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Latest issue consulted: 2007/2008.

Spray dried combinations of lactoferrin with antibiotics appear superior to monotherapy for reducing biofilm formation by pseudomonas aeruginosa

SD Apo Lactoferrin-Tobramycin/Gentamicin Combinations are superior to monotherapy in the eradication of Pseudomonas aeruginosa Biofilm in the lungs Wilson Oguejiofor1, Lindsay J. Marshall1, Andrew J. Ingham1, Robert Price2, Jag. Shur2 1School of Life and Health Sciences, Aston University, Birmingham, UK. 2School of Pharmacy and Pharmacology, University of Bath, Bath, UK. KEYWORDS: lactoferrin, apo lactoferrin, spray drying, biofilm, cystic fibrosis Introduction Chronic lung infections from the opportunistic pathogeen Pseudomonas aeruginosa has been recognised as a major contributor to the incidences of high morbidity and mortality amongst cystic fibrosis (CF) patients (1,2). Currently, strategies for managing lung infections in CF patients involves the aggressive use of aerosolised antibiotics (3), however, increasing evidence suggests that the biofilm component of P. aeruginosa in the lower airway remains unperturbed and is associated with the development of antibiotic resistance. If this is so then, there is an urgent need to suitably adjust the current treatment strategy so that it includes compounds that prevent biofilm formation or disrupt established biofilms. It is well understood that biofilm formation is strongly dependent on iron (Fe3+) availability (4), therefore aerosolised anti-infective formulations which has the ability to chelate iron may essentially be a well suited therapy for eliminating P. aeruginosa biofilms on CF airway epithelial cells (5). In this study, we report the use of combination therapy; an aminoglycosides (tobramycin and gentamicin) and an antimicrobial peptide (lactoferrin) to significantly deplete P. aeruginosa biofilms. We demonstrate that lactoferrin-tobramycin and lactoferrin-gentamicin combinations are superior to the single antibiotic regime currently being employed to combat P. aeruginosa biofilms. MATERIALS AND METHOD Antibiotics: The antibiotics used in this study included gentamicin and tobramycin supplied by Fagron, UK. Bacterial strain and growth conditions: Pseudomonas aeruginosa strain PAO1 was provided by Prof. Peter Lambert of Aston University, Birmingham UK. The Strains were routinely grown from storage in a medium supplemented with magnesium chloride, glucose and casamino acids. Dialysis of lactoferrin: Apo lactoferrin was prepared by dialyzing a suspension of lactoferrin for 24 hrs at 4 °C against 20 mmol/L sodium dihydrogen phosphate, 20 mmol/L sodium acetate and 40 mmol/L EDTA (pH 3.5). Ferric ion (Fe3+) removal was verified by atomic absorption spectroscopy measurements. Spray drying of combinations of lactoferrin and apo lactoferrin with the different aminoglycosides: Combinations of tobramycin and gentamicin with the different preparations of lactoferrin were spray dried (SD) as a 2% (w/v) aqueous suspension. The spray drying parameters utilized for the production of suitable micron-sized particles includes: Inlet temperature, 180°C, spray flow rate, 606 L/hr; pump setting, 10%; aspirator setting, 85% (34m3/hr) to produce various outlet temperatures ranging from 99 - 106°C. Viability assay: To test the bactericidal activity of the various combinations, a viability assay was performed as previously described by Xu, Xiong et al. (6) with some modifications. Briefly, 10µL of ~ c. 6.6 x 107 CFU mL-1 P. aeruginosa strain PAO1 suspension were incubated (37°C, 60 mins) with 90 µL of a 2 µg/mL concentration of the various combinations and sampled every 10 mins. After incubation, the cells were diluted in deionised water and plated in Mueller hinton agar plates. Following 24 h incubation of the plates at 37°C, the percentage of viable cells was determined relative to incubation without added antibiotics. Biofilm assay: To test the susceptibility of the P. aeruginosa strain to various antibiotics in the biofilms mode of growth, overnight cultures of P. aeruginosa were diluted 1:100 into fresh medium supplemented with magnesium chloride, glucose and casamino acids. Aliquots of the dilution were dispensed into a 96 well dish and incubated (37°C, 24 h). Excess broth was removed and the number of colony forming units per milliliter (CFU/mL) of the planktonic bacteria was quantified. The biofilms were then washed and stained with 0.1% (w/v) crystal violet for 15 mins at room temperature. Following