Measuring the Morphologic Response of Braided Rivers to Lateral Constriction

The aim of this thesis is to understand the morphologic changes to a set of historically braided rivers that have been narrowed. Braided rivers from the agriculturally developed Canterbury Plains, New Zealand, were studied from a period prior to much development (mid-1900s) to the present. Narrowing of channels, decreased braiding intensity, and loss of braided planforms were determined based on aerial imagery, changing the geography of braiding along all rivers. Channel width and count were statistically correlated and show the predictability of braiding change based on narrowing. Reaches with initially wide channels require more narrowing to induce a simplification of braiding, while narrower reaches may be closer to a threshold of change and require less narrowing to transition. The implications of the results can be used in river management to create wide enough river corridors that allow the rivers to maintain their naturally braided planforms while mitigating flood risk

Development of an integrated cognitive behavioral therapy for anxiety and opioid use disorder: Study protocol and methods

Opioid use disorder is a highly disabling psychiatric disorder, and is associated with both significant functional disruption and risk for negative health outcomes such as infectious disease and fatal overdose. Even among those who receive evidence-based pharmacotherapy for opioid use disorder, many drop out of treatment or relapse, highlighting the importance of novel treatment strategies for this population. Over 60% of those with opioid use disorder also meet diagnostic criteria for an anxiety disorder; however, efficacious treatments for this common co-occurrence have not be established. This manuscript describes the rationale and methods for a behavioral treatment development study designed to develop and test an integrated cognitive-behavioral therapy for those with co-occurring opioid use disorder and anxiety disorders. The aims of the study are (1) to develop and pilot test a new manualized cognitive behavioral therapy for co-occurring opioid use disorder and anxiety disorders, (2) to test the efficacy of this treatment relative to an active comparison treatment that targets opioid use disorder alone, and (3) to investigate the role of stress reactivity in both prognosis and recovery from opioid use disorder and anxiety disorders. Our overarching aim is to investigate whether this new treatment improves both anxiety and opioid use disorder outcomes relative to standard treatment. Identifying optimal treatment strategies for this population are needed to improve outcomes among those with this highly disabling and life-threatening disorder.This study was funded by NIDA grant DA035297. The funding source had no involvement in the study design, analysis and interpretation of data, writing of the report, or the decision to submit the article for publication. (DA035297 - NIDA)Accepted manuscrip

Identification of bacterial posttranslational modifications that regulate antimicrobial resistance

Rapidly spreading antimicrobial resistance is one of the greatest threats to human health of our time. There exists many mechanisms by which bacteria evade destruction by antibiotics and various research is being conducted globally in an attempt to combat this alarming situation. A so far under-researched area is the hypothesised role of protein post-translational modifications in the regulation of antimicrobial resistance responses. This thesis identifies two proteins involved in antimicrobial resistance and known to undergo post-translational modification, then uses genetic code expansion and unnatural amino acid incorporation to characterise the effect of these modifications on protein function. The first protein is AdeT1, a component of a multi-drug efflux pump from the multidrug resistant pathogen Acinetobacter baumannii, which undergoes lysine propionylation at Lys280 in vivo. Here, such propionylation is shown to modulate function of the efflux pump in living cells, with the propionylated protein complex demonstrating increased ethidium bromide efflux. Further, cells producing propionylated AdeT1 required a 6-fold higher concentration of erythromycin to inhibit their growth compared to cells producing unmodified AdeT1. The second protein investigated is the DNA-binding protein HU from Escherichia coli, which undergoes acetylation at five lysine residues in vivo. The protein was characterised in DNA-binding assays in vitro, which revealed that protein acetylated at Lys86 has significantly lower affinity for DNA compared to its wildtype counterpart. The consequences of such altered binding on bacterial gene expression and antimicrobial resistance are contemplated. Together, the results presented here are in agreement with the known roles of post-translational modifications in modulating protein function and provide evidence toward a novel analysis of posttranslational modifications as regulatory components in antimicrobial resistance response

Planejamento de educação parental on-line para um estilo de vida de peso saudável em pacientes pediátricos de cuidados primários de 6 a 12 anos

