Sensitization of Staphylococcus aureus to Methicillin and Other Antibiotics In Vitro and In Vivo in the Presence of HAMLET
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{"title"=>"Sensitization of Staphylococcus aureus to Methicillin and Other Antibiotics In Vitro and In Vivo in the Presence of HAMLET", "type"=>"journal", "authors"=>[{"first_name"=>"Laura R.", "last_name"=>"Marks", "scopus_author_id"=>"55326767300"}, {"first_name"=>"Emily A.", "last_name"=>"Clementi", "scopus_author_id"=>"57190085163"}, {"first_name"=>"Anders P.", "last_name"=>"Hakansson", "scopus_author_id"=>"56404191100"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84877032645", "doi"=>"10.1371/journal.pone.0063158", "sgr"=>"84877032645", "isbn"=>"1932-6203", "pmid"=>"23650551", "issn"=>"19326203", "pui"=>"368847412"}, "id"=>"57d193d0-e083-392d-b33f-af5d43fde88f", "abstract"=>"HAMLET (human alpha-lactalbumin made lethal to tumor cells) is a protein-lipid complex from human milk with both tumoricidal and bactericidal activities. HAMLET exerts a rather specific bactericidal activity against some respiratory pathogens, with highest activity against Streptococcus pneumoniae, but lacks activity against most other bacterial pathogens, including Staphylococci. Still, ion transport associated with death in S. pneumoniae is also detected to a lower degree in insensitive organisms. In this study we demonstrate that HAMLET acts as an antimicrobial adjuvant that can increase the activity of a broad spectrum of antibiotics (methicillin, vancomycin, gentamicin and erythromycin) against multi-drug resistant Staphylococcus aureus, to a degree where they become sensitive to those same antibiotics, both in antimicrobial assays against planktonic and biofilm bacteria and in an in vivo model of nasopharyngeal colonization. We show that HAMLET exerts these effects specifically by dissipating the proton gradient and inducing a sodium-dependent calcium influx that partially depolarizes the plasma membrane, the same mechanism induced during pneumococcal death. These effects results in an increased cell associated binding and/or uptake of penicillin, gentamicin and vancomycin, especially in resistant stains. Finally, HAMLET inhibits the increased resistance of methicillin seen under antibiotic pressure and the bacteria do not become resistant to the adjuvant, which is a major advantageous feature of the molecule. These results highlight HAMLET as a novel antimicrobial adjuvant with the potential to increase the clinical usefulness of antibiotics against drug resistant strains of S. aureus.", "link"=>"http://www.mendeley.com/research/sensitization-staphylococcus-aureus-methicillin-other-antibiotics-vitro-vivo-presence-hamlet", "reader_count"=>58, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>17, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>3, "Student > Master"=>12, "Other"=>4, "Student > Bachelor"=>6, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>17, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>3, "Student > Master"=>12, "Other"=>4, "Student > Bachelor"=>6, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>5, "Materials Science"=>1, "Agricultural and Biological Sciences"=>27, "Medicine and Dentistry"=>12, "Arts and Humanities"=>1, "Chemistry"=>4, "Social Sciences"=>1, "Computer Science"=>1, "Immunology and Microbiology"=>5, "Energy"=>1}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>12}, "Chemistry"=>{"Chemistry"=>4}, "Social Sciences"=>{"Social Sciences"=>1}, "Energy"=>{"Energy"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>27}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Arts and Humanities"=>{"Arts and Humanities"=>1}}, "reader_count_by_country"=>{"United States"=>3, "United Kingdom"=>2, "Mexico"=>1, "Germany"=>1}, "group_count"=>3}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1050980"], "description"=>"<p>Mice were colonized with <i>S. aureus</i> NRS 70 for 24 hours, treated intranasally with various doses of gentamicin in the presence (blue) or absence (black) of HAMLET (100 µg) for 12 hours, and the bacterial burden associated with the nasal wash (A) and the nasopharyngeal tissue (B) was determined. The graph shows colonization data for individual mice, with the mean recovered bacteria depicted with a line. The results are based on experiments using groups of 6 mice. Statistics was performed using the unpaired Student t-test. Significance was indicated as follows: *  =  <i>P</i><0.05.</p>", "links"=>[], "tags"=>["microbiology", "bacteriology", "Bacterial biochemistry", "Bacterial biofilms", "Host-pathogen interaction", "Medical microbiology", "Microbial physiology", "Global health", "Infectious diseases", "Bacterial diseases", "Methicillin-resistant Staphylococcus aureus", "Staphylococcus aureus", "Vancomycin-resistant staphylococcus aureus", "Skin infections", "staphylococcal", "nasopharyngeal"], "article_id"=>695788, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Laura R. Marks", "Emily A. Clementi", "Anders P. Hakansson"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063158.g003", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_HAMLET_Methicillin_combination_treatment_reduces_Staphylococcal_nasopharyngeal_colonization_/695788", "title"=>"HAMLET/Methicillin combination treatment reduces Staphylococcal nasopharyngeal colonization.