Publications by authors named "Sara Smith"

116 Publications

Latent Class Analysis Reveals Distinct Groups Based on Executive Function and Socioemotional Traits, Developmental Conditions, and Stuttering: A Population Study.

Child Psychiatry Hum Dev 2021 Mar 30. Epub 2021 Mar 30.

Rightpath Research and Innovation Center, University of South Florida, 13301 Bruce B. Downs Blvd., MHC 1721, Tampa, FL, 33602, USA.

A growing body of research has reported associations between weaker Executive Functions (EF), the set capacities that are needed to manage and allocate one's cognitive resources during cognitively challenging activities and various neurodevelopmental conditions, including stuttering. The majority of this research has been based on variable-centered approaches, which have the potential to obscure within-population heterogeneity. Person-centered analyses are essential to understanding multifactorial disorders where relationships between indicators have been elusive, such as stuttering. The current study addressed gaps in the literature by using latent class analysis (LCA), a person-centered approach, to identify homogenous subgroups within the National Health Interview Survey (2004-2018) publicly available data set. Using this exploratory approach, we examined the hypothesis that there exist distinct classes (or subgroups) of children based on parent reports of EF, Socioemotional (SE) traits, developmental atypicality, and stuttering. Our analyses revealed distinct subgroups with substantially different likelihoods of parent-reported stuttering behaviors and developmental atypicality. For children with both EF and SE difficulties, the likelihood of parental report of stuttering and atypical development was even higher, in fact this likelihood (of stuttering and not-typically developing) was highest among all subgroups. In contrast, children without difficulties were the least likely to be reported with stuttering or not-typically developing. Our findings are consistent with theoretical frameworks for stuttering, which cite EF as a crucial component in the disorder. Additionally, our findings suggest within-population heterogeneity among children with EF difficulties and, specifically, EF and SE heterogeneity among children who stutter.
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http://dx.doi.org/10.1007/s10578-021-01160-3DOI Listing
March 2021

A systematic analysis of hypermucoviscosity and capsule reveals distinct and overlapping genes that impact Klebsiella pneumoniae fitness.

PLoS Pathog 2021 Mar 15;17(3):e1009376. Epub 2021 Mar 15.

Department of Microbiology & Immunology, University of Michigan, Ann Arbor, Michigan, United States of America.

Hypervirulent K. pneumoniae (hvKp) is a distinct pathotype that causes invasive community-acquired infections in healthy individuals. Hypermucoviscosity (hmv) is a major phenotype associated with hvKp characterized by copious capsule production and poor sedimentation. Dissecting the individual functions of CPS production and hmv in hvKp has been hindered by the conflation of these two properties. Although hmv requires capsular polysaccharide (CPS) biosynthesis, other cellular factors may also be required and some fitness phenotypes ascribed to CPS may be distinctly attributed to hmv. To address this challenge, we systematically identified genes that impact capsule and hmv. We generated a condensed, ordered transposon library in hypervirulent strain KPPR1, then evaluated the CPS production and hmv phenotypes of the 3,733 transposon mutants, representing 72% of all open reading frames in the genome. We employed forward and reverse genetic screens to evaluate effects of novel and known genes on CPS biosynthesis and hmv. These screens expand our understanding of core genes that coordinate CPS biosynthesis and hmv, as well as identify central metabolism genes that distinctly impact CPS biosynthesis or hmv, specifically those related to purine metabolism, pyruvate metabolism and the TCA cycle. Six representative mutants, with varying effect on CPS biosynthesis and hmv, were evaluated for their impact on CPS thickness, serum resistance, host cell association, and fitness in a murine model of disseminating pneumonia. Altogether, these data demonstrate that hmv requires both CPS biosynthesis and other cellular factors, and that hmv and CPS may serve distinct functions during pathogenesis. The integration of hmv and CPS to the metabolic status of the cell suggests that hvKp may require certain nutrients to specifically cause deep tissue infections.
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http://dx.doi.org/10.1371/journal.ppat.1009376DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7993769PMC
March 2021

Increasing Median Time between Interruptions in a Busy Reading Room.

Radiographics 2021 Mar-Apr;41(2):E47-E56

From the Department of Radiology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Ave, MLC 5031, Cincinnati, OH 45229-3026 (E.A.S., A.H.S., R.S., S.E.O., S.N.S., A.L.E., A.J.T.); and Department of Radiology, University of Cincinnati College of Medicine, Cincinnati, Ohio (E.A.S., A.H.S., A.J.T.).

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http://dx.doi.org/10.1148/rg.2021200094DOI Listing
March 2021

Affecting Length of Stay in Well-appearing Febrile Infants.

Pediatr Qual Saf 2020 Nov-Dec;5(6):e359. Epub 2020 Oct 23.

Department of Pediatric and Adolescent Medicine, University of Illinois Hospital and Health Sciences System, Chicago, Ill.

The management of infants under 2 months of age presenting with fever has perplexed pediatricians for decades. The University of Illinois at Chicago was selected as one of the primary sites for the REVISE (Reducing Variation in Infant Sepsis Evaluations) study through the American Academy of Pediatrics. Our primary objective was to decrease the length of stay (LOS) for well-appearing febrile infants by 20% over 8 months from December 2016 to August 2017.

Methods: We introduced the use of a decision support smartphone application to providers caring for febrile infants. Monthly retrospective chart review of patients 7-59 days old with fever seen in the emergency department or the inpatient setting was performed from September 2015 to August 2016 for baseline data, from December 2016 to August 2017 for intervention data, and from September 2017 to December 2018 for surveillance data.

Results: A total of 1013 patients of ages 7-59 days seen in the emergency department or inpatient unit between September 2015 to December 2018 were screened for study inclusion. Forty-one febrile, well-appearing infants of ages 7-59 days met inclusion criteria. During the baseline period, there was a mean LOS of 48 hours. Intervention and surveillance data did not change the mean from baseline.

