Penicillin-resistant Bacteria Research Articles

Cbf-14, a cationic peptide derived from cathelin-domain, exhibits anti-inflammation activity via inhibiting PI3K- Akt /ROS/ NF-κB signaling pathway

Cbf-14 with the sequence RLLRKFFRKLKKSV, is an effective antimicrobial peptide derived from a cathelin-like domain. Previous reports have demonstrated that Cbf-14 not only exerts antimicrobial activity against penicillin-resistant bacteria but also alleviates bacterial-induced inflammation in E. coli BL21 (DE3)-NDM-1-infected mice. In this article, we demonstrated that Cbf-14 can effectively reduce RAW 264.7 intracellular infection caused by clinical strain E. coli and alleviate the inflammatory response of cells and improve cell survival after infection. Therefore, we established the LPS-stimulated RAW 264.7 cell inflammation model to uncover the molecular mechanisms of the peptide Cbf-14 in anti-inflammatory activity. The results reveal that Cbf-14 can decrease LPS-induced ROS secretion by blocking the membrane translocation of p47-phox subunits and suppressing p47-phox protein phosphorylation. Meanwhile, this peptide can down-regulate the over-expression of iNOS, and finally inhibit the NO excessive secretion from RAW 264.7 macrophages stimulated by LPS. Moreover, Cbf-14 also down-regulates the expression levels of p-IκB and p-p65 and inhibits the nuclear translocation of NF-κB through blocking MAPK- and/or PI3K-Akt signaling pathways. Overall, Cbf-14 exhibits anti-inflammatory activity through inhibiting NF-κB activity and ROS production via PI3K- Akt signaling pathway.

Parabens Increase Sulfamethoxazole-, Tetracycline- and Paraben-Resistant Bacteria and Reshape the Nitrogen/Sulfur Cycle-Associated Microbial Communities in Freshwater River Sediments.

Backgrounds Parabens are pollutants of emerging concern in aquatic environments. Extensive studies regarding the occurrences, fates and behavior of parabens in aquatic environments have been reported. However, little is known about the effects of parabens on microbial communities in freshwater river sediments. This study reveals the effects of methylparaben (MP), ethylparaben (EP), propylparaben (PP) and butylparaben (BP) on antimicrobial-resistant microbiomes, nitrogen/sulfur cycle-associated microbial communities and xenobiotic degrading microbial communities in freshwater river sediments. Methods The river water and sediments collected from the Wai-shuangh-si Stream in Taipei City, Taiwan were used to construct a model system in fish tanks to test the effects of parabens in laboratory. Results Tetracycline-, sulfamethoxazole- and paraben-resistant bacteria increased in all paraben treated river sediments. The order of the overall ability to produce an increment in sulfamethoxazole-, tetracycline- and paraben-resistant bacteria was MP > EP > PP > BP. The proportions of microbial communities associated with xenobiotic degradation also increased in all paraben-treated sediments. In contrast, penicillin-resistant bacteria in both the aerobic and anaerobic culture of paraben-treated sediments decreased drastically at the early stage of the experiments. The proportions of four microbial communities associated with the nitrogen cycle (anammox, nitrogen fixation, denitrification and dissimilatory nitrate reduction) and sulfur cycle (thiosulfate oxidation) largely increased after the 11th week in all paraben-treated sediments. Moreover, methanogens and methanotrophic bacteria increased in all paraben-treated sediments. In contrast, the nitrification, assimilatory sulfate reduction and sulfate-sulfur assimilation associated to microbial communities in the sediments were decreased by the parabens. The results of this study uncover the potential effects and consequences of parabens on microbial communities in a freshwater river environment.

Bacterial Profile of External Ocular Infections, Its Associated Factors, and Antimicrobial Susceptibility Pattern among Patients Attending Karamara Hospital, Jigjiga, Eastern Ethiopia.

