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  • 1
    In: The Journal of Bacteriology, 2008, Vol. 190(3), p.1036
    Description: We have identified a homologue to the staphylococcal biofilm-associated protein (Bap) in a bloodstream isolate of Acinetobacter baumannii. The fully sequenced open reading frame is 25,863 bp and encodes a protein with a predicted molecular mass of 854 kDa. Analysis of the nucleotide sequence reveals a repetitive structure consistent with bacterial cell surface adhesins. Bap-specific monoclonal antibody (MAb) 6E3 was generated to an epitope conserved among 41% of A. baumannii strains isolated during a recent outbreak in the U.S. military health care system. Flow cytometry confirms that the MAb 6E3 epitope is surface exposed. Random transposon mutagenesis was used to generate A. baumannii bap1302::EZ-Tn5, a mutant negative for surface reactivity to MAb 6E3 in which the transposon disrupts the coding sequence of bap. Time course confocal laser scanning microscopy and three-dimensional image analysis of actively growing biofilms demonstrates that this mutant is unable to sustain biofilm thickness and volume, suggesting a role for Bap in supporting the development of the mature biofilm structure. This is the first identification of a specific cell surface protein directly involved in biofilm formation by A. baumannii and suggests that Bap is involved in intercellular adhesion within the mature biofilm.
    Keywords: Biology;
    ISSN: 0021-9193
    ISSN: 00219193
    E-ISSN: 10985530
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  • 2
    Language: English
    In: The Journal of infectious diseases, 15 February 2009, Vol.199(4), pp.513-21
    Description: Acinetobacter baumannii is a bacterial pathogen of increasing medical importance. Little is known about genes important for its survival in vivo. Screening of random transposon mutants of the model pathogen AB307-0294 identified the mutant AB307.27. AB307.27 contained its transposon insertion in pbpG, which encodes the putative low-molecular-mass penicillin-binding protein 7/8 (PBP-7/8). AB307.27 was significantly killed in ascites (P〈.001), but its growth in Luria-Bertani broth was similar to that of its parent, AB307-0294 (P=.13). The survival of AB307.27 was significantly decreased in a rat soft-tissue infection model (P〈.001) and a rat pneumonia model (P=.002), compared with AB307-0294. AB307.27 was significantly killed in 90% human serum in vitro, compared with AB307-0294 (P〈.001). Electron microscopy demonstrated more coccobacillary forms of AB307.27, compared with AB307-0294, suggesting a possible modulation in the peptidoglycan, which may affect susceptibility to host defense factors. These findings demonstrate that PBP-7/8 contributes to the pathogenesis of A. baumannii. PBP-7/8 either directly or indirectly contributes to the resistance of AB307-0294 to complement-mediated bactericidal activity. An understanding of how PBP-7/8 contributes to serum resistance will lend insight into the role of this low-molecular-mass PBP whose function is poorly understood.
    Keywords: Acinetobacter Infections -- Immunology ; Acinetobacter Baumannii -- Growth & Development ; Penicillin-Binding Proteins -- Physiology
    ISSN: 0022-1899
    E-ISSN: 15376613
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  • 3
    In: Clinical Orthopaedics and Related Research, 2015, Vol.473(9), pp.2856-2864
    Description: BACKGROUND: Effective treatments for implant-associated infections are often lacking. Cathodic voltage-controlled electrical stimulation has shown potential as a treatment of implant-associated infections of methicillin-resistant Staphylococcus aureus (MRSA). QUESTIONS/PURPOSES: The primary purpose of this study was to (1) determine if cathodic voltage-controlled electrical stimulation combined with vancomycin therapy is more effective at reducing the MRSA bacterial burden on the implant, bone, and synovial fluid in comparison to either treatment alone or no treatment controls. We also sought to (2) evaluate the histologic effects of the various treatments on the surrounding bone; and to (3) determine if the cathodic voltage-controlled electrical stimulation treatment had an effect on the mechanical properties of the titanium implant as a result of possible hydrogen embrittlement. METHODS: Thirty-two adult male Long-Evans rats (Harlan Laboratories, Indianapolis, IN, USA) with surgically placed shoulder titanium implants were infected with a clinical strain of MRSA (NRS70). One week after infection, eight animals received a treatment of cathodic voltage-controlled