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Biomedical subjects

David R Johnson

Publications and source records attributed to David R Johnson.

At least 19 recordsLinked to original sources

Soil erosion and landscape elevation as unnoticed determinants of environmental antibiotic resistance distribution.

Climate change is reshaping the global antibiotic resistance gene (ARG) landscape through geomorphological processes that remain largely overlooked in the One Health framework. This critical review synthesises evidence on how soil erosion and landscape elevation gradients redistribute, select for, and disseminate ARGs across terrestrial and aquatic ecosystems. Erosion physically removes and transports ARG-bearing microbes, depletes nutrients, and co-selects for resistance via heavy metal exposure and horizontal gene transfer, creating source-sink dynamics that connect eroding hillslopes to downstream water bodies and food systems. Elevation gradients impose abiotic stressors-declining temperature, elevated UV radiation, and shifting pH-that drive microbial community reassembly through environmental selection and dispersal limitation, with emerging evidence linking bacterial competition at high altitude to enhanced multidrug efflux and resistome complexity. The review identifies critical knowledge gaps, including unquantified ARG mass fluxes across erosion-deposition gradients, unresolved dispersal-versus-selection mechanisms along elevation transects, and the absence of integrated One Health surveillance linking environmental ARG reservoirs to clinical outcomes. A synthesis of global case studies illustrates how these processes converge across diverse landscapes. The review concludes with a mechanistic research agenda-including reciprocal transplant experiments, landscape connectivity modelling, and cross-sectoral surveillance-needed to translate these emerging drivers into actionable climate-AMR mitigation policy.

Drug Resistance, Microbial↗

Suppression of fibroblast metalloproteinases by ajulemic acid, a nonpsychoactive cannabinoid acid.

Production of matrix metalloproteinases (MMP) in joint tissue of patients with inflammatory arthritis facilitates cartilage degradation and bone erosion, and leads to joint deformities and crippling. Thus, MMPs are important targets for agents designed to treat inflammatory arthritis. Oral administration of ajulemic acid (AjA), a synthetic, nonpsychoactive cannabinoid acid, prevents joint tissue injury in rats with adjuvant arthritis. AjA binds to and activates PPARgamma directly. Therefore, we investigated the influence of AjA on MMP production in human fibroblast-like synovial cells (FLS), and examined the role of PPARgamma in the mechanism of action of AjA. FLS, treated or not with a PPARgamma antagonist, were treated with AjA then stimulated with TNFalpha or IL-1alpha. Release of MMPs-1, 3, and 9 was measured by ELISA. The influence of AjA on MMP-3 release from stimulated PPARgamma positive (PPAR+/-) and PPARgamma null (PPAR-/-) mouse embryonic fibroblasts (MEF) was also examined. Addition of AjA to FLS suppressed production of MMPs whether or not PPARgamma activation was blocked. Secretion of MMP-3 was also suppressed by AjA in both TNFalpha- and IL-1alpha-stimulated PPARgamma+/- and PPARgamma-/- MEF. Suppression of MMP secretion from FLS by AjA appears to be PPARgamma independent. Prevention by AjA of joint tissue injury and crippling in the rat adjuvant arthritis model may be explained in large part by inhibition of MMPs. These results suggest that AjA may be useful for treatment of patients with rheumatoid arthritis and osteoarthritis.

Animals↗

Development and confirmatory factor analysis of the community norms of child neglect scale.

This article describes the development of the Community Norms of Child Neglect Scale (CNCNS), a new measure of perceptions of child neglect, for use in community samples. The CNCNS differentiates among four subtypes of neglect (failure to provide for basic needs, lack of supervision, emotional neglect, and educational neglect). Scenarios ranging in seriousness for each subtype were presented to a large community sample (N = 3,809). Confirmatory factor analyses indicated that a four-factor model provided a better fit to the data than did a model specifying only one overall neglect factor, suggesting this sample distinguished among the four subtypes of neglect. The authors tested measurement equivalence across individuals who work with children and lay community respondents and across rural and urban respondents, with results indicating a very similar structure across these groups. These initial reliability and validity data suggest that the CNCNS may be of use in comparing perceptions of child neglect among individuals and across communities.