Conclusions: Infants with a negative diagnostic evaluation for urinary tract infection, bacteremia, or meningitis drove our LOS. Further study is needed to affect the LOS in febrile infants with diagnoses of urinary tract infection, bacteremia, or meningitis.
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http://dx.doi.org/10.1097/pq9.0000000000000359DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7870218PMC
October 2020

Phonetic discrimination, phonological awareness, and pre-literacy skills in Spanish-English dual language preschoolers.

J Child Lang 2021 Feb 11:1-34. Epub 2021 Feb 11.

McGill University, Canada.

The current study explores variation in phonemic representation among Spanish-English dual language learners (DLLs, n = 60) who were dominant in English or in Spanish. Children were given a phonetic discrimination task with speech sounds that: 1) occur in English and Spanish, 2) are exclusive to English, and 3) are exclusive to Russian, during Fall (age m = 57 months) and Spring (age m = 62 months, n = 42). In Fall, English-dominant DLLs discriminated more accurately than Spanish-dominant DLLs between English-Spanish phones and English-exclusive phones. Both groups discriminated Russian phones at or close to chance. In Spring, however, groups no longer differed in discriminating English-exclusive phones and both groups discriminated Russian phones above chance. Additionally, joint English-Spanish and English-exclusive phonetic discrimination predicted children's phonological awareness in both groups. Results demonstrate plasticity in early childhood through diverse language exposure and suggest that phonemic representation begins to emerge driven by lexical restructuring.
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http://dx.doi.org/10.1017/S0305000920000768DOI Listing
February 2021

Associations between stuttering, comorbid conditions and executive function in children: a population-based study.

BMC Psychol 2020 Oct 31;8(1):113. Epub 2020 Oct 31.

Department of Educational Policy Studies, Georgia State University, 30 Pryor St SW, Atlanta, GA, 30303, USA.

Background: The aim of this study was to investigate the relationship between executive function (EF), stuttering, and comorbidity by examining children who stutter (CWS) and children who do not stutter (CWNS) with and without comorbid conditions. Data from the National Health Interview Survey were used to examine behavioral manifestations of EF, such as inattention and self-regulation, in CWS and CWNS.

Methods: The sample included 2258 CWS (girls = 638, boys = 1620), and 117,725 CWNS (girls = 57,512; boys = 60,213). EF, and the presence of stuttering and comorbid conditions were based on parent report. Descriptive statistics were used to describe the distribution of stuttering and comorbidity across group and sex. Regression analyses were to determine the effects of stuttering and comorbidity on EF, and the relationship between EF and socioemotional competence.

Results: Results point to weaker EF in CWS compared to CWNS. Also, having comorbid conditions was also associated with weaker EF. CWS with comorbidity showed the weakest EF compared to CWNS with and without comorbidity, and CWS without comorbidity. Children with stronger EF showed higher socioemotional competence. A majority (60.32%) of CWS had at least one other comorbid condition in addition to stuttering. Boys who stutter were more likely to have comorbid conditions compared to girls who stutter.

Conclusion: Present findings suggest that comorbidity is a common feature in CWS. Stuttering and comorbid conditions negatively impact EF.
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http://dx.doi.org/10.1186/s40359-020-00481-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7603732PMC
October 2020

The Gene Expression Profile of Uropathogenic Escherichia coli in Women with Uncomplicated Urinary Tract Infections Is Recapitulated in the Mouse Model.

mBio 2020 08 11;11(4). Epub 2020 Aug 11.

Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan, USA

Uropathogenic (UPEC) is the primary causative agent of uncomplicated urinary tract infections (UTIs). UPEC fitness and virulence determinants have been evaluated in a variety of laboratory settings, including a well-established mouse model of UTI. However, the extent to which bacterial physiologies differ between experimental models and human infections remains largely understudied. To address this important issue, we compared the transcriptomes of three different UPEC isolates in human infection and under a variety of laboratory conditions, including LB culture, filter-sterilized urine culture, and the UTI mouse model. We observed high correlation in gene expression between the mouse model and human infection in all three strains examined (Pearson correlation coefficients of 0.86 to 0.87). Only 175 of 3,266 (5.4%) genes shared by all three strains had significantly different expression levels, with the majority of them (145 genes) downregulated in patients. Importantly, gene expression levels of both canonical virulence factors and metabolic machinery were highly similar between the mouse model and human infection, while the conditions displayed more substantial differences. Interestingly, comparison of gene expression between the mouse model and human infection hinted at differences in bladder oxygenation as well as nutrient composition. In summary, our work strongly validates the continued use of this mouse model for the study of the pathogenesis of human UTI. Different experimental models have been used to study UPEC pathogenesis, including cultures in different media, tissue culture, and mouse models of infection. The last is especially important since it allows evaluation of mechanisms of pathogenesis and potential therapeutic strategies against UPEC. Bacterial physiology is greatly shaped by environment, and it is therefore critical to understand how closely bacterial physiology in any experimental model relates to human infection. In this study, we found strong correlation in bacterial gene expression between the mouse model and human UTI using identical strains, suggesting that the mouse model accurately mimics human infection, definitively supporting its continued use in UTI research.
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http://dx.doi.org/10.1128/mBio.01412-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7439467PMC
August 2020

Overcoming barriers to adolescent vaccination: perspectives from vaccine providers in North Carolina.

Women Health 2020 Nov-Dec;60(10):1129-1140. Epub 2020 Aug 9.

Department of Epidemiology, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina, USA.