electrical stimulation at −1.8 V versus Ag/AgCl for 1 hour (STIM), eight received vancomycin twice daily for 1 week (VANCO), eight received the cathodic voltage-controlled electrical stimulation and vancomycin therapy combined (STIM + VANCO), and eight served as controls with no treatment (CONT). Two weeks after initial infection, the implant, bone, and synovial fluid were collected for colony-forming unit (CFU) enumeration, qualitative histological analysis by a pathologist blinded to the treatments each animal received, and implant three-point bend testing. RESULTS: The implant-associated CFU enumerated from the STIM + VANCO (mean, 3.7 × 10; SD, 6.3 × 10) group were less than those from the CONT (mean, 1.3 × 10; SD, 2.8 × 10; 95% confidence interval [CI] of difference, −4.3 × 10 to −9.9 × 10; p 〈 0.001), STIM (mean, 1.4 × 10; SD, 2.0 × 10; 95% CI of difference, −2.1 × 10 to −1.8 × 10; p = 0.002), and VANCO (mean, 5.8 × 10; SD, 5.7 × 10; 95% CI of difference, −6.4 × 10 to −1.7 × 10; p 〈 0.001) group. The bone-associated CFU enumerated from the STIM + VANCO group (6.3 × 10; SD, 1.1 × 10) were less than those from the CONT (mean, 2.8 × 10; SD, 4.8 × 10; 95% CI of difference, −9.4 × 10 to −5.0 × 10; p 〈 0.001) and STIM (mean, 2.6 × 10; SD, 2.5 × 10; 95% CI of difference, −4.1 × 10 to −1.6 × 10; p 〈 0.001) groups. The VANCO group (4.3 × 10; SD, 6.3 × 10) also had lower bone-associated CFU as compared with the CONT (mean 95% CI of difference, −9.3 × 10 to −4.5 × 10; p 〈 0.001) and STIM (95% CI of difference, −4.0 × 10 to −1.5 × 10; p 〈 0.001) groups. In comparison to the synovial fluid CFU enumerated from the CONT group (mean, 3.3 × 10; SD, 6.0 × 10), lower synovial CFU were reported for both the STIM + VANCO group (mean, 4.6 × 10; SD, 1.2 × 10; 95% CI of difference, −4.9 × 10 to −3.0 × 10; p 〈 0.001) and the VANCO group (mean, 6.8 × 10; SD, 9.2 × 10; 95% CI of difference, −4.9 × 10 to −2.8 × 10; p = 0.007). The histological analysis showed no discernable deleterious effects on the surrounding tissue as a result of the treatments. No brittle fracture occurred during mechanical testing and with the numbers available, no differences in implant flexural yield strength were detected between the groups. CONCLUSIONS: In this rodent model, cathodic voltage-controlled electrical stimulation combined with vancomycin is an effective treatment for titanium implant-associated infections showing greater than 99.8% reduction in bacterial burden on the implant, surrounding bone, and synovial fluid as compared with the controls and the stimulation alone groups. CLINICAL RELEVANCE: Cathodic voltage-controlled electrical stimulation combined with vancomycin may enable successful treatment of titanium orthopaedic implant-associated infections with implant retention. Future studies will focus on optimization of the stimulation parameters for complete eradication of infection and the ability to promote beneficial host tissue responses.
    Keywords: Staphylococcus Aureus Infections – Care and Treatment ; Staphylococcus Aureus Infections – Analysis ; Staphylococcus Aureus Infections – Health Aspects ; Vancomycin – Analysis ; Vancomycin – Health Aspects ; Staphylococcus Aureus – Analysis ; Staphylococcus Aureus – Health Aspects ; Microbial Drug Resistance – Care and Treatment ; Microbial Drug Resistance – Analysis ; Microbial Drug Resistance – Health Aspects;
    ISSN: 0009-921X
    E-ISSN: 15281132
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  • 4
    Language: English
    In: The Elementary School Journal June 2011, Vol.111(4), pp.641-661
    Description: Abstract Two experimental studies at one urban middle school investigated the effects of the combination of Tier I and Tier II evidence-based reading instruction compared to Tier I alone on struggling sixth-grade readers ( N = 109). All participants received free or reduced-price lunch, and 95% were considered English learners at some point in their school history. In both studies, Tier II intervention consisted of intensive instruction in word analysis, fluency building, comprehension, and vocabulary for 30 hours across 10 weeks. Results of both studies taken individually and combined indicated significant differences in favor of the intervention groups on oral reading fluency. The second study indicated significantly stronger performances for the intervention group on the Woodcock Reading Mastery Test—Revised (WRMT-R/NU) passage comprehension subtest. Tier II interventions and Response to Intervention (RTI) for older struggling readers are discussed related to educational implications and future research.