External ocular infection is a global public health problem. Frequently, bacteria cause an ocular infection that ranges from morbidity to loss of vision. The increasing bacterial resistance in ocular infections leads to the risk of treatment failure with possibly serious consequences. The study aimed to assess the bacterial profile of external ocular infections, their associated factors, and antimicrobial susceptibility pattern among patients admitted to Karamara hospital, Jigjiga, Eastern Ethiopia. Institutional-basedcross-sectional study was conducted on 288 conveniently selected patients among patients admitted to Karamara hospital from May 1 to June 30, 2020. Data were collected using a structured questionnaire. The ocular sample was collected and cultured in the appropriate culture media and identified using a series of biochemical tests. Antimicrobial susceptibility testing of isolates was performed by using the disk diffusion method. Data were double entered onto EpiData version 3.1 then exported to SPSS version 20 and analyzed to calculate descriptive frequency and odds ratio, and p value ≤0.05 was taken as the significant value. The prevalence of bacterial infection in external ocular samples was 62.2% (95% CI: 56.6%, 68.4%). Out of the 179 isolates, the majority of the bacterial isolates (87.7%) were Gram-positive. Staphylococcus aureus (53.1%) was the predominant isolate. Using soap for washing the face (AOR = 0.43; 95% CI: 0.29, 0.95), having diabetes mellitus (AOR = 3.11; 95% CI: 1.45, 6.75), and history of hospitalization (AOR = 2.82; 95% CI: 1.44, 5.54) were significantly associated with external ocular infection. Most (95.5%) of the Gram-positive bacteria showed resistance to penicillin, but they were susceptible to vancomycin, clindamycin, and ciprofloxacin. The study showed a high prevalence of bacterial infections with the predominant isolate was S. aureus. Penicillin-resistant bacteria were identified among Gram-positive bacterial isolates. Soap usage, hospitalization, and diabetes mellitus were associated with the infection. Antibiotics that were susceptible to the specific bacteria should be used as a drug of choice and using soap for washing the face is advisable to protect against external ocular infection.

Hugh John Forster Cairns. 21 November 1922—12 November 2018

John Cairns was a magnet for scientists (and non-scientists) of all types, from undergraduates to Nobel laureates. For him science was fun, and discussing science with others even more fun. Soon after finishing medical school (Oxford) at the end of the Second World War he gravitated towards research, his first publication linking the incidence of penicillin-resistant bacteria with long hospital stays. He then studied virology with Macfarlane Burnet FRS in Melbourne, Australia, after which he repaid his two years of National Service from the Second World War at the Virus Research Institute in Entebbe, Uganda. He returned to Australia (Canberra), continuing influenza virus research. However, while visiting Caltech in the 1950s, he was exposed to the emerging field of molecular biology, of which he became a pioneer. After returning to Canberra from a sabbatical with Alfred Hershey at Cold Spring Harbor in 1961, he showed that the Escherichia coli genome is circular, and demonstrated the replication fork. In 1963 he was appointed director of the Cold Spring Harbor Laboratory (CSHL). He successfully saved CSHL from bankruptcy before handing over directorship to Jim Watson (ForMemRS 1981). At CSHL he proved that the so-called Kornberg DNA polymerase was not required for replication of the E. coli genome. In the 1970s Cairns turned his attention to understanding cancer and moved to North London to direct a small research institute in Mill Hill, a satellite of the Imperial Cancer Research Fund based in central London. Cairns wrote two influential reviews on cancer plus the book Cancer: science and society , seamlessly weaving together information across disciplines, from human population levels to molecular levels, plus everything in between. He fostered multi-disciplinary basic research at the Mill Hill labs, recruiting scientists who used model organisms such as bacteria, fruit flies, slime moulds, amphibians and mice as models to explore the determinants of cell fate during development; such pathways seemed likely to be involved in the transformation of normal cells into cancer cells. Cairns created and fostered an exceptionally stimulating, interactive, nurturing and cutting-edge research environment that launched the careers of many scientists. In his own lab at Mill Hill, Cairns discovered new pathways for DNA alkylation repair in E. coli , and subsequently as professor of cancer biology at the Harvard School of Public Health he discovered what is now known as stress-induced mutagenesis, also in E. coli .