To capture strategies for achieving high adolescent coverage of tetanus-diphtheria-acellular pertussis (Tdap), meningococcal conjugate (MenACWY), and human papillomavirus (HPV) vaccination, we surveyed employees of 20 North Carolina (N.C.) clinics that achieved adolescent vaccination coverage higher than the state average. One employee per clinic completed a surveysummarizing clinic practices regarding adolescent vaccination; perceived barriers and facilitators to Tdap/MenACWY/HPV vaccination; and the role of "champions" who made special efforts to promote adolescent vaccination. Common perceived barriers for all vaccinations were parental opposition and logistical barriers to receiving vaccination. For HPV vaccination, employees cited parental concerns about sexual behavior and injection site pain; no school vaccination requirement; and low-perceived benefit in boys. Most clinics (80%) implemented successful changes to increase adolescent vaccination: consistently offering vaccination, tracking vaccination status using existing data, providing appointment reminders, updating providers on vaccination recommendations, and expanding vaccination hours. Strategies to improve HPV vaccination included co-administration with Tdap and MenACWY, and providing reminders to complete the vaccination series. Vaccine champions strongly recommended vaccination to parents (55%) and educated parents on vaccination recommendations (36%). Clinics in N.C.and similar settings can implement these and other low-resource strategies to overcome adolescent vaccination barriers.

Abbreviations: CDC=Centers for Disease Control and Prevention; EHR=Electronic health record; HPV=Human papillomavirus; Tdap=Tetanus-diphtheria-acellular pertussis vaccine; MenACWY=Meningococcal Conjugate Vaccine; NCIB=North Carolina Immunization Branch; NCIR=North Carolina Immunization Registry; ACIP=Advisory Committee on Immunization Practices.
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http://dx.doi.org/10.1080/03630242.2020.1802639DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7556355PMC
March 2021

Pediatric Hepatitis C.

Adv Pediatr 2020 08 15;67:47-56. Epub 2020 May 15.

Department of Pediatric Gastroenterology, Hepatology, and Nutrition, University of California, San Francisco, 550 16th Street, 5th Floor, Mail Code 0136, San Francisco, CA 94158, USA.

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http://dx.doi.org/10.1016/j.yapd.2020.03.008DOI Listing
August 2020

Transposon Insertion Site Sequencing of Providencia stuartii: Essential Genes, Fitness Factors for Catheter-Associated Urinary Tract Infection, and the Impact of Polymicrobial Infection on Fitness Requirements.

mSphere 2020 05 27;5(3). Epub 2020 May 27.

Department of Microbiology and Immunology, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, New York, USA

is a common cause of polymicrobial catheter-associated urinary tract infection (CAUTI), and yet literature describing the molecular mechanisms of its pathogenesis is limited. To identify factors important for colonization during single-species infection and during polymicrobial infection with a common cocolonizer, , we created a saturating library of ∼50,000 transposon mutants and conducted transposon insertion site sequencing (Tn-Seq) in a murine model of CAUTI. strain BE2467 carries 4,398 genes, 521 of which were identified as essential for growth in laboratory medium and therefore could not be assessed for contribution to infection. Using an input/output fold change cutoff value of 20 and values of <0.05, 340 genes were identified as important for establishing single-species infection only and 63 genes as uniquely important for polymicrobial infection with , and 168 genes contributed to both single-species and coinfection. Seven mutants were constructed for experimental validation of the primary screen that corresponded to flagella ( mutant), twin arginine translocation (), an ATP-dependent protease (), d-alanine-d-alanine ligase (), type 3 secretion ( and ), and type VI secretion (). Infection-specific phenotypes validated 6/7 (86%) mutants during direct cochallenge with wild-type and 3/5 (60%) mutants during coinfection with , for a combined validation rate of 9/12 (75%). Tn-Seq therefore successfully identified genes that contribute to fitness of within the urinary tract, determined the impact of coinfection on fitness requirements, and added to the identification of a collection of genes that may contribute to fitness of multiple urinary tract pathogens. is a common cause of polymicrobial catheter-associated urinary tract infections (CAUTIs), particularly during long-term catheterization. However, little is known regarding the pathogenesis of this organism. Using transposon insertion site sequencing (Tn-Seq), we performed a global assessment of fitness factors for CAUTI while simultaneously determining how coinfection with another pathogen alters fitness requirements. This approach provides four important contributions to the field: (i) the first global estimation of genes essential for growth in laboratory medium, (ii) identification of novel fitness factors for colonization of the catheterized urinary tract, (iii) identification of core fitness factors for both single-species and polymicrobial CAUTI, and (iv) assessment of conservation of fitness factors between common uropathogens. Genomewide assessment of the fitness requirements for common uropathogens during single-species and polymicrobial CAUTI thus elucidates complex interactions that contribute to disease severity and will uncover conserved targets for therapeutic intervention.
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http://dx.doi.org/10.1128/mSphere.00412-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7253602PMC
May 2020

The Serratia marcescens Siderophore Serratiochelin Is Necessary for Full Virulence during Bloodstream Infection.

Infect Immun 2020 07 21;88(8). Epub 2020 Jul 21.

University of Michigan Medical School, Department of Microbiology and Immunology, Ann Arbor, Michigan, USA

is a bacterium frequently found in the environment, but over the last several decades it has evolved into a concerning clinical pathogen, causing fatal bacteremia. To establish such infections, pathogens require specific nutrients; one very limited but essential nutrient is iron. We sought to characterize the iron acquisition systems in isolate UMH9, which was recovered from a clinical bloodstream infection. Using RNA sequencing (RNA-seq), we identified two predicted siderophore gene clusters ( and ) that were regulated by iron. Mutants were constructed to delete each iron acquisition locus individually and in conjunction, generating both single and double mutants for the putative siderophore systems. Mutants lacking the gene cluster lost their iron-chelating ability as quantified by the chrome azurol S (CAS) assay, whereas the mutant retained wild-type activity. Mass spectrometry-based analysis identified the chelating siderophore to be serratiochelin, a siderophore previously identified in Serratiochelin-producing mutants also displayed a decreased growth rate under iron-limited conditions created by dipyridyl added to LB medium. Additionally, mutants lacking serratiochelin were significantly outcompeted during cochallenge with wild-type UMH9 in the kidneys and spleen after inoculation via the tail vein in a bacteremia mouse model. This result was further confirmed by an independent challenge, suggesting that serratiochelin is required for full pathogenesis in the bloodstream. Nine other clinical isolates have at least 90% protein identity to the UMH9 serratiochelin system; therefore, our results are broadly applicable to emerging clinical isolates of causing bacteremia.
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http://dx.doi.org/10.1128/IAI.00117-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7375758PMC
July 2020

Conjunctival staining with lissamine green as a predictor of tear film deficiency in dogs.

