Influence of plasma on the efficacy of ozone in aqueous medium.
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Objective To study the influence of plasma on the sterilization effect of ozone in aqueous medium and to provide data for further evaluation on the efficacy of ozone for sterilization of body fluid such as ascites. Method Escherichia coli(ATCC25922)and Staphylococcus areus(ATCC25923)were exposed to various concentrations of normal saline diluted plasma. The killing rate of ozone (90 μg / mL) was calculated. The dose of ozone and the kinetic of bacterial cell killing were determined in normal saline with 10.0% human plasma. Result The efficacy of ozone in normal saline was dose dependently decreased, especially at the plasma concentration of 10% (P0.05). Under the conditions of 10% plasma, 90 μg / mL and the flow rate of 12 ml / min, the killing rate for Escherichia coli and Staphylococcus aureus was 100% and 99.05%, respectively. Differences were observed in groups treated with 40 μg / mL, 60 μg / mL and 90 μg / mL of ozone (P0.05). The efficacy was also varied with the exposure time (5 min,10 min and 20 min). The best bactericidal effect was observed when the cells were treated with 90 μg / mL of ozone and the exposure time was 20 min. Conclusion The efficacy of ozone was reduced at the plasma concentration of 10%. The efficacy can be improved by increasing both the concentration and the exposure time to ozone.Keywords:
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To study the efficacy and safety of adsorption of endotoxin and proinflammatory cytokines with HB-H-7 resin in vitro.Static adsorption experiment: HB-H-7 resin was added into plasma of endotoxin-positive patients according to the ratio of 1;10. Then, they were put in a constant temperature water bath oscillator, and oscillated for 2 hours. Before and after the experiment, plasma endotoxin, proinflammatory cytokines, protein and electrolytes were detected, and the rate of 2-hour absorption was calculated. The experiment was repeated 10 times. Dynamic adsorption experiment: 5 ml resin was put into a self-made perfusion unit. Endotoxin-positive patients plasma (50 ml) was perfused for 2 hours. The flow rate of plasma was controlled at 4 ml/min with the infusion pump, and the plasma endotoxin was determined at 0, 30, 60 and 120 minutes after plasma hemoperfusion, and the absorption rates were calculated. The results were compared with static adsorption. The influence of temperature during adsorption was determined as follows. Perfusion method was similar with dynamic adsorption experiment. The perfusion units were either placed in a 37 centigrade water path or 25 centigrade (room temperature). Then, the plasma endotoxin was measured 2 hours after plasma perfusion, and the absorption rates were calculated.In static adsorption experiment, the plasma endotoxin and proinflammatory cytokines were significantly lower after adsorption with HB-H-7 resin adsorption (all P<0.05). The adsorption rates of endotoxin, tumor necrosis factor-alpha and interleukin-1, 6, 8 were 99.2%, 55.0%, 57.0%, 75.0%, 42.0%, respectively. Changes in protein were small (all P<0.05) , and there was no significant change in plasma electrolytes (all P>0.05). Dynamic adsorption rate was higher than that of static, but the differences were not significant (99.8% vs. 99.1%, P>0.05). There was no statistically significant difference between difference in temperature (37 centigrade vs. 25 centigrade, 99.8% vs. 99.9%, P>0.05).HB-H-7 resin effectively absorbs endotoxin and proinflammatory cytokines in vitro. Its adsorption rate for protein is lower, and it has no obvious effects on electrolytes.
