Objective: To evaluate the acute toxicity of carboxymethyl chitosan-2, 2’ ethylenedioxy bis-ethylamine-folate (CMC-EDBE-FA) and as well as possible effect on microbial growth and in vitro cell cyto-toxicity. Methods: CMC-EDBE-FA was prepared on basis of carboxymethyl chitosan tagged with folic acid by covalently linkage through 2, 2’ ethylenedioxy bis-ethylamine. In vivo acute toxicity, in vitro cyto-toxicity and antimicrobial activity of CMC-EDBE-FA nanoparticle were determined. Results: Vancomycin exhibited the antibacterial activity against vancomycin sensitive Staphylococcus aureus, but CMC-EDBE-FA nanoparticle did not give any antibacterial activity as evidenced by minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), disc agar diffusion (DAD) and killing kinetic assay. Further, the CMC-EDBE-FA nanoparticle showed no signs of in vivo acute toxicity up to a dose level of 1000 mg/kg p.o., and as well as in vitro cyto-toxicity up to 250 μg/mL. Conclusions: These findings suggest that CMC-EDBE-FA nanoparticle is expected to be safe for biomedical applications.

OBJECTIVETo evaluate the free radical generation and antioxidant enzymes status in murine peritoneal macrophage during in vitro amikacin resistant Pseudomonas aeruginosa (ARPA) treatment with different time interval.

METHODSPeritoneal macrophages were treated with 1×10(8) CFU/mL ARPA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and super oxide anion generation, NO generation, reduced glutathione level and antioxidant enzymes status were analyzed.

RESULTSSuper oxide anion generation and NO generation got peak at 12 h, indicating maximal free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during ARPA transfection. Reduced glutathione level and antioxidant enzymes status were decreased significantly (P<0.05) with increasing time of ARPA transfection. All the changes in peritoneal macrophages after 12 h in vitro ARPA transfection had significant difference (P<0.05).

CONCLUSIONSFrom this study, it may be summarized that in vitro ARPA infection not only generates excess free radical but also affects the antioxidant system and glutathione cycle in murine peritoneal macrophage.

Amikacin ; pharmacology ; Animals ; Anti-Bacterial Agents ; pharmacology ; Antioxidants ; analysis ; Cells, Cultured ; Drug Resistance, Bacterial ; Free Radicals ; analysis ; Glutathione ; analysis ; Macrophages, Peritoneal ; immunology ; microbiology ; physiology ; Male ; Mice ; Oxidative Stress ; Pseudomonas aeruginosa ; growth & development ; immunology ; Time Factors

OBJECTIVETo observe the biochemical characters and antibiotic susceptibility of isolated Staphylococcus aureus (S. auerus) strains against some conventional and traditional antibiotics.

METHODSThirty post operative pathogenic isolated S. aureus strains were used in this study. Bacterial culture was done in Mueller-Hinton broth at 37 °C. Characters of these strains were determined by traditional biochemical tests such as hydrolysis test of gelatin, urea, galactose, starch and protein, and fermentation of lactose and sucrose. Antibiotic susceptibility were carried out by minimum inhibitory concentration test, minium bactericidal concentration test, disc agar diffusion test and brain heart infusion oxacillin screening agar.

RESULTSFrom this study, it was observed that 100% S. aureus isolates showed positive results in gelatin, urea and galactose hydrolysis test, 50% isolates were positive in starch hydrolysis test, 35% in protein hydrolysis test, 100% isolates in lactose fermenting test, but no isolate was positive in sucrose fermenting test. Antibiotic susceptibility testing suggested that 20% of isolates were resistant to kanamycin and 46.67% were resistant to oxacillin.

CONCLUSIONSThese findings show that all these isolates have gelatin, urea, galactose hydrolysis and lactose fermenting activity. 20% of these isolates were resistant to kanamycin and 46.67% were resistant to oxacillin.

Anti-Bacterial Agents ; pharmacology ; Disk Diffusion Antimicrobial Tests ; Galactose ; metabolism ; Gelatin ; metabolism ; Hydrolysis ; Microbial Sensitivity Tests ; Staphylococcus aureus ; drug effects ; isolation & purification ; metabolism ; Starch ; metabolism ; Urea ; metabolism

OBJECTIVETo evaluate the potency of carboxymethyl chitosan-2, 2' ethylenedioxy bis-ethylamine-folate (CMC-EDBE-FA) on tissue injury, antioxidant status and glutathione system in tissue mitochondria and serum against nicotine-induced oxidative stress in mice.

METHODSCMC-EDBE-FA was prepared on basis of carboxymethyl chitosan tagged with folic acid by covalently linkage through 2, 2' ethylenedioxy bis-ethylamine. Animals were divided into four groups, i.e., control, nicotine (1 mg/kg bw/day), CMC-EDBE-FA (1 mg/kg bw/day) and nicotine (1 mg/kg bw/day) and CMC-EDBE-FA (1 mg/kg bw/day) for 7 days. Levels of lipid peroxidation, oxidized glutathione level, antioxidant enzyme status and DNA damage were observed and compared.

RESULTSThe significantly increase of lipid peroxidation, oxidized glutathione levels and DNA damage was observed in nicotine treated group as compared with control group; those were significantly reduced in CMC-EDBE-FA supplemented group. Moreover, significantly reduced antioxidant status in nicotine treated group was effectively ameliorated by the supplementation of CMC-EDBE-FA. Only CMC-EDBE-FA treated groups showed no significant change as compared with control group; rather than it repairs the tissue damage of nicotine treated group.

CONCLUSIONSThese findings suggest that CMC-EDBE-FA is non-toxic and ameliorates nicotine-induced toxicity.

Animals ; Antioxidants ; chemistry ; pharmacology ; Chitosan ; analogs & derivatives ; chemistry ; pharmacology ; DNA Fragmentation ; drug effects ; Folic Acid ; chemistry ; pharmacology ; Glutathione ; analysis ; metabolism ; Glutathione Transferase ; metabolism ; Male ; Mice ; Nanoparticles ; chemistry ; Nicotine ; toxicity ; Organ Specificity ; Oxidoreductases ; metabolism

Objective: To evaluate phytopharmacologically eugenol and two extract products of Ocimum gratissimum Linn. (O. gratissimum) (Labiaceae) on free radical scavenging and antioxidant activity. Methods: Aqueous and methanol extract of fresh aerial part of O. gratissimum were prepared and eugenol (1-allyl-4-hydroxy-3-methoxybenzene) was isolated from fresh leaves and characterized by high performance liquid chromatography, fourier transform infrared spectroscopy, 1 h nuclear magnetic resonance. To establish the antioxidant potentiality of aqueous extract, methanol extract and eugenol, 1, 1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, nitric oxide scavenging activity, antioxidant activity by ferric thiocyanate and reducing power were measured in chemical system in vitro. Results: Significant (P<0.05) concentration-dependent free radical scavenging activity, antioxidant activity, and reducing power was observed by O. gratissimum products. Moreover, eugenol is more potent than the two extract products of O. gratissimum, but lower than potent antioxidant ascorbic acid. Conclusions: Hence, O. gratissimum presents a potentially valuable source of natural antioxidant and bioactive material.

OBJECTIVETo evaluate the free radical generation and status of the antioxidant enzymes in murine peritoneal macrophage during in vitro vancomycin sensitive Staphylococcus aureus (VSSA) treatment with different time interval.

METHODSPeritoneal macrophages were treated with 5×10(6) CFU/mL VSSA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and superoxide anion generation, NADPH oxidase activity, myeloperoxidase activity, nitric oxide generation, antioxidant enzyme status and components of glutathione cycle were analyzed.

RESULTSSuperoxide anion generation, NADPH oxidase activity, myeloperoxidase activity and nitric oxide generation got peak at 3 h, indicating maximum free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during VSSA infection. Reduced glutathione level, glutathione peroxidase, glutathione reductase, and glutathione-s-transferase activity were decreased significantly (P<0.05) with increasing time of VSSA infection. But the oxidized glutathione level was time dependently increased significantly (P<0.05) in murine peritoneal macrophages. All the changes in peritoneal macrophages after 3 h in vitro VSSA treatment had no significant difference.