Vet Ophthalmol 2020 Jul 9;23(4):624-631. Epub 2020 May 9.

Section of Ophthalmology, Department of Clinical Sciences & Advanced Medicine, University of Pennsylvania School of Veterinary Medicine, Philadelphia, Pennsylvania.

Objective: To evaluate a grading scheme for conjunctival staining patterns with lissamine green ocular dye in the diagnosis of tear film deficiencies in dogs.

Procedures: Client-owned and research colony dogs were enrolled in a prospective study between February and October 2018 in which slit-lamp biomicroscopy, Schirmer tear test (STT), tear film breakup time (TFBUT), conjunctival lissamine green staining (LGS), and intraocular pressure (IOP) measurement were performed in both eyes of all dogs. Lissamine green staining of the temporal bulbar conjunctiva was graded from 0-3, with a higher grade corresponding to an increased stain intensity.

Results: Fifty-four dogs (107 eyes), comprising 31 males and 23 females with a mean age of 5.0 ± 3.9 years (range 0.5-14.3), were enrolled in the study. STT was <15 mm/min in 21 eyes and ≥15 mm/min in 86 eyes. Lissamine green staining grade for eyes with a STT of <15 mm/min (2.0 ± 0.9) was significantly higher than for eyes with a STT ≥15 mm/min (0.2 ± 0.7) (P < .001). TFBUT for eyes with a STT <15 mm/min (6.5 ± 4.4 seconds) was significantly shorter than for eyes with a STT ≥ 15 mm/min (16.1 ± 3.6 seconds) (P < .001). As LGS grade increased, both STT (P < .001) and TFBUT (P < .001) significantly decreased.

Conclusions: A higher LGS grade was significantly associated with a lower STT and more rapid TFBUT in dogs. Lissamine green ocular dye can be considered as an adjunctive diagnostic test when evaluating tear film deficiency in dogs.
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http://dx.doi.org/10.1111/vop.12762DOI Listing
July 2020

Achieving Congruence among Reference Laboratories for Absolute Abundance Measurement of Analytes for Rare Diseases: Psychosine for Diagnosis and Prognosis of Krabbe Disease.

Int J Neonatal Screen 2020 Jun 31;6(2). Epub 2020 Mar 31.

Departments of Chemistry and Biochemistry, University of Washington, Seattle, WA 98195, USA.

Measurement of the absolute concentration of the biomarker psychosine in dried blood spots (DBS) is useful for diagnosis and prognosis of Krabbe disease and to support newborn screening of this leukodystrophy. As for assays for more common diseases, it is important to achieve congruence when multiple clinical laboratories provide testing. Four clinical laboratories, one research laboratory, and a commercial vendor collaborated with the goal to achieve congruence in quantitative psychosine measurement in DBS. A set of DBS calibrators was prepared by a single vendor and used in each reference laboratory to make a standard curve of measured psychosine in DBS versus the stated calibrator psychosine level. Congruence between the participating five laboratories was achieved using the psychosine DBS calibrators. This allowed application of disease-specific reference ranges obtained by the reference laboratory with the most extensive data by the other reference laboratories. Congruence between multiple reference laboratories in the measurement of the absolute concentration of biomarkers in DBS (and by extension other samples) is possible by the use of a common set of DBS calibrators.
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http://dx.doi.org/10.3390/ijns6020029DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7199469PMC
June 2020

Escherichia coli CFT073 Fitness Factors during Urinary Tract Infection: Identification Using an Ordered Transposon Library.

Appl Environ Microbiol 2020 06 17;86(13). Epub 2020 Jun 17.

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, USA

Urinary tract infections (UTI), the second most diagnosed infectious disease worldwide, are caused primarily by uropathogenic (UPEC), placing a significant financial burden on the health care system. High-throughput transposon mutagenesis combined with genome-targeted sequencing is a powerful technique to interrogate genomes for fitness genes. Genome-wide analysis of requires random libraries of at least 50,000 mutants to achieve 99.99% saturation; however, the traditional murine model of ascending UTI does not permit testing of large mutant pools due to a bottleneck during infection. To address this, an CFT073 transposon mutant ordered library of 9,216 mutants was created and insertion sites were identified. A single transposon mutant was selected for each gene to assemble a condensed library consisting of 2,913 unique nonessential mutants. Using a modified UTI model in BALB/c mice, we identified 36 genes important for colonizing the bladder, including , , and Screening of the condensed library identified and to be essential for growth in human urine. Additionally, we developed a novel quantitative PCR (qPCR) technique to identify genes with fitness defects within defined subgroups of related genes (e.g., genes encoding fimbriae, toxins, etc.) following UTI. The number of mutants within these subgroups circumvents bottleneck restriction and facilitates validation of multiple mutants to generate individual competitive indices. Collectively, this study investigates the bottleneck effects during UTI, provides two techniques for evading those effects that can be applied to other disease models, and contributes a genetic tool in prototype strain CFT073 to the field. Uropathogenic strains cause most uncomplicated urinary tract infections (UTI), one of the most common infectious diseases worldwide. Random transposon mutagenesis techniques have been utilized to identify essential bacterial genes during infection; however, this has been met with limitations when applied to the murine UTI model. Conventional high-throughput transposon mutagenesis screens are not feasible because of inoculum size restrictions due to a bottleneck during infection. Our study utilizes a condensed ordered transposon library, limiting the number of mutants while maintaining the largest possible genome coverage. Screening of this library , and in human urine , identified numerous candidate fitness factors. Additionally, we have developed a novel technique using qPCR to quantify bacterial outputs following infection with small subgroups of transposon mutants. Molecular approaches developed in this study will serve as useful tools to probe models that are restricted by anatomical, physiological, or genetic bottleneck limitations.
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http://dx.doi.org/10.1128/AEM.00691-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7301846PMC
June 2020

Optimization of an Experimental Vaccine To Prevent Escherichia coli Urinary Tract Infection.

mBio 2020 04 28;11(2). Epub 2020 Apr 28.