Bacterial Resistance to β-Lactam Antibiotics in Municipal Wastewater: Insights from a Full-Scale Treatment Plant in Poland.

This study investigated enzymatic and genetic determinants of bacterial resistance to β-lactam antibiotics in the biocenosis involved in the process of biological treatment of wastewater by activated sludge. The frequency of bacteria resistant to selected antibiotics and the activity of enzymes responsible for resistance to β-lactam antibiotics were estimated. The phenomenon of selection and spread of a number of genes determining antibiotic resistance was traced using PCR and gene sequencing. An increase in the percentage of bacteria showing resistance to β-lactam antibiotics in the microflora of wastewater during the treatment process was found. The highest number of resistant microorganisms, including multi-resistant strains, was recorded in the aeration chamber. Significant amounts of these bacteria were also present in treated wastewater, where the percentage of penicillin-resistant bacteria exceeded 50%, while those resistant to the new generation β-lactam antibiotics meropenem and imipenem were found at 8.8% and 6.4%, respectively. Antibiotic resistance was repeatedly accompanied by the activity of enzymes such as carbapenemases, metallo-β-lactamases, cephalosporinases and β-lactamases with an extended substrate spectrum. The activity of carbapenemases was shown in up to 97% of the multi-resistant bacteria. Studies using molecular biology techniques showed a high frequency of genes determining resistance to β-lactam antibiotics, especially the blaTEM1 gene. The analysis of the nucleotide sequences of blaTEM1 gene variants present in bacteria at different stages of wastewater treatment showed 50-100% mutual similarity of.

Molecular detection of antibiotic resistance genes in multidrug-resistant Staphylococcus aureus isolates from dog dental plaque

Multidrug resistant Staphylococcus aureus strains are a major health care problem both in humans and animals. In this work we described three multiplex PCR assays for detection of clinically relevant antibiotic resistance genes in S. aureus isolated from dog dental plaques. Thirty dental plaque samples were collected; then cultural, biochemical and molecular tests performed for isolation and identification of S. aureus from samples. The antibiotic susceptibility of the isolates were checked by Kirby Bauer disc diffusion method and the prevalence of antibiotic resistance genes determined using multiplex PCR assay. As a result S. aureus was isolated from 18 dog plaque samples. Fifteen of these isolates were resistant to penicillin. The mecA gene was more prevalent than blaZ among penicillin-resistant bacteria. Ten of the isolates were resistant to tetracycline. The percentage of tetM was higher than tetK among them. Also, 10 of the isolates were resistant to cefazolin among them bla TEM detected in higher rate than blaSHV and blaOXA-1. Hence multiplex PCR assay is a suitable method for detection of antibiotic resistance patterns of S. aureus isolates.

Semi-synthesis, antibacterial activity, and molecular docking study of novel pleuromutilin derivatives bearing cinnamic acids moieties.

To develop new antibiotics owning a special mechanism, we used the molecular assembly method to synthesize a series of novel pleuromutilin derivatives containing a cinnamic acid scaffold at the C-14 side chain. We evaluated their antibacterial activity and used in silico molecular docking to study their binding mode with the target. The structure-activity relationship (SAR) study suggested that compounds with NO2 (13e), OH (13u), and NH2 (13y) appeared more active (0.0625-2 µg/mL) in vitro against several penicillin-resistant Gram-positive bacteria and the position of the substituent on the benzene ring would affect the activity. The in vivo efficacy investigation of 13e, 13u, and 13y with once daily intragastric (i.g.) administration at 40 mg/kg for 3 consecutive days in a mouse systemic infection model showed that 13u had equal activity as valnemulin providing the mice with 60% survival, while 13e and 13y gave 30 and 40% survival, respectively. The molecular docking studies indicated that π-π stacking and hydrogen bond formation played important roles in improving the antibacterial activity.

