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In 1973-74, a polybrominated biphenyl (PBB) flame retardant mixture was shipped to Michigan livestock feed mills in place of a nutritional supplement and contaminated the food supply. Following the accident, the Michigan PBB Registry was established to study the long-term health effects of halogenated compounds and is now led by a community-academic partnership. PBB exposure is associated with altered DNA methylation in sperm, which may lead to adverse birth outcomes in children whose fathers have increased levels of serum PBB or polychlorinated biphenyl (PCB). Paternal PBB and PCB levels of men enrolled in the Michigan PBB Registry (n = 155) were analyzed against matched offspring birthweight and gestational age (n = 336). Birthweight and gestational age were dichotomized at the 25th percentile and 37 weeks, respectively, and paternal PBB and PCB levels were examined as continuous measures and divided into tertiles. Associations of offspring birthweight and gestational age with paternal PBB and PCB serum concentrations were modeled using multivariable linear spline and log-risk regression, adjusting for family clustering, paternal health and lifestyle factors, maternal PBB, and PCB serum concentrations, sex, and offspring gestational age (for birthweight). Fathers in the middle and upper PBB and PCB tertiles had increased risks for lowest quartile birthweight compared to the first tertile, with adjusted risk ratios (aRR) = 1.67 (95% CI: 0.93, 2.99) and aRR = 2.06 (95% CI: 1.12, 3.79) for PBB, and aRR = 1.47 (95% CI: 0.79, 2.75) and aRR = 1.34 (95% CI: 0.70, 2.54) for PCB, respectively. Elevated paternal PBB levels were not associated with an increased risk for preterm birth, while PCB levels were associated with a small, but not significant, decrease in gestational age, β = -0.37 (95% CI: -0.76, 0.03) weeks per log unit increase PCB. The findings suggest that increased paternal PBB and PCB levels negatively impact offspring birthweight, and paternal PCB levels may negatively impact gestational age.

Community-engaged approaches to research can increase trust, enhance the relevance and use of research, address issues of equity and justice, and increase community knowledge and capacity. The HERCULES Exposome Research Center sought to engage local Atlanta communities to learn about and address their self-identified environmental health concerns. To do this, HERCULES and their stakeholder partners collaboratively developed a community grant program. The program was evaluated using mixed qualitative methods that included document review and semi-structured interviews. This paper presents the development, implementation, and evaluation of the grant program. HERCULES awarded one-year grants of $2,500 to 12 organizations within the Atlanta region, for a total 13 grants and $32,500 in funding. Grantees reported accomplishments related to community knowledge, awareness, and engagement in addition to material accomplishments. All grantees planned to sustain their programs, and some received additional funding to do so. Some grantees remained actively involved with HERCULES beyond the grant program. The HERCULES Community Grant Program was able to increase awareness of HERCULES among applicant communities, establish or enhance relationships with community-based organizations, and identify local environmental health concerns while providing tangible results for grantees and the communities they serve. Mini-grant programs are a feasible approach to address community environmental health and establish new relationships. This model may benefit others who aim to establish community-academic relationships while addressing community health concerns.

IntroductionThe exposome concept provides a framework to better incorporate the environment into the study of health and disease and has been defined by academics to encompass all lifetime exposures including toxicants, diet, and lifestyle choices. However, initial applications of the exposome concept have been less apt at measuring social determinants of health, focusing primarily on conventional environmental exposures and lifestyle choices that do not reflect the complex lived experience of many communities. To bring community voice into the exposome concept, the HERCULES Exposome Research Center and its Stakeholder Advisory Board co-developed the Exposome Roadshow. We present and discuss the resulting community-exposome definition to inform and improve exposome research.Materials and MethodsFour communities from distinct areas across metro-Atlanta participated in separate 2-day Exposome Roadshow workshops with concept mapping. Aligned with a popular education approach in which community knowledge is used to work collectively for change, concept mapping provided a systematic method to collect and visualize community members' knowledge and create a shared understanding to take action. Community members brainstormed, sorted, and rated their responses to the prompt: “What in your environment is affecting your and your community's health?” Responses were analyzed and visually depicted by concept maps consisting of separate but interrelated clusters of ideas. Community members discussed and validated the maps, selecting a final map illustrating their community's exposome.ResultsA total of 118 community members completed concept mapping. On average communities identified 7 clusters to define their exposome. The resulting concept maps offer a community definition of the exposome. Five major themes arose across all four communities: conventional environmental concerns, built environment, social relationships, crime and safety, and individual health and behaviors.DiscussionThe resulting community-exposome definition demonstrates the importance of expanding the scope of exposures beyond traditional environmental influences to include the lived experience of individuals and communities. While newer exposome definitions align more closely with this community definition, traditional exposome methods do not routinely include these factors. To truly capture the totality of lifetime exposures and improve human health, researchers should incorporate community perspectives into exposome research.

The Michigan PBB Oral History Project documented community residents’ descriptions of a large-scale chemical contamination—the PBB contamination—that occurred in Michigan in 1973. These oral histories document residents’ and others’ experiences during and after the contamination. We conducted a grounded theory qualitative analysis of 31 oral histories to examine the experiences of community members, researchers, lawyers, and others who actively sought out and contributed essential information about the contamination and its impacts. Our findings highlight several challenges encountered in the development of this knowledge including four central themes—contested knowledge, community skills, inaction, and uncertainty. Integrating environmental health literacy, community capacity, and contested illness frameworks, we examine the contributions of community residents, scientists (from inside and outside the community), and others to the development of knowledge to inform decisions and sustain action regarding this large-scale contamination. We close with a discussion of lessons learned regarding efforts to build environmental health knowledge within uncertain and often contested contexts and for promoting environmental health and action related to large-scale chemical contaminations. Our findings suggest the importance of integrated frameworks for examining and promoting the critical role of community skills, leadership, participation, sense of community, and community power in promoting environmental health.