    Keywords: Reading Deficiencies (70900) ; Reading Instruction (70950) ; Junior High School Students (40210) ; English As a Second Language Learning (22130) ; Reading Tests (71550) ; Applied Linguistics; Reading Readiness/Acquisition/Achievement ; Article;
    ISSN: 00135984
    E-ISSN: 15548279
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  • 5
    Language: English
    In: Gene, 2011, Vol.477(1), pp.19-23
    Description: is a Gram-negative aerobic diplococcus that is a mucosal pathogen of the upper and lower respiratory tracts in humans. In order to colonize the human host and establish an infection, must be able to effectively attach to the respiratory mucosal epithelia. Although little is known about pathogenesis, our laboratory has previously shown that expression of type IV pili (TFP) contributes to mucosal colonization. TFP are filamentous surface appendages primarily composed of a single protein subunit termed pilin, which is encoded by in . These surface structures play a crucial role in the initiation of disease by a wide range of pathogenic bacteria. Our studies also indicate that unlike the pilin of the pathogenic species, which exhibit both phase and antigenic variation, the pilin subunit of appears to be more highly conserved as there are no major pilin variants produced by a single strain and only two major PilA antigenic variants, termed clade 1 and clade 2, have been observed between strains. Moreover, we have determined that these highly conserved bacterial surface structures are expressed by all clinical isolates evaluated. Therapeutic or vaccine-based interventions that prevent or diminish nasopharyngeal colonization will likely decrease acute and recurrent infections in prone populations. Thus, our data indicate that additional studies aimed at elucidating the role of PilA in the pathogenesis and host response to infections are warranted.
    Keywords: Pili ; Pilin ; Antigenic Clades ; Engineering ; Biology ; Anatomy & Physiology
    ISSN: 0378-1119
    E-ISSN: 1879-0038
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  • 6
    Language: English
    In: Microbiology (Reading, England), April 2005, Vol.151(Pt 4), pp.1151-8
    Description: Moraxella catarrhalis is a leading cause of acute otitis media in children and is a cause of respiratory disease in adults with underlying lung disease. This organism is a strict human pathogen that has an absolute requirement for iron in order to grow and cause disease. Previous studies identified transferrin and lactoferrin receptors used by M. catarrhalis to obtain iron from the human host, yet other iron-acquisition systems remain undefined. In this study, it is demonstrated that this strict mucosal pathogen can utilize haemoglobin (Hb) as a sole source of iron for growth. A novel 107 kDa outer-membrane protein involved in Hb utilization by this pathogen was also identified. An isogenic mutant defective in this Moraxella Hb-utilization protein (MhuA), 7169 : : mhuA, showed a significant lag during growth in the presence of Hb as the sole iron source. This protein appears to be expressed constitutively, regardless of growth conditions, and a mAb directed to MhuA demonstrated that this protein contains highly conserved, surface-exposed epitopes. Data demonstrating that expression of MhuA may be highly specific to isolates of M. catarrhalis are also presented, suggesting a potential role as a diagnostic marker. To our knowledge, this is the first report demonstrating that M. catarrhalis expresses an Hb-binding protein and that this bacterium can utilize Hb as a sole iron source for growth.
    Keywords: Bacterial Proteins -- Metabolism ; Hemoglobins -- Metabolism ; Moraxella (Branhamella) Catarrhalis -- Metabolism
    ISSN: 1350-0872
    E-ISSN: 14652080
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  • 7
    Language: English
    In: Microbial Pathogenesis, 2006, Vol.40(3), pp.110-115
    Description: express two porin proteins, termed OmpP2A and OmpP2B. To test whether expression of OmpP2A and OmpP2B was necessary for virulence in humans, eight volunteers were experimentally infected with the parent (35000HP) in one arm and a double OmpP2A OmpP2B mutant (35000HP::P2AB) in the other arm. The pustule formation rates were 58.3% (95% CI, 33.2–83.5%) for the parent and 41.7% (95% CI, 19.3–64.0%) for the mutant ( =0.25). Biopsy of 35000HP and 35000HP::P2AB-infected sites yielded similar amounts of bacteria in quantitative culture. These results indicate that expression of OmpP2A and OmpP2B is not necessary to initiate disease or to progress to pustule formation in humans.
    Keywords: Pustule Formation ; Haemophilus Ducreyi ; Porin Proteins ; Uirulence ; Human Challenge Model ; Biology ; Chemistry
    ISSN: 0882-4010
    E-ISSN: 1096-1208
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  • 8
    Language: English
    In: Clinical Orthopaedics and Related Research®, 2016, Vol.474(7), pp.1668-1675
    Description: To access, purchase, authenticate, or subscribe to the full-text of this article, please visit this link: http://dx.doi.org/10.1007/s11999-016-4705-7 Byline: Scott R. Nodzo (1), Menachem Tobias (1), Richard Ahn (1), Lisa Hansen (2), Nicole R. Luke-Marshall (2), Craig Howard (1), Linda Wild (3), Anthony A. Campagnari (2), Mark T. Ehrensberger (4) Abstract: Background Cathodic voltage-controlled electrical stimulation (CVCES) of titanium implants, either alone or combined with a short course of vancomycin, has previously been shown to reduce the bone and implant bacterial burden in a rodent model of methicillin-resistant Staphylococcus aureus (MRSA) implant-associated infection (IAI). Clinically, the goal is to achieve complete eradication of the IAI therefore, the rationale for the present study was to evaluate the antimicrobial effects of combining CVCES with prolonged antibiotic therapy with the goal of decreasing the colony-forming units (CFUs) to undetectable levels. Questions/purposes (1) In an animal MRSA IAI model, does combining CVCES with prolonged vancomycin therapy decrease bacteria burden on the implant and surrounding bone to undetectable levels? (2) When used with prolonged vancomycin therapy, are two CVCES treatments more effective than one? (3) What are the longer term histologic effects (inflammation and granulation tissue) of CVCES on the surrounding tissue? Methods Twenty adult male Long-Evans rats with surgically placed shoulder titanium implants were infected with a clinical strain of MRSA (NRS70). One week after infection, the rats were randomly divided into four groups of five: (1) VANCO: only vancomycin treatment (150 mg/kg, subcutaneous, twice daily for 5 weeks) (2) VANCO + 1STIM: vancomycin treatment (same as the VANCO group) coupled with one CVCES treatment (-1.8 V for 1 hour on postoperative day [POD] 7) (3) VANCO + 2STIM: vancomycin treatment (same as the VANCO group) coupled with two CVCES treatments (-1.8 V for 1 hour on POD 7 and POD 21) or (4) CONT: no treatment. On POD 42, the implant, bone, and peripheral blood were collected for CFU enumeration and histological analysis, where we compared CFU/mL on the implants and bone among the groups. A pathologist, blinded to the experimental conditions, performed a semiquantitative analysis of inflammation and granulation tissue present in serial sections of the humeral head for animals in each experimental group. Results The VANCO + 1STIM decreased the implant bacterial burden (median = 0, range = 0--10 CFU/mL) when compared with CONT (median = 5.7 x 10.sup.4, range = 4.0 x 10.sup.3-8.0 x 10.sup.5 CFU/mL difference of medians = -5.6 x 10.sup.4 p 〈 0.001) and VANCO (median = 4.9 x 10.sup.3, range = 9.0 x 10.sup.2-2.1 x 10.sup.4 CFU/mL difference of medians = -4.9 x 10.sup.3 p 〈 0.001). The VANCO + 1STIM decreased the bone bacterial burden (median = 0, range = 0--0 CFU/mL) when compared with CONT (median = 1.3 x 10.sup.2, range = 0--9.4 x 10.sup.2 CFU/mL difference of medians = -1.3 x 10.sup.2 p 〈 0.001) but was not different from VANCO (median = 0, range = 0--1.3 x 10.sup.2 CFU/mL difference of medians = 0 p = 0.210). The VANCO + 2STIM group had implant CFU (median = 0, range = 0--8.0 x 10.sup.1 CFU/mL) and bone CFU (median = 0, range = 0--2.0 x 10.sup.1 CFU/mL) that were not different from the VANCO + 1STIM treatment group implant CFU (median = 0, range = 0--10 CFU/mL difference of medians = 0 p = 0.334) and bone CFU (median = 0, range = 0--0 CFU/mL difference of medians = 0 p = 0.473). The histological analysis showed no deleterious effects on the surrounding tissue as a result of the treatments. Conclusions Using CVCES in combination with prolonged vancomycin resulted in decreased MRSA bacterial burden, and it may be beneficial in treating biofilm-related implant infections. Clinical Relevance CVCES combined with clinically relevant lengths of vancomycin therapy may be a treatment option for IAI and allow for component retention in certain clinical scenarios. However, more animal research and human trials confirming the efficacy of this approach are needed before such a clinical recommendation could be made. Author Affiliation: (1) Department of Orthopedics, State University of New York at Buffalo, Buffalo, NY, USA (2) Department of Microbiology and Immunology, State University of New York at Buffalo, Buffalo, NY, USA (3) Department of Pathology and Anatomical Sciences, State University of New York at Buffalo, Buffalo, NY, USA (4) Department of Biomedical Engineering, State University of New York at Buffalo, 162 Farber Hall, 3435 Main Street, Buffalo, NY, 14214, USA Article History: Registration Date: 08/01/2016 Online Date: 22/01/2016 Article note: The institution of one or more of the authors (MTE) has received, during the study period, funding from the Congressionally Directed Medical Research Program, Peer Reviewed Orthopedic Research Program, and the Bruce Holm Memorial Catalyst Fund. All ICMJE Conflict of Interest Forms for authors and Clinical Orthopaedics and Related Research [R] editors and board members are on file with the publication and can be viewed on request. Clinical Orthopaedics and Related Research [R] neither advocates nor endorses the use of any treatment, drug, or device. Readers are encouraged to always seek additional information, including FDA-approval status, of any drug or device prior to clinical use. Each author certifies that his or her institution approved the animal protocol for this investigation and that all investigations were conducted in conformity with ethical principles of research. A comment to this article is available at http://dx.doi.org/10.1007/s11999-016-4744-0.
    Keywords: Vancomycin – Health Aspects ; Vancomycin – Analysis;
    ISSN: 0009-921X
    E-ISSN: 1528-1132
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  • 9
    In: Lasers in Surgery and Medicine, November 2014, Vol.46(9), pp.712-717
    Keywords: Antimicrobial Photodynamic Therapy ; Biofilm ; Gram‐Negative Bacteria ; Otitis Media ; Photofrin
    ISSN: 0196-8092
    E-ISSN: 1096-9101
    Source: John Wiley & Sons, Inc.
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  • 10
    Language: English
    In: Journal of Early and Intensive Behavior Intervention, 2008, Vol.5(1), pp.23-39
    Description: We tested the effect of a Rotated Protocol Immersion package on the emergence of observing responses as prerequisites for more complex verbal developmental capabilities. Three elementary aged students between the ages of 6 and 7 participated. They were diagnosed with autism spectrum disabilities. The treatment condition consisted of total immersion in a rotation of six pre-listener Protocols ( Greer & Ross, 2008 ), designed to induce foundations for verbal developmental capabilities. The participants were selected for their demonstrated lack of early observing responses ( Keohane, Delgado & Greer, in press ). The y did not respond when their names were called, orient toward voices in the environment, or follow instructions. They did not seek out the attention of others unless it was to fill an immediate need. The dependent variables in the study were observing responses; learn units to criterion, instructional objectives met, and incidental performances across instructional and non-instructional settings. We used a time-lagged multiple probe design and found significant increases in the dependent variables. Additionally, the post-probes demonstrated a range of increases in the number and level of complexity of students' observing responses. The results are discussed in terms of theoretical implications, as well as, in the context of behavioral research on child development, and the hierarchy of verbal developmental capabilities.
    Keywords: Development ; Verbal Developmental Capabilities ; Behavioral Developmental Cusps ; Learn Units ; Observing Responses ; Developmental Delays ; Verbal Behavior
    E-ISSN: 1554-4893
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