Insights into the antibacterial mechanism of PEGylated nano-bacitracin A against Streptococcus pneumonia: both penicillin-sensitive and penicillin-resistant strains.

BackgroundMultidrug-resistant (MDR) Streptococcus pneumonia constitute a major worldwide public health concern.Materials and methodsIn our preliminary study, PEGylated nano-self-assemblies of bacitracin A (PEGylated Nano-BA12K) showed strong antibacterial potency against reference S. pneumonia strain (ATCC 49619). In this study, the possibility of applying PEGylated Nano-BA12K against penicillin-resistant S. pneumonia was further investigated. In addition, the underlying antibacterial mechanism of PEGylated Nano-BA12K against both sensitive and resistant S. pneumonia was also clarified systematically, since S. pneumonia was naturally resistant to its unassembled counterpart bacitracin A (BA).ResultsPEGylated Nano-BA12K showed strong antibacterial potency against 13 clinical isolates of S. pneumonia, including five penicillin-resistant strains. Structural changes, partial collapse, and even lysis of both penicillin-sensitive and penicillin-resistant bacteria were observed after incubation with PEGylated Nano-BA12K via transmission electron microscopy and atomic force microscopy. Thus, the cell wall or/and cell membrane might be the main target of PEGylated Nano-BA12K against S. pneumonia. PEGylated Nano-BA12K exhibited limited effect on the permeabilization and peptidoglycan content of cell wall. Surface pressure measurement suggested that PEGylated Nano-BA12K was much more tensioactive than BA, which was usually translated into a good membranolytic effect, and is helpful to permeabilize the cell membrane and damage membrane integrity, as evidenced by depolarization of the membrane potential, permeabilization of membrane and leakage of calcein from liposomes.ConclusionCollectively, great cell membrane permeability and formidable membrane disruption may work together for the strong antibacterial activity of PEGylated Nano-BA12K against S. pneumonia. Taken together, PEGylated Nano-BA12K has excellent potential against both penicillin-sensitive and penicillin-resistant S. pneumonia and might be suitable for the treatment of S. pneumonia infectious diseases.

Penicillium Antibiotic Effect

In this lesson students will use the Penicillium chrysogenum fungus, which naturally produces the antibiotic penicillin, to investigate the effect of naturally produced antibiotics on bacteria in laboratory cultures. Students co-culture P. chrysogenum with three species of bacteria to observe differences between penicillin-resistant and penicillin-sensitive bacteria. They will normalize fungal spore suspension and bacterial culture concentrations before inoculating co-cultures. After bacteria have been exposed to the antibiotic, students will quantify culture density to determine antibiotic effect in liquid culture and on solid media. Students will learn about natural product antibiotics as well as experimental design and application.

Effective antimicrobial activity of a peptide mutant Cbf-14-2 against penicillin-resistant bacteria based on its unnatural amino acids

Broad spectrum activities, a unique mode of actions and rare resistant variants make antimicrobial peptide (AMP) a potential alternative to antibiotics. However, AMPs still have limitations in clinical development due to their physiological stability, toxicity and manufacturing costs. Cbf-14, derived from cathelicidin-BF has been proven to be effective against drug-resistant bacteria. Herein, a series of Cbf-14 mutants were designed to overcome these limitations. Design strategies included substitutions of lysine (Lys) or leucine (Leu) with similar residues such as ornithine (Orn) and norleucine (Ile), which are unnatural amino acids, to generate AMPs with enhanced therapeutic potential. Antimicrobial activity, hemolytic activity and cytotoxicity against mouse spleen cells of the peptide mutants were investigated. Among all of the mutants, Cbf-14-2 was regarded as the most potent candidate with the amino acid sequence of RLLR-Orn-FFR-Orn-LKKSV-NH2, which exhibited a superior antimicrobial activity with a minimum inhibitory concentration (MIC) of 4–32μg/ml. Meanwhile, Cbf-14-2 displayed low levels of hemolysis in sheep red blood cells (sRBCs) and negligible cytotoxicity against mouse spleen cells, suggesting low toxicity against mammalian cells. A circular dichroism (CD) study indicated that Cbf-14-2 has a higher alpha-helix content than Cbf-14 (68.3% vs 35.1%) in SDS, which may contribute to its superior activity. Time-killing curves showed Cbf-14-2 can eliminate all tested bacteria within 240min, suggesting its rapid bactericidal effect. Transmission electron microscopy (TEM), flow cytometry and calcein release assays revealed its excellent antimicrobial potency by inducing membrane permeation and disruption. In addition, Cbf-14-2 (10mg/kg) could significantly elevate the survival rate of clinical strain infected mice, with a survival rate of 70.0%. Taken together, the data suggest that Cbf-14-2 possesses effective antimicrobial activity against penicillin-resistant bacteria in vitro and in vitro, thus rendering it as a potential anti-infective agent in clinical settings.