Background: Childhood obesity is a pandemic with immediate and future health risks that demand holistic, personalized, and innovative treatments. Healthcare providers struggle to provide such treatment within the constraints of medical visits. A theoretical and evidence-based eHealth intervention could engage parents and caregivers to improve health behaviors and increase reach to low-income vulnerable families, a high-risk chort often neglected in clinical research. This study aims to identify needs and preferences of potential users of an eHealth intervention. Methods: Focus groups and interviews of ethnically diverse, low-income parents and semi-structured interviews of primary healthcare staff obtained perspectives on the content and structure of a eHealth intervention intended to improve lifestyle habits to reduce childhood obesity. Research team members analyzed the data to identify specific needs and communication strategies for the target population.   Results:  Parents endorsed learning more about healthy eating, physical activity, and the effects of sugar-sweetened beverages on their child’s weight.  Parents reported a preference for bright colors, short texts, videos, summary quizzes, and additional resources to explore on their own and with their child. Healthcare staff reported time limitations during clinical visits and an interest in looping family progress into their EHR. Using an eHealth intervention, parents expect an improved relationship and increased trust with their child’s pediatrician.   Conclusion:  Parents and staff saw value in this tool and provided useful recommendations for its design. This eHealth intervention could create a health alliance between the healthcare provider and family, provide personalized content, and be scaled to address health disparities broadly.Antecedentes. la obesidad infantil es una pandemia con riesgos para la salud inmediatos y futuros que demandan tratamientos holísticos, personalizados e innovadores. Los proveedores de atención médica luchan por brindar dicho tratamiento dentro de las limitaciones de las visitas médicas. Una intervención de eSalud teórica y basada en la evidencia podría involucrar a los padres y cuidadores para mejorar los comportamientos de salud y aumentar el alcance a las familias vulnerables de bajos ingresos, un grupo de alto riesgo que a menudo se descuida en la investigación clínica. Este estudio tiene como objetivo identificar las necesidades y preferencias de los usuarios potenciales de una intervención de eSalud. Métodos. Grupos focales y entrevistas de padres étnicamente diversos y de bajos ingresos y entrevistas semiestructuradas del personal de atención primaria de la salud obtuvieron perspectivas sobre el contenido y la estructura de una intervención de eSalud destinada a mejorar los hábitos de estilo de vida para reducir la obesidad infantil. Los miembros del equipo de investigación analizaron los datos para identificar necesidades específicas y estrategias de comunicación para la población objetivo. Resultados. Los padres aprobaron aprender más sobre alimentación saludable, actividad física y los efectos de las bebidas azucaradas en el peso de sus hijos. Los padres informaron una preferencia por los colores brillantes, los textos breves, los videos, los cuestionarios resumidos y los recursos adicionales para explorar por su cuenta y con sus hijos. El personal de atención médica informó limitaciones de tiempo durante las visitas clínicas y un interés en incluir el progreso de la familia en su EHR. Con una intervención de eSalud, los padres esperan una mejor relación y una mayor confianza con el pediatra de su hijo. Conclusión. Los padres y el personal vieron valor en esta herramienta y brindaron recomendaciones útiles para su diseño. Esta intervención de eSalud podría crear una alianza de salud entre el proveedor de atención médica y la familia, brindar contenido personalizado y ampliarse para abordar las disparidades de salud de manera amplia.Antecedentes. A obesidade infantil é uma pandemia com riscos à saúde imediatos e futuros que demandam tratamentos holísticos, personalizados e inovadores. Os prestadores de cuidados de saúde lutam para fornecer esse tratamento dentro das limitações das consultas médicas. Uma intervenção teórica e baseada em evidências de eSaúde poderia envolver pais e cuidadores para melhorar os comportamentos de saúde e aumentar o alcance de famílias vulneráveis de baixa renda, um grupo de alto risco que muitas vezes é negligenciado em pesquisas clínicas. Este estudo visa identificar as necessidades e preferências dos potenciais utilizadores de uma intervenção de eSaúde. Métodos. