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Biomedical subjects

X Z Li

Publications and source records attributed to X Z Li.

At least 55 records · Page 3Linked to original sources
PubMed1999

Performance of fibrous bed bioreactor for treating odorous gas. Scientific note.

A fibrous bed bioreactor was used for treatment of odorous volatile fatty acid (VFA). The effect of gaseous VFA (acetic, propionic, and butyric acids) mass loading on the bioreactor performance was investigated. The VFA degrading microbial culture was selected from activated sludge by the three VFAs using a shake-flask culture. The selected microorganisms were then immobilized in a biofilter using cotton fabric as packing material. In the biofiltration experiment, the inlet gas flow rates ranged from 1 to 4 L/min, the total VFA concentrations ranged from 0.10 to 0.43 g/m3, and the resulting total mass loadings of VFA studied ranged from 9.7 to 104.3 g/m3/h. At total mass loading of 104.3 g/m3/h, the VFA removal efficiency was 87.7%. Higher removal efficiencies (>90%) were achieved at mass loadings below 50.3 g/m3/h.

Journal Article↗
PubMed1998

Beta-lactamase inhibitors are substrates for the multidrug efflux pumps of Pseudomonas aeruginosa.

The MexAB-OprM multidrug efflux system exports a number of antimicrobial compounds, including beta-lactams. In an attempt to define more fully the range of antimicrobial compounds exported by this system, and, in particular, to determine whether beta-lactamase inhibitors were also accommodated by the MexAB-OprM pump, the influence of pump status (its presence or absence) on the intrinsic antibacterial activities of these compounds and on their abilities to enhance beta-lactam susceptibility in intact cells was assessed. MIC determinations clearly demonstrated that all three compounds tested, clavulanate, cloxacillin, and BRL42715, were accommodated by the pump. Moreover, by using beta-lactams which were readily hydrolyzed by the Pseudomonas aeruginosa class C chromosomal beta-lactamase, it was demonstrated that elimination of the mexAB-oprM-encoded efflux system greatly enhanced the abilities of cloxacillin and BRL42715 (but not clavulanate) to increase beta-lactam susceptibility. With beta-lactams which were poorly hydrolyzed, however, the inhibitors failed to enhance beta-lactam susceptibility in MexAB-OprM+ strains, although BRL42715 did enhance beta-lactam susceptibility in MexAB-OprM- strains, suggesting that even with poorly hydrolyzed beta-lactams this inhibitor was effective when it was not subjected to efflux. MexEF-OprN-overexpressing strains, but not MexCD-OprJ-overexpressing strains, also facilitated resistance to beta-lactamase inhibitors, indicating that these compounds are also substrates for the MexEF-OprN pump. These data indicate that an ability to inactivate MexAB-OprM (and like efflux systems in other bacteria) will markedly enhance the efficacies of beta-lactam-beta-lactamase inhibitor combinations in treating bacterial infections.

Anti-Bacterial Agents↗
PubMed1998

Contribution of outer membrane efflux protein OprM to antibiotic resistance in Pseudomonas aeruginosa independent of MexAB.

A Pseudomonas aeruginosa strain carrying an insertion of an omega Hg interposon in the mexB gene (mexB::omega Hg; strain K879) produced markedly reduced but still detectable levels of OprM, the product of the third gene of the mexAB-oprM multidrug efflux operon. By using a lacZ transcriptional fusion vector, promoter activity likely responsible for OprM expression in the mexB::omega Hg mutant was identified upstream of oprM. Introduction of the oprM gene, but not the mexAB genes, into a P. aeruginosa multidrug-susceptible delta mexAB-oprM mutant increased resistance to quinolones, cephalosporins, erythromycin, and tetracycline. A delta mexAB-oprM strain carrying the oprM gene accumulated markedly less antibiotic than the deletion strain without oprM. Antibiotic accumulation by the MexAB- OprM+ strain was markedly enhanced upon treatment of cells with the uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP), indicating that MexAB-independent OprM function likely involves an efflux process. Moreover, pretreatment of cells with CCCP prior to the accumulation assay abrogated any differences in accumulation levels between the MexAB- OprM+ and MexAB- OprM- strains, indicating that reduced drug accumulation by the OprM+ strain (in the absence of CCCP) cannot be due to OprM-mediated reduction in outer membrane permeability. It appears, therefore, the OprM can be expressed and function in a drug efflux capacity independent of MexAB.