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-01 01:36:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/1050977"], "description"=>"<p><i>S. aureus</i> strains 11090306 (MSSA) (left) and NRS 384 (MRSA) (right) were grown in broth for 16 hours in the presence of 2 µg/ml methicillin (5 µM) with and without the addition of 100 µg/mL (6 µM) HAMLET. The figure shows representative growth curves for the lowest concentration of antibiotic and HAMLET that inhibited bacterial growth by combination treatment without either agent alone affecting growth.</p>", "links"=>[], "tags"=>["microbiology", "bacteriology", "Bacterial biochemistry", "Bacterial biofilms", "Host-pathogen interaction", "Medical microbiology", "Microbial physiology", "Global health", "Infectious diseases", "Bacterial diseases", "Methicillin-resistant Staphylococcus aureus", "Staphylococcus aureus", "Vancomycin-resistant staphylococcus aureus", "Skin infections", "lowers", "methicillin"], "article_id"=>695785, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Laura R. Marks", "Emily A. Clementi", "Anders P. Hakansson"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063158.g001", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_HAMLET_lowers_the_methicillin_MIC_/695785", "title"=>"HAMLET lowers the methicillin MIC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-01 01:36:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1050985"], "description"=>"*<p>HL  =  HAMLET. Concentration used was 100 µg/ml (6 µM).</p>**<p>The last column describes the concentration of HAMLET required to make each strain sensitive to the respective antibiotic.</p>", "links"=>[], "tags"=>["microbiology", "bacteriology", "Bacterial biochemistry", "Bacterial biofilms", "Host-pathogen interaction", "Medical microbiology", "Microbial physiology", "Global health", "Infectious diseases", "Bacterial diseases", "Methicillin-resistant Staphylococcus aureus", "Staphylococcus aureus", "Vancomycin-resistant staphylococcus aureus", "Skin infections", "mbc", "strains", "exposed", "antibiotics"], "article_id"=>695793, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Laura R. Marks", "Emily A. Clementi", "Anders P. Hakansson"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063158.t001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MIC_and_MBC_values_for_S_aureus_strains_exposed_to_various_antibiotics_in_the_absence_and_presence_of_HAMLET_/695793", "title"=>"MIC and MBC values for <i>S. aureus</i> strains exposed to various antibiotics in the absence and presence of HAMLET.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-05-01 01:36:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1050983"], "description"=>"<p>(A) Staphylococci were incubated with Bocillin FL or (B) with vancomycin FL in the presence or absence of 100 µg/mL (6 µM) HAMLET. The results are based on three individual experiments with duplicate samples. Statistics was performed using the unpaired Student's t-test. Significance was indicated as follows: **  =  P<0.01, ***  =  P<0.001, ns  =  not significant.</p>", "links"=>[], "tags"=>["microbiology", "bacteriology", "Bacterial biochemistry", "Bacterial biofilms", "Host-pathogen interaction", "Medical microbiology", "Microbial physiology", "Global health", "Infectious diseases", "Bacterial diseases", "Methicillin-resistant Staphylococcus aureus", "Staphylococcus aureus", "Vancomycin-resistant staphylococcus aureus", "Skin infections", "hamlet", "uptake", "binding", "bocillin", "fl", "vancomycin"], "article_id"=>695791, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Laura R. Marks", "Emily A. Clementi", "Anders P. Hakansson"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063158.g005", "stats"=>{"downloads"=>2, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Impact_of_HAMLET_on_uptake_and_binding_of_Bocillin_FL_and_vancomycin_FL_/695791", "title"=>"Impact of HAMLET on uptake and binding of Bocillin FL and vancomycin FL.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-01 01:36:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/1050984"], "description"=>"<p>Methicillin adaptation of the MRSA strain NRS 384 after exposure to stepwise increasing concentrations of methicillin alone (1–512 µg/mL or 2.5–1,350 µM) or methicillin in combination with 100 µg/mL (6 µM) HAMLET. The filled blue circles show the methicillin MICs after each cycle when no HAMLET was used. The addition of 100 µg/mL of HAMLET reduced methicillin-induced resistance (blue unfilled circles). The unfilled green circle represents the MIC of methicillin of the bacteria grown in presence of HAMLET, when HAMLET was also present during the MIC assay to potentiate the effect of the antibiotic. Reintroduction of HAMLET to these isolates again returned the methicillin MIC back to the levels denoted by the green unfilled circle.</p>", "links"=>[], "tags"=>["microbiology", "bacteriology", "Bacterial biochemistry", "Bacterial biofilms", "Host-pathogen interaction", "Medical microbiology", "Microbial physiology", "Global health", "Infectious diseases", "Bacterial diseases", "Methicillin-resistant Staphylococcus aureus", "Staphylococcus aureus", "Vancomycin-resistant staphylococcus aureus", "Skin infections"], "article_id"=>695792, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Laura R. Marks", "Emily A. Clementi", "Anders P. Hakansson"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063158.g006", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_HAMLET_and_resistance_development_/695792", "title"=>"HAMLET and resistance development.