The effect of nitric oxide combined with fluoroquinolones against Salmonella enterica serovar Typhimurium in vitro

Two regulons, soxRS and marRAB, are associated with resistance to quinolones or multiple antibiotic in Salmonella enterica serovar Typhimurium. These regulons are activated by nitric oxide and redox-cycling drugs, such as paraquat and cause on activation of the acrAB-encoded efflux pump. In this study, we investigated the effect of nitric oxide (NO) alone and in combination with ofloxacin, ciprofloxacin, and pefloxacin against S. typhimurium clinical isolates and mutant strains in vitro. We did not observe synergistic effect against clinical isolates and SH5014 (parent strain of acr mutant), while we found synergistic effect against PP120 (soxRS mutant) and SH7616 (an acr mutant) S. typhimurium for all quinolones. Our results suggest that the efficiencies of some antibiotics, including ofloxacin, ciprofloxacin, and pefloxacin are decreased via activation of soxRS and marRAB regulons by NO in S. enterica serovar Typhimurium. Further studies are warranted to establish the interaction of NO with the genes of Salmonella and, with multiple antibiotic resistance.     

Evaluation of hydroxyl radical-scavenging property of garlic

While antibiotics are broadly used in dental and medical therapy, little attention has been directed towards the potential toxic side effects of antibiotics on tissue regeneration. Here we examined the effect of a quinolone antibiotic, pefloxacin (Rhone Poulenc) on rat parotid gland responses to chronic isoproterenol treatment. Groups of rats received injections of isoproterenol to induce glandular growth, saline (controls), pefloxacin, or isoproterenol and pefloxacin in combination. Parotid gland weight decreased significantly after pefloxacin treatment for 7 days as well as inhibiting glandular enlargement provoked by isoproterenol. The same trend was observed for the rates of DNA synthesis, with the incorporation of [³H]-thymidine in isoproterenol/pefloxacin-treated rats reduced to 49% of isoproterenol treatment alone levels. Saline-treated animals were 42% of the rate of [³H]-thymidine incorporation into DNA observed in isoproterenol treated rats. While isoproterenol treatment increased steady-state mRNA levels for fos, jun, myc, src, c-erbB-2, ras and topo II, inclusion of pefloxacin with the isoproterenol regimen blocked these increases. Pefloxacin treatment by itself did not alter proto-oncogene mRNA levels in the parotid gland. Glandular amylase activity was decreased in the pefloxacin treated group, while the combination of isoproterenol with pefloxacin did not decrease glandular amylase levels to the extent of that observed with -agonist treatment alone. In acute experiments, pefloxacin significantly decreased the volume of saliva secreted by the parotid gland. These results suggest that quinolone-based antibiotics disturb the secretory function of the parotid gland and can inhibit cell proliferation and regeneration. (Mol Cell Biochem 165: 55–63, 1996)     

红球菌HX-2所产胞外多糖的特性和对Cu2+的吸附

【背景】目前,微生物所产胞外多糖(exopolysaccharide,EPS)的理化性质及其在重金属吸附中的应用受到了广泛关注。【目的】研究红球菌HX-2所产胞外多糖的理化性质,并探究其对重金属的吸附情况。【方法】使用离子交换和凝胶色谱分离法对胞外多糖粗品进行纯化;利用苯酚硫酸法测胞外多糖中糖含量;用Bradford试剂盒检测胞外多糖中蛋白含量;使用甲醇萃取法检测胞外多糖中脂质含量;用高效液相色谱(high performance liquid chromatography,HPLC)法分析胞外多糖中单糖组成;用扫描电镜(scanningelectronmicroscopy,SEM)法观察多糖表面形态;通过等温吸附模型和动力学模型探究胞外多糖对重金属的吸附效果。【结果】测得胞外多糖主要成分EPS-G-1中总糖含量为78.43%,蛋白含量为8.31%,脂质含量为8.22%;纯化后胞外多糖中单糖组成为葡萄糖、甘露糖、半乳糖、葡萄糖醛酸和岩藻糖,质量比为27.31:26.67:24.83:15.85:4.80;通过等温吸附模型拟合得到HX-2所产胞外多糖对Cu~(2+)的最大吸附量为144.93 mg/g。【结论】红球菌HX-2所产胞外多糖对水体中Cu~(2+)具有良好的吸附作用,可用于工业废水中重金属离子的处理。     

