSF, carrying the cytosine deaminase (CD) and granulocyte-macrophage colony-stimulating factor (GM-CSF) transcription units driven by a cytomegalovirus promoter was constructed, and the in vitro efficacy of the vector was tested in tumor cell lines and a syngeneic mouse model of colon cancer. The tumor cells infected with Ad-CD-GMCSF vector were found to produce a substantial amount of GM-CSF in tumor cell lines. Accordingly, the vector carrying CD and GM-CSF transcription units together induced a potent antitumor immunity with a significantly increased number of tumor-specific T cells and tumor-specific T-cell cytotoxicity (p < 0.001). The tumor growth rate of Ad-CD-GMCSF-treated mice was significantly lower when compared to the control and an adenoviral vector carrying only the CD transcription unit (Ad-CD; p < 0.05). Likewise, the median overall survival of the Ad-CD-GMCSF vector group was significantly higher than that of the control and Ad-CD groups (34.0 ± 12.8 vs. 14.0 ± 0.5 and 23.0 ± 2.8 days, respectively; p < 0.001). In conclusion, along with its cytotoxic effect, the high immunostimulatory effect of the bicistronic Ad-CD-GMCSF vector has excellent potential in the treatment of cancers.
Asunto(s)
Adenoviridae/genética , Citosina Desaminasa/genética , Terapia Genética , Vectores Genéticos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Neoplasias/genética , Neoplasias/terapia , Transgenes , Animales , Citocinas/metabolismo , Citosina Desaminasa/metabolismo , Citotoxicidad Inmunológica , Modelos Animales de Enfermedad , Expresión Génica , Orden Génico , Terapia Genética/métodos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Inmunomodulación , Ratones , Neoplasias/inmunología , Neoplasias/patología , Resultado del Tratamiento , Microambiente Tumoral , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
This study was carried out to determine the in vitro antimicrobial and antioxidant activities of the essential oil, seed oil, and methanolic extract of seed oil obtained from Laurus nobilis L. (Lauraceae). The methanolic extract of seed oil exhibited more effective antibacterial activity comparing to essential oil and seed oil, GC-MS analyses of the essential oil resulted in the identification of 25 compounds. 1.8-Cineol (44.72%), a-Terpinyl acetate (12.95%), Sabinene (12.82%) were the main components. The fatty acid composition was characterized with the high content of linoleic acid (40.79%) and lauric acid (38.08%). The 50% (IC50) inhibition activity of the essential oil on the free radical DPPH was determined as 94.655 mg ml(-1), whereas IC50 value of methanolic extract of seed oil was found unstable. In the case of the linoleic acid system, oxidation of linoleic acid was inhibited by essential oil and methanolic extract of seed oil, which showed 64.28 and 88.76% inhibition, respectively. The inhibition value of the methanolic extract of seed oil was quite close to the synthetic antioxidant BHT, 92.46% inhibition.
Asunto(s)
Antibacterianos/farmacología , Antioxidantes/farmacología , Laurus/química , Aceites de Plantas/farmacología , Semillas/química , Antibacterianos/química , Antioxidantes/química , Aceites de Plantas/químicaRESUMEN
The diversity of archaeal strains from six hypersaline environments in Turkey was analyzed by comparing their phenotypic characteristics and 16S rDNA sequences. Thirty-three isolates were characterized in terms of their phenotypic properties including morphological and biochemical characteristics, susceptibility to different antibiotics, and total lipid and plasmid contents, and finally compared by 16S rDNA gene sequences. The results showed that all isolates belong to the family Halobacteriaceae. Phylogenetic analyses using approximately 1,388 bp comparisions of 16S rDNA sequences demonstrated that all isolates clustered closely to species belonging to 9 genera, namely Halorubrum (8 isolates), Natrinema (5 isolates), Haloarcula (4 isolates), Natronococcus (4 isolates), Natrialba (4 isolates), Haloferax (3 isolates), Haloterrigena (3 isolates), Halalkalicoccus (1 isolate), and Halomicrobium (1 isolate). The results revealed a high diversity among the isolated halophilic strains and indicated that some of these strains constitute new taxa of extremely halophilic archaea.
Asunto(s)
Halobacteriaceae/clasificación , Microbiología del Agua , Halobacteriaceae/efectos de los fármacos , Halobacteriaceae/genética , Lípidos/análisis , Filogenia , Plásmidos , ARN Ribosómico 16S/genética , Cloruro de SodioRESUMEN
To determine the antibiotic resistance pattern and resistance plasmids, we studied 23 antibiotic-resistant clinical isolates of Enterococcus spp. which caused infection in Bayindir-Ankara Hospital, Turkey. Biochemical and physiological identification tests were applied by the Vitek system and compared with the results of protein profiles by SDS-PAGE. From 23 isolates, 20 were identified as E. faecalis, 2 as E. faecium and 1 as E. gallinarum. Twenty four antibiotics belong to 10 different groups were used in susceptibility tests. Multiple antibiotic resistance was determined in 10 of 23 Enterococcus spp. Overall resistance to the used antibiotics was 47.3% and low level resistance was 16.6%. Among the isolates tested, 8.7% demonstrated high level gentamicin resistance, 17.4% demonstrated high level streptomycin resistance, and 43.5% demonstrated penicillin resistance. High level vancomycin resistant Enterococcus spp. rate was 34.8%, and 60.9% exhibited low level resistance to vancomycin and teicoplanin. They contain plasmids which varied in numbers between 1 and 11 and the plasmid sizes ranged from 2.08 to 56.15 kb. In curing experiments with acriflavine, two different plasmids were shown in different molecular sizes of 33.49 and 13.6 kb while the first determined glycopeptide and penicillin resistance, the second one determined either glycopeptide or penicillin resistance in two different E. faecalis strains. On the other hand, a 22.58 kb plasmid, determining kanamycin resistance, was detected in an E. faecium strain. After the curing experiments, an elimination of 37.17 and 44.47 kDa protein bands was shown in E. faecium EFA1 and E. faecalis EFA13 in SDS-PAGE, respectively. This survey indicates the increase of antibiotic-resistant enterococci, especially to vancomycin in our hospital isolates.