vigorous washing with water, the stained biofilms were solubilized in 30% acetic acid and the absorbance at 550nm of a 125 µL aliquot was determined in a microplate reader (Multiskan spectrum, Thermo Scientific) using 30% acetic acid in water as the blank. Aliquots of the broth prior to staining were used as an indicator of the level of planktonic growth. RESULTS AND DISCUSSION Following spray drying, the mean yield, volume weighted mean diameter and moisture content of lactoferrin powder were measured and were as follows (Table 1 and table 2); Table 1: Spray drying parameters FormulationInlet temp (°C)Outlet temp (°C)Airflow rate (L/hr)Mean yield (%)Moisture content (%) SD Lactoferrin18099 - 10060645.2 ±2.75.9 ±0.4 SD Apo Lactoferrin180100 - 10260657.8 ±1.85.7 ±0.2 Tobramycin180102 - 10460682.1 ±2.23.2 ±0.4 Lactoferrin + Tobramycin180104 - 10660687.5 ±1.43.7 ±0.2 Apo Lactoferrin + Tobramycin180103 - 10460676.3 ±2.43.3 ±0.5 Gentamicin18099 - 10260685.4 ±1.34.0 ±0.2 Lactoferrin + Gentamicin180102 - 10460687.3 ±2.13.9 ±0.3 Apo Lactoferrin + Gentamicin18099 -10360680.1±1.93.4 ±0.4 Table 2: Particle size distribution d10 d50d90 SD Lactoferrin1.384.9111.08 SD Apo Lactoferrin1.284.7911.04 SD Tobramycin1.254.9011.29 SD Lactoferrin + Tobramycin1.175.2715.23 SD Apo Lactoferrin + Tobramycin1.115.0614.31 SD Gentamicin1.406.0614.38 SD Lactoferrin + Gentamicin1.476.2314.41 SD Apo Lactoferrin + Gentamicin1.465.1511.53 The bactericidal activity of the various combinations were tested against P. aeruginosa PAO1 following a 60 minute incubation period (Figure 1 and Figure 2). While 2 µg/mL of a 1:1 combination of spray dried apo lactoferrin and Gentamicin was able to completely kill all bacterial cells within 40 mins, the same concentration was not as effective for the other antibiotic combinations. However, there was an overall reduction of bacterial cells by over 3 log units by the other combinations within 60 mins. Figure 1: Logarithmic plot of bacterial cell viability of various combinations of tobramycin and lactoferrin preparations at 2µg/mL (n = 3). Figure 2: Logarithmic plot of bacterial cell viability of various combinations of gentamicin and lactoferrin preparations at 2µg/mL (n = 3). Crystal violet staining showed that biofilm formation by P. aeruginosa PAO1 was significantly (ANOVA, p < 0.05) inhibited in the presence of the different lactoferrin preparations. Interestingly, apo lactoferrin and spray dried lactoferrin exhibited greater inhibition of both biofilm formation and biofilm persistence (Figure 2). Figure 2: Crystal violet staining of residual biofilms of P. aeruginosa following a 24hr incubation with the various combinations of antibiotics and an exposure to 48 hr formed biofilms. CONCLUSION In conclusion, combination therapy comprising of an antimicrobial peptide (lactoferrin) and an aminoglycosides (tobramycin or gentamicin) provides a feasible and alternative approach to monotherapy since the various combinations are more efficient than the respective monotherapy in the eradication of both planktonic and biofilms of P. aeruginosa. ACKNOWLEDGEMENT The authors would like to thank Mr. John Swarbrick and Friesland Campina for their generous donation of the Lactoferrin. REFERENCES 1.Hassett, D.J., Sutton, M.D., Schurr, M.J., Herr, A.B., Caldwell, C.C. and Matu, J.O. (2009), "Pseudomonas aeruginosa hypoxic or anaerobic biofilm infections within cystic fibrosis airways". Trends in Microbiology, 17, 130-138. 2.Trust, C.F. (2009), "Antibiotic treatment for cystic fibrosis". Report of the UK Cystic Fibrosis Trust Antibiotic Working Group. Consensus document. London: Cystic Fibrosis Trust. 3.Garcia-Contreras, L. and Hickey, A.J. (2002), "Pharmaceutical and biotechnological aerosols for cystic fibrosis therapy". Advanced Drug Delivery Reviews, 54, 1491-1504. 4.O'May, C.Y., Sanderson, K., Roddam, L.F., Kirov, S.M. and Reid, D.W. (2009), "Iron-binding compounds impair Pseudomonas aeruginosa biofilm formation, especially under anaerobic conditions". J Med Microbiol, 58, 765-773. 5.Reid, D.W., Carroll, V., O'May, C., Champion, A. and Kirov, S.M. (2007), "Increased airway iron as a potential factor in the persistence of Pseudomonas aeruginosa infection in cystic fibrosis". European Respiratory Journal, 30, 286-292. 6.Xu, G., Xiong, W., Hu, Q., Zuo, P., Shao, B., Lan, F., Lu, X., Xu, Y. and Xiong, S. (2010), "Lactoferrin-derived peptides and Lactoferricin chimera inhibit virulence factor production and biofilm formation in Pseudomonas aeruginosa". J Appl Microbiol, 109, 1311-1318.