Effects of subinhibitory concentrations of antibiotics on biological properties ofSalmonella typhimurium

We followed the effects of subinhibitory concentrations (sub-MICs) of 7 antibiotics (ticarcilin, cefotaxim, streptomycin, gentamicin, ciprofloxacin, pefloxacin, mitomycin C) on the sensitivity of aSalmonella typhimurium strain to standard bacteriophages, on the phage DNA as well as on the factors of virulence (permeability and cytotoxic activity). The phage type was not changed by the sub-MICs of the tested antibiotics. However, differences were found in culture filtrates prepared from the bacterial suspensions of the strain cultivated with the sub-MICs. Marked inducing effects on phage DNA were exhibited by mitomycin C (1/2, 1/4, 1/8 of the MIC), pefloxacin (1/2, 1/4, 1/8 of the MIC) and ciprofloxacin (1/2, 1/4, weakly also 1/8 of the MIC). Ticarcilin (1/2 of the MIC), like the aminoglycosides streptomycin and gentamicin (1/2, 1/4, 1/8 of the MIC), had a weak effect. Sub-MICs of the studied antibiotics (with the exception of 1/8 of the MIC of ciprofloxacin and 1/4 of the MIC of ticarcilin) decreased the permeability reaction in rabbit skin. Most effective was streptomycin (1/2 of the MIC). Sub-MICs of the tested antibiotics (with the exception of 1/4 and 1/8 of the MIC of ciprofloxacin and 1/4 of the MIC of pefloxacin) caused also an inhibition of the factor responsible for morphological changes on Vero cells. Gentamicin and streptomycin were effective at all the sub-MICs tested.     

Optimization of a medium and the role of its various components for the biosynthesis of Mycobacterium cyaneum exopolysaccharide

The effect of some components of the medium on the growth of Mycobacterium cyaneum B-646 and on the biosynthesis of an exoglycan by the culture was studied. A medium in which the M. cyaneum M variant produced up to 2.2 g of the exopolysaccharide per litre, which was nearly 4 times more than in the original medium, was proposed. The new medium differed from the original one in an elevated content of carbon, iron and potassium (11.2, 5.0 and 1.4 times, respectively) and in a lower phosphorus content (6.7 times). The exopolysaccharide produced by the culture in this medium contained glucose, galactose, fucose and uronic acid. Therefore, its monomeric composition did not depend on the medium used for growing the culture which produced the exopolysaccharide.     

Quinolone therapy in the prevention of mortality after irradiation

The effect of oral therapy with three quinolones (ofloxacin, ciprofloxacin, and pefloxacin) in the prevention of postirradiation bacteremia and mortality was tested in B6D2F1 mice given 9.5 Gy 60Co gamma radiation. Only 8 of 60 (13%) untreated mice survived for 30 days, compared to 47 of 60 (78%) mice treated with ofloxacin, 44 of 60 (74%) mice treated with ciprofloxacin, and 42 of 60 (70%) mice treated with pefloxacin (P less than 0.05). The organisms recovered from the mice were Streptococcus spp. and Enterobacteriaceae. More Enterobacteriaceae were recovered from the livers of untreated animals than from the mice treated with the quinolones. However, no reduction in the number of Streptococcus spp. was noted in the animals given quinolones when compared to controls. This study shows that quinolones prolonged survival and decreased systemic spread of Enterobacteriaceae up to 30 days after exposure of mice to lethal irradiation.     

Decreased Exopolysaccharide Synthesis by Anaerobic and Symbiotic Cells of Bradyrhizobium japonicum

Experiments were conducted to determine whether symbiotic bacteroids of Bradyrhizobium japonicum produce exopolysaccharide within soybean (Glycine max [L.] Merr. cv `Lee 74') nodules. B. japonicum strains RT2, a derivative of USDA 110 with resistance to streptomycin and rifampicin, and RT176-1, a mutant deficient in exopolysaccharide synthesis, were used. Although aerobically cultured RT2 produced 1550 micrograms of exopolysaccharide per 10¹⁰ cells, root nodules formed by RT2 contained only 55.7 micrograms of polysaccharide per 10¹⁰ bacteroids, indicating that little exopolysaccharide synthesis occurred within the nodules. The polysaccharide level of RT2 nodules was about equal to that of nodules containing the exopolysaccharide mutant RT176-1 (61.0 micrograms per 10¹⁰ bacteroids). Gas chromatographic analysis showed that the sugar composition of polysaccharide from nodules of RT2 or RT176-1 was almost the same as that of polysaccharide from unnodulated root tissue, but differed strikingly from that of rhizobial exopolysaccharide from aerobic cultures. Thus, the host plant and not the bacteroids was probably the source of most or all of the polysaccharide in the nodule extracts. Also, bacteroids from nodules failed to bind soybean lectin, confirming the absence of an exopolysaccharide capsule.     

