@article{recht_comen_fine_fleming_hardenbergh_ho_hudis_hwang_kirshner_morrow_et al._2017, title={Postmastectomy radiotherapy: an american society of clinical oncology, american society for radiation oncology, and society of surgical oncology focused guideline update}, volume={24}, number={1}, journal={Annals of Surgical Oncology}, author={Recht, A. and Comen, E. A. and Fine, R. E. and Fleming, G. F. and Hardenbergh, P. H. and Ho, A. Y. and Hudis, C. A. and Hwang, E. S. and Kirshner, J. J. and Morrow, M. and et al.}, year={2017}, pages={38–51} }
 @misc{ligibel_barry_alfano_hershman_irwin_neuhouser_thomson_delahanty_frank_spears_et al._2017, title={Randomized phase III trial evaluating the role of weight loss in adjuvant treatment of overweight and obese women with early breast cancer (Alliance A011401): study design}, volume={3}, journal={npj Breast Cancer}, author={Ligibel, J. A. and Barry, W. T. and Alfano, C. and Hershman, D. L. and Irwin, M. and Neuhouser, M. and Thomson, C. A. and Delahanty, L. and Frank, E. and Spears, P. and et al.}, year={2017} }
 @article{recht_comen_fine_fleming_hardenbergh_ho_hudis_hwang_kirshner_morrow_et al._2017, title={Regarding current recommendations for postmastectomy radiation therapy in patients with one to three positive axillary lymph nodes reply}, volume={35}, number={11}, journal={Journal of Clinical Oncology}, author={Recht, A. and Comen, E. A. and Fine, R. E. and Fleming, G. F. and Hardenbergh, P. H. and Ho, A. Y. and Hudis, C. A. and Hwang, E. S. and Kirshner, J. J. and Morrow, M. and et al.}, year={2017}, pages={1258–1258} }
 @article{spears_havell_hamrick_goforth_levine_abraham_heiss_azadi_orndorff_2016, title={Listeria monocytogenes wall teichoic acid decoration in virulence and cell-to-cell spread}, volume={101}, ISSN={["1365-2958"]}, DOI={10.1111/mmi.13353}, abstractNote={Wall teichoic acid (WTA) comprises a class of glycopolymers covalently attached to the peptidoglycan of gram positive bacteria. In Listeria monocytogenes, mutations that prevent addition of certain WTA decorating sugars are attenuating. However, the steps required for decoration and the pathogenic process interrupted are not well described. We systematically examined the requirement for WTA galactosylation in a mouse oral-virulent strain by first creating mutations in four genes whose products conferred resistance to a WTA-binding bacteriophage. WTA biochemical and structural studies indicated that galactosylated WTA was directly required for bacteriophage adsorption and that mutant WTA lacked appreciable galactose in all except one mutant - which retained a level ca. 7% of the parent. All mutants were profoundly attenuated in orally infected mice and were impaired in cell-to-cell spread in vitro. Confocal microscopy of cytosolic mutants revealed that all expressed ActA on their cell surface and formed actin tails with a frequency similar to the parent. However, the mutant tails were significantly shorter - suggesting a defect in actin based motility. Roles for the gene products in WTA galactosylation are proposed. Identification and interruption of WTA decoration pathways may provide a general strategy to discover non-antibiotic therapeutics for gram positive infections. © 2016 John Wiley & Sons Ltd.}, number={5}, journal={MOLECULAR MICROBIOLOGY}, author={Spears, Patricia A. and Havell, Edward A. and Hamrick, Terri S. and Goforth, John B. and Levine, Alexandra L. and Abraham, S. Thomas and Heiss, Christian and Azadi, Parastoo and Orndorff, Paul E.}, year={2016}, month={Sep}, pages={714–730} }
 @article{recht_comen_fine_fleming_hardenbergh_ho_hudis_hwang_kirshner_morrow_et al._2016, title={Postmastectomy radiotherapy: An American Society of Clinical Oncology, American Society for Radiation Oncology, and Society of Surgical Oncology focused guideline update}, volume={6}, number={6}, journal={Practical Radiation Oncology}, author={Recht, A. and Comen, E. A. and Fine, R. E. and Fleming, G. F. and Hardenbergh, P. H. and Ho, A. Y. and Hudis, C. A. and Hwang, E. S. and Kirshner, J. J. and Morrow, M. and et al.}, year={2016}, pages={E219–234} }