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-01 01:36:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/1050982"], "description"=>"<p>(A) Representative growth curves for <i>S. aureus</i> strain NRS 123 (MRSA) grown in broth for 16 hours (960 min) in the presence of methicillin with and without the addition of HAMLET and the inhibitors Ruthenium Red (RuR) or Amiloride (Amil). (B) Mid-log phase grown NRS 123 Staphylococci re-suspended in PBS alone or PBS plus Amiloride or Ruthenium Red, were incubated with the fluorescent indicator dye DiBAC<sub>4</sub>(3) and membrane depolarization was detected by measuring fluorescence over time. HAMLET was added at twenty minutes (arrow). The detergent Triton X-100 (0.1%) was included as a positive control. The results presented are from one representative experiment. (C) Mid-log phase grown NRS 384 Staphylococci were incubated with the radioisotope <sup>45</sup>Ca<sup>2+</sup> (2.5 µCi/mL) in PBS or PBS + Ruthenium Red (30 µM). After recording baseline readings, PBS (untreated), or HAMLET was added (Time  = 0 min) to the bacteria and radioactivity was measured over time. Results from a representative experiment are shown. (D) NRS 384 Staphylococci were loaded with the pH sensitive dye BCECF-AM, and were washed and resuspended in PBS + 25 mM glucose. After recording baseline readings, at the first arrow, PBS (untreated), the protonophore CCCP (100 µM), HAMLET (100 µg/mL or 6 µM), HAMLET + RuR (30 µM), or HAMLET + Amiloride (1 mM) were added to the bacteria and fluorescence was measured over time. At the second arrow 20 µM each of nigericin and valinomycin was added to completely dissipate the transmembrane proton gradient.</p>", "links"=>[], "tags"=>["microbiology", "bacteriology", "Bacterial biochemistry", "Bacterial biofilms", "Host-pathogen interaction", "Medical microbiology", "Microbial physiology", "Global health", "Infectious diseases", "Bacterial diseases", "Methicillin-resistant Staphylococcus aureus", "Staphylococcus aureus", "Vancomycin-resistant staphylococcus aureus", "Skin infections", "hamlet", "membrane"], "article_id"=>695790, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Laura R. Marks", "Emily A. Clementi", "Anders P. Hakansson"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063158.g004", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_HAMLET_on_membrane_potential_/695790", "title"=>"Effect of HAMLET on membrane potential.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-01 01:36:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1050979"], "description"=>"<p>(A) The activity of methicillin (250 μg/mL or 660 µM), HAMLET (200 μg/mL or 12 µM), or the combination of both agents were tested on <i>in vitro</i> biofilms of the methicillin-resistant strain NRS 70 (MRSA) or the methicillin-sensitive strain 11090306 (MSSA) by determining the bacterial death (in log<sub>10</sub>) after culturing dilutions overnight on blood agar. (B) The activity of vancomycin (32 μg/mL or 21 µM), HAMLET (500 μg/mL or 30 µM), or the combination of both agents on <i>in vitro</i> biofilms of the vancomycin-resistant strain NRS 1 (VISA) and the vancomycin-sensitive strain NRS384 (VSSA) were tested in a similar fashion as was the activity of (C) 50 µg/ml (105 µM) gentamicin alone or in combination with 500 µg/ml (30 µM) HAMLET for the gentamicin-resistant strain. The results are based on three independent experiments with duplicate samples. Statistics was performed using the paired Student t-test. Significance was indicated as follows: ns  =  not significant, *  =  <i>P</i><0.05, **  =  <i>P</i><0.01.</p>", "links"=>[], "tags"=>["microbiology", "bacteriology", "Bacterial biochemistry", "Bacterial biofilms", "Host-pathogen interaction", "Medical microbiology", "Microbial physiology", "Global health", "Infectious diseases", "Bacterial diseases", "Methicillin-resistant Staphylococcus aureus", "Staphylococcus aureus", "Vancomycin-resistant staphylococcus aureus", "Skin infections", "biofilm"], "article_id"=>695787, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Laura R. Marks", "Emily A. Clementi", "Anders P. Hakansson"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0063158.g002", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_effect_of_HAMLET_antibiotic_combination_treatment_on_in_vitro_biofilm_viability_/695787", "title"=>"The effect of HAMLET/antibiotic combination treatment on <i>in vitro</i> biofilm viability.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-05-01 01:36:27"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Microbiology", "average_usage"=>[293, 503, 638, 755, 861, 960, 1056, 1146, 1239, 1323, 1403, 1491, 1568]}, {"subject_area"=>"/Ecology and environmental sciences", "average_usage"=>[284, 475, 603, 722, 826, 928, 1026, 1129, 1225, 1310, 1390, 1468, 1549]}, {"subject_area"=>"/Ecology and environmental sciences/Ecology", "average_usage"=>[298, 487, 610, 722, 827, 929, 1029, 1125, 1217, 1306, 1388, 1464, 1535]}]}
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