Bacterial Outer Membrane Proteins↗
PubMed1998

Influence of the TonB energy-coupling protein on efflux-mediated multidrug resistance in Pseudomonas aeruginosa.

TonB couples the energized state of the cytoplasmic membrane to the operation of outer membrane receptors responsible for Fe(III) siderophore uptake across the outer membrane of gram-negative bacteria. A tonB mutant of Pseudomonas aeruginosa deficient in iron siderophore uptake was shown in the present study to be hypersusceptible to a wide variety of antibiotics, reminiscent of the phenotype of mutants defective in the mexAB-oprM antibiotic efflux operon. This was not related to influences of a tonB mutation on the iron status of the cell, and indeed, intrinsic antibiotic susceptibility and mexAB-oprM expression were unaffected by iron levels in the growth medium. The presence of tonB on a multicopy plasmid increased the level of resistance of a MexAB-OprM+ strain but not that of a MexAB-OprM- strain to a variety of antimicrobial agents. mexAB-oprM expression was not, however, altered in a tonB deletion mutant, indicating that any influence of TonB on MexAB-OprM-mediated multidrug resistance was at the level of pump activity. Consistent with this, drug accumulation assays revealed that the tonB deletion mutant exhibited decreased levels of drug efflux. Still, the multidrug resistance of a nalB strain was not wholly abrogated by a tonB mutation, indicating that it is likely not an essential component of the efflux apparatus. Similarly, elimination of tonB from an nfxB strain only partially compromised MexCD-OprJ-mediated multidrug resistance. Intriguingly, the drug susceptibility of a mexAB-oprM deletion strain was increased following deletion of tonB, suggesting that TonB may also influence antibiotic resistance mediated by determinants other than MexAB-OprM (and MexCD-OprJ). Thus, TonB plays an important role in both intrinsic and acquired antibiotic resistance in P. aeruginosa.

Anti-Bacterial Agents↗
PubMed1998

Role of the multidrug efflux systems of Pseudomonas aeruginosa in organic solvent tolerance.

Multidrug efflux pumps with a broad substrate specificity make a major contribution to intrinsic and acquired multiple antibiotic resistance in Pseudomonas aeruginosa. Using genetically defined efflux pump mutants, we investigated the involvement of the three known efflux systems, MexA-MexB-OprM, MexC-MexD-OprJ, and MexE-MexF-OprN, in organic solvent tolerance in this organism. Our results showed that all three systems are capable of providing some level of tolerance to organic solvents such as n-hexane and p-xylene. Expression of MexAB-OprM was correlated with the highest levels of tolerance, and indeed, this efflux system was a major contributor to the intrinsic solvent tolerance of P. aeruginosa. Intrinsic organic solvent tolerance was compromised by a protonophore, indicating that it is substantially energy dependent. These data suggest that the efflux of organic solvents is a factor in the tolerance of P. aeruginosa to these compounds and that the multidrug efflux systems of this organism can accommodate organic solvents, as well as antibiotics.

Bacterial Outer Membrane Proteins↗
PubMed1997

Silver-resistant mutants of Escherichia coli display active efflux of Ag+ and are deficient in porins.

Silver-resistant mutants were selected by stepwise exposure of silver-susceptible clinical strains of Escherichia coli, two of which did not contain any plasmids, to either silver nitrate or silver sulfadiazine. These mutants showed complete cross-resistance to both compounds. They showed low-level cross-resistance to cephalosporins and HgCl2 but not to other heavy metals. The Ag-resistant mutants had decreased outer membrane (OM) permeability to cephalosporins, and all five resistant mutants tested were deficient in major porins, either OmpF or OmpF plus OmpC. However, the well-studied OmpF- and/or OmpC-deficient mutants of laboratory strains K-12 and B/r were not resistant to either silver compound. Resistant strains accumulated up to fourfold less (110m)AgNO3 than the parental strains. The treatment of cells with carbonyl cyanide m-chlorophenylhydrazone increased Ag accumulation in Ag-susceptible and -resistant strains, suggesting that even the wild-type Ag-susceptible strains had an endogenous Ag efflux activity, which occurred at higher levels in Ag-resistant mutants. The addition of glucose as an energy source to starved cells activated the efflux of Ag. The results suggest that active efflux, presumably coded by a chromosomal gene(s), may play a major role in silver resistance, which is likely to be enhanced synergistically by decreases in OM permeability.