 @misc{wolff_hammond_hicks_allison_bartlett_bilous_fitzgibbons_hanna_jenkins_mangu_et al._2015, title={National guidelines and level of evidence: Comments on some of the new recommendations in the American Society of Clinical Oncology and the College of American Pathologists human epidermal growth factor receptor 2 guidelines for breast cancer reply}, volume={33}, number={11}, journal={Journal of Clinical Oncology}, author={Wolff, A. C. and Hammond, M. E. H. and Hicks, D. G. and Allison, K. H. and Bartlett, J. M. S. and Bilous, M. and Fitzgibbons, P. and Hanna, W. and Jenkins, R. B. and Mangu, P. B. and et al.}, year={2015}, pages={1302–1304} }
 @article{wolff_hammond_hicks_dowsett_mcshane_allison_allred_bartlett_bilous_fitzgibbons_et al._2014, title={Recommendations for human epidermal growth factor receptor 2 testing in breast cancer American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update}, volume={138}, number={2}, journal={Archives of Pathology & Laboratory Medicine}, author={Wolff, A. C. and Hammond, M. E. H. and Hicks, D. G. and Dowsett, M. and McShane, L. M. and Allison, K. H. and Allred, D. C. and Bartlett, J. M. S. and Bilous, M. and Fitzgibbons, P. and et al.}, year={2014}, pages={241–256} }
 @article{suyemoto_hamrick_spears_horton_washington_havell_borst_orndorff_2013, title={Extrauterine Listeriosis in the gravid mouse influences embryonic growth and development}, volume={8}, number={8}, journal={PLoS One}, author={Suyemoto, M. M. and Hamrick, T. S. and Spears, P. A. and Horton, J. R. and Washington, I. M. and Havell, E. A. and Borst, L. B. and Orndorff, P. E.}, year={2013} }
 @article{wolff_hammond_hicks_dowsett_mcshane_allison_allred_bartlett_bilous_fitzgibbons_et al._2013, title={Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update}, volume={31}, number={31}, journal={Journal of Clinical Oncology}, author={Wolff, A. C. and Hammond, M. E. H. and Hicks, D. G. and Dowsett, M. and McShane, L. M. and Allison, K. H. and Allred, D. C. and Bartlett, J. M. S. and Bilous, M. and Fitzgibbons, P. and et al.}, year={2013}, pages={3997-} }
 @article{spears_suyemoto_hamrick_wolf_havell_orndorff_2011, title={In Vitro Properties of a Listeria monocytogenes Bacteriophage-Resistant Mutant Predict Its Efficacy as a Live Oral Vaccine Strain}, volume={79}, ISSN={["1098-5522"]}, DOI={10.1128/iai.05700-11}, abstractNote={A Listeria monocytogenes glcV mutation precludes the binding of certain listerial phages and produces a profound attenuation characterized by the absence of detectable mutants in the livers and spleens of orally inoculated mice. In vitro, we found that the mutant formed plaques on mouse enterocyte monolayers as efficiently as the parent but the plaques formed were smaller. Intracellular growth rate determinations and examination of infected enterocytes by light and fluorescence microscopy established that the mutant was impaired not in intracellular growth rate but in cell-to-cell spreading. Because this property is shared by other immunogenic mutants (e.g., actA mutants), our glcV mutant was tested for vaccine efficacy. Oral immunization with the mutant and subsequent oral challenge (22 days postvaccination) with the parent revealed a ca. 10,000-fold increase in protection afforded by the mutant compared to sham-vaccinated controls. The glcV mutant did not stimulate innate immunity under the dose and route employed for vaccination, and an infectivity index time course experiment revealed pronounced mutant persistence in Peyer's patches. The immunogenicity of the glcV mutant compared to an isogenic actA mutant reference strain was next tested in an experiment with a challenge given 52 days postvaccination. Both mutant strains showed scant vital organ infectivity and high levels of protection similar to those seen using the glcV mutant in the 22-day postvaccination challenge. Our results indicate that oral administration of a profoundly attenuated listerial mutant can safely elicit solid protective immunity.}, number={12}, journal={INFECTION AND IMMUNITY}, author={Spears, Patricia A. and Suyemoto, M. Mitsu and Hamrick, Terri S. and Wolf, Rebecca L. and Havell, Edward A. and Orndorff, Paul E.}, year={2011}, month={Dec}, pages={5001–5009} }
 @article{lee_wyse_lesher_everett_lou_holzknecht_whitesides_spears_bowles_lin_et al._2010, title={Adaptation in a Mouse Colony Monoassociated with Escherichia coli K-12 for More than 1,000 Days}, volume={76}, ISSN={["0099-2240"]}, DOI={10.1128/aem.00358-10}, abstractNote={Although mice associated with a single bacterial species have been used to provide a simple model for analysis of host-bacteria relationships, bacteria have been shown to display adaptability when grown in a variety of novel environments. In this study, changes associated with the host-bacterium relationship in mice monoassociated with Escherichia coli K-12 over a period of 1,031 days were evaluated. After 80 days, phenotypic diversification of E. coli was observed, with the colonizing bacteria having a broader distribution of growth rates in the laboratory than the parent E. coli. After 1,031 days, which included three generations of mice and an estimated 20,000 generations of E. coli, the initially homogeneous bacteria colonizing the mice had evolved to have widely different growth rates on agar, a potential decrease in tendency for spontaneous lysis in vivo, and an increased tendency for spontaneous lysis in vitro. Importantly, mice at the end of the experiment were colonized at an average density of bacteria that was more than 3-fold greater than mice colonized on day 80. Evaluation of selected isolates on day 1,031 revealed unique restriction endonuclease patterns and differences between isolates in expression of more than 10% of the proteins identified by two-dimensional electrophoresis, suggesting complex changes underlying the evolution of diversity during the experiment. These results suggest that monoassociated mice might be used as a tool for characterizing niches occupied by the intestinal flora and potentially as a method of targeting the evolution of bacteria for applications in biotechnology.}, number={14}, journal={APPLIED AND ENVIRONMENTAL MICROBIOLOGY}, author={Lee, Sean M. and Wyse, Aaron and Lesher, Aaron and Everett, Mary Lou and Lou, Linda and Holzknecht, Zoie E. and Whitesides, John F. and Spears, Patricia A. and Bowles, Dawn E. and Lin, Shu S. and et al.}, year={2010}, month={Jul}, pages={4655–4663} }
 @article{suyemoto_spears_hamrick_barnes_havell_orndorff_2010, title={Factors Associated with the Acquisition and Severity of Gestational Listeriosis}, volume={5}, ISSN={["1932-6203"]}, DOI={10.1371/journal.pone.0013000}, abstractNote={Gravid mammals are more prone to listeriosis than their nongravid counterparts. However, many features of the disease in gravid animals are not well defined. We determined, in mice, that increased susceptibility to lethal infection following oral inoculation begins surprisingly early in pregnancy and extends through embryonic development. Pregnancy did not demonstrably increase the spread of listeriae from the intestine to the liver and spleen in the initial 96 h period post inoculation. Consequently, it appeared that gravid animals were competent to contain an enteric infection, but in those instances where escape did occur, a lethal outcome was more likely. Interestingly, colonic colonization level and prevalence, measured 96 h post inoculation, was significantly higher in gravid individuals. In terms of human risk factors for listeriosis, our results suggest that the window of listeriosis susceptibility afforded by pregnancy may be open longer than previously appreciated. Our results also suggest that while gravid animals are competent to contain an enteric infection, enteric carriage rate may be more of a factor in defining disease incidence than previously considered.}, number={9}, journal={PLOS ONE}, author={Suyemoto, M. Mitsu and Spears, Patricia A. and Hamrick, Terri S. and Barnes, Jill A. and Havell, Edward A. and Orndorff, Paul E.}, year={2010}, month={Sep} }
 @article{temple_miyamoto_mehta_capitini_von stetina_barnes_christensen_horton_spears_orndorff_2010, title={Identification and Characterization of Two Bordetella avium Gene Products Required for Hemagglutination}, volume={78}, ISSN={["1098-5522"]}, DOI={10.1128/iai.00140-10}, abstractNote={ABSTRACT Bordetella avium causes bordetellosis in birds, a disease similar to whooping cough caused by Bordetella pertussis in children. B. avium agglutinates guinea pig erythrocytes via an unknown mechanism. Loss of hemagglutination ability results in attenuation. We report the use of transposon mutagenesis to identify two genes required for hemagglutination. The genes ( hagA and hagB ) were adjacent and divergently oriented and had no orthologs in the genomes of other Bordetella species. Construction of in-frame, unmarked mutations in each gene allowed examination of the role of each in conferring erythrocyte agglutination, explanted tracheal cell adherence, and turkey poult tracheal colonization. In all of the in vitro and in vivo assays, the requirement for the trans -acting products of hagA and hagB (HagA and HagB) was readily shown. Western blotting, using antibodies to purified HagA and HagB, revealed proteins of the predicted sizes of HagA and HagB in an outer membrane-enriched fraction. Antiserum to HagB, but not HagA, blocked B. avium erythrocyte agglutination and explanted turkey tracheal ring binding. Bioinformatic analysis indicated the similarity of HagA and HagB to several two-component secretory apparatuses in which one product facilitates the exposition of the other. HagB has the potential to serve as a useful immunogen to protect turkeys against colonization and subsequent disease.}, number={6}, journal={INFECTION AND IMMUNITY}, author={Temple, Louise M. and Miyamoto, David M. and Mehta, Manju and Capitini, Christian M. and Von Stetina, Stephen and Barnes, H. John and Christensen, Vern L. and Horton, John R. and Spears, Patricia A. and Orndorff, Paul E.}, year={2010}, month={Jun}, pages={2370–2376} }
 @article{spears_suyemoto_palermo_horton_hamrick_havell_orndorff_2008, title={A Listeria monocytogenes mutant defective in bacteriophage attachment is attenuated in orally inoculated mice and impaired in enterocyte intracellular growth}, volume={76}, ISSN={["0019-9567"]}, DOI={10.1128/IAI.00283-08}, abstractNote={A Listeria monocytogenes bacteriophage was used to identify a phage-resistant Tn917 insertion mutant of the mouse-virulent listerial strain F6214-1. The mutant was attenuated when it was inoculated orally into female A/J mice and failed to replicate efficiently in cultured mouse enterocytes. Phage binding studies indicated that the mutant had a cell surface alteration that precluded phage attachment. All phenotypes associated with the mutation could be complemented in trans by a single open reading frame (ORF) that corresponded to the ORF interrupted by the Tn917 insertion. The complementation effected was, in all cases, at a level indistinguishable from that of the parent. The Tn917 insertion interrupted a gene that is predicted to encode a group 2 glycosyl transferase (provisionally designated glcV). A similar glcV gene is present in Listeria welshimeri and Listeria innocua and in some serotypes of L. monocytogenes. We speculate that the loss of the glcV product results in a defective phage receptor and that this alteration coincidentally influences a feature of the normal host-pathogen interaction required for virulence. Interestingly, the glcV lesion, while preventing phage attachment, did not alter the mutant's ability to bind to cultured mouse enterocyte monolayers. Rather, the mutation appeared to alter a subsequent step in intracellular replication measured by a reduction in plaque-forming efficiency and plaque size. In vivo, the mutant was undetectable in the liver and spleen 48 h after oral inoculation. The mutation is significant in part because it is one of the few that produce attenuation when the mutant is delivered orally.}, number={9}, journal={INFECTION AND IMMUNITY}, author={Spears, Patricia A. and Suyemoto, M. Mitsu and Palermo, Angela M. and Horton, John R. and Hamrick, Terri S. and Havell, Edward A. and Orndorff, Paul E.}, year={2008}, month={Sep}, pages={4046–4054} }
 @article{hamrick_horton_spears_havell_smoak_orndorff_2003, title={Influence of pregnancy on the pathogenesis of listeriosis in mice inoculated intragastrically}, volume={71}, ISSN={["1098-5522"]}, DOI={10.1128/IAI.71.9.5202-5209.2003}, abstractNote={Pregnancy increases the risk of listeriosis, a systemic disease caused by Listeria monocytogenes. However, there is incomplete agreement on the reasons for this increased risk. We examined two features of listeriosis in gravid and nongravid female mice following intragastric (gavage) inoculation, namely, (i) disease severity (measured by lethality) and (ii) listerial infectivity (measured by liver and spleen colonization levels up to 120 h postinoculation). Two listerial strains of differing serotype (1/2a and 4nonb) were initially employed. Neither strain produced a lethal infection in nonpregnant female mice (dose range, 10(6) to 10(9) CFU/mouse), and only the 4nonb strain produced lethalities in pregnant mice (dose range, 10(6) to 10(8) CFU/mouse). The 4nonb strain also produced a higher level of liver and spleen colonization than the 1/2a strain following gavage administration. (The two strains showed similar levels of colonization if parenterally administered.) Both strains were equally capable of binding to and forming plaques upon cultured mouse enterocytes. The ability of the 4nonb strain to produce a lethal infection in pregnant animals did not correlate with an increased incidence or level of liver and spleen colonization over that in nonpregnant females. However, the lethality rate did correlate well with the rate at which embryos and their surrounding decidual covering became infected, suggesting that intrauterine infection could be responsible for the increased disease severity in the gravid females.}, number={9}, journal={INFECTION AND IMMUNITY}, author={Hamrick, TS and Horton, JR and Spears, PA and Havell, EA and Smoak, IW and Orndorff, PE}, year={2003}, month={Sep}, pages={5202–5209} }
 @article{valenski_harris_spears_horton_orndorff_2003, title={The product of the fimI gene is necessary for Escherichia coli type 1 pilus biosynthesis}, volume={185}, ISSN={["0021-9193"]}, DOI={10.1128/JB.185.16.5007-5011.2003}, abstractNote={ABSTRACT Site-directed mutagenesis was employed to create lesions in fimI , a gene of uncertain function located in the chromosomal gene cluster ( fim ) involved in Escherichia coli type 1 pilus biosynthesis. Chromosomal fimI mutations produced a piliation-negative phenotype. Complementation analysis indicated that a fimI ′ -kan insertion mutation and a fimI frameshift mutation produced polarity-like effects not seen with an in-frame fimI deletion mutation. Minicell analysis associated fimI with a 16.4-kDa noncytoplasmic protein product (FimI). We conclude that FimI has a required role in normal pilus biosynthesis.}, number={16}, journal={JOURNAL OF BACTERIOLOGY}, author={Valenski, ML and Harris, SL and Spears, PA and Horton, JR and Orndorff, PE}, year={2003}, month={Aug}, pages={5007–5011} }
 @article{spears_temple_miyamoto_maskell_orndorff_2003, title={Unexpected similarities between Bordetella avium and other pathogenic bordetellae}, volume={71}, ISSN={["0019-9567"]}, DOI={10.1128/IAI.71.5.2591-2597.2003}, abstractNote={ABSTRACT Bordetella avium causes an upper respiratory tract disease (bordetellosis) in avian species. Commercially raised turkeys are particularly susceptible. Like other pathogenic members of the genus Bordetella ( B. pertussis and B. bronchiseptica ) that infect mammals, B. avium binds preferentially to ciliated tracheal epithelial cells and produces similar signs of disease. These similarities prompted us to study bordetellosis in turkeys as a possible nonmammalian model for whooping cough, the exclusively human childhood disease caused by B. pertussis . One impediment to accepting such a host-pathogen model as relevant to the human situation is evidence suggesting that B. avium does not express a number of the factors known to be associated with virulence in the other two Bordetella species. Nevertheless, with signature-tagged mutagenesis, four avirulent mutants that had lesions in genes orthologous to those associated with virulence in B. pertussis and B. bronchiseptica ( bvgS , fhaB , fhaC , and fimC ) were identified. None of the four B. avium genes had been previously identified as encoding factors associated with virulence, and three of the insertions (in fhaB , bvgS , and fimC ) were in genes or gene clusters inferred as being absent or incomplete in B. avium , based upon the lack of DNA sequence