Betts, Jonathan W.;Wareham, David W.;Haswell, Stephen J.;Kelly, Stephen M.
Journal of Microbiology and Biotechnology
/
v.23
no.9
/
pp.1322-1326
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2013
New antifungal agents are required to compensate for the increase in resistance to standard antifungal agents of Candida albicans, which is an important opportunistic fungal pathogen that causes minor infections in many individuals but very serious infections in those who are immune-compromised. In this study, combinations of theaflavin and epicatechin are investigated as potential antifungal agents and also to establish whether antifungal synergy exists between these two readily accessible and cost-effective polyphenols isolated from black and green tea. The results of disc diffusion assays showed stronger antibacterial activity of theaflavin:epicatechin combinations against C. albicans NCTC 3255 and NCTC 3179, than that of theaflavin alone. Minimum inhibitory concentrations (MICs) of 1,024 ${\mu}g/ml$ with theaflavin and 128-256 ${\mu}g/ml$ with theaflavin:epicatechin combinations were found. The fractional inhibitory concentration indexes were calculated, and the synergy between theaflavin and epicatechin against both isolates of C. albicans was confirmed. Theaflavin:epicatechin combinations show real potential for future use as a treatment for infections caused by C. albicans.
The uterus receives adrenergic terminals from the mesenteric ganglia and considerably large amount of catecholamines have been shown to be contained in this organ. On the other hand, the activities of epinephrine, norepinephrine or adrenergic nerve on uterine motility is so complicated that many controversial results have been reporter. Recently, a large number of reports concerning the changes of uterine catecholamines content have appeared, but little is known about the role of uterine catecholamines in their activities on uterine motility. The present experiments were undertaken to determine the significance of the intrinsic uterine catecholamines in the physiology of uterus. Female albino rabbits weighing approximately 2 kg were employed in this experiment. uterine strip3 were prepared and suspended in a constant temperature $bath(38^{\circ}C)$ containing 100 ml of Locke's solution aerated with 95% oxygen and 5% carbon dioxide. Spontaneous motility was recorded on a smoked drum with an isotonic lever. The catecholamines concentration of the uterus was determined according to the Procedure described of Shore and Olin (1958). Human uterus obtained from patients was also used to determine the catecholam ines content of myometrium. Followings are summarized results. 1) On the non-pregnant rabbit uterine strips, epinephrine and norepinephrine significantly elevated the tonus and stimulated the spontaneous motility. Pretreatment with dichloroisoproterenol(DCI), an adrenergic beta-receptor blocker, enhanced the stimulatory activity of epinephrine or norepinephrine. On the other hand, pretreatment with dibenamine, an adrenergic alpha-receptor blocker, rendered the uterine muscle to exhibit inhibition after the administration of epinephrine or norepinephrine. Following the treatment with both DCI and dibenamine, epinephrine or norepinephrine produced no appreciable effects on the spontaneous motility of the uterus. These results suggest there exist both alpha and beta-adrenergic receptors in the uterine muscle and the response to epinephrine of the former is predominant over that of latter in the non-pregnant uterus of rabbits. The total catecholamines concentration of the non-pregnant uterus was $351\;m{\mu}g/g$ and the fractional concentrations of epinephrine and norepinephrine were $125\;m{\mu}g/g(35.7%)$ and $226\;m{\mu}g/g$ respectively. It is interesting to note that the catecholamines content of uterus was characterized by a high fractional corcentration of epinephrine relative to norepinephrine. 2) On the pregnant rabbit uterine strips, the effects of epinephrine and norepinephrine varied according to the period of pregnancy. The response to epinephrine of adrenergic beta receptor of uterus increased during pregnancy, and the effect of catecholamine was inhibitory in the early pregnancy but became stimulatory as the pregnancy progressed. This stimulating action on the uterine motility was found to occur through the action of norepinephrine. The uterine catecholamines concentration was markedly reduced during pregnancy. The catecholamines concentration was started to decrease in the early pregnancy, reached the lowest level in the mid-pregnancy and then started to increaae again in the late pregnancy when the total catecholamines content became the highest level of all. This increase of catefholamines in late pregnancy was chiefly due to the increase of norepinephrine. These results suggest that the uterine motility may be related to the catecholamines content, especially norepinephrine content in the uterus. 