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, USA

Urinary tract infections (UTI) affect half of all women at least once during their lifetime. The rise in the numbers of extended-spectrum beta-lactamase-producing strains and the potential for carbapenem resistance within uropathogenic (UPEC), the most common causative agent of UTI, create an urgent need for vaccine development. Intranasal immunization of mice with UPEC outer membrane iron receptors FyuA, Hma, IreA, and IutA, conjugated to cholera toxin, provides protection in the bladder or kidneys under conditions of challenge with UPEC strain CFT073 or strain 536. On the basis of these data, we sought to optimize the vaccination route (intramuscular, intranasal, or subcutaneous) in combination with adjuvants suitable for human use, including aluminum hydroxide gel (alum), monophosphoryl lipid A (MPLA), unmethylated CpG synthetic oligodeoxynucleotides (CpG), polyinosinic:polycytidylic acid (polyIC), and mutated heat-labile enterotoxin (dmLT). Mice intranasally vaccinated with dmLT-IutA and dmLT-Hma displayed significant reductions in bladder colonization (86-fold and 32-fold, respectively), with 40% to 42% of mice having no detectable CFU. Intranasal vaccination of mice with CpG-IutA and polyIC-IutA significantly reduced kidney colonization (131-fold) and urine CFU (22-fold), respectively. dmLT generated the most consistently robust antibody response in intranasally immunized mice, while MPLA and alum produced greater concentrations of antigen-specific serum IgG with intramuscular immunization. On the basis of these results, we conclude that intranasal administration of Hma or IutA formulated with dmLT adjuvant provides the greatest protection from UPEC UTI. This report advances our progress toward a vaccine against uncomplicated UTI, which will significantly improve the quality of life for women burdened by recurrent UTI and enable better antibiotic stewardship. Urinary tract infections (UTI) are among the most common bacterial infection in humans, affecting half of all women at least once during their lifetimes. The rise in antibiotic resistance and health care costs emphasizes the need to develop a vaccine against the most common UTI pathogen, Vaccinating mice intranasally with a detoxified heat-labile enterotoxin and two surface-exposed receptors, Hma or IutA, significantly reduced bacterial burden in the bladder. This work highlights progress in the development of a UTI vaccine formulated with adjuvants suitable for human use and antigens that encode outer membrane iron receptors required for infection in the iron-limited urinary tract.
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http://dx.doi.org/10.1128/mBio.00555-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7188996PMC
April 2020

Role of school nurses in identifying and supporting children and young people with mental health issues.

Nurs Child Young People 2020 03 11;32(2):23-28. Epub 2020 Feb 11.

School of Health and Social Care, Bournemouth University, Bournemouth, England.

School nurses have an important role in the provision of mental health services because of their expertise in healthcare and education. The aim of this literature review was to explore research about school nurses' ability to identify and support children and young people in secondary education with mental health issues. A search of healthcare-related databases was undertaken using search terms such as 'specialist community public health nurse' (SCPHN), 'school nurses', 'young people', mental health' and 'adolescent mental health' to identify relevant research. The literature review found that school nurses perform various activities for children and young people, for example promoting optimal mental health, identifying concerns and initiating early interventions. However, the literature review also suggests that unless school nurses receive further education in mental health they will be unable to develop the necessary skills required to improve outcomes for children and young people in secondary education.
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http://dx.doi.org/10.7748/ncyp.2020.e1253DOI Listing
March 2020

College students' disclosure of mental health problems on campus.

J Am Coll Health 2020 Jan 16:1-8. Epub 2020 Jan 16.

Department of Teaching and Learning, University of South Florida, Tampa, FL, USA.

To explore, in two studies, student experiences with disclosing diagnosed and undiagnosed mental health problems to faculty, staff, and peers. Participants for both studies were college students at two 4-year public universities. Study 1 was conducted in June 2016 and Study 2 in August 2017. In Study 1, participants had a formal diagnosis of a mental health condition, were registered for accommodations, and completed semi-structured interviews ( = 6). In Study 2, participants ( = 66) were mostly non-diagnosed and completed an online survey. Study 1 interviews were coded for interactions with faculty (52% positive, 40% negative), staff (100% positive), and peers (31% positive, 54% negative). In Study 2, participants reported better experiences disclosing to peers than faculty. Disclosure experience varied by type of mental health problem. Collaboration across campus and community agencies will better support students with diagnosed and undiagnosed mental health problems.
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http://dx.doi.org/10.1080/07448481.2019.1706533DOI Listing
January 2020

Bilingual children who stutter: Convergence, gaps and directions for research.

J Fluency Disord 2020 03 9;63:105741. Epub 2019 Dec 9.

Department of Teaching and Learning, University of South Florida, Tampa, FL 33620, United States.

Purpose: The aim of this systematic review is to examine the early interactions between bilingualism and stuttering to synthesize knowledge that could inform diagnosis and treatment for bilingual children who stutter.

Method: Scopus, Science Direct, PubMed, ERIC Ebsco, and Google Scholar were searched with no limits placed on the year of publication. Search terms consisted of: ("stuttering" [MeSH] OR "stutter") AND ("child" [MeSH] OR "children") AND ("multilingualism" [MeSH] OR "bilingualism"). Inclusion criteria were children who stutter, bilinguals who stutter, empirical research articles, and published in peer review journals. Exclusion criteria were studies that reported on only adults, only monolinguals, or were not published in English.