similarities in hybridization studies and/or the lack of immunological cross-reactivity of the putative products. We further found that the genotypic arrangements of most of the B. avium orthologues were very similar in all three Bordetella species. In vitro tests, including hemagglutination, tracheal ring binding, and serum sensitivity, helped further define the phenotypes conferred by the mutations. Our findings strengthen the connection between the causative agents and the pathogenesis of bordetellosis in all hosts and may help explain the striking similarities of the histopathologic characteristics of this upper airway disease in avian and mammalian species.}, number={5}, journal={INFECTION AND IMMUNITY}, author={Spears, PA and Temple, LM and Miyamoto, DM and Maskell, DJ and Orndorff, PE}, year={2003}, month={May}, pages={2591–2597} }
 @article{spears_2002, title={Breast cancer prevention through the eyes of a survivor}, volume={39}, ISSN={["1098-2280"]}, DOI={10.1002/em.10056}, abstractNote={Environmental and Molecular MutagenesisVolume 39, Issue 2-3 p. 108-111 Meeting Report/PerspectiveFree Access Breast cancer prevention through the eyes of a survivor Patricia A. Spears, Corresponding Author Patricia A. Spears paspears@unity.ncsu.edu Department of Anatomy, Physiological Sciences, and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaNorth Carolina State University, College of Veterinary Medicine, 4700 Hillsborough Street, Raleigh, NC 27606Search for more papers by this author Patricia A. Spears, Corresponding Author Patricia A. Spears paspears@unity.ncsu.edu Department of Anatomy, Physiological Sciences, and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North CarolinaNorth Carolina State University, College of Veterinary Medicine, 4700 Hillsborough Street, Raleigh, NC 27606Search for more papers by this author First published: 29 March 2002 https://doi.org/10.1002/em.10056AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Volume39, Issue2-32002Pages 108-111 ReferencesRelatedInformation}, number={2-3}, journal={ENVIRONMENTAL AND MOLECULAR MUTAGENESIS}, author={Spears, PA}, year={2002}, pages={108–111} }
 @article{harris_spears_havell_hamrick_horton_orndorff_2001, title={Characterization of Escherichia coli type 1 pilus mutants with altered binding specificities}, volume={183}, ISSN={["0021-9193"]}, DOI={10.1128/JB.183.13.4099-4102.2001}, abstractNote={ABSTRACT PCR mutagenesis and a unique enrichment scheme were used to obtain two mutants, each with a single lesion in fimH , the chromosomal gene that encodes the adhesin protein (FimH) of Escherichia coli type 1 pili. These mutants were noteworthy in part because both were altered in the normal range of cell types bound by FimH. One mutation altered an amino acid at a site previously shown to be involved in temperature-dependent binding, and the other altered an amino acid lining the predicted FimH binding pocket.}, number={13}, journal={JOURNAL OF BACTERIOLOGY}, author={Harris, SL and Spears, PA and Havell, EA and Hamrick, TS and Horton, JR and Orndorff, PE}, year={2001}, month={Jul}, pages={4099–4102} }
 @article{spears_temple_orndorff_2000, title={A role for lipopolysaccharide in turkey tracheal colonization by Bordetella avium as demonstrated in vivo and in vitro}, volume={36}, ISSN={["0950-382X"]}, DOI={10.1046/j.1365-2958.2000.01963.x}, abstractNote={We isolated two insertion mutants of Bordetella avium that exhibited a peculiar clumped‐growth phenotype and found them to be attenuated in turkey tracheal colonization. The mutants contained transposon insertions in homologues of the wlbA and wlbL genes of Bordetella pertussis . The wlb genetic locus of B. pertussis has been previously described as containing 12 genes involved in lipopolysaccharide (LPS) biosynthesis. Polyacrylamide gel analysis of LPS from B. avium wlbA and wlbL insertion mutants confirmed an alteration in the LPS profile. Subsequent cloning and complementation of the wlbA and wlbL mutants in trans with a recombinant plasmid containing the homologous wlb locus from B. avium eliminated the clumped‐growth phenotype and restored the LPS profile to that of wild‐type B. avium . Also, a parental level of tracheal