Effective antimicrobial activity of Cbf-14, derived from a cathelin-like domain, against penicillin-resistant bacteria

Cbf-14, a cationic peptide derived from a cathelin-like domain, was designed by inserting the highly α-helical sequence RLLR into an antibacterial sequence and deleting the inactive amino acids in Cbf-K16. Clinical penicillin-resistant isolates as well as NDM-1-carrying Escherichia coli and a correspondingly infected mice model were employed to evaluate Cbf-14 antibacterial activity. The results showed that Cbf-14 possessed potent antimicrobial effects with an MIC of 8–64 μg/ml, and killed almost all bacteria within 240 min. Cbf-14-treated mice achieved an 80% survival rate and approximate 2.5 log unit reduction in CFU in tissues; additionally, this peptide significantly suppressed the production of pro-inflammatory cytokines by the disaggregation of lipopolysaccharide (LPS), suggesting its anti-inflammatory effects. Furthermore, Cbf-14, concentration higher than 2 × MIC value, increased membrane uptake to NPN and PI dye by 96.2% and 63.7%, respectively, neutralised the negative zeta potential of LPS and bacteria surface, and induced 100% leakage of liposome-entrapped calcein and cytoplasmic membrane disruption of E. coli, indicating obvious membrane permeation. Finally, it bound to DNA and respectively evoked 85.0% and 63.3% inhibition of gene replication and protein expression of NDM-1 at sub-MIC concentration in E. coli BL21 (DE3)-NDM-1. These data indicated that Cbf-14 possessed effective antimicrobial activity against penicillin-resistant bacteria in vitro/vivo through membrane disruption, DNA binding, down-regulating NDM-1 expression by plasmid replication inhibition, and anti-inflammatory activity by LPS disaggregation, suggesting a potential anti-infective clinical agent.

Antibiotic regimens for postpartum endometritis.