Pharmacy Education and Accreditation Reviews (PEARs) project:final report

Report commissioned by the Pharmaceutical Society of Ireland.

Corporate Social Reporting (CSR) in emerging economies:a review and future direction

Purpose – Previous reviews of Corporate Social Reporting (CSR) literature have tended to focus on developed economies. The aim of this study is to extend reviews of CSR literature to emerging economies. Design/methodology/approach – A desk-based research method, using a classification framework of three categories. Findings – Most CSR studies in emerging economies have concentrated on the Asia-Pacific and African regions and are descriptive in nature, used content analysis methods and measured the extent and volume of disclosures contained within the annual reports. Such studies provide indirect explanation of the reasons behind CSR adoption, but of late, a handful of studies have started to probe managerial motivations behind CSR directly through in-depth interviews finding that CSR agendas in emerging economies are largely driven by external forces, namely pressures from parent companies, international market and international agencies. Originality/value – This is the first review and analysis of CSR studies from the emerging economy perspective. Following this analysis, the authors have identified some important future research questions.

Leveraging product adopter information from online reviews for product recommendation

The availability of the sheer volume of online product reviews makes it possible to derive implicit demographic information of product adopters from review documents. This paper proposes a novel approach to the extraction of product adopter mentions from online reviews. The extracted product adopters are the ncategorise into a number of different demographic user groups. The aggregated demographic information of many product adopters can be used to characterize both products and users, which can be incorporated into a recommendation method using weighted regularised matrix factorisation. Our experimental results on over 15 million reviews crawled from JINGDONG, the largest B2C e-commerce website in China, show the feasibility and effectiveness of our proposed frame work for product recommendation.

So what is Capital in the Twenty-First Century? Some notes on Piketty’s book Working paper, forthcoming in Capitalism & Society

This study was inspired by Piketty’s excellent and important book. Its title and numerous statements in it arouse in readers expectations of a comprehensive analysis of capitalism. By comparison the author of this paper felt important aspects were lacking. The capitalist system has numerous immanent traits and innate tendencies, of which the paper takes a closer look at three properties. 1. One basic feature is dynamism, innovation, and creative destruction. No picture of capitalism can be full if this basic aspect is ignored. 2. Capitalism inevitably brings about a high degree of inequality; this must be eased by reforms, but cannot be entirely overcome. 3. The basic characteristics of capitalism – private ownership and the dominance of market coordination – give rise to strong incentive mechanisms that encourage but owners and enterprise executives to innovate and to cooperate effectively. One of the main incentives is competition, especially oligopolistic competition. There are strong mutual effects among these three important tendencies. It is impossible to understand well Piketty’s main subject, the distribution of income and wealth, if it is divorced from the other two tendencies. The study ends with its author’s own value judgements on the favourable and harmful, unjust attributes of the capitalist system.

The representation of motherhood in contemporary Catalan fiction

This dissertation analyzes four twenty-first-century Catalan novels which present the complex positions occupied by mothers in the last seven decades. Its conceptual framework posits motherhood as both a changing social construction and a political institution in a constant state of flux. In Inma Monsó´s Todo un carácter (2001), Eva Piquer´s Una victoria diferente (2002), Carme Riera´s La mitad del alma (2004), and Najat El Hachmi´s El último patriarca (2008) motherhood is explored as a metaphorical act, a gender-constructing experience, as well as the locus of expression with regard to gender and power relations. During the dictatorship of Francisco Franco (1939–1975), the majority of women were excluded from public spaces, and forced to stay home to care for their husbands and children. Furthermore, the state criminalized abortion, made contraception and divorce illegal, and promoted an ideal of femininity based on silence, sacrifice, and self-denial. The political changes of the late 1970s allowed women greater personal autonomy, and many women writers began to challenge stereotypical views of women’s social roles. Yet in the 70s and 80s, the narratives of Esther Tusquets, Ana María Moix, and Montserrat Roig represent the mother as a repressive figure whom the daughter must reject in order to liberate herself and regain her voice. It is not until the 90s when the novelists Mercedes Abad, Maruja Torres, Carme Riera, Imma Monsó, Eva Piquer, and María Barbal rehumanize the mother figure, recovering their matrilineal heritage. However, far from suggesting a unified trend in representations of motherhood in Catalan fiction, the diverse points of view of the novels under discussion here reveal that differences in attitudes among women authors about mother-daughter conflict are far from resolved. The theoretical background for this dissertation draws mainly on the work of Adrienne Rich, Nancy Chodorow, and Julia Kristeva. It includes psychoanalytic studies as well as sociologically based essays by Anna López Puig, Amparo Acereda, Jacqueline Cruz, Barbara Zecchi, Ángeles de la Concha, and Raquel Osborne, among others.