Results: A total of 50 articles met the criteria. There was convergence with monolingual studies reporting sexually dimorphic and familial trends in the prevalence of stuttering and rates of recovery. Findings surrounding language proficiency, cross-linguistic stuttering severity, and development were ambivalent. Results point to the difficulty in identifying stuttering in bilingual children, and the need for culturally competent research and interpretations.

Conclusion: Current findings offer a fragmented view of bilingual development and echoes a recurring theme, i.e., the current understanding of bilingualism and stuttering is limited and more research is warranted.
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http://dx.doi.org/10.1016/j.jfludis.2019.105741DOI Listing
March 2020

Hydrogen peroxide triggers an increase in cell surface expression of system x in cultured human glioma cells.

Neurochem Int 2020 03 23;134:104648. Epub 2019 Dec 23.

Department of Neurology, Department of Neuroscience, Johns Hopkins University, Baltimore, MD, 21287, USA.

System x exchanges extracellular cystine for intracellular glutamate across the plasma membrane of many cell types. One of the physiological roles of System x is to provide cystine for synthesis of the antioxidant glutathione. Here we report that hydrogen peroxide (HO) triggers the translocation of System x to the plasma membrane within 10 min of the initial exposure. Specifically, we observed a three-fold increase in S-l-cystine uptake following a 10 min exposure to 0.3 mM HO. This effect was dose-dependent with an EC for HO of 65 μM. We then used cell surface biotinylation analysis to test the hypothesis that the increase in activity is due to an increased number of transporters on the plasma membrane. We demonstrated that the amount of transporter protein, xCT, localized to the plasma membrane doubles within 10 min of HO exposure as a result of an increase in its delivery rate and a reduction in its internalization rate. In addition, we demonstrated that HO triggered a rapid decrease in total cellular glutathione which recovered within 2 h of the oxidative insult. The kinetics of glutathione recovery matched the time course for the recovery of xCT cell surface expression and System x activity following removal of the oxidative insult. Collectively, these results suggest that oxidants acutely modulate the activity of System x by increasing its cell surface expression, and that this process may serve as an important mechanism to increase de novo glutathione synthesis during periods of oxidative stress.
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http://dx.doi.org/10.1016/j.neuint.2019.104648DOI Listing
March 2020

Genetically diverse uropathogenic adopt a common transcriptional program in patients with UTIs.

Elife 2019 10 21;8. Epub 2019 Oct 21.

Department of Microbiology and Immunology, University of Michigan, Ann Arbor, United States.

Uropathogenic (UPEC) is the major causative agent of uncomplicated urinary tract infections (UTIs). A common virulence genotype of UPEC strains responsible for UTIs is yet to be defined, due to the large variation of virulence factors observed in UPEC strains. We hypothesized that studying UPEC functional responses in patients might reveal universal UPEC features that enable pathogenesis. Here we identify a transcriptional program shared by genetically diverse UPEC strains isolated from 14 patients during uncomplicated UTIs. Strikingly, this in vivo gene expression program is marked by upregulation of translational machinery, providing a mechanism for the rapid growth within the host. Our analysis indicates that switching to a more specialized catabolism and scavenging lifestyle in the host allows for the increased translational output. Our study identifies a common transcriptional program underlying UTIs and illuminates the molecular underpinnings that likely facilitate the fast growth rate of UPEC in infected patients.
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http://dx.doi.org/10.7554/eLife.49748DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6802966PMC
October 2019

Communication skills: placement reflections of a children's nursing student.

Nurs Child Young People 2018 Dec 24. Epub 2018 Dec 24.

Children and young people's nursing, Faculty of Health and Social Sciences, Bournemouth University, Dorset, England.

Reflection is a natural human activity. It is important for nursing students during practice placements when they are making sense of clinical surroundings and making decisions about care. This article uses a reflective framework to consider the placement experiences of a third-year undergraduate children's nursing student in an acute general setting, caring for an adolescent with anorexia. It focuses on the decisions made when assisting an inexperienced junior doctor during a venepuncture procedure. Reflecting on placements provides nursing students with important transferable skills to use in practice as registered nurses.
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http://dx.doi.org/10.7748/ncyp.2018.e1112DOI Listing
December 2018

Vaccination to Protect Against Proteus mirabilis Challenge Utilizing the Ascending Model of Urinary Tract Infection.

Methods Mol Biol 2019 ;2021:201-215

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA.

Proteus mirabilis is a major cause of complicated urinary tract infections (UTIs). P. mirabilis' urease activity hydrolyzes urea and raises urine pH levels, which can catalyze bladder and kidney stone formation. This urolithiasis leads to harder-to-treat infections, possible urinary blockage, and subsequent septicemia. Development of a mucosal vaccine against P. mirabilis urinary tract infections is critical to protect against this potentially deadly infection process. Here, we describe the methodology necessary to produce a vaccine candidate conjugated to cholera toxin, administer the vaccine via the intranasal route, and test efficacy in a murine transurethral P. mirabilis infection model.
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http://dx.doi.org/10.1007/978-1-4939-9601-8_18DOI Listing
March 2020

Indwelling Urinary Catheter Model of Proteus mirabilis Infection.

Methods Mol Biol 2019 ;2021:187-200

Department of Microbiology and Immunology, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, USA.

Uncomplicated urinary tract infections, especially those caused by Escherichia coli, have historically been widely studied. However, complicated urinary tract infections are presenting ever increasing healthcare challenges, particularly with Proteus mirabilis. P. mirabilis is often found on indwelling urinary catheters causing monomicrobial and polymicrobial catheter-associated urinary tract infection (CAUTI). Widespread antibiotic resistance, combined with the ability of P. mirabilis to form urinary calculi during infection, warrants further investigation of this pathogen and its host interaction in an infection model that more closely mimics the presence of an indwelling urinary catheter. Here, we describe the methods necessary to establish a murine model of P. mirabilis CAUTI.
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http://dx.doi.org/10.1007/978-1-4939-9601-8_17DOI Listing
March 2020

Cochallenge Inoculation with Proteus mirabilis in a Murine Transurethral Urinary Tract Model of Ascending Infection.