Adenosine Triphosphate↗
PubMed1997

Inner membrane efflux components are responsible for beta-lactam specificity of multidrug efflux pumps in Pseudomonas aeruginosa.

A major feature of the MexAB-OprM multidrug efflux pump which distinguishes it from the MexCD-OprJ and MexEF-OprN multidrug efflux systems in Pseudomonas aeruginosa is its ability to export a wide variety of beta-lactam antibiotics. Given the periplasmic location of their targets it is feasible that beta-lactams exit the cell via the outer membrane OprM without interaction with MexA and MexB, though the latter appear to be necessary for OprM function. To test this, chimeric MexAB-OprJ and MexCD-OprM efflux pumps were reconstituted in delta mexCD delta oprM and delta mexAB delta oprJ strains, respectively, and the influence of the exchange of outer membrane components on substrate (i.e., beta-lactam) specificity was assessed. Both chimeric pumps were active in antibiotic efflux, as evidenced by their contributions to resistance to a variety of antimicrobial agents, although there was no change in resistance profiles relative to the native pumps, indicating that OprM is not the determining factor for the beta-lactam specificity of MexAB-OprM. Thus, one or both of inner membrane-associated proteins MexA and MexB are responsible for drug recognition, including recognition of beta-lactams.

Bacterial Outer Membrane Proteins↗
PubMed1996

Overexpression of the mexC-mexD-oprJ efflux operon in nfxB-type multidrug-resistant strains of Pseudomonas aeruginosa.

OprJ, overproduced in nfxB multidrug-resistant strains of Pseudomonas aeruginosa, and OprK, overproduced in the multidrug-resistant strain K385, were demonstrated to be immunologically cross-reactive using an OprJ-specific monoclonal antibody. Treatment of the purified proteins with trypsin or chymotrypsin yielded virtually indistinguishable digestion patterns, and the N-terminal sequence of two trypsin fragments was identical for both proteins, indicating that OprJ and OprK share identity. The N-terminal amino acid sequences were used to facilitate cloning of the oprJ gene on a 5kbp Kpnl fragment and a 10 kbp BamHl fragment. Nucleotide sequencing of portions of these fragments revealed that oprJ was the terminal gene in a putative three-gene operon, mexC-mexD-oprJ. The predicted mexC-mexD-oprJ gene products exhibit homology to the MexA-MexB-OprM components of the multidrug-resistance efflux pump of P. aeruginosa (43-46% identity). Consistent with an implied role for mexC-mexD-oprJ in drug efflux, the mexC-mexD-oprJ-hyperexpressing strain K385 showed reduced accumulation of a variety of antibiotics as compared with its parent strain, and this drug 'exclusion' was abrogated by energy inhibitors. The mexC and oprJ products are putative lipoproteins of a molecular mass of 40,707 and 51,742 Da, respectively, while mexD was predicted to encode a protein of 111 936 Da. Sequencing upstream of mexC revealed the presence of the nfxB gene transcribed divergently from the efflux genes. Overproduction of OprJ and the attendant multiple-antibiotic resistance of strain K385 was shown to result from a point mutation in nfxB, resulting in a H87-->R change in the predicted NfxB polypeptide. OprJ overproduction and multidrug resistance in K385 was reversed by the cloned nfxB gene, suggesting that nfxB encodes a repressor of mexC-mexD-oprJ expression. Consistent with this, the cloned nfxB gene repressed synthesis of a mexC-lacZ fusion in Escherichia coli. nfxB also repressed expression of a nfxB-lacZ fusion, indicating that NfxB negatively regulates its own expression. These data indicate that the multidrug resistance of nfxB strains is due to overexpression of an efflux operon, mexC-mexD-oprJ, encoding components of a second efflux pump in P. aeruginosa.