CONCLUSIONSFrom this study, it may be summarized that in vitro VSSA infection not only generates excess free radical but also affects the antioxidant status and glutathione cycle in murine peritoneal macrophages.

OBJECTIVETo test the survival of Staphylococcus aureus (S. aureus) inside lymphocyte that contributes to the pathogenesis of infection and possible anti-inflammatory and antioxidative effect of nanoconjugated vancomycin against in vivo S. aureus infection in a dose and duration dependent manner.

METHODS5×10(6) CFU/mL vancomycin-sensitive S. aureus (VSSA) and vancomycin-resistive S. aureus (VRSA) were challenged in Swiss male mice for 3 days, 5 days, 10 days and 15 days, respectively. Bacteremia and inflammatory parameters were observed to evaluate the duration for development of VSSA and VRSA infection. 100 mg/kg bw/day and 500 mg/kg bw/day nanoconjugated vancomycin were administrated to VSSA and VRSA infected group for 5 days. Bacteremia, inflammatory parameters and oxidative stress related parameters were tested to observe the effective dose of nanoconjugated vancomycin against VSSA and VRSA infection. Nanoconjugated vancomycin was treated at a dose of 100 mg/kg bw/day and 500 mg/kg bw/day, respectively, to VSSA and VRSA infected group for successive 5 days, 10 days and 15 days. Bacteremia, inflammatory parameters and oxidative stress related parameters were observed to assess the effective duration of nanoconjugated vancomycin against VSSA and VRSA infection.

RESULTSThe result revealed that in vivo VSSA and VRSA infection developed after 5 days of challenge by elevating the NO generation in lymphocyte and serum inflammatory markers. Administration with nanoconjugated vancomycin to VSSA and VRSA infected group at a dose of 100 mg/kg bw/day and 500 mg/kg bw/day, respectively, for successive 10 days eliminated bacterimia, decreased NO generation in lymphocyte, serum inflammatory markers and increased antioxidant enzyme status.

CONCLUSIONSThese findings suggest, in vivo challenge of VSSA and VRSA for 5 days can produce the highest degree of damage in lymphocyte which can be ameliorated by treatment with nanoconjugated vancomycin for 10 successive days.

Animals ; Anti-Bacterial Agents ; administration & dosage ; chemistry ; Bacteremia ; drug therapy ; immunology ; microbiology ; Drug Delivery Systems ; Humans ; Male ; Mice ; Nanoparticles ; chemistry ; Nitric Oxide ; immunology ; Staphylococcal Infections ; drug therapy ; immunology ; microbiology ; Staphylococcus aureus ; drug effects ; pathogenicity ; physiology ; Vancomycin ; administration & dosage ; chemistry ; Vancomycin Resistance

Objective: To compare the protective effects of chitosan-trypolyphosphate (CS-TPP) nanoparticle conjugated chloroquine(CQ) with effect of CQ alone on the reversal of splenic damages and induction of apoptosis. Methods: Different researches have been carried out to explore the potential role of chitosan based drug delivery system against parasitic diseases. After successive Plasmodium berghei NK65 parasiste infection by intraperitoneal injection in Swiss mice and subsequent parasite development, the ROS generation, anti-apoptotic and pro apoptotic protein levels in spleen were measured. To analyze caspases, flow cytometry study was performed with annexin V-FITC and with PI staining. Results: The results revealed that ROS mediated caspase 3 and 9 activation and the induction of apoptosis occurred during the parasitic infection. However, CS-TPP conjugated CQ was relatively better in reversing the splenic damage compared with similar effects of CQ alone. Conclusions: This study indicates that Plasmodium berghei NK65 induces apoptosis in the spleen. The study further shows that CS-TPP nanoparticles conjugation with CQ have positive influence on the recovery of damaged host's system towards maintenance of normal homeostasis, and this is shown to be selective to CS-TPP conjugated CQ treated animals only.