colonization was restored to both mutants by the recombinant plasmid. Interestingly, complementation of the wlbA and wlbL mutants with a recombinant plasmid containing the heterologous wlb locus from B. pertussis , B. bronchiseptica, or Bordetella parapertussis eliminated the clumped‐growth phenotype and resulted in a change in the LPS profile, although not to that of wild‐type B. avium . The mutants also acquired resistance to a newly identified B. avium ‐specific bacteriophage, Ba1. Complementation of both wlbA and wlbL mutants with the homologous wlb locus of B. avium , but not the heterologous B. pertussis locus, restored sensitivity to Ba1. Complementation of the wlbL mutant, but not the wlbA mutant, with the heterologous wlb locus of Bordetella bronchiseptica or B. parapertussis restored partial sensitivity to Ba1. Comparisons of the LPS profile and phage sensitivity of the mutants upon complementation by wlb loci from the heterologous species and by B. avium suggested that phage sensitivity required the presence of O‐antigen. At the mechanistic level, both mutants showed a dramatic decrease in serum resistance and a decrease in binding to turkey tracheal rings in vitro . In the case of serum resistance, complementation of both mutants with the homologous wlb locus of B. avium restored serum resistance to wild‐type levels. However, in the case of epithelial cell binding, only complementation of the wlbA mutant completely restored binding to wild‐type levels (binding was only partially restored in the wlbL mutant). This is the first characterization of LPS mutants of B. avium at the genetic level and the first report of virulence changes by both in vivo and in vitro measurements}, number={6}, journal={MOLECULAR MICROBIOLOGY}, author={Spears, PA and Temple, LM and Orndorff, PE}, year={2000}, month={Jun}, pages={1425–1435} }
 @article{hamrick_harris_spears_havell_horton_russell_orndorff_2000, title={Genetic characterization of Escherichia coli type 1 pilus adhesin mutants and identification of a novel binding phenotype}, volume={182}, ISSN={["0021-9193"]}, DOI={10.1128/JB.182.14.4012-4021.2000}, abstractNote={ABSTRACT Five Escherichia coli type 1 pilus mutants that had point mutations in fimH , the gene encoding the type 1 pilus adhesin FimH, were characterized. FimH is a minor component of type 1 pili that is required for the pili to bind and agglutinate guinea pig erythrocytes in a mannose-inhibitable manner. Point mutations were located by DNA sequencing and deletion mapping. All mutations mapped within the signal sequence or in the first 28% of the predicted mature protein. All mutations were missense mutations except for one, a frameshift lesion that was predicted to cause the loss of approximately 60% of the mature FimH protein. Bacterial agglutination tests with polyclonal antiserum raised to a LacZ-FimH fusion protein failed to confirm that parental amounts of FimH cross-reacting material were expressed in four of the five mutants. The remaining mutant, a temperature-sensitive (ts) fimH mutant that agglutinated guinea pig erythrocytes after growth at 31°C but not at 42°C, reacted with antiserum at both temperatures in a manner similar to the parent. Consequently, this mutant was chosen for further study. Temperature shift experiments revealed that new FimH biosynthesis was required for the phenotypic change. Guinea pig erythrocyte and mouse macrophage binding experiments using the ts mutant grown at the restrictive and permissive temperatures revealed that whereas erythrocyte binding was reduced to a level comparable to that of a fimH insertion mutant at the restrictive temperature, mouse peritoneal macrophages were bound with parental efficiency at both the permissive and restrictive temperatures. Also, macrophage binding by the ts mutant was insensitive to mannose inhibition after growth at 42°C but sensitive after growth at 31°C. The ts mutant thus binds macrophages with one receptor specificity at 31°C and another at 42°C.}, number={14}, journal={JOURNAL OF BACTERIOLOGY}, author={Hamrick, TS and Harris, SL and Spears, PA and Havell, EA and Horton, JR and Russell, PW and Orndorff, PE}, year={2000}, month={Jul}, pages={4012–4021} }