Authors:
Sara N Smith

Methods Mol Biol 2019 ;2021:173-186

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA.

Determining virulence factors which contribute to the ability of Proteus mirabilis to establish itself as a prominent cause of urinary tract infections is key in developing targets for potential therapies and treatments. Constructing mutants and testing them in a head-to-head in vivo challenge with the parental strain or other mutants enables determination of what, if any, competitive advantage may exist. Here we describe the methods necessary to ensure successful mouse transurethral inoculation and colonization in a cochallenge infection.
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http://dx.doi.org/10.1007/978-1-4939-9601-8_16DOI Listing
March 2020

Independent Transurethral Urinary Tract Inoculation in a Murine Model of Ascending Infection with Proteus mirabilis.

Authors:
Sara N Smith

Methods Mol Biol 2019 ;2021:159-172

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA.

Live animal modeling enables more in-depth realistic methods for studying the pathogenesis of urinary tract infections. Initiating a transurethral urinary tract infection in the female mouse is a challenging endeavor. However, when done with consistency and care, this infection model yields irreplaceable data. The methods necessary to ensure successful mouse transurethral inoculation and colonization in a single strain infection model with Proteus mirabilis are presented here.
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http://dx.doi.org/10.1007/978-1-4939-9601-8_15DOI Listing
March 2020

Twin arginine translocation, ammonia incorporation, and polyamine biosynthesis are crucial for Proteus mirabilis fitness during bloodstream infection.

PLoS Pathog 2019 04 22;15(4):e1007653. Epub 2019 Apr 22.

Department of Microbiology and Immunology; University of Michigan Medical School; Ann Arbor, MI, United States of America.

The Gram-negative bacterium Proteus mirabilis is a common cause of catheter-associated urinary tract infections (CAUTI), which can progress to secondary bacteremia. While numerous studies have investigated experimental infection with P. mirabilis in the urinary tract, little is known about pathogenesis in the bloodstream. This study identifies the genes that are important for survival in the bloodstream using a whole-genome transposon insertion-site sequencing (Tn-Seq) approach. A library of 50,000 transposon mutants was utilized to assess the relative contribution of each non-essential gene in the P. mirabilis HI4320 genome to fitness in the livers and spleens of mice at 24 hours following tail vein inoculation compared to growth in RPMI, heat-inactivated (HI) naïve serum, and HI acute phase serum. 138 genes were identified as ex vivo fitness factors in serum, which were primarily involved in amino acid transport and metabolism, and 143 genes were identified as infection-specific in vivo fitness factors for both spleen and liver colonization. Infection-specific fitness factors included genes involved in twin arginine translocation, ammonia incorporation, and polyamine biosynthesis. Mutants in sixteen genes were constructed to validate both the ex vivo and in vivo results of the transposon screen, and 12/16 (75%) exhibited the predicted phenotype. Our studies indicate a role for the twin arginine translocation (tatAC) system in motility, translocation of potential virulence factors, and fitness within the bloodstream. We also demonstrate the interplay between two nitrogen assimilation pathways in the bloodstream, providing evidence that the GS-GOGAT system may be preferentially utilized. Furthermore, we show that a dual-function arginine decarboxylase (speA) is important for fitness within the bloodstream due to its role in putrescine biosynthesis rather than its contribution to maintenance of membrane potential. This study therefore provides insight into pathways needed for fitness within the bloodstream, which may guide strategies to reduce bacteremia-associated mortality.
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http://dx.doi.org/10.1371/journal.ppat.1007653DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6497324PMC
April 2019

CpaA Is a Glycan-Specific Adamalysin-like Protease Secreted by Acinetobacter baumannii That Inactivates Coagulation Factor XII.

mBio 2018 12 18;9(6). Epub 2018 Dec 18.

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, USA

Antibiotic-resistant is increasingly recognized as a cause of difficult-to-treat nosocomial infections, including pneumonia, wound infections, and bacteremia. Previous studies have demonstrated that the metalloprotease CpaA contributes to virulence and prolongs clotting time when added to human plasma as measured by the activated partial thromboplastin time (aPTT) assay. Here, we show that CpaA interferes with the intrinsic coagulation pathway, also called the contact activation system, in human as well as murine plasma, but has no discernible effect on the extrinsic pathway. By utilizing a modified aPTT assay, we demonstrate that coagulation factor XII (fXII) is a target of CpaA. In addition, we map the cleavage by CpaA to two positions, 279-280 and 308-309, within the highly glycosylated proline-rich region of human fXII, and show that cleavage at the 308-309 site is responsible for inactivation of fXII. At both sites, cleavage occurs between proline and an O-linked glycosylated threonine, and deglycosylation of fXII prevents cleavage by CpaA. Consistent with this, mutant fXII (fXII-Thr309Lys) from patients with hereditary angioedema type III (HAEIII) is protected from CpaA inactivation. This raises the possibility that individuals with HAEIII who harbor this mutation may be partially protected from infection if CpaA contributes to human disease. By inactivating fXII, CpaA may attenuate important antimicrobial defense mechanisms such as intravascular thrombus formation, thus allowing to disseminate. Ventilator-associated pneumonia and catheter-related bacteremia are the most common and severe infections caused by Besides the capsule, lipopolysaccharides, and the outer membrane porin OmpA, little is known about the contribution of secreted proteins to survival Here we focus on CpaA, a potentially recently acquired virulence factor that inhibits blood coagulation We identify coagulation factor XII as a target of CpaA, map the cleavage sites, and show that glycosylation is a prerequisite for CpaA-mediated inactivation of factor XII. We propose adding CpaA to a small, but growing list of bacterial proteases that are specific for highly glycosylated components of the host defense system.
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http://dx.doi.org/10.1128/mBio.01606-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6299215PMC
December 2018

Pharmacist insights into adolescent human papillomavirus vaccination provision in the United States.