Effect of pefloxacin on immune response]

The influence on cellular immune response of different doses of the pefloxacin was studied in vivo as well as in vitro experiments. The pefloxacin in super bactericidal concentrations (2.0 mg/ml and 0.4 mg/ml) possess pronounced supressing effect the T-lymphocyte proliferation in blast transformation reaction. While in concentration 0.08 mg/ml pefloxacin does not show such activity. The pefloxacin in maximal effective concentration (200 mg/kg) suppressed activity in delayed hypersensitivity skin reaction of intact mice towards sheep erythrocytes on 20.3 percent only.     

Influence of phosphate and ammonia on the growth, exopolysaccharide production and symbiosis of Rhizobium leguminosarum bv. trifolii TA1 with clover (Trifolium pratense)

The Rhizobium-legume interaction is sensitive to a number of environmental factors, among which phosphate (Pi) and ammonium availability are the most important. We investigated the effect of Pi and ammonia concentration on exopolysaccharide production and symbiosis Trifolium pratense with Rhizobium leguminosarum bv. trifolii TA1 (RtTA1). The optimal Pi concentration in the bacterial growth medium for RtTA1 growth and exopolysaccharide production was in the range from 0.9 mM to 8.1 mM. Independently of Pi concentration, ammonium (NH4Cl) concentration above 8.1 mM in the culture medium significantly decreased EPS production, indicating a regulatory role of this nutrient on the EPS production in the RtTA1 strain. Pi availability has a beneficial effect on both partners of symbiosis. Pi concentration in the plant medium in the range from 1.7 mM to 5 mM was optimal for nodule formation, nodule occupancy and nitrogen fixation ability. Despite of T. pratense cv. Bryza tolerance on high Pi concentration, 20 mM Pi occurs to be nearly phytotoxic, which negatively affects almost all symbiotic parameters. Large amounts of starch were accumulated in the nodules formed by clover grown on medium containing high Pi concentration.     

Efficacy of pefoxacin (abactal) in the complex treatment of patients with pancreonecrosis]

The efficacy of pefloxacin in the complex treatment of 28 patients with pancreatonecrosis of various etiology was estimated in a prospective trial. The diagnosis of pancreatonecrosis was verified by the data of the disease clinical progress, laboratory findings and instrumental examination. Pefloxacin (Abactal; LEK) was administered intravenously in a dose of 400 mg every 8 hours (1200 mg) in combination with metronidazole in a dose of 1.5-2.0 g a day intravenously. When indicated 3 days after the start of the pefloxacin therapy, the treatment was switched to the oral use of the drug in the same dosage. The positive clinical effect (cure and improvement) at the end of the treatment with pefloxacin was stated in 78 per cent of the patients in spite of the initial severity state of above 15 APACHE II. It was shown that in the treatment of patients with pancreatonecrosis when the severity state was not above 12 APACHE II the antibacterial therapy with pefloxacin in combination with metronidazole was optimal.     

The experimental validation of the advantages of combined emergency (fluoroquinolones) and specific (EV Nalr) prevention of plague versus their sequential use]

Mice immunization with reference vaccine at the early stage of plague infection provided animals survival and prolonged mean survival period up to 2-5 days. Ciprofloxacin, ofloxacin and pefloxacin prevents development of post vaccine immunity at white mice, immunized by reference vaccine strain EV. Nalidixic acid and norfloxacin effect on post vaccine immunity was lower. Use of immunogenic strain EV Nafr (resistant to nalidixic acid and fluoroquinolones) provided antiplague immunity formation at the background of fluoroquinolones prophylaxis. Ciprofloxacin, ofloxacin and pefloxacin used for plague prophylaxis at white mice infected with Yersinia pestis (about 1000 LD50) inhibited postinfective immunity development. Nalidixic acid and norfloxacin didn't demonstrate such effect. Urgent (fluoroquinolones) and specific (EV Nalr) combined prophylaxis was evaluated as more effective for a 5-day period and provided the development of antiplague immunity.     