Experts recommend reporting environmental exposure results back to research participants and communities, yet environmental health researchers need further guidance to improve the practice of reporting back. We present the results of a workshop developed to identify pertinent issues and areas for action in reporting back environmental health research results. Thirty-five attendees participated, brainstorming responses to the prompt: &ldquo, What are some specific issues that are relevant to reporting back research results to individuals or the larger community?&rdquo, and then grouping responses by similarity and rating their importance. Based on a combined theoretical foundation of grounded theory and qualitative content analysis, we used concept mapping to develop a collective understanding of the issues. Visual maps of the participants&rsquo, responses were created using nonmetric multidimensional scaling and hierarchical cluster analysis. The resulting concept map provided a spatial depiction of five issue areas: Effective Communication Strategies, Community Knowledge and Concerns, Uncertainty, Empowering Action, and Institutional Review and Oversight (listed from highest to lowest rating). Through these efforts, we disentangled the complex issues affecting how and whether environmental health research results are reported back to participants and communities, by identifying five distinct themes to guide recommendations and action. Engaging community partners in the process of reporting back emerged as a unifying global theme, which could improve how researchers report back research results by understanding community context to develop effective communication methods and address uncertainty, the ability to act, and institutional concerns about beneficence and justice.

The important uropathogenProteus mirabilisencodes a record number of chaperone/usher-pathway adhesive fimbriae. Such fimbriae, which are used for adhesion to cell surfaces/tissues and for biofilm formation, are typically important virulence factors in bacterial pathogenesis. Here, the structures of the receptor-binding domains of the tip-located two-domain adhesins UcaD (1.5 Å resolution) and AtfE (1.58 Å resolution) from twoP. mirabilisfimbriae (UCA/NAF and ATF) are presented. The structures of UcaD and AtfE are both similar to the F17G type of tip-located fimbrial receptor-binding domains, and the structures are very similar despite having only limited sequence similarity. These structures represent an important step towards a molecular-level understanding ofP. mirabilisfimbrial adhesins and their roles in the complex pathogenesis of urinary-tract infections.

Proteus mirabilis is a model organism for urease-producing uropathogens. These diverse bacteria cause infection stones in the urinary tract and form crystalline biofilms on indwelling urinary catheters, frequently leading to poly-microbial infection. Recent work has elucidated how P. mirabilis causes all of these disease states. Particularly exciting is the discovery that this bacterium forms large clusters in the bladder lumen that are sites for stone formation. These clusters, and other steps of infection, require two virulence factors in particular: urease and MR/P fimbriae. Highlighting the importance of MR/P fimbriae is the cotranscribed regulator, MrpJ, which globally controls virulence. Overall, P. mirabilis exhibits an extraordinary lifestyle, and further probing will answer exciting basic microbiological and clinically relevant questions.

Although best management practices have been recommended by government agencies and non-profit organizations to reduce community gardeners' potential exposure to soil contaminants such as lead, some gardeners do not perform these practices. Understanding gardeners' beliefs and motivations is critical for effective promotion of safer gardening practices. This study, grounded in the Theory of Planned Behavior (TPB), employed five focus groups to investigate Atlanta community garden leaders' perspectives concerning three gardening practices: composting, hygiene behaviors, and mulching. These general practices are also considered safe gardening practices in that they can reduce exposure to toxicants in urban gardens. Qualitative analysis identified advantages and disadvantages; supporters and non-supporters; and barriers and facilitators that might influence gardeners' opinions regarding these behaviors. Gardeners expressed that more funding, volunteers, and training are needed to promote these behaviors. Gardeners noted that mulch and compost provided advantages such as improving soil quality, but a primary barrier was concern about contamination of source materials. Focus group participants did not directly associate composting and mulching with reduction of exposure to soil contaminants. Behavioral challenges related to hygiene included concerns about decreased exposure to salubrious bacteria, inadequate access to potable water, and limited availability of gloves and wipes. These study findings characterize factors that community garden stakeholders should consider when promoting safe gardening practices and interventions.

Widespread polybrominated biphenyls (PBBs) contamination occurred in Michigan from 1973 to 1974, when PBBs were accidentally substituted for a nutritional supplement in livestock feed. People who lived in the state were exposed to PBBs via several routes including ingestion, inhalation and skin absorption. PBBs sequestered in lipid-rich matrices such as adipose tissue, are slowly eliminated after entering the human body, and can also be transferred from a mother to her offspring through the placenta and breastfeeding. Due to the long biological half-lives of PBBs, as well as concerns from the exposed community, biomonitoring measurements were conducted from 2012 to 2015. Because of their similar structures, serum PBBs, polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) were all measured 40 years after the PBB contamination incident (N = 862). The serum PBB-153 levels among the original highly-exposed groups (i.e., chemical workers, the family of chemical workers, and individuals who lived on or received food from the contaminated farms) remains significantly higher than other Michigan residents. Several predictors such as sampling age, sex, and smoking status were significantly associated with the serum levels of some persistent organic pollutants (POPs). Higher average values and also wider ranges of serum POP levels were found in this study compared to the National Health and Nutrition Examination Survey (NHANES), with the most substantial difference in serum PBB-153. This was true for all groups of Michigan residents including those who were not part of the above-described highly-exposed groups. Moreover, the people born after the contamination incident began also have higher serum PBB-153 levels when compared with more recent NHANES data (2010–2014), which suggests potential intergenerational exposure and/or continued environmental exposure following the contamination period. Keywords: Polybrominated biphenyls (PBBs), Polychlorinated biphenyls (PCBs), Polybrominated diphenyl ethers (PBDEs), Persistent organic pollutants (POPs), Michigan PBB Registry, Biomonitoring

Genetic mutation enables the study of the function of specific genes, particularly when a mutant is compared against its isogenic parent. In Proteus mirabilis bacteria, traditional allelic exchange mutation is labor-intensive and has a high failure rate in some strains. Likewise, there is no working protocol for lambda red recombinase-based mutation in P. mirabilis. Here we describe an alternative method of insertional mutagenesis based on retargeting of group II introns. The protocol includes steps to generate single or multiple mutations, with the possibility to delete intervening sequences of DNA.