Amino Acid Sequence↗
PubMed1995

Role of mexA-mexB-oprM in antibiotic efflux in Pseudomonas aeruginosa.

We have earlier described mexA-mexB-oprK, an operon involved in pyoverdine export in Pseudomonas aeruginosa, and suggested that the products of these genes also contribute to the active efflux of several antibiotics (K. Poole, K. Krebes, C. McNally, and S. Neshat, J. Bacteriol. 175:7363-7372, 1993). Recently the outer membrane component of this efflux system was shown to be OprM, rather than OprK (N. Gotoh and K. Poole, unpublished results). In the present study, the conclusion concerning the efflux activity of this system was confirmed and extended by the measurement of drug accumulation in intact cells. Thus, the steady-state accumulation levels of tetracycline and norfloxacin were increased in mexA and oprM null mutants. mexA and oprM null mutants also showed an increase in susceptibility to a wide variety of beta-lactam antibiotics and an increase in the steady-state accumulation level of benzylpenicillin, indicating that the MexA-MexB-OprM pump also effluxes beta-lactams. Furthermore, deenergization of the cytoplasmic membrane with a proton conductor always produced a strong increase in the accumulation level. Finally, a single-step mutant over-producing MexAB-OprM accumulated less tetracycline and chloramphenicol than the parent strain and was more resistant to a wide range of antimicrobial compounds, including beta-lactams. These results support the notion that these proteins contribute to the intrinsic resistance of P. aeruginosa through the multidrug active efflux process.

Anti-Bacterial Agents↗
PubMed1994

Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa: resistance to tetracycline, chloramphenicol, and norfloxacin.

Most strains of Pseudomonas aeruginosa are significantly more resistant, even in the absence of R plasmids, to many antimicrobial agents, including beta-lactams, tetracycline, chloramphenicol, and fluoroquinolones, than most other gram-negative rods. This broad-range resistance has so far been assumed to be mainly due to the low permeability of the P. aeruginosa outer membrane. The intrinsic-resistance phenotype becomes further enhanced in "intrinsically carbenicillin-resistant" isolates, which were often assumed to produce outer membranes of even lower permeability. It has been shown, however, that this hypothesis cannot explain the beta-lactam resistance of these isolates (D.M. Livermore and K.W.M. Davy, Antimicrob. Agents Chemother. 35:916-921, 1991). In this study, we examined the uptake of tetracycline, chloramphenicol, and norfloxacin by intact cells using strains showing widely different levels of intrinsic resistance. Their accumulation and the response to the addition of a proton conductor showed that even relatively susceptible strains of P. aeruginosa actively pump out these compounds from the cell and that the efflux activity becomes much stronger in strains showing higher levels of intrinsic resistance. We conclude that the efflux mechanism(s) are likely to contribute significantly to the intrinsic resistance of P. aeruginosa isolates to tetracycline, chloramphenicol, and fluoroquinolones, as does the low permeability of the outer membrane. This conclusion is supported by the observation that the hypersusceptibility to various agents of the mutant K799/61 (W. Zimmermann, Antimicrob. Agents Chemother. 18:94-100, 1980) was apparently caused by the lack of active efflux. Although the hypersusceptibility of this mutant has hitherto been assumed to be solely due to its higher outer membrane permeability, its outer membrane was shown to have a coefficient of permeability to cephaloridine that was not significantly different from that of the parent, resistant strain K799/WT. The strains with elevated intrinsic resistance overproduced two cytoplasmic membrane proteins and one outer membrane protein; at least two of these proteins appeared different from the proteins overproduced in the recently described mutant with a derepressed multidrug efflux system, MexA-MexB-OprK (K. Poole, K. Krebes, C. McNally, and S. Neshat, J. Bacteriol. 175:7363-7372, 1993).

Chloramphenicol↗
PubMed1994

Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa: active efflux as a contributing factor to beta-lactam resistance.