Postpartum endometritis occurs when vaginal organisms invade the endometrial cavity during the labor process and cause infection. This is more common following cesarean birth. The condition warrants antibiotic treatment. Systematically, to review treatment failure and other complications of different antibiotic regimens for postpartum endometritis. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (30 November 2014) and reference lists of retrieved studies. We included randomized trials of different antibiotic regimens after cesarean birth or vaginal birth; no quasi-randomized trials were included. Two review authors independently assessed trials for inclusion and risk of bias, extracted data and checked them for accuracy. The review includes a total of 42 trials, and 40 of these trials contributed data on 4240 participants.Regarding the primary outcomes, seven studies compared clindamycin plus an aminoglycoside versus penicillins and showed fewer treatment failures (risk ratio (RR) 0.65, 95% confidence interval (CI) 0.46 to 0.90). There were more treatment failures in those treated with an aminoglycoside plus penicillin when compared to those treated with gentamycin/clindamycin (RR 2.57, 95% CI 1.48 to 4.46). There were more treatment failures (RR 1.66, 95% CI 1.01 to 2.74) and wound infections (RR 1.88, 95% CI 1.08 to 3.28) in those treated with second or third generation cephalosporins (excluding cephamycins) versus those treated with clindamycin plus gentamycin. In four studies comparing once-daily with thrice-daily dosing of gentamicin, there were fewer failures with once-daily dosing. There were more treatment failures (RR 1.94, 95% CI 1.38 to 2.72) and wound infections (RR 1.88, 95% CI 1.17 to 3.02) in those treated with a regimen with poor activity against penicillin-resistant anaerobic bacteria as compared to those treated with a regimen with good activity against penicillin-resistant anaerobic bacteria. There were no differences between groups with respect to severe complications and no trials reported any maternal deaths.Regarding the secondary outcomes, three studies that compared continued oral antibiotic therapy after intravenous therapy with no oral therapy, found no differences in recurrent endometritis or other outcomes. Four trials that compared clindamycin plus aminoglycoside versus cephalosporins identified fewer wound infections in those treated with clindamycin plus an aminoglycoside (RR 0.53, 95% CI 0.30 to 0.93). There were no differences between groups for the outcomes of allergic reactions. The overall risk of bias was unclear in the most of the studies. The quality of the evidence using GRADE comparing clindamycin and an aminoglycoside with another regimen (compared with cephalosporins or penicillins) was low to very low for therapeutic failure, severe complications, wound infection and allergic reaction. The combination of clindamycin and gentamicin is appropriate for the treatment of endometritis. Regimens with good activity against penicillin-resistant anaerobic bacteria are better than those with poor activity against penicillin-resistant anaerobic bacteria. There is no evidence that any one regimen is associated with fewer side-effects. Following clinical improvement of uncomplicated endometritis which has been treated with intravenous therapy, the use of additional oral therapy has not been proven to be beneficial.

Improvement of 7α-methoxycephalosporins production by overexpression of cmcJ and cmcI controlled by promoter ermEp* in Streptomyces clavuligerus.

This study aimed to improve the efficiency of bioconversion of cephalosporin C to 7α-methoxycephalosporin C. We performed the genetic modulation of the expression level of tailoring genes cmcI and cmcJ, which are responsible for the methoxylation of cephalosporin C, to improve the efficiency of bioconversion in the recombinant strains. The results showed that the coordinate adjustment of the enzyme amounts of CmcI and CmcJ in the host strain by placing the ermEp* promoter in the upstream of both cmcI and cmcJ genes in their expression cassette can improve the efficiency of bioconversion from cephalosporin C to 7α-methoxycephalosporin C. Under the optimized cultivation condition, 2·0gl(-1) of the substrate cephalosporin C can be transformed into 0·78gl(-1) of 7α-methoxycephalosporin C by the extract of the recombinant strain Streptomyces clavuligerus DYG3003, in which the transcript ratio of cmcJ/cmcI has been engineered to 1·7:1. Coordinate adjustment of the enzyme amounts of CmcI and CmcJ in the host strain can greatly enhance the efficiency of bioconversion from cephalosporin C (CPC) to 7a-methoxycephalosporin (MCPC). As one kind of the most important β-lactam antibiotics, 7α-methoxycephalosporins show stronger activity against penicillin-resistant bacteria than cephalosporins because of the higher stability of 7α-methoxycephalosporins to β-lactamase. Our study provides a practical strategy to perform the industrial production of the clinically useful 7α-methoxycephalosporin C through biotransformation.

Neutral β-Lactams Inactivate High Molecular Mass Penicillin-Binding Proteins of Class B1, Including PBP2a of MRSA.

The targets of β-lactam antibiotics are bacterial DD-peptidases (penicillin-binding proteins). β-Lactam SAR studies over many years have demonstrated the importance of a specifically placed negative charge, usually carboxylate, on these molecules. We show here that neutral analogues of classical β-lactam antibiotics are of comparable activity to the originals against β-lactam-resistant high molecular mass DD-peptidases of the B1 class, a group that includes PBP2a of methicillin-resistant Staphylococcus aureus. These neutral β-lactams may direct new development of antibiotics against certain penicillin-resistant bacteria. These molecules do have antibiotic activity against Gram-positive bacteria.