Proteus mirabilis is generally easy to culture, but its tendency to swarm on a wide variety of media can interfere with isolation of single colonies or identification of other species in a sample. Therefore, specialized media may be needed to control swarming or to study the bacteria under chemically defined conditions. Here, methods are described for routine culture of P. mirabilis, isolation of P. mirabilis from mixed cultures, and culture of P. mirabilis on physiologically relevant media.

Mannose-resistant Proteus-like (MR/P) fimbriae, produced by the uropathogen Proteus mirabilis, are major contributors to urinary tract infection. Expression of mrp genes is controlled by an invertible element in the mrp operon promoter, and promoter orientation is controlled by a single recombinase, MrpI, which determines whether the element is in the on or off orientation. Detailed here is a simple assay to determine the orientation of the invertible element in a population of P. mirabilis, a rapid screen to detect element-locked mrpI mutants, and quantification of mixtures of on and off bacteria.

Proteus mirabilis is well known for using its flagella to swim through liquids or swarm across solid surfaces. Both phenomena are easy to observe. Described here are two agar-based assays for studying both swimming and swarming behavior, and considerations that affect the outcome.

Community gardens offer numerous benefits, but there are also potential risks from exposure to chemical contaminants in the soil. Through the lens of the Theory of Planned Behavior, this mixed methods study examined community gardeners&rsquo, beliefs and intentions to conduct heavy metal soil testing. The qualitative component involved five focus groups of community garden leaders in Atlanta, Georgia. Qualitative analysis of the focus group data revealed that heavy metal soil contamination was not frequently identified as a common gardening hazard and several barriers limited soil testing in community gardens. The focus group results informed the development of a questionnaire that was administered to 500 community gardeners across the United States. Logistic regression analysis revealed that the soil testing intention was associated with attitude (aOR = 2.46, 95% CI: 1.34, 4.53), subjective norms (aOR = 3.39 95% CI: 2.07, 5.57), and perceived behavioral control (aOR = 1.81, 95% CI: 1.10, 2.99). Study findings have implications for interventions involving community garden risk mitigation, particularly gardens that engage children and vulnerable populations.

Não recebi financiamento Children with Autism Spectrum Disorder (ASD) may present deficits in communication and social interaction, as well as restricted and repetitive behaviors and interests. Research has been carried out in the area of music, however the majority of studies for this population contemplate the use of music as a therapeutic and not an educational activity. Were found studies that evaluated the teaching of rudimentary reading of musical notes through stimulus equivalence and mostly with individuals with typical development. This study had as a general objective to verify the effects of conditional discriminations between figures (each composed of a background color and a specific musical symbol) and keys in the recognition of musical notes to the piano in a boy with 4 years and 10 months of age diagnosed with ASD. As specific objectives, verify if the control of the answer would be given by the color or symbol of the musical figures, examine the formation of classes of equivalent stimuli and test the recombination of the notes learned in new musical sequences. The method had an experimental design with multiple baseline between sets of musical notes. The teaching, which had procedures of the Applied Behavior Analysis (ABA), was divided in three phases, in which seven musical notes written in the treble clef, from central C to central B, were taught in configurations that ranged from one to 38 combined and recombined symbols (units). A pre and post-test were applied at all phases. The connection between symbol and color cards and the corresponding keys (AB) and between cards with symbol and color and the name of the corresponding note (AD) were taught. The procedure of teaching discrimination of musical notes promoted the acquisition of repertoire referring to musical reading, as measured by the response of playing the corresponding key. The results suggest efficacy in teaching with an achievement of over 70% in the teaching and testing phases. Emerging relations between teaching classes could also be noticed. It is intended with this research, expand previous studies found In music and equivalence with adults for a child with ASD. Crianças com Transtorno do Espectro do Autismo (TEA) podem apresentar déficits na comunicação e na interação social, bem como comportamentos e interesses restritos e repetitivos. Pesquisas vem sendo realizadas na área da música, entretanto a maioria dos estudos para esta população contempla o uso da música como atividade terapêutica e não educacional. Foram encontrados estudos que avaliaram o ensino de leitura e rudimentos musicais por meio da equivalência de estímulo e em sua maioria com indivíduos com desenvolvimento típico. Este estudo teve como objetivo geral verificar os efeitos do ensino com discriminações condicionais entre figuras (cada uma composta por uma cor de fundo e um símbolo musical específico) e teclas no reconhecimento de notas musicais ao piano em um menino com 4 anos e 10 meses de idade diagnosticado com TEA. Como objetivos específicos, verificar se o controle da resposta se daria pela cor ou pelo símbolo das figuras musicais, verificar a formação de classes de estímulos equivalentes e testar a recombinação das notas aprendidas em novas sequências musicais. O método contou com um delineamento experimental com linha de base múltipla entre conjuntos de notas musicais. O ensino, que contou com procedimentos da Análise do Comportamento Aplicada (ABA), foi dividido em três fases, em que sete notas musicais grafadas na clave de sol, de dó central a si, foram ensinadas em configurações que variaram de 1 a 38 símbolos (unidades) combinadas e recombinadas. Todas as fases contaram com pré e pós-teste. Foram ensinadas as relações entre cartões com símbolo e cor e as teclas correspondentes (AB) e entre cartões com símbolo e cor e o nome da nota correspondente (AD). O procedimento de ensino de discriminação de notas musicais promoveu a aquisição de repertório referente a leitura musical, medido pela resposta de tocar a tecla correspondente. Os resultados sugerem eficácia no ensino com um aproveitamento superior a 70% nas fases de ensino e teste. Relações emergentes entre as classes de ensino também puderam ser notadas. Pretende-se com esta pesquisa, expandir os estudos anteriores encontrados sobre música e equivalência com adultos para uma criança com TEA.