The effects of Anabaena spiroides (Cyanophyceae) exopolysaccharide on copper toxicity to Simocephalus serrulatus (Cladocera, Daphnidae)

1. This work investigated the consumption of a microalgal (Anabaena spiroides) exopolysaccharide by the cladoceran Simocephalus serrulatus (Cladocera, Daphnidae) and its effect on copper toxicity. 2. Total organic carbon concentration was used to quantify the microalgal exopolysaccharide. Both copper‐complexed exopolysaccharides and copper‐free exopolysaccharides were taken up by S. serrulatus. 3. A reduction of free copper ions was obtained in the presence of A. spiroides exopolysaccharide. Copper toxicity (EC₅₀) and zooplankton concentrations were inversely correlated to exopolysaccharide concentration in experimental media. 4. An increase of free copper ions in experimental media was obtained after exopolysaccharide consumption, suggesting that S. serrulatus excreted copper to the surrounding environment.     

Effect of dissolved oxygen on growth and production of exopolysaccharide by Rhizobium trifolii

Batch fermentations of Rhizobium trifolii 0403 were performed with dissolved oxygen (DO) control to determine the effect of DO on growth rate, and the production of exopolysaccharide and biomass. When DO was maintained between 4 and 20% of saturation, growth was exponential. The apparent specific growth rate, biomass and exopolysaccharide production did not vary significantly over this DO range. At oxygen transfer rates less than the minimum required for exponential growth, DO fell to below 1%. Growth under these conditions was linear and in direct relation to the oxygen transfer rate. Exopolymer production increased and biomass production decreased under oxygen-limited conditions.     

Comparative efficacy and safety of ciprofloxacin, ofloxacin, and pefloxacin in treatment of respiratory infections in children with cystic fibrosis]

Data on comparative investigation of the clinical and bacteriological efficacy and tolerability of monofluoroquinolones ciprofloxacin, ofloxacin and pefloxacin are given. The results confirm good clinical efficacy of all three monofluoroquinolones and high antistaphylococcal activity of pefloxacin. Efficacy of monofluoroquinolones against P. aeruginosa and S. maltophilia was moderate--only isolation of bacteria from spetrum became lower. Tolerability of monofluoroquinolones was good. Only at 5 patients the drugs use was stopped (4 in the ciprofloxacin group and 1--in the pefloxacin group). At 2 patients it was caused by arthropathy which was drug- and age-dependent. Quinolone-arthropathy was more often in the pefloxacin group and was registered only at the children elder than 10 years old with arthrological anamnesis. This arthropathy differed from experimental one by positive outcome and full recovery in the period from 7 days to 3 months. Results of morphological investigation confirmed clinical data--no invalidizing cartilage damage was revealed.     

Pefloxacin in the treatment of chronic urinary infection]

This study aimed at determining sensitivity of the hospital isolates to pefloxacin compared with other urinary antiseptics and assessment of its efficiency in the treatment of the chronic urinary infections. Disc diffusion technique was used to determine bacterial sensitivity to pefloxacin and other agents. Bacteria were isolated from the urine of the hospitalized patients. All tested strains were sensitive to pefloxacin. Its sensitivity exceeded in vitro 80%. Fifteen patients full sensitive to pefloxacin were treated. No bacteria were seen in the urine of 8 patients in two bacteriological cultures following the treatment, 3 patients stopped therapy due to adverse reactions, and no sterile urine was noted in 4 patients despite following a complete cycle of therapy.     

Evaluation of tolerance and efficacy of pefloxacin in the treatment and prevention of severe infections in children with mucoviscidosis and aplastic anemia]

Pefloxacin (Abaktal) efficacy and safety were investigated at 21 children (7-16 years old) randomized in 2 groups: children with mucoviscidosis and children with aplastic anemia. The drug was used at the dose 15-20 mg/kg per day bid for 14-28 days. Pefloxacin was used in combination with ceftazidime and amikacin. Combined therapy demonstrated good clinical efficacy. Bacteriological efficacy was not uniform: staphylococci were not isolated from sputum since the 7th day of treatment, but pseudomonads were cultured even on the 14th day of the treatment (the sensitivity to pefloxacin remained). The only but frequent side-effect was arthropathy. The background and some peculiarities of arthropathy development were analyzed. This phenomenon is called quinolone-induced synovitis. The risk group for quinolone-induced synovitis was estimated--children elder than 10 years with allergic anamnesis. Good clinical efficacy and tolerability of pefloxacin at the children with mucoviscidosis or aplastic anemia is a reason and base to cancel the limits to its use in pediatrics.     