Management of secondary immune deficiencies

Management of secondary immune deficiencies

Resistance Index of Penicillin-Resistant Bacteria to Various Physicochemical Agents

Widespread use of various antimicrobial agents resulted in the emergence of bacterial resistance. Mechanisms like direct efflux, formation, and sequestration of metals and drugs in complexes and antiporter pumps are some examples. This investigation aims to investigate the resistance pattern of penicillin-resistant bacterial strains to some physicochemical agents. Sensitivity/resistance pattern of common bacterial strains to antimicrobial agents were evaluated by disk diffusion assay. Broth and agar dilution method were used for determination of minimum inhibitory concentration and minimal bactericidal concentration. The impact of UV ray on the bacterial growth under laminar flow hood was measured using photonmeter. Our data demonstrates that the most prevalent metal resistance was against arsenate (95.92%), followed by cadmium (52.04%) and mercury (36.73%). There was significant difference between cetrimide resistances among studied microbial strains especially for P. aeruginosa (P < 0.05). High rate of pathogen resistance to various antibacterial agents in our study supports previously published data. This great rate of bacterial resistance is attributed to the emergence of defense mechanisms developed in pathogens. The higher general bacterial resistance rate among Staphylococcus strains rather than E. coli and P. aeruginosa strains draws attention towards focusing on designing newer therapeutic compounds for Staphylococcus strains.

New Noncovalent Inhibitors of Penicillin-Binding Proteins from Penicillin-Resistant Bacteria

BackgroundPenicillin-binding proteins (PBPs) are well known and validated targets for antibacterial therapy. The most important clinically used inhibitors of PBPs β-lactams inhibit transpeptidase activity of PBPs by forming a covalent penicilloyl-enzyme complex that blocks the normal transpeptidation reaction; this finally results in bacterial death. In some resistant bacteria the resistance is acquired by active-site distortion of PBPs, which lowers their acylation efficiency for β-lactams. To address this problem we focused our attention to discovery of novel noncovalent inhibitors of PBPs.Methodology/Principal FindingsOur in-house bank of compounds was screened for inhibition of three PBPs from resistant bacteria: PBP2a from Methicillin-resistant Staphylococcus aureus (MRSA), PBP2x from Streptococcus pneumoniae strain 5204, and PBP5fm from Enterococcus faecium strain D63r. Initial hit inhibitor obtained by screening was then used as a starting point for computational similarity searching for structurally related compounds and several new noncovalent inhibitors were discovered. Two compounds had promising inhibitory activities of both PBP2a and PBP2x 5204, and good in-vitro antibacterial activities against a panel of Gram-positive bacterial strains.ConclusionsWe found new noncovalent inhibitors of PBPs which represent important starting points for development of more potent inhibitors of PBPs that can target penicillin-resistant bacteria.

Inactivation and reactivation of antibiotic-resistant bacteria by chlorination in secondary effluents of a municipal wastewater treatment plant

Reports state that chlorination of drinking water and wastewater affects the proportions of antibiotic-resistant bacteria by potentially assisting in microbial selection. Studies on the effect of chlorination on like species of antibiotic-resistant bacteria, however, have shown to be conflicting; furthermore, few studies have inspected the regrowth or reactivation of antibiotic-resistant bacteria after chlorination in wastewater. To understand the risks of chlorination resulting from potentially selecting for antibiotic-resistant bacteria, inactivation and reactivation rates of both total heterotrophic bacteria and antibiotic-resistant bacteria (including penicillin-, ampicillin-, tetracycline-, chloramphenicol-, and rifampicin-resistant bacteria) were examined after chlorinating secondary effluent samples from a municipal wastewater treatment plant in this study. Our experimental results indicated similar inactivation rates of both total heterotrophic bacteria and antibiotic-resistant bacteria. Microbial community composition, however, was affected by chlorination: treating samples with 10 mg Cl 2/L for 10 min resulted in chloramphenicol-resistant bacteria accounting for nearly 100% of the microbial population in contrast to 78% before chlorination. This trend shows that chlorination contributes to selection of some antibiotic-resistant strains. Reactivation of antibiotic-resistant bacteria occurred at 2.0 mg Cl 2/L for 10 min; specifically, chloramphenicol-, ampicillin-, and penicillin-resistant bacteria were the three prevalent groups present, and the reactivation of chloramphenicol-resistant bacteria exceeded 50%. Regrowth and reactivation of antibiotic-resistant bacteria in secondary effluents after chlorination with a long retention time could threaten public health security during wastewater reuse.