Wild-type strains of Pseudomonas aeruginosa are more resistant to various beta-lactam antibiotics as well as other agents than most enteric bacteria. Although resistance to compounds of earlier generations is explained by the synergism between the outer membrane barrier and the inducible beta-lactamase, it was puzzling to see significant levels of resistance to compounds that do not act as inducers or are not hydrolyzed rapidly by the chromosomally encoded enzyme. This intrinsic-resistance phenotype becomes enhanced in those strains with the so-called intrinsic carbenicillin resistance. In the accompanying paper (X.-Z. Li, D. M. Livermore, and H. Nikaido, Antimicrob. Agents Chemother. 38:1732-1741, 1994), we showed that active efflux played a role in the resistance, to various non-beta-lactam agents, of P. aeruginosa strains in general and that the efflux was enhanced in intrinsically carbenicillin-resistant strains. We show in this paper that, in comparison with the drug-hypersusceptible mutant K799/61, less benzylpenicillin was accumulated in wild-type strains of P. aeruginosa and that the accumulation levels were even lower in intrinsically carbenicillin-resistant strains. Deenergization by the addition of a proton conductor increased the accumulation level to that expected for equilibration across the cytoplasmic membrane. In intrinsically carbenicillin-resistant isolates, there was no evidence that either nonspecific or specific permeation rates of beta-lactams across the outer membrane were lowered in comparison with those of the more susceptible isolates. Furthermore, these carbenicillin-resistant isolates were previously shown to have no alteration in the level or the inducibility of beta-lactamase and in the affinity of penicillin-binding proteins. These data together suggest the involvement of an active efflux mechanism also in the resistance to beta-lactams. Hydrophilic beta-lactams with more than one charged group did not cross the cytoplasmic membrane readily. Yet one such compound, ceftriaxone, appeared to be extruded from the cells of more-resistant strains, although with this compound effects of proton conductors could not be shown. We postulate that wild-type strains of P. aeruginosa pump out such hydrophilic beta-lactams either from the periplasm or from the outer leaflet of the lipid bilayer of the cytoplasmic membrane, in a manner analogous to that hypothesized for multidrug resistance protein of human cancer cells (M.M. Gottesman and I. Pastan, Annu. Rev. Biochem. 62:385-427, 1993).

Anti-Bacterial Agents↗
PubMed1994

Gene amplification and high-level expression of human pro-urokinase cDNA in Chinese hamster ovary cells.

Human Pro-Urokinase (Pro-UK) is expressed in CHO-DHFR- cells at high efficiency by co-transfecting the mouse dhfr gene and Pro-UK cDNA under the control of the SV40 late promoter. After gene co-amplification, the product level could reach 2-3 micrograms/10(6) cells/24 h in the presence of 5 x 10(-6) mol/L MTX, and the levels can be further raised to 3.5-4 micrograms/10(6) cells/24 h by PMA superinduction. The copy number of Pro-UK cDNA in the genomes of host cells is about 200-300 copies/cell. The Western blot analysis shows that the recombinant Pro-UK has similar molecular weight to its natural counterpart, and also the amidolytic activity of the product is determined by S-2444 assay.

Animals↗
PubMed1993

[A study of cerebral vasospasm after subarachnoid hemorrhage with transcranial Doppler].

Transcranial Doppler (TCD) was performed in 64 patients with subarachnoid hemorrhage (SAH) to detect the blood velocity in middle cerebral artery (MCA). Based on Poiseuille's principle, the increase of blood velocity was used as an indicator of cerebral vasospasm (CVS). It was found that 61.67% of the patients with SAH developed CVS, especially in the second week after SAH. The incidence of CVS in patients below 50 years old was higher than that in patients above 51 years old. The results showed that TCD is valuable in evaluation of CVS after SAH.

Adult↗
PubMed1991

Expression of pro-urokinase cDNA in Chinese hamster ovary cell line.