尖顶羊肚菌胞外多糖提取物对皮肤成纤维细胞增殖和衰老的影响

不同浓度尖顶羊肚菌胞外多糖提取物作用人皮肤成纤维细胞（human skin fibroblasts,HSF）,检测对HSF细胞形态、细胞增殖、衰老相关β-半乳糖苷酶活性、羟脯氨酸含量的影响,探究尖顶羊肚菌胞外多糖提取物对HSF增殖和衰老的影响。结果显示,125μg/mL尖顶羊肚菌胞外多糖提取物使HSF细胞活力增加了25.2%,羟脯氨酸含量增加了12.1%,β-半乳糖苷酶活性降低了48.1%。说明适宜浓度尖顶羊肚菌胞外多糖提取物具有促进HSF细胞增殖、胶原蛋白合成,延缓细胞衰老的作用。     

Effect of Different Antibiotics and Non-Steroidal Anti-Inflammatory Drugs on the Growth of Lactobacillus casei Shirota

The purpose of this study was to investigate whether some non-steroidal anti-inflammatory drugs (NSAIDs) could cause inhibition of the growth of Lactobacillus casei Shirota (LcS) or whether this microorganism is able to use some of them as the sole carbon source, considering that the simultaneous consumption of NSAIDs and a dairy drink fermented with LcS could help to prevent the appearance or improve the healing of gastric ulcers. Acetylsalicylic acid (ASA), sodium acetylsalicylate (SAS), acetaminophen, sodium naproxen, and sodium ibuprofen were added as the sole carbon source to a basal medium and tested for biodegradation by LcS. The same NSAIDs were added in different concentrations to disks and plated on MRS Agar to test the possible inhibitory effect of these compounds on LcS. Also, the resistance of LcS to 12 different antibiotics was studied on MRS agar. None of the NSAIDs tested could be used by LcS as the sole carbon source at the assayed concentrations. In the case of the disk diffusion method, sodium naproxen showed inhibition zones for the 500-μg disks and sodium ibuprofen was inhibitory for the 250- and 500-μg disks. However, when the macrobroth dilution method was used, the growth of LcS was inhibited by ASA, SAS, acetaminophen, and sodium ibuprofen. This strain showed resistance to the antibiotics sulfamethoxazole with trimethoprim, pefloxacin, and gentamicin. This is the first study on the effect of NSAIDs on probiotic bacteria. The results of the biodegradation test indicate that the simultaneous consumption of NSAIDs and a dairy beverage with LcS is not likely to change the bioavailability of the drugs.     

Ssg, a putative glycosyltransferase, functions in lipo- and exopolysaccharide biosynthesis and cell surface-related properties in Pseudomonas alkylphenolia

In the presence of vaporized p-cresol, Pseudomonas alkylphenolia KL28 forms specialized aerial structures (SAS). A transposon mutant of strain KL28 (C23) incapable of forming mature SAS was isolated. Genetic analysis of the C23 mutant revealed the transposon insertion in a gene (ssg) encoding a putative glycosyltransferase, which is homologous to the Pseudomonas aeruginosa PAO1 PA5001 gene. Deletion of ssg in KL28 caused the loss of lipopolysaccharide O antigen and altered the composition of the exopolysaccharide. Wild-type KL28 produced a fucose-, glucose- and mannose-rich exopolysaccharide, while the mutant exopolysaccharide completely lacked fucose and mannose, resulting in an exopolysaccharide with glucose as the major component. The mutant strain showed reduced surface spreading, pellicle and biofilm formation, probably due to the cumulative effect of lipopolysaccharide truncation and altered exopolysaccharide composition. Our results show that the ssg gene of KL28 is involved in both lipopolysaccharide and exopolysaccharide biosynthesis and thus plays an important role in cell surface properties and cell-cell interactions of P. alkylphenolia.