Nonlinear acoustics in ultrasound metrology and other selected applications

A succinct background explaining why, initially, both the scientific community and industry were skeptical about the existence of the nonlinear (NL) wave propagation in tissue will be given and the design of an adequately wideband piezoelectric polymer hydrophone probe that was eventually used to verify that the 1–5 MHz probing wave then used in diagnostic ultrasound imaging was undergoing nonlinear distortion and generated harmonics in tissue will be discussed. The far-reaching implications of the advent of the piezoelectric PVDF polymer material will be reviewed and the advances in ultrasound metrology prompted by the regulatory agencies such as US Food and Drug Administration (FDA) and International Electrotechnical Commission (IEC) will be presented. These advances include the development of absolute calibration techniques for hydrophones along with the methods of accounting for spatial averaging corrections up to 100 MHz and the development of “point-receiver” hydrophone probes utilizing acousto-optic sensors. Next, selected therapeutic applications of nonlinear ultrasonics (NLU), including lithotripters will be briefly discussed. Also, the use of shock waves as pain relief tool and in abating penicillin resistant bacteria that develop rock hard “biofilm” that can be shattered by the finite amplitude wave will also be mentioned. The growing applications of NLU in cosmetic industry where it is used for redistribution and reduction of fatty tissue within the body will be briefly reviewed, and, finally, selected examples of NLU applications in retail and entertainment industry will also be pointed out.

Nationwide survey of the development of drug-resistant pathogens in the pediatric field: drug sensitivity of Streptococcus pneumoniae in Japan

We evaluated the resistance to 20 different antibacterial agents of 362 clinically isolated strains of Streptococcus pneumoniae accumulated from October 2000 to July 2001 (phase 1) and of 332 different strains accumulated from January to June 2004 (phase 2), from institutions throughout Japan that participated in the surveys carried out by the Drug-Resistant Pathogen Surveillance Group in Pediatric Infectious Disease. In phase 1, the proportions of penicillin-sensitive S. pneumoniae (PSSP), penicillin-insensitive S. pneumoniae (PISP), and penicillin-resistant S. pneumoniae (PRSP) were 35.4%, 34.8%, and 29.8%, respectively, and the proportions were almost the same in phase 2: 33.1%, 37.0%, and 29.8%, respectively. Comparison of the MIC(90) values of the antibacterial agents for PRSP in phase 1 and phase 2 revealed that these values for cefditoren, cefpodoxime, cefdinir, faropenem, ceftriaxone, cefotaxime, meropenem, and vancomycin increased by twofold to fourfold during the 3 years between phase 1 and phase 2. However the MIC(90) of rokitamycin increased more than fourfold. The proportion of S. pneumoniae that were PISP + PRSP remained almost constant over the 3 years between phase 1 and phase 2. The background factors of patient age, previous administration of antibacterial agents, and attendance at a day nursery were examined; we found that in phase 1, the proportion of PISP + PRSP was significantly higher than that of PSSP in patients under 4 years old who had previously received antibacterial agents, but no significant differences were found in any of these background factors in the phase 2 survey. No significant difference was found in the proportions of penicillin-resistant bacteria according to whether or not the child had attended a day nursery.