3) Bilateral oophorectomy of rabbits results in a marked shrink of the uterus in size. The spontaneous motility of the uterine segment of these animals was very weak and irregular. Norepinephrine produced inhibitory effect, whereas epinephrine was stimulatory or inhibitory effect on the uterine segment. The total catecholamines tontent in whole uterus was markedly reduced. The injection of estrogen into the oophorectornized rabbit increased the weight of uterus to approximately three times of that of oophorectornized animal. The apontaneous motility and the response to epinephrine and norepinephrine of the uterine segment were greatly enhanced. Both epinephrine and norepinephrine produced a marked stimulatory effects of the uterine motility. The uterine content of catecholamines, particularly epinephrine, was markedly increased. The injection of progesterone into the oophorectornized rabbit increaeed the weight of uterus to approximately 2.5 times of that of eophorectornized animal. The spontaneous motility of the uterine segment was weak and irregular. Epinephrine produced stimulatory effect at high concentrations but norepinephrine always prcdnced inhibitory effect on the uterine segment. The uterine content of catecholamines, particularly of norepinephrine, was markedly reduced. These results suggested that ovarian hormones play an important role not only on the growth and spontaneous norepinephrine of uterus but also on the catecholamines content and responee to epinephrine and norepinephrine of the uterus. 4) The intraperitoneal injection of reserpine(3 mg/kg) into the non-pregnant, pregnant and oophorectornieed rabbits markedly decreased the uterine content of catecholamines, particularly of the norepinephrine. The stimulatory response to epinephrine and. norepinephrine of the uterine segment of these reserpinized ratbits was markedly reduced whereas the inhibitory response to these catecholamines was enhanced. This finding further support the close relationship between the uterine catecholamines content and uterine response to epineptrire and norepinephrine. 5) In the human uterus, the concentration of epinephrine was actrally greater than that of norepinephrine and it was significantly greater during the proliferative phase of the menstrtal cycle. In the human pregnant uterus, the concentrations of toth epinephrine and ncrefinephrine were markedly reduced and showed about 45 percent rednction after 6-8 weeks of ectopic Pregnancy. At full term ana during labor, the concentrations of epinephrine and norepinephrine at placental sites were less than those found in the non-pregnant group. Of interest was the finding that the norepinephrine concentration of uterus from toxemic patients was two and half times higher than that of lower uterine segment of the nontoxemic pregnant individuals. Also the epinephrine concentraticn was slightly increaeed.
The effects of Cymbopogon citratus essential oil (Lemongrass oil) and its main component, citral (84.30%), on antibiotic-susceptible and -resistant strains of Salmonella enteritidis and S. typhimurium were assessed. C. citratus oil and citral significantly inhibited all strains of the two Salmonella species examined, with minimum inhibiting concentrations (MICs) ranging from 0.5 mg/ml to 8.0 mg/ml. The combined effects of C. citratus oil and citral (84.30%) were evaluated using a checkerboard microtiter assay. Essential oil fractions of C. citratus and citral exhibited strong synergistic or additive effects with streptomycin or kanamycin against S. typhimurium strains with fractional inhibitory concentration (FIC) indices in the range of 0.28 to 1.00. In conclusion, a combination of streptomycin and lemongrass oil or its main component, citral, may be useful for reducing the minimum effective dose of antibiotic required for the treatment of resistant S. typhimurium infections.
Objectives: This study aims to determine the Fractional Inhibitory Concentration Index (FICI) of combinations of Melastoma malabathricum leaf fraction with ciprofloxacin or gentamicin against pathogenic bacteria, Escherichia coli, Staphylococcus aureus, and Bacillus cereus, isolated from Diabetic Foot Ulcer (DFU) patients. Methods: We tested concentrations of 45%, 55%, 65%, and 75% of gentamicin and ciprofloxacin using dilution and agar diffusion methods. The combination of M. malabathricum leaf extract with these antibiotics was tested in vitro against all three bacteria. Results: The combination of M. malabathricum leaf extract and ciprofloxacin gave a FICI value of 0.5, indicating synergistic antibacterial activity against the test bacteria. Conclusion: The results show that the antibacterial effect of a combination of high doses of the leaf extract with either antibiotic is greater than that of the leaf extract and the antibiotics in single use.
As part of our search for less toxic antimicrobial agents from natural resources, the antimicrobial activity of Elfvingia applanata $(P_{ers.})\;K_{arst.}$ extract was examined alone and in combination with naringenin. EA, the aqueous extract from the carpophores of E. applanata, was lyophilized and a dark brownish powder was obtained. Antimicrobial activity of EA was tested in vitro against nineteen strains of bacteria and eleven strains of fungi by serial broth dilution method, and expressed by minimal inhibitory concentration (MIC). Among nineteen strains of bacteria tested, the antimicrobial activity of EA was the most potent against Proteus vulgaris showing MIC of 1.125 mg/ml. EA also inhibited the growth of the selected fungi at higher concentrations ranging from 7.5 mg/ml to 15.0 mg/ml. To investigate the effect of antimicrobial combinations of EA with naringenin, the fractional inhibitory concentration index (FICI) was determined by checkerboard assay for each strain. The antimicrobial combinations of EA with naringenin resulted in partial synergism against Staphylococcus aureus only, and showed additive effect in two strains including Klebsiella pneumoniae and Salmonella typhi. Antagonism was not found.
Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.
The essential oil fraction from the aerial parts of A. annua was analyzed by GC-MS. As the results, caryophyllene oxide (11.7%), caryophyllene (7.54%), camphor (7.32%), 1,8-cineol (4.98%), and borneol (3.99%) were confirmed as the main components of the oil fraction. The effects of this oil and its main components on antibiotic-susceptible and -resistant strains of Salmonella enteritidis and S. typhimurium were assessed. A. annua oil fraction significantly inhibited all strains of the two Salmonella species examined, with minimum inhibiting concentrations (MICs) ranging from 2.0 mg/ml to 8.0 mg/ml. Among the main components of the oil, borneol and camphor showed relatively strong inhibiting activity with MICs between 1.0 mg/ml and 4.0 mg/ml. The MICs of caryophyllene and caryophyllene oxide were higher than 16 mg/ml. The combination effects of the oils with kanamycin were evaluated using a checkerboard microtiter assay. Against S. typhimurium KCCM11862 and CCARM8009 strains, the oil fraction of A. annua, camphor, and 1,8-cineol exhibited significant synergistic with kanamycin with fractional inhibitory concentration (FIC) indices in the range of 0.085 to 0.375. In conclusion, a combination of kanamycin and A. annua oil or its main component, camphor, and cineol, may be useful for reducing the minimum effective dose of antibiotic required for the treatment of resistant S. typhimurium infections.
Kim, Ki-Hyun;Eom, Sung-Hwan;Kim, Hyo-Jung;Lee, Dae-Sung;Nshimiyumukiza, Ossiniel;Kim, Dongsoo;Kim, Young-Mog;Lee, Myung-Suk
Fisheries and Aquatic Sciences
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v.17
no.2
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pp.209-214
/
2014
With the continuing demand for new solutions in the development of effective and safe candidiasis therapies, we investigated the efficacy of an antifungal agent from the marine brown alga Eisenia bicyclis. The methanolic extract of E. bicyclis evinced potential antifungal activity against Candida species. The ethyl acetate (EtOAc)-soluble extract from E. bicyclis demonstrated the strongest antifungal activity against Candida species among five solvent-soluble extracts. Indeed, the EtOAc-soluble extract showed minimum inhibitory concentrations (MICs) ranging from 4 to 8 mg/mL. Furthermore, the EtOAc-soluble extract considerably reversed high-level fluconazole resistance of Candida species. The MIC values of fluconazole against Candida species decreased substantially (from 64 to $4{\mu}g/mL$) in combination with the MIC of the EtOAc-soluble extract (4 mg/mL). The fractional inhibitory concentration indices of fluconazole ranged from 0.531 to 0.625 in combination with 4, 2, or 1 mg/mL of the EtOAc-soluble extract against Candida isolates, indicating that these combinations exert a marked synergistic effect against Candida isolates. These findings imply that compounds derived from E. bicyclis can be a potential source of natural antifungal agents against Candida species.
This study was investigated the synergistic effects from the combination of cefalexin and gentamicin (CCG) against Staphylococcus spp. isolated from bovine milk. Minimum inhibitory concentrations (MICs) and fractional inhibitory concentration (FIC) index were determined for 16 bacteria from 13 bovine farms in Gyeongsangnam province by the National Committee for Clinical Laboratory Standards broth microdilution procedure. Antimicrobial agents tested included cefalexin, gentamicin and CCG. Against Staphylococcus spp. tested, the MIC ranges of cefalexin, gentamicin and CCG were 0.25~1.0, 0.5~0.125 and $0.06{\sim}0.125{\mu}g/mL$, respectively. In addition, the FIC index ranges of CCG against most of the isolates was 0.28~0.43 and the FIC index of CCG against 2 strains of Staphylococcus haemolyticus (S. haemolyticus) was 0.59. In conclusion, CCG has a high antibacterial activity against Staphylococcus isolates, then the combination may be applied for the treatment of bovine mastitis caused by Staphylococcus spp.
Objectives In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA). Methods The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively. Results The MIC of MEH was 25~1,600 ㎍/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 ㎍/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH. Conclusions These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.
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