Hum Vaccin Immunother 2019 6;15(7-8):1839-1850. Epub 2019 Feb 6.

a Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina at Chapel Hill , Chapel Hill , NC , USA.

HPV vaccination coverage in the United States (US) falls short of the Healthy People 2020 goal of 80% coverage among 13-15 year-old adolescents. Pharmacies are a promising alternative vaccine delivery site that may increase access to HPV vaccination. Our objective was to assess pharmacists' insights into HPV vaccination provision to adolescents. We recruited 40 licensed pharmacists in eight states with different pharmacy vaccination laws: Alabama, California, Indiana, Kentucky, Maine, Tennessee, Texas, and Washington. Eligible pharmacists either previously provided or were currently providing HPV, tetanus-diphtheria-pertussis, or meningococcal vaccines to adolescents aged 9-17 years. Pharmacists were administered a semi-structured survey to explore insights into HPV vaccination provision. Forty-five percent of surveyed pharmacies offered HPV vaccination to adolescents. Pharmacists' reported challenges to providing HPV vaccination were parental consent (28%), tracking and patient recall (17%), perceived stigma of vaccination (17%), and education about or promotion of vaccination (17%). Pharmacists offering HPV vaccination sent patient reminders for vaccines with multiple doses (89%) and utilized telephone reminders (72%). Pharmacists informed patients' primary care providers of HPV vaccination doses most commonly through fax (72%) and updating electronic medical records (22%). One-third of pharmacists reported vaccination provision using the state immunization information system (IIS). Seventy-five percent reported vaccination rates could be increased at their respective pharmacy. Pharmacies are underutilized, although highly accessible, for HPV vaccination in the US. National efforts should expand educational programs to improve public awareness of in-pharmacy HPV vaccination, and improve the utilization of state IIS for reporting immunization coverage of adolescents by pharmacists.
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http://dx.doi.org/10.1080/21645515.2018.1556077DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6746488PMC
March 2020

Detecting cervical precancer and reaching underscreened women by using HPV testing on self samples: updated meta-analyses.

BMJ 2018 12 5;363:k4823. Epub 2018 Dec 5.

Global Coalition Against Cervical Cancer, Durham, NC, USA.

Objective: To evaluate the diagnostic accuracy of high-risk human papillomavirus (hrHPV) assays on self samples and the efficacy of self sampling strategies to reach underscreened women.

Design: Updated meta-analysis.

Data Sources: Medline (PubMed), Embase, and CENTRAL from 1 January 2013 to 15 April 2018 (accuracy review), and 1 January 2014 to 15 April 2018 (participation review).

Review Methods: Accuracy review: hrHPV assay on a vaginal self sample and a clinician sample; and verification of the presence of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) by colposcopy and biopsy in all enrolled women or in women with positive tests. Participation review: study population included women who were irregularly or never screened; women in the self sampling arm (intervention arm) were invited to collect a self sample for hrHPV testing; women in the control arm were invited or reminded to undergo a screening test on a clinician sample; participation in both arms was documented; and a population minimum of 400 women.

Results: 56 accuracy studies and 25 participation trials were included. hrHPV assays based on polymerase chain reaction were as sensitive on self samples as on clinician samples to detect CIN2+ or CIN3+ (pooled ratio 0.99, 95% confidence interval 0.97 to 1.02). However, hrHPV assays based on signal amplification were less sensitive on self samples (pooled ratio 0.85, 95% confidence interval 0.80 to 0.89). The specificity to exclude CIN2+ was 2% or 4% lower on self samples than on clinician samples, for hrHPV assays based on polymerase chain reaction or signal amplification, respectively. Mailing self sample kits to the woman's home address generated higher response rates to have a sample taken by a clinician than invitation or reminder letters (pooled relative participation in intention-to-treat-analysis of 2.33, 95% confidence interval 1.86 to 2.91). Opt-in strategies where women had to request a self sampling kit were generally not more effective than invitation letters (relative participation of 1.22, 95% confidence interval 0.93 to 1.61). Direct offer of self sampling devices to women in communities that were underscreened generated high participation rates (>75%). Substantial interstudy heterogeneity was noted (I>95%).

Conclusions: When used with hrHPV assays based on polymerase chain reaction, testing on self samples was similarly accurate as on clinician samples. Offering self sampling kits generally is more effective in reaching underscreened women than sending invitations. However, since response rates are highly variable among settings, pilots should be set up before regional or national roll out of self sampling strategies.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6278587PMC
http://dx.doi.org/10.1136/bmj.k4823DOI Listing
December 2018

Water-soluble cranberry extract inhibits Vibrio cholerae biofilm formation possibly through modulating the second messenger 3', 5' - Cyclic diguanylate level.

PLoS One 2018 7;13(11):e0207056. Epub 2018 Nov 7.

Department of Biological Sciences, Clemson University, Clemson, South Carolina, United States of America.

Quorum sensing (QS) and nucleotide-based second messengers are vital signaling systems that regulate bacterial physiology in response to changing environments. Disrupting bacterial signal transduction is a promising direction to combat infectious diseases, and QS and the second messengers are undoubtedly potential targets. In Vibrio cholerae, both QS and the second messenger 3', 5'-cyclic diguanylate (c-di-GMP) play a central role in controlling motility, motile-to-sessile life transition, and virulence. In this study, we found that water-soluble extract from the North American cranberry could significantly inhibit V. cholerae biofilm formation during the development/maturation stage by reducing the biofilm matrix production and secretion. The anti-biofilm effect by water-soluble cranberry extract was possibly through modulating the intracellular c-di-GMP level and was independent of QS and the QS master regulator HapR. Our results suggest an opportunity to explore more functional foods to fight stubborn infections through interference with the bacterial signaling systems.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0207056PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6221352PMC
April 2019