"Unfair'' Restaurant Reviews:To Sue Or Not To Sue

In their discussion entitled - “Unfair” Restaurant Reviews: To Sue Or Not To Sue - by John Schroeder and Bruce Lazarus, Assistant Professors, Department of Restaurant, Hotel and Institutional Management at Purdue University, the authors initially state: “Both advantages and disadvantages exist on bringing lawsuits against restaurant critics who write “unfair” reviews. The authors, both of whom have experience with restaurant criticism, offer practical advice on what realistically can be done by the restaurateur outside of the courtroom to combat unfair criticism.” Well, this is going to be a sticky wicket no matter how you try to defend it, reviews being what they are; very subjective pieces of opinionated journalism, especially in the food industry. And, of course, unless you can prove malicious intent there really is no a basis for a libel suit. So, a restaurateur is at the mercy of written opinion and the press. “Libel is the written or published form of slander which is the statement of false remarks that may damage the reputation of others. It also includes any false and malicious publication which may damage a person's business, trade, or employment,” is the defined form of the law provided by the authors. Anecdotally, Schroeder and Lazarus offer a few of the more scathing pieces reviewers have written about particular eating establishments. And, yes, they can be a bit comical, unless you are the owner of an establishment that appears in the crosshairs of such a reviewer. A bad review can kneecap even a popular eatery. “Because of the large readership of restaurant reviews in the publication (consumer dining out habits indicate that nearly 50 percent of consumers read a review before visiting a new restaurant) your business begins a very dangerous downward tailspin,” the authors reveal, with attribution. “Many restaurant operators contend that a bad review can cost them an immediate trade loss of upward of 50 percent,” Schroeder and Lazarus warn. “The United States Supreme Court has ruled that a restaurant owner can collect damages only if he proves that the statement or statements were made with “actual malice,” even if the statements were untrue,” the authors say by way of citation. And that last portion of the statement cannot be over-emphasized. The first amendment to the U.S. Constitution does wield a heavy hammer, indeed, and it should. So, what recourse does a restaurateur have? The authors cautiously give a guarded thumbs-up to a lawsuit, but you better be prepared to prove a misstatement of fact, as opposed to the distinguishable press protected right of opinion. For the restaurateur the pitfalls are many, the rewards few and far between, Schroeder and Lazarus will have you know. “…after weighing the advantages and disadvantages of a lawsuit against a critic...the disadvantages are overwhelming,” the authors say. “Chicago restaurant critic James Ward said that someone dumped a load of manure on his yard accompanied by a note that read - Stop writing that s--t! - after he wrote a review of a local restaurant.” Such is a novel if not legally measurable tack against an un-mutual review.

Influence of Restaurant Reviews Upon Consumers

It is generally believed that restaurant reviews can influence consumers' decisions in choosing a restaurant. A survey administered to a sample of 420 college faculty and staff members suggests that while most restaurant patrons may read reviews, they are not used as the sole selection criterion. Recommendations of friends, the restaurant's current reputation, and perceived value may have greater influence upon the choice than does a re- view. The authors discuss the implications of both favorable and unfavorable reviews.

Tatyana Mamonova holding her underground unpublished book on Russian Women and feminism

General note: Title and date provided by Bettye Lane.

Florence Rush, writer of a recent book on incest, at Pornography and Self-image Meeting

General note: Title and date provided by Bettye Lane.

Pat Moore poses with her book

General note: Title and date provided by Bettye Lane.

Phyllis Chesler, author of "Women and Madness" reads from her new book at Dalton Book Store

General note: Title and date provided by Bettye Lane.