Mannose-resistant Proteus-like (MR/P) fimbriae, produced by the uropathogen Proteus mirabilis, are major contributors to urinary tract infection. Expression of mrp genes is controlled by an invertible element in the mrp operon promoter, and promoter orientation is controlled by a single recombinase, MrpI, which determines whether the element is in the on or off orientation. Detailed here is a simple assay to determine the orientation of the invertible element in a population of P. mirabilis, a rapid screen to detect element-locked mrpI mutants, and quantification of mixtures of on and off bacteria.

Proteus mirabilis is generally easy to culture, but its tendency to swarm on a wide variety of media can interfere with isolation of single colonies or identification of other species in a sample. Therefore, specialized media may be needed to control swarming or to study the bacteria under chemically defined conditions. Here, methods are described for routine culture of P. mirabilis, isolation of P. mirabilis from mixed cultures, and culture of P. mirabilis on physiologically relevant media.

Genetic mutation enables the study of the function of specific genes, particularly when a mutant is compared against its isogenic parent. In Proteus mirabilis bacteria, traditional allelic exchange mutation is labor-intensive and has a high failure rate in some strains. Likewise, there is no working protocol for lambda red recombinase-based mutation in P. mirabilis. Here we describe an alternative method of insertional mutagenesis based on retargeting of group II introns. The protocol includes steps to generate single or multiple mutations, with the possibility to delete intervening sequences of DNA.

A critical first step in bacterial virulence and colonization is adherence to mucosal surfaces, often mediated by fimbriae and other protein adhesins. Here are described three short methods for studying these surface proteins and their behaviors, using protocols developed for the opportunistic pathogen Proteus mirabilis. Unlike the mannose-binding type 1 fimbriae produced by Escherichia coli, most P. mirabilis strains produce mannose-resistant/Proteus-like (MR/P) fimbriae. Both types of fimbrial production and adhesion can be easily demonstrated by a simple and economical hemagglutination assay which uses a model system of erythrocytes. The second and third fimbrial methods presented here show how to shear surface-exposed proteins and use acid treatment to separate interlocked fimbrial subunits into monomers.

Proteus mirabilis is well known for using its flagella to swim through liquids or swarm across solid surfaces. Both phenomena are easy to observe. Described here are two agar-based assays for studying both swimming and swarming behavior, and considerations that affect the outcome.

Proteus mirabilis is a leading cause of catheter-associated urinary tract infections (CAUTIs) and urolithiasis. The transcriptional regulator MrpJ inversely modulates two critical aspects of P. mirabilis UTI progression: fimbria-mediated attachment and flagellum-mediated motility. Transcriptome data indicated a network of virulence-associated genes under MrpJ9s control. Here, we identify the direct gene regulon of MrpJ and its contribution to P. mirabilis pathogenesis, leading to the discovery of novel virulence targets. Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) was used for the first time in a CAUTI pathogen to probe for in vivo direct targets of MrpJ. Selected MrpJ-regulated genes were mutated and assessed for their contribution to UTI using a mouse model. ChIP-seq revealed a palindromic MrpJ binding sequence and 78 MrpJ-bound regions, including binding sites upstream of genes involved in motility, fimbriae, and a type VI secretion system (T6SS). A combinatorial mutation approach established the contribution of three fimbriae (fim8A, fim14A, and pmpA) to UTI and a new pathogenic role for the T6SS in UTI progression. In conclusion, this study (i) establishes the direct gene regulon and an MrpJ consensus binding site and (ii) led to the discovery of new virulence genes in P. mirabilis UTI, which could be targeted for therapeutic intervention of CAUTI.

Proteus mirabilis , a Gram-negative rod-shaped bacterium most noted for its swarming motility and urease activity, frequently causes catheter-associated urinary tract infections (CAUTIs) that are often polymicrobial. These infections may be accompanied by urolithiasis, the development of bladder or kidney stones due to alkalinization of urine from urease-catalyzed urea hydrolysis. Adherence of the bacterium to epithelial and catheter surfaces is mediated by 17 different fimbriae, most notably MR/P fimbriae. Repressors of motility are often encoded by these fimbrial operons. Motility is mediated by flagella encoded on a single contiguous 54-kb chromosomal sequence. On agar plates, P. mirabilis undergoes a morphological conversion to a filamentous swarmer cell expressing hundreds of flagella. When swarms from different strains meet, a line of demarcation, a “Dienes line,” develops due to the killing action of each strain’s type VI secretion system. During infection, histological damage is caused by cytotoxins including hemolysin and a variety of proteases, some autotransported. The pathogenesis of infection, including assessment of individual genes or global screens for virulence or fitness factors has been assessed in murine models of ascending urinary tract infections or CAUTIs using both single-species and polymicrobial models. Global gene expression studies performed in culture and in the murine model have revealed the unique metabolism of this bacterium. Vaccines, using MR/P fimbria and its adhesin, MrpH, have been shown to be efficacious in the murine model. A comprehensive review of factors associated with urinary tract infection is presented, encompassing both historical perspectives and current advances.