Proinflammatory cytokine
Hemoperfusion
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Objective:To study the efficacy and safety of HB-H-7 resin for the adsorption of endotoxin by plasmaperfusion in vitro.Methods:Static adsorption experiment:HB-H-7 resin was added into endotoxin-positive patients plasma according to the volume ratio of 1:10.Then,they were putted into constant temperature water bath oscillator at 37 ℃,and had been vibrated for 2 hours.Before and after the experiment,plasma endotoxin,protein and electrolytes were detected,and the rate of 2 hour adsorption was calculated.The experiment was repeated 10 times.Dynamic adsorption experiment.5 ml resin was putted into a selfmade perfusion unit.50 ml endotoxin-positive patients plasma had been plasmaperfused for 2 hours in the flow rate of plasma 4 ml/min with the infusion pump.The plasma endotoxin were detected at 0,30,60 and 120 minutes' plasma hemoperfusion,and the adsorption rates were calculated.The results were compared with those from static adsorption experiment.The supernatent of resin extracts was screened by ultraviolet light sepetrophotometer.Results:The plasma endotoxin was significantly lower after HB-H-7 resin adsorption (P 0.05);Protien in plasma was lower (P 0.05);and plasma electrolytes were not significant changes (P0.05).Dynamic adsorption rate was higher than static's.No dissolved matters of resins were found.Conclusion:There is obvious endotoxin absorption efficiency in plasma with HB-H-7 resin.It has not obvious effects on electrolytes adsorption,but it has effects on protein adsorption.Further endotoxemia treatment experiments by plasmaperfusion in vivo should be carried out.
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Abstract The amount of the cold trichloroacetic acid soluble fraction (metabolic pool), expressed as a percentage of its 32P content, released from labelled cells of Escherichia coli treated with chlorhexidine has been compared with the percentage of cells killed by the chlorhexidine. Treatment was with amounts varying from 0–64 μg/ml for 5 min at 20°. Approximately 100% of the cells were killed and 100% of the metabolic pool released by treatment with 64μg/ml of chlorhexidine. At 4, 8 and 16 μg/ml the percentage of cells killed by chlorhexidine is significantly lower than the percentage of the metabolic pool released.
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Abstract: Percutaneous absorption of lipophilic substances has major implications for therapeutical use or toxicological effects. We, therefore, using dermal microdialysis, measured local toluene concentrations and assessed the effects of duration of exposure, skin barrier disruption and the use of skin‐care products. Three microdialysis membranes (3000 kDa) were inserted intradermally at a length of 2 cm in the abdominal skin of 82 anaesthetized male Wistar rats. They were perfused with albumin solution (5%) at 10 µl/min. A skin area of 1.5 × 0.6 cm above the membranes was exposed to toluene (100%, 200 µl) for 15 or 240 min. Dialysate was sampled at 20‐min intervals. Using GC‐FPD (gas charomotography flame photometric detector), it was analysed for toluene. In addition, the effects of tape stripping and pretreatment with topical products were assessed. In each of the 12 permutations of exposure time, pretreatments and tape stripping, five to eight animals were investigated. Maximum toluene concentrations were reached at 60 min after exposure (3.07 ± 0.40 µg/ml, 15 min; 5.38 ± 0.92 µg/ml, 240 min). In 15‐min exposure experiments, dermal toluene concentrations decreased slowly to reach baseline values after 240 min. After 240‐min exposure, a plateau of approximately 6 µg/ml was reached after 60 min. Neither tape stripping nor the pretreatment with barrier cream induced a significant change on dermal toluene concentrations. The slow kinetics of toluene penetration results in a steep concentration gradient in the skin with very‐high local toluene concentrations and a delayed wash out, which might be relevant not only toxicologically, but also therapeutically.
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In concentrations of 6 microgram/ml Fosfomycin acted bactericidal against E. coli ATCC 10536. The sensitivity of E. coli was evaluated by turbidity measurement (Table 1) and by counting colony forming units (CFU) (Table 2). Thus the bactericidal action began at different times in respect to the concentration and the method of documentation: turbidity fell 30-120 min after the administration of 6 microgram/ml and 10-30 min after 60 microgram/ml; CFU were reduced 10-30 min after 6 microgram/ml and 3-10 min after 60 microgram/ml. Before the cytoplasm and DNA-region were disorganized with reduced electron density, some elongated (up to more than 20 micron) cells occurred (Fig. 1,2). More prominent alterations in shape and ultrastructure were obvious 120 min after 6 microgram/ml (Fig. 5) and after 30 min when 60 microgram Fosfomycin per ml were administered (Fig. 3, 4), i.e. considerably later than the reduction of reproductivity.