Adolescent↗

A real-time PCR method for quantifying viable ascaris eggs using the first internally transcribed spacer region of ribosomal DNA.

Worldwide, 1.4 billion people are infected with the intestinal worm Ascaris lumbricoides. As a result, Ascaris eggs are commonly found in wastewater and sludges. The current microscopy method for detecting viable Ascaris eggs is time- and labor-intensive. The goal of this study was to develop a real-time quantitative PCR (qPCR) method to determine the levels of total and viable Ascaris eggs in laboratory solutions using the first internally transcribed spacer (ITS-1) region of ribosomal DNA (rDNA) and rRNA. ITS-1 rDNA levels were proportional to Ascaris egg cell numbers, increasing as eggs developed from single cells to mature larvae and ultimately reaching a constant level per egg. Treatments causing >99% inactivation (high heat, moderate heat, ammonia, and UV) eliminated this increase in ITS-1 rDNA levels and caused decreases that were dependent on the treatment type. By taking advantage of this difference in ITS-1 rDNA level between viable, larvated eggs and inactivated, single-celled eggs, qPCR results were used to develop inactivation profiles for the different treatments. No statistical difference from the standard microscopy method was found in 75% of the samples (12 of 16). ITS-1 rRNA was detected only in samples containing viable eggs, but the levels were more variable than rDNA levels and ITS-1 rRNA could not be used for quantification. The detection limit of the rDNA-based method was approximately one larvated egg or 90 single-celled eggs; the detection limit for the rRNA-based method was several orders of magnitude higher. The rDNA qPCR method is promising for both research and regulatory applications.

Animals↗

Effect of carbon starvation on toluene degradation activity by toluene monooxygenase-expressing bacteria.

Subsurface bacteria commonly exist in a starvation state with only periodic exposure to utilizable sources of carbon and energy. In this study, the effect of carbon starvation on aerobic toluene degradation was quantitatively evaluated with a selection of bacteria representing all the known toluene oxygenase enzyme pathways. For all the investigated strains, the rate of toluene biodegradation decreased exponentially with starvation time. First-order deactivation rate constants for TMO-expressing bacteria were approximately an order of magnitude greater than those for other oxygenase-expressing bacteria. When growth conditions (the type of growth substrate and the type and concentration of toluene oxygenase inducer) were varied in the cultures prior to the deactivation experiments, the rate of deactivation was not significantly affected, suggesting that the rate of deactivation is independent of previous substrate/inducer conditions. Because TMO-expressing bacteria are known to efficiently detoxify TCE in subsurface environments, these findings have significant implications for in situ TCE bioremediation, specifically for environments experiencing variable growth-substrate exposure conditions.

Biodegradation, Environmental↗

Reductive dehalogenase gene expression as a biomarker for physiological activity of Dehalococcoides spp.

This study characterizes the transcriptional expression of the reductive dehalogenase (RDase)-encoding tceA and vcrA genes and evaluates their applicability as potential biological markers of Dehalococcoides activity. When Dehalococcoides ethenogenes 195 was provided with trichloroethene (TCE) as the electron acceptor, the expression of the tceA gene increased by 90-fold relative to that in cells starved of chlorinated ethenes, demonstrating that tceA gene expression is indicative of the active physiological state of this strain. In a Dehalococcoides-containing enrichment culture that contains both the tceA and vcrA genes, the tceA gene was up-regulated in response to TCE and cis-1,2-dichloroethene (cDCE) exposure, while the vcrA gene was up-regulated in response to TCE, cDCE, and vinyl chloride (VC). When chlorinated ethenes were depleted, the RDase-encoding gene transcripts decayed exponentially, with a half-life between 4.8 and 6.1 h, until they reached a stable background level after 2 days. We found that while gene expression correlated generally to the presence of chlorinated ethenes, there was no apparent direct relationship between RDase-encoding transcript numbers and respective rates of TCE, cDCE, and VC dechlorination activities. However, elevated tceA and vcrA expression did correlate with chlorinated-ethene reduction beyond cDCE, suggesting that elevated RDase-encoding transcript numbers could serve as a biomarker for the physiological ability of Dehalococcoides spp. to dechlorinate beyond cDCE.