The enteric bacterium Proteus mirabilis is associated with a significant number of catheter-associated urinary tract infections (UTIs). Strict regulation of the antagonistic processes of adhesion and motility, mediated by fimbriae and flagella, respectively, is essential for disease progression. Previously, the transcriptional regulator MrpJ, which is encoded by the mrp fimbrial operon, has been shown to repress both swimming and swarming motility. Here we show that MrpJ affects an array of cellular processes beyond adherence and motility. Microarray analysis found that expression of mrpJ mimicking levels observed during UTIs leads to differential expression of 217 genes related to, among other functions, bacterial virulence, type VI secretion, and metabolism. We probed the molecular mechanism of transcriptional regulation by MrpJ using transcriptional reporters and chromatin immunoprecipitation (ChIP). Binding of MrpJ to two virulence-associated target gene promoters, the promoters of the flagellar master regulator flhDC and mrp itself, appears to be affected by the condensation state of the native chromosome, although both targets share a direct MrpJ binding site proximal to the transcriptional start. Furthermore, an mrpJ deletion mutant colonized the bladders of mice at significantly lower levels in a transurethral model of infection. Additionally, we observed that mrpJ is widely conserved in a collection of recent clinical isolates. Altogether, these findings support a role of MrpJ as a global regulator of P. mirabilis virulence.

Proteus mirabilis, a Gram-negative uropathogen, is a major causative agent in catheter-associated urinary tract infections (CAUTI). Mannose-resistant Proteus-like fimbriae (MR/P) are crucially important for P. mirabilis infectivity and are required for biofilm formation and auto-aggregation, as well as for bladder and kidney colonization. Here, the X-ray crystal structure of the MR/P tip adhesin, MrpH, is reported. The structure has a fold not previously described and contains a transition metal center with Zn2+ coordinated by three conserved histidine residues and a ligand. Using biofilm assays, chelation, metal complementation, and site-directed mutagenesis of the three histidines, we show that an intact metal binding site occupied by zinc is essential for MR/P fimbria-mediated biofilm formation, and furthermore, that P. mirabilis biofilm formation is reversible in a zinc-dependent manner. Zinc is also required for MR/P-dependent agglutination of erythrocytes, and mutation of the metal binding site renders P. mirabilis unfit in a mouse model of UTI. The studies presented here provide important clues as to the mechanism of MR/P-mediated biofilm formation and serve as a starting point for identifying the physiological MR/P fimbrial receptor., Author summary Many bacteria use fimbriae to adhere to surfaces, and this function is often essential for pathogens to gain a foothold in the host. In this study, we examine the major virulence-associated fimbrial protein, MrpH, of the bacterial urinary tract pathogen Proteus mirabilis. This species is particularly known for causing catheter-associated urinary tract infections, in which it forms damaging urinary stones and crystalline biofilms that can block the flow of urine through indwelling catheters. MrpH resides at the tip of mannose-resistant Proteus-like (MR/P) fimbriae and is required for MR/P-dependent adherence to surfaces. Although MR/P belongs to a well-known class of adhesive fimbriae encoded by the chaperone-usher pathway, we found that MrpH has a dramatically different structure compared with other tip-located adhesins in this family. Unexpectedly, MrpH was found to bind a zinc cation, which we show is essential for MR/P-mediated biofilm formation and adherence to red blood cells. Furthermore, MR/P-mediated adherence can be modified by controlling zinc levels. These findings have the potential to aid development of better anti-biofilm urinary catheters or other methods to prevent P. mirabilis infection of the urinary tract.

Brain aging is characterized by changes in both hemodynamic and neuronal responses, which may be influenced by the cardiorespiratory fitness of the individual. To investigate the relationship between neuronal and hemodynamic changes, we studied the brain activity elicited by visual stimulation (checkerboard reversals at different frequencies) in younger adults and in older adults varying in physical fitness. Four functional brain measures were used to compare neuronal and hemodynamic responses obtained from BA17: two reflecting neuronal activity (the event-related optical signal, EROS, and the C1 response of the ERP), and two reflecting functional hemodynamic changes (functional magnetic resonance imaging, fMRI, and near-infrared spectroscopy, NIRS). The results indicated that both younger and older adults exhibited a quadratic relationship between neuronal and hemodynamic effects, with reduced increases of the hemodynamic response at high levels of neuronal activity. Although older adults showed reduced activation, similar neurovascular coupling functions were observed in the two age groups when fMRI and deoxy-hemoglobin measures were used. However, the coupling between oxy- and deoxy-hemoglobin changes decreased with age and increased with increasing fitness. These data indicate that departures from linearity in neurovascular coupling may be present when using hemodynamic measures to study neuronal function.

Alcohol Myopia Theory (AMT) posits that alcohol restricts the focus of attention, such that behaviors are determined only by highly salient environmental cues. While AMT is most commonly understood in terms of spatial attention, the present study tested the effects of alcohol in the temporal domain of attention. Seventy-one participants consumed either a placebo beverage or one of two doses of alcohol (0.40 g/kg or 0.80 g/kg ETOH) before performing an auditory discrimination task while event-related potentials (ERPs) were recorded. Consistent with typical sequential effects, placebo participants showed increased P300 amplitude and slowed behavioral responses when the current target differed from the two-back tone. In contrast, alcohol caused increased P300 and response slowing when the target tone differed from the one-back tone. These findings suggest that alcohol increases the salience of more recently-encountered information, consistent with the general tenets of AMT.