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The immediate killing effect of atmospheric-pressure plasma (APP) has been frequently investigated, but its sustained killing activity is poorly understood. The goal of the present study is to evaluate both the immediate and sustained killing effects of APP on Enterococcus faecalis. The APP jet was evaluated by optical emission spectroscopy (OES) and laser-induced fluorescence spectroscopy (LIF). Hydroxyapatite (HA) discs coated with bovine dermal type I collagen were used as substrates for bacterial growth. After the formation of E. faecalis biofilms on the HA discs for seven days or three weeks, the samples were treated with (A) 2 ml of saline, (B) APP, or (C) 2 ml of 2% chlorhexidine digluconate (CHX) for 5 min. The treated samples were then cultured for three or seven days, after which they were examined by scanning electron microscopy and confocal laser scanning microscopy. The OES results showed that typical reactive oxygen and nitrogen species were included in the full spectrum. The fitted curve indicated that the rotation temperature of N2 was close to room temperature. The LIF results showed that the maximal O and OH intensities occurred at 5 mm from the nozzle. For both the seven-day and three-week biofilms, the CHX and APP treatments had significant sterilization results (P < 0.05) compared to the saline group in terms of immediate and sustained killing effects. APP demonstrated excellent potential for use as an alternative approach for the treatment of periapical diseases.
Enterococcus faecalis
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Infections after epidural and spinal blocks are rare. The topical anesthetic liclocaine used in these procedures has been found to have antibacterial effects on various microorganisms.The aim of this study was to assess the antibacterial effects of alkalinized liclocaine on Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa.Lidocaine 2%, alkalinized lidocaine, and physiologic saline (as a control solution) were added to standard bacterial preparations. The final concentration of the lidocaine was 10 mg/mL (1%). At baseline and 3 and 6 hours after incubation at 37°C, 3-mL aliquots were vortexed and pipetted into sterile polystyrene spectrophotometer cuvettes. Baseline referred to the end of the period of preparation of the solution (≤20 minutes). Growth was measured as the optical density at a wavelength of 540 nm.Compared with the control, lidocaine significantly inhibited the growth of S aureus, E coli, and P aeruginosa at baseline and 3 and 6 hours after incubation (all, P < 0.05). Alkalinized lidocaine significantly inhibited the growth of S aureus at baseline and 3 and 6 hours (all, P < 0.05), while it significantly inhibited the growth of E coli and P aeruginosa only at 6 hours (both, P < 0.05). The growth of E coli was significantly less in lidocaine than in alkalinized lidocaine at 0 and 3 hours (both, P < 0.05).The antibacterial effect of lidocaine 1% on S aureus was not changed after alkalinization. The effect of alkalinized lidocaine on E coli and P aeruginosa was significant only at 6 hours. Lidocaine significantly inhibited the growth of these 3 microorganisms at all study periods.
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Sodium polyanethol sulfonate (SPS) at 500 microgram/ml, but not sodium amylosulfate (SAS) at 500 microgram/ml, precipitated egg white lysozyme (1 mg and 50 microgram of lysozyme per ml) as determined with the assay strain Micrococcus lysodeikticus ATCC 4698. Fresh and heat-inactivated (56 degrees C, 30 min) human serum (80%, vol/vol) killed M. lysodeikticus (10(4) bacteria per ml at zero time) within 1 to 2 h after exposure. Addition of 250 to 500 microgram of SPS per ml to fresh human serum protected M. lysodeikticus for 22 h as effectively as absorption of either fresh or heat-inactivated human serum with bentonite (10 mg/ml of serum, 10 min, 37 degrees C); the latter procedure is known to remove serum lysozyme. In contrast, SAS at 250 and 500 microgram/ml of serum retarded killing of the assay bacteria for periods of 4 h; after overnight (22 h) incubation, however, the number of M. lysodeikticus survivors had decreased significantly. The finding that SPS, but not SAS, at 250 to 500 microgram/ml effectively neutralized serum lysozyme-mediated killing of a lysozyme-sensitive assay strain may be of relevance with respect to laboratory processing of human blood culture specimens.
Microgram
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