Base Sequence↗

An adolescent and adult formulation combined tetanus, diphtheria and five-component pertussis vaccine.

Pertussis causes substantial morbidity among adolescents and adults, as well as other persons in contact with affected individuals. In the context of widespread childhood immunization, vaccination of adolescents and adults is a relatively new strategy for reducing the disease in all age groups. ADACEL is a tetanus-diphtheria-acellular pertussis combination vaccine (incorporating pertussis toxoid, filamentous hemagglutinin, pertactin and fimbriae type 2 and 3 antigens) formulated for use in adolescents and adults based on similar products designed for infants and children. ADACEL vaccine is approved for use in Australia, Canada, Germany and the USA, and has been demonstrated to induce robust immune responses and acceptable levels of reactogenicity in clinical trials. Surveillance data from Canada, where ADACEL vaccine has been used in adolescents and adults in conjunction with ongoing childhood vaccination for several years, indicate an enhanced overall reduction in pertussis disease.

Adolescent↗

A study of the efficacy of various home filtration substrates in the removal of microcystin-LR from drinking water.

This study was conducted to determine whether common water filtration and purification systems bought by consumers and used in the home would remove cyanotoxins from water. Commonly used universal filter housings and filter sizes were utilized to identify filter media that may be effective in the removal of microcystin-LR in deionized water. Results suggest that the efficacy of home filtration devices in removing microcystin-LR varies considerably with the type of device being used. Carbon filters successfully removed microcystin-LR allowing only 0.05-0.3% of the toxin load to pass through the filter. On the other hand, pleated paper and string wound filters allowed > 90% of microcystin-LR present in the sample to pass through the filters. Theoretically, the use of carbon home filtration devices tested in this study may provide protection against human exposure to cyanotoxin in addition to protection provided by water treatment methodologies utilized in water treatment facilities. Further studies need to be done to assess the efficacy of home filtration devices for various cyanotoxins and for other filtering conditions such as increased toxin load, the presence of other contaminants in drinking water, and the repetitive use of the same filter over longer time intervals.

Bacterial Toxins↗

Infertility: testing a helpseeking model.

This paper uses data from a study of 196 infertile women from the Midwestern US to examine a general theory of helpseeking behavior applied to infertility. All of these women report meeting the medical definition of infertility--12 months or more of regular intercourse without conception--at some point in their lives. Only 35 percent of this sample of infertile women identified themselves as having had fertility problems and only 40 percent had sought medical treatment. Drawing on prior theories of helpseeking, we examine the effects of symptom salience, life course cues, attitudes, predisposing factors, and enabling conditions on helpseeking. We posit a model in which a cognitive dimension (perceived infertility) mediates between these predictors and medical helpseeking. Symptom salience (experienced infertility while actively trying to get pregnant), low parity, and poor subjective health are significantly related to perceived infertility, which is, in turn, significantly associated with helpseeking for infertility. Supporting the conclusion that the cognitive dimension of identifying oneself as infertile is critical to helpseeking, the relationship of symptom salience to helpseeking is partially mediated by perceived problems. Internal health locus of control is associated with lower odds of helpseeking but not to perceived infertility.

Adult↗

Regulation and binding of pregnane X receptor by nuclear receptor corepressor silencing mediator of retinoid and thyroid hormone receptors (SMRT).