A citizen-science study was conducted in two low-income, flood-prone communities in Atlanta, Georgia, in order to document environmental exposures and the prevalence of occupant asthma. Teams consisting of a public-health graduate student and a resident from one of the two communities administered a questionnaire, inspected residences for mold growth, and collected a dust sample for quantifying mold contamination. The dust samples were analyzed for the 36 molds that make up the Environmental Relative Moldiness Index (ERMI). Most residents (76%) were renters. The median duration of residence was 2.5 years. Although only 12% of occupants reported a history of flooding, 46% reported at least one water leak. Homes with visible mold (35%) had significantly (P<0.05) higher mean ERMI values compared to homes without (14.0 versus 9.6). The prevalence of self-reported, current asthma among participants was 14%. In logistic regression models controlling for indoor smoking, among participants residing at their current residence for two years or less, a positive association was observed between asthma and the homes’ ERMI values (adjusted odds ratio per unit increase in ERMI = 1.12, 95% confidence intervals (CI): 1.01–1.25; two-tailedP=0.04). Documentation of the exposures and asthma prevalence has been presented to the communities and public officials. Community-based organizations have taken responsibility for planning and implementing activities in response to the study findings.

Proteus mirabilis contributes to a significant number of catheter-associated urinary tract infections, where coordinated regulation of adherence and motility is critical for ascending disease progression. Previously, the mannose-resistant Proteus -like (MR/P) fimbria-associated transcriptional regulator MrpJ has been shown to both repress motility and directly induce the transcription of its own operon; in addition, it affects the expression of a wide range of cellular processes. Interestingly, 14 additional mrpJ paralogs are included in the P. mirabilis genome. Looking at a selection of MrpJ paralogs, we discovered that these proteins, which consistently repress motility, also have nonidentical functions that include cross-regulation of fimbrial operons. A subset of paralogs, including AtfJ (encoded by the ambient temperature fimbrial operon), Fim8J, and MrpJ, are capable of autoinduction. We identified an element of the atf promoter extending from 487 to 655 nucleotides upstream of the transcriptional start site that is responsive to AtfJ, and we found that AtfJ directly binds this fragment. Mutational analysis of AtfJ revealed that its two identified functions, autoregulation and motility repression, are not invariably linked. Residues within the DNA-binding helix-turn-helix domain are required for motility repression but not necessarily autoregulation. Likewise, the C-terminal domain is dispensable for motility repression but is essential for autoregulation. Supported by a three-dimensional (3D) structural model, we hypothesize that the C-terminal domain confers unique regulatory capacities on the AtfJ family of regulators. IMPORTANCE Balancing adherence with motility is essential for uropathogens to successfully establish a foothold in their host. Proteus mirabilis uses a fimbria-associated transcriptional regulator to switch between these antagonistic processes by increasing fimbrial adherence while simultaneously downregulating flagella. The discovery of multiple related proteins, many of which also function as motility repressors, encoded in the P. mirabilis genome has raised considerable interest as to their functionality and potential redundancy in this organism. This study provides an important advance in this field by elucidating the nonidentical effects of these paralogs on a molecular level. Our mechanistic studies of one member of this group, AtfJ, shed light on how these differing functions may be conferred despite the limited sequence variety exhibited by the paralogous proteins.

Background The data presented here are a response to calls for more direct measurements of pesticide residues in foods consumed by children and provide an opportunity to compare direct measures of pesticide residues in foods representing actual consumption with those reported by the U.S. Department of Agriculture Pesticide Data Program. Objective We measured pesticide residues in 24-hr duplicate food samples collected from a group of 46 young children participating in the Children’s Pesticide Exposure Study (CPES). Methods Parents were instructed to collect 24-hr duplicate food samples of all conventional fruits, vegetables, and fruit juices equal to the quantity consumed by their children, similarly prewashed/prepared, and from the same source or batch. Individual or composite food items were analyzed for organophosphate (OP) and pyrethroid insecticide residues. Results We collected a total of 239 24-hr duplicate food samples collected from the 46 CPES children. We found 14% or 5% of those food samples contained at least one OP or pyrethroid insecticide, respectively. We measured a total of 11 OP insecticides, at levels ranging from 1 to 387 ng/g, and three pyrethroid insecticides, at levels ranging from 2 to 1,133 ng/g, in children’s food samples. We found that many of the food items consumed by the CPES children were also on the list of the most contaminated food commodities reported by the Environmental Working Group. Conclusions The frequent consumption of food commodities with episodic presence of pesticide residues that are suspected to cause developmental and neurological effects in young children supports the need for further mitigation.

Swarming motility by the urinary tract pathogen Proteus mirabilis has been a long-studied but little understood phenomenon. On agar, a P. mirabilis colony grows outward in a bull's-eye pattern formed by consecutive waves of rapid swarming followed by consolidation into shorter cells. To examine differential gene expression in these growth phases, a microarray was constructed based on the completed genome sequence and annotation. RNA was extracted from broth-cultured, swarming, and consolidation-phase cells to assess transcription during each of these growth states. A total of 587 genes were differentially expressed in broth-cultured cells versus swarming cells, and 527 genes were differentially expressed in broth-cultured cells versus consolidation-phase cells (consolidate). Flagellar genes were highly upregulated in both swarming cells and consolidation-phase cells. Fimbriae were downregulated in swarming cells, while genes involved in cell division and anaerobic growth were upregulated in broth-cultured cells. Direct comparison of swarming cells to consolidation-phase cells found that 541 genes were upregulated in consolidate, but only nine genes were upregulated in swarm cells. Genes involved in flagellar biosynthesis, oligopeptide transport, amino acid import and metabolism, cell division, and phage were upregulated in consolidate. Mutation of dppA , oppB , and cysJ , upregulated during consolidation compared to during swarming, revealed that although these genes play a minor role in swarming, dppA and cysJ are required during ascending urinary tract infection. Swarming on agar to which chloramphenicol had been added suggested that protein synthesis is not required for swarming. These data suggest that the consolidation phase is a state in which P. mirabilis prepares for the next wave of swarming.

MR/P fimbriae of uropathogenic Proteus mirabilis undergo invertible element-mediated phase variation whereby an individual bacterium switches between expressing fimbriae (phase ON) and not expressing fimbriae (phase OFF). Under different conditions, the percentage of fimbriate bacteria within a population varies and could be dictated by either selection (growth advantage of one phase) or signaling (preferentially converting one phase to the other in response to external signals). Expression of MR/P fimbriae increases in a cell-density dependent manner in vitro and in vivo. However, rather than the increased cell density itself, this increase in fimbrial expression is due to an enrichment of fimbriate bacteria under oxygen limitation resulting from increased cell density. Our data also indicate that the persistence of MR/P fimbriate bacteria under oxygen-limiting conditions is a result of both selection (of MR/P fimbrial phase variants) and signaling (via modulation of expression of the MrpI recombinase). Furthermore, the mrpJ transcriptional regulator encoded within the mrp operon contributes to phase switching. Type 1 fimbriae of Escherichia coli , which are likewise subject to phase variation via an invertible element, also increase in expression during reduced oxygenation. These findings provide evidence to support a mechanism for persistence of fimbriate bacteria under oxygen limitation, which is relevant to disease progression within the oxygen-restricted urinary tract.

Urinary contaminant concentrations are commonly adjusted by creatinine to account for the variability in urinary output. This approach may not be optimal among children due to developmental growth of muscle mass and the associated increase in creatinine formation. An alternative approach is to measure the specific gravity of the urine sample, which reflects the solute concentration of the urine. We compare the appropriateness of urinary creatinine and urinary-specific gravity as factors for correcting morning and evening spot urine samples collected from 23 children (3-11 years) for a total of 41 days in four different seasons. Two linear mixed-effects models were fit using age, sex, season, and sample collection time (morning/evening) as predictors with specific gravity and creatinine as dependent variables. Specific gravity was significantly associated with the sample collection time (P

Despite the widespread use of synthetic pyrethroid insecticides that led to common exposure in the population, very few studies have been conducted to quantitatively assess human, particularly, children's, long-term exposures to pyrethroid insecticides. The objective of the Children Pesticide Exposure Study - Washington (CPES-WA) was to establish the longitudinal exposure profiles for pyrethroid insecticides in a cohort of children living in an urban and suburban community using urinary pyrethroid metabolites as exposure biomarkers. The results from this analysis will allow us to examine potential risk factors in relation to the elevated pyrethroid insecticide exposure in children. A total of 23 children, aged 3-11 years, who only consumed conventional diets were enrolled in this 1-year study. We provided organic food items to children for 5 consecutive days in the summer and fall sampling seasons. We measured urinary metabolites for the synthetic pyrethroid insecticides in urine samples that were collected twice daily during each of the four sampling seasons. 3-phenoxybenzoic acid was frequently detected in the urine samples with mean and median daily volume-weighted average levels of 1.5 and 1.2 microg/l, followed by trans-2,2-(dichloro)-2-dimethylvinylcyclopropane carboxylic acid (1.4 and 1.0 microg/l) and cis-2,2-(dichloro)-2-dimethylvinylcyclopropane carboxylic acid (0.5 microg/l, and non-detected). When we took into account season, age, sex, diet, and self-reported residential use of pyrethroid insecticides in a linear mixed-effects model, the results suggested that the combination of the use of pyrethroid insecticides in the household, dietary intake, and seasonal differences play a significant role in predicting children's exposure to synthetic pyrethroid insecticides. We found CPES-WA children were continuously exposed to pyrethroid insecticides through their diets all year long, and this chronic exposure pattern was periodically modified by episodes of relatively high exposures from residential uses. Future research should be devoted to enhancing our understanding of the complexity of pyrethroid insecticide exposure patterns.

Proteus mirabilis alternates between motile and adherent forms. MrpJ, a transcriptional regulator previously reported to repress motility, is encoded at the 3' end of the mrp fimbrial operon in P. mirabilis. Sequencing of the P. mirabilis genome revealed 14 additional paralogues of mrpJ, 10 of which are associated with fimbrial operons. Twelve of these genes, when overexpressed, repressed motility; several distinct patterns of swarming motility were noted. Expression of 10 of the 14 mrpJ paralogues repressed flagellin (FlaA) synthesis. Alignment of the predicted amino acid sequences of MrpJ and its 14 paralogues revealed a conserved consensus motif (SQQQFSRYE) within the helix-turn-helix domain. Site-directed mutagenesis of these residues coupled with linker insertion mutagenesis of MrpJ confirmed the importance of this domain for repression of motility. Gel shift assays demonstrated that MrpJ and another paralogue UcaJ bind directly to the promoter region of the flagellar master regulator flhDC. Thus, P. mirabilis appears to use a related mechanism to inhibit motility during the production of at least 10 of its predicted fimbriae.