Expression vectors containing the pro-urokinase (pro-UK) cDNA (pSV2-proUK) and a dihydrofolate reductase cDNA (pSV2-dhfr or MMTV-dhfr) were cotransfected into CHO-dhfr- cells by the calcium phosphate precipitation technique. The dhfr+ transformants were selected by fibrinolytic agarose plate assay. Two colonies, named CLF-14 and CLF-8, exhibited significantly high expression levels of the biological activity of urokinase-type plasminogen activator (mu-Pa). They reached more than 24 IU/10(6) cells/48 h and 16 IU/10(6) cells/48 h, respectively. Examination of the cell supernatants for mu-Pa antigenicity using ELISA method also showed strong positive results, and the quantities of expression were about 0.14-0.22 micrograms/10(6) cells/48 h and 0.08-0.14 micrograms/10(6) cells/48 h, respectively. The mu-Pa secreted by stable transformed cells could be completely inhibited by UK anti-serum, but not by tissue-type plasminogen activator (t-PA) antiserum nor by normal rabbit serum.

Animals↗
PubMed1990

The differences of serum cholesterol and lipoproteins in animals susceptible and nonsusceptible to atherosclerosis.

The differences of serum lipid and lipoprotein (LP, profiles of animals susceptible (rabbit) and nonsusceptible (Beijing duck) to atherosclerosis as well as the distribution of apolipoprotein (apo) A-I and its catabolism in vivo were studied. Eight items, i.e. total cholesterol (TC), high density lipoprotein (HDL) cholesterol, triglyceride, percentage of alpha-LP and beta-LP, beta-LP concentration, lecithin cholesterol acyl transferase (LCAT), and agarose and polyacrylamide gel electrophoresis of both species were assayed before and after feeding a high fat, high cholesterol diet. Results indicate that the exogenous cholesterol consumed by Beijing ducks was carried and transported by HDL, while that in rabbits was transported by low density lipoprotein (LDL). The biological half lives of apo A-I in serum and in HDL were 41.1 +/- 2.4 and 42.8 +/- 1.7 h respectively, and its distribution in different organs was in the order of liver greater than kidney greater than spleen greater than lung greater than heart greater than intestine greater than muscles greater than aorta. These results show that the liver is the major organ for metabolizing HDL apo A-I, and the kidney is also important. The results also imply that in Beijing ducks the cholesterol carried by HDL may be catabolized through apo A-I receptors in the liver and kidney. The differences in cholesterol binding and in lipoprotein and apolipoprotein metabolism of the two species provide important clues for the elucidation of the pathogenesis of atherosclerosis.

Animals↗
PubMed1990

[Ovarian sex cord tumor with annular tubules--a report of 7 cases].

Seven cases of ovarian sex cord tumor with annular tubules are reported in this paper. All lesions occurred in the young (6-25 years of age with a mean of 15 years) without Poutz-Jeghers syndrome. Four cases had estrogenic hormonal imbalance and abdominal mass. Gross examination revealed that the tumors were unilateral with smooth surface. Except one, all showed varying degrees of cystic degeneration on section. The sections appeared as roe-like granular substance in a pale yellowish brown shade in the parenchymal region. Microscopic examination showed single or complex annular tubules with prominent basement membrane-like material around the tubules. Lipid in the cytoplasm and granulosa cell tumor-like differentiation were observed in isolated area of some cases. Ultrastructurally, Charcot-Battcher filaments were found in 1 case, indicating differentiation towards Sertoli's cells. The malignant potential of this tumor had been elucidated in the literature.

Adult↗
PubMed1989

The nucleotide sequence of a human cDNA encoding the highly conserved nucleolar phosphoprotein B23.

A cDNA clone containing the complete coding sequence for the human nucleolar phosphoprotein B23 was isolated from a Burkitt's lymphoma cDNA library by immunoscreening with human autoantibodies. The B23 clone contained a 1.3 kb cDNA insert encoding a polypeptide of 294 amino acids with a predicted molecular mass of 32,539 daltons. The deduced B23 amino acid sequence contained 2 acidic domains rich in aspartic and glutamic acid, a feature shared by a number of nuclear and nucleolar proteins. The human B23 amino acid sequence showed 98% homology with rat B23 and 68% homology with the Xenopus laevis nucleolar phosphoprotein, NO38 showing that the primary structure of B23 is highly conserved among these species.

Amino Acid Sequence↗