The pregnane X receptor (PXR) is an orphan nuclear receptor predominantly expressed in liver and intestine. PXR coordinates hepatic responses to prevent liver injury induced by environmental toxins. PXR activates cytochrome P450 3A4 gene expression upon binding to rifampicin (Rif) and clotrimazole (CTZ) by recruiting transcriptional coactivators. It remains unclear whether and how PXR regulates gene expression in the absence of ligand. In this study, we analyzed interactions between PXR and the silencing mediator of retinoid and thyroid hormone receptors (SMRT) and determined the role of SMRT in regulating PXR activity. We show that SMRT interacts with PXR in glutathione S-transferase pull-down, yeast two-hybrid, and mammalian two-hybrid assays. The interaction is mediated through the ligand-binding domain of PXR and the SMRTs' nuclear receptor-interacting domain 2. The PXR-SMRT interaction is sensitive to species-specific ligands, and Rif causes an exchange of the corepressor SMRT with the p160 coactivator known as receptor-associated coactivator 3 (RAC3). Deletion of the PXR's activation function 2 helix enhances SMRT binding and abolishes ligand-dependent dissociation of SMRT. Coexpression of PXR with SMRT results in colocalization at discrete nuclear foci. Finally, transient transfection assays show that overexpression of SMRT inhibits PXR's transactivation of the Cyp3A4 promoter, whereas silencing of SMRT enhances the reporter expression. Taken together, our results suggest that the corepressor SMRT may bind to and regulate the transcriptional activity of PXR.

Base Sequence↗

Transcriptional expression of the tceA gene in a Dehalococcoides-containing microbial enrichment.

Dynamic changes in the transcriptional expression of the tceA gene, which encodes a trichloroethene reductive dehalogenase, were characterized in a Dehalococcoides-containing microbial enrichment culture. Expression was quantified by real-time PCR as the number of tceA transcripts per tceA gene. Expression of tceA increased 40-fold after chlorinated ethene-starved cells were exposed to trichloroethene (TCE), cis-dichloroethene (DCE), or 1,1-DCE but did not increase after exposure to tetrachloroethene or vinyl chloride. Surprisingly, tceA expression also increased 30-fold after cellular exposure to the nonmetabolic substrate trans-DCE, indicating that expression of tceA is induced by both growth-supporting and non-growth-supporting chlorinated ethenes. Additional experiments revealed that the level of tceA expression was independent of the concentration of chlorinated ethenes (sum concentrations of TCE and DCEs of 2.2 to 333 microM), the concentration of the electron donor hydrogen (concentrations of 12 nM to 17 microM), and the presence of alternate bacterial electron acceptors (5 mM concentrations of fumarate, sulfate, sulfite, thiosulfate, nitrate, or nitrite) but was highly dependent on incubation temperature.

Chloroflexi↗

An internal reference technique for accurately quantifying specific mRNAs by real-time PCR with application to the tceA reductive dehalogenase gene.

The accuracy of mRNA quantification by reverse transcription (RT) in conjunction with real-time PCR (qPCR) is limited by mRNA losses during sample preparation (cell lysis, RNA isolation, and DNA removal) and by inefficiencies in reverse transcription. To control for these losses and inefficiencies, a technique was developed that utilizes an exogenous internal reference mRNA (ref mRNA) along with mRNA absolute standard curves. The technique was applied to quantify mRNA of the trichloroethene (TCE) reductive dehalogenase-encoding tceA gene in an anaerobic TCE-to-ethene dechlorinating microbial enrichment. Compared to RT-qPCR protocols that utilize DNA absolute standard curves, application of the new technique increased measured quantities of tceA mRNA by threefold, demonstrating a substantial improvement in quantification. The technique was also effective for quantifying the loss of mRNA during specific steps of the sample processing protocol. Analysis revealed that the efficiency of the RNA isolation (56%) step was significantly less than that of the cell lysis (84%), DNA removal (93%), and RT (88%) steps. The technique was applied to compare the effects of cellular exposure to different chlorinated ethenes on tceA expression. Results show that exposure to TCE or cis-1,2-dichloroethene resulted in 25-fold-higher quantities of tceA mRNA than exposure to vinyl chloride or chlorinated ethene starvation.

Anaerobiosis↗

Mediator analysis of Moms on the move.

This study examined whether improvements in physical activity discovered in a test of Moms on the Move were mediated by the behavioral constructs from which the intervention was derived. Identifying mediator variables is vital for intervention planning. The intervention was derived from the Transtheoretical Model (TTM) of behavior change and promoted moderately intense activities like walking. Sedentary mothers with children enrolled in the Women, Infants, and Children program (N = 44) were randomly assigned to the experimental intervention or attention control. Large effect sizes were reported for improvements in physical activity and changes in TTM constructs. This study examined whether the physical activity improvements were mediated by the behavioral constructs. Statistical analyses used bivariate correlation coefficients and two-stage multiple linear regression. These exploratory findings did not support the hypothesis that increased physical activity, which was associated with the experimental intervention, was mediated by the TTM constructs.

Adolescent↗

NuRD complex component Mi-2beta binds to and represses RORgamma-mediated transcriptional activation.

RORgamma is a nuclear receptor that binds to DNA motifs as a monomer to constitutively activate target genes. RORgamma plays an important role in thymocyte development and lymph node organogenesis, while the regulation of RORgamma-mediated transcriptional activation is currently unclear. The purpose of this study was to identify other nuclear proteins that interact with RORgamma. A yeast two-hybrid screen with Y190 yeast cells under stringent conditions resulted in the identification of CHD4, also known as Mi-2beta, as a RORgamma-interacting protein. This interaction was confirmed by GST pull-down assays. This interaction occurred within the middle regulatory region (amino acids 719-1164) of Mi-2beta. Transfection of Gal4-RORgamma into HeLa cells resulted in constitutive transactivation of the MH100-tk-luc reporter. The addition of Mi-2beta resulted in a dramatic 50% decrease in Gal4-RORgamma-mediated transactivation. These data demonstrate that RORgamma forms a protein-protein interaction with the regulatory region of Mi-2beta, resulting in inhibition of RORgamma transcriptional activity. These results may provide evidence as to how RORgamma-mediated transactivation is regulated by other nuclear proteins.

Animals↗

Endothelial cells present antigens in vivo.

BACKGROUND: Immune recognition of vascular endothelial cells (EC) has been implicated in allograft rejection, protection against pathogens, and lymphocyte recruitment. However, EC pervade nearly all tissues and predominate in none, complicating any direct test of immune recognition. Here, we examined antigen presentation by EC in vivo by testing immune responses against E. coli beta-galactosidase (beta-gal) in two lines of transgenic mice that express beta-gal exclusively in their EC. TIE2-lacZ mice express beta-gal in all EC and VWF-lacZ mice express beta-gal in heart and brain microvascular EC. RESULTS: Transgenic and congenic wild type FVB mice immunized with beta-gal expression vector DNA or beta-gal protein generated high titer, high affinity antisera containing comparable levels of antigen-specific IgG1 and IgG2a isotypes, suggesting equivalent activation of T helper cell subsets. The immunized transgenic mice remained healthy, their EC continued to express beta-gal, and their blood vessels showed no histological abnormalities. In response to beta-gal in vitro, CD4+ and CD8+ T cells from immunized transgenic and FVB mice proliferated, expressed CD25, and secreted IFN-gamma. Infection with recombinant vaccinia virus encoding beta-gal raised equivalent responses in transgenic and FVB mice. Hearts transplanted from transgenic mice into FVB mice continued to beat and the graft EC continued to express beta-gal. These results suggested immunological ignorance of the transgene encoded EC protein. However, skin transplanted from TIE2-lacZ onto FVB mice lost beta-gal+ EC and the hosts developed beta-gal-specific antisera, demonstrating activation of host immune effector mechanisms. In contrast, skin grafted from TIE2-lacZ onto VWF-lacZ mice retained beta-gal+ EC and no antisera developed, suggesting a tolerant host immune system. CONCLUSION: Resting, beta-gal+ EC in transgenic mice tolerize specific lymphocytes that would otherwise respond against beta-gal expressed by EC within transplanted skin. We conclude that EC effectively present intracellular "self" proteins to the immune system. However, antigen presentation by EC does not delete or anergize a large population of specific lymphocytes that respond to the same protein following conventional immunization with protein or expression vector DNA. These results clearly demonstrate striking context sensitivity in the immune recognition of EC, a subtlety that must be better understood in order to treat immune diseases and complications involving the vasculature.

Animals↗

Trauma and coping in Somali and Oromo refugee youth.

PURPOSE: To describe war-related trauma history, immigration factors, problems, and coping of Somali and Oromo refugee youth. METHODS: Analysis of a subset of participants (N = 338) aged 18-25 years from a population-based survey of Somali and Oromo refugees conducted in 2000-2002. Data included trauma history, life situation, and scales for physical (Cronbach alpha =.69), psychological (alpha =.56), and social problems (alpha =.69). Data were analyzed using Chi-square and Mann-Whitney U tests. RESULTS: Average emigration age was 14.8 years, with 4.2 years in transit and 2.0 years in the United States; 60% reported plans to return home to live. Two-thirds (66%) had less than a high school education, 49% had English language problems, 49% were employed (38% female vs. 57% male); 70% were single, with Somali females more likely than Oromo to be partnered and mothers (39% vs. 19%). There were significant ethnicity/gender differences for all problem scales. More females reported feeling alone (24% vs. 61%, p <.001). Youth with symptoms of posttraumatic stress syndrome reported more traumatic events (mean number of events: 28 vs.16). Trauma history was strongly associated with physical, psychological, and social problems. Most frequent strategies to combat sadness were praying (55.3%), sleeping (39.9%), reading (32.3%), and talking to friends (27.8%). CONCLUSIONS: Many young Somali and Oromo immigrants to the United States experience life problems associated with war trauma and torture, but many others are coping well. The findings suggest a need to develop age-appropriate strategies to promote the health of refugee youth to facilitate their successful adaptation to adult life in the United States.

Adaptation, Psychological↗

A randomized pilot test of "Moms on the Move": a physical activity intervention for WIC mothers.

BACKGROUND: Sedentary mothers are important to reach with physical activity promotion. PURPOSE: This study pilot tested "Moms on the Move," a Transtheoretical Model (TTM)-derived physical activity (PA) intervention for low-income mothers enrolled in the Women, Infants, and Children (WIC) program. METHOD: Sedentary mothers (N = 44) were randomized to (a). Moms on the Move (PA intervention) or (b). counseling on self-breast examination (control). Pre- and posttest measurement (baseline and 2 weeks after the 8-week interventions) included (a). stage of PA behavior change, (b). PA behavior, (c). selected TTM constructs, and (d). social support. Pre- and postdifference scores, chi-square, and one-way analyses of variance (ANOVAs) were used. RESULTS: The experimental group progressed in stage of change more than control, chi2(1, N = 44) = 20.50, p <.001. The experimental group had greater PA behavior: weekly minutes of PA, F(1, 42) = 46.85, p <.001; daily energy expenditure (EE), F(1, 42) = 23.01, p <.001; and weekly moderate PA EE, F(1, 42) = 32.63, p <.001. Experimental subgroup (n = 11) step counts increased pre-post, t(10) = 6.16, p <.001. An ANOVA showed that the experimental group had greater improvements in all TTM constructs and social support, ps <.001. CONCLUSIONS: WIC mothers are at risk for sedentary living and have not been targeted for PA behavior change using a provider-counseled approach. Although further testing is needed, Moms on the Move appears to be efficacious.

Adult↗

Impact of PCV7 on invasive pneumococcal disease among children younger than 5 years: a population-based study.

We examined the impact of the heptavalent pneumococcal conjugate vaccine (PCV7) on hospital discharge rates for invasive pneumococcal disease among Michigan children younger than 5 years old. After the introduction of PCV7, the hospital discharge rate for children younger than 1 years old was significantly lower than before introduction. We correlated the decreased rates with the introduction and rapid uptake of PCV7. Lack of change in the hospital discharge rates for other age groups likely represents a slower uptake of PCV7 compared with that for children aged younger than 1 year.

Female↗