The gram-negative enteric bacterium Proteus mirabilis is a frequent cause of urinary tract infections in individuals with long-term indwelling catheters or with complicated urinary tracts (e.g., due to spinal cord injury or anatomic abnormality). P. mirabilis bacteriuria may lead to acute pyelonephritis, fever, and bacteremia. Most notoriously, this pathogen uses urease to catalyze the formation of kidney and bladder stones or to encrust or obstruct indwelling urinary catheters. Here we report the complete genome sequence of P. mirabilis HI4320, a representative strain cultured in our laboratory from the urine of a nursing home patient with a long-term (≥30 days) indwelling urinary catheter. The genome is 4.063 Mb long and has a G+C content of 38.88%. There is a single plasmid consisting of 36,289 nucleotides. Annotation of the genome identified 3,685 coding sequences and seven rRNA loci. Analysis of the sequence confirmed the presence of previously identified virulence determinants, as well as a contiguous 54-kb flagellar regulon and 17 types of fimbriae. Genes encoding a potential type III secretion system were identified on a low-G+C-content genomic island containing 24 intact genes that appear to encode all components necessary to assemble a type III secretion system needle complex. In addition, the P. mirabilis HI4320 genome possesses four tandem copies of the zapE metalloprotease gene, genes encoding six putative autotransporters, an extension of the atf fimbrial operon to six genes, including an mrpJ homolog, and genes encoding at least five iron uptake mechanisms, two potential type IV secretion systems, and 16 two-component regulators.

The objective of this study was to determine the feasibility of using salivary biomarkers to assess chlorpyrifos exposure using data collected from laboratory controlled animal study, as well as from farmers in Thailand and Nicaragua who applied chlorpyrifos in the field. Time-matched saliva and arterial blood samples were collected from rats and adult agricultural workers, while spot saliva samples were collected from children. Specimen samples were analyzed for chlorpyrifos using a commercially available enzyme-linked immunosorbent assay. The results from both animal and farmer studies show that chlorpyrifos is excreted into saliva. Nevertheless, salivary excretion of chlorpyrifos seems to differ from other pesticides, as evidenced by the lack of correspondence of chlorpyrifos levels between saliva and plasma samples. The lower chlorpyrifos concentrations in saliva collected from rats, and from farmers and their children, may have resulted from the rapid hydrolysis of chlorpyrifos during the intracellul...

The nucleotide sequence from the genome of Moraxella catarrhalis ATCC 43617 was annotated and used both to assess the metabolic capabilities and limitations of this bacterium and to design probes for a DNA microarray. An absence of gene products for utilization of exogenous carbohydrates was noteworthy and could be correlated with published phenotypic data. Gene products necessary for aerobic energy generation were present, as were a few gene products generally ascribed to anaerobic systems. Enzymes for synthesis of all amino acids except proline and arginine were present. M. catarrhalis DNA microarrays containing 70-mer oligonucleotide probes were designed from the genome-derived nucleotide sequence data. Analysis of total RNA extracted from M. catarrhalis ATCC 43617 cells grown under iron-replete and iron-restricted conditions was used to establish the utility of these DNA microarrays. These DNA microarrays were then used to analyze total RNA from M. catarrhalis cells grown in a continuous-flow biofilm system and in the planktonic state. The genes whose expression was most dramatically increased by growth in the biofilm state included those encoding a nitrate reductase, a nitrite reductase, and a nitric oxide reductase. Real-time reverse transcriptase PCR analysis was used to validate these DNA microarray results. These results indicate that growth of M. catarrhalis in a biofilm results in increased expression of gene products which can function not only in energy generation but also in resisting certain elements of the innate immune response.

Moraxella catarrhalis ETSU-9 was subjected to random transposon insertion mutagenesis to identify genes encoding products involved in the ability of the organism to form biofilms in vitro. Screening of approximately 3,000 transposon insertion mutants in the crystal violet-based biofilm assay system yielded six mutants that exhibited greatly reduced abilities to form biofilms. Three of these mutants had transposon insertions in the uspA2H gene, which encodes a surface protein previously shown to be involved in the ability of M. catarrhalis to both attach to human cell lines in vitro and resist killing by normal human serum. Random insertion mutagenesis of the uspA2H gene, involving the introduction of a 15-nucleotide fragment encoding 5 amino acids, was used to attempt to identify the domain(s) necessary for biofilm formation. Most of these insertions adversely affected biofilm formation, whereas the abilities of these same mutants to attach to Chang conjunctival epithelial cells in vitro were usually not reduced. Gain-of-function experiments showed that introduction of the M. catarrhalis ETSU-9 uspA2H gene into Escherichia coli conferred biofilm formation ability on this recombinant strain. Two of the other three M. catarrhalis ETSU-9 transposon insertion mutants that had greatly reduced abilities to form biofilms were shown to have insertions in genes encoding products predicted to be directly or indirectly involved in cell wall metabolism.

Proteus mirabilis is a Gram-negative bacterium and is well known for its ability to robustly swarm across surfaces in a striking bulls’-eye pattern. Clinically, this organism is most frequently a pathogen of the urinary tract, particularly in patients undergoing long-term catheterization. This review covers P. mirabilis with a focus on urinary tract infections (UTI), including disease models, vaccine development efforts, and clinical perspectives. Flagella-mediated motility, both swimming and swarming, is a central facet of this organism. The regulation of this complex process and its contribution to virulence is discussed, along with the type VI-secretion system-dependent intra-strain competition, which occurs during swarming. P. mirabilis uses a diverse set of virulence factors to access and colonize the host urinary tract, including urease and stone formation, fimbriae and other adhesins, iron and zinc acquisition, proteases and toxins, biofilm formation, and regulation of pathogenesis. While significant advances in this field have been made, challenges remain to combatting complicated UTI and deciphering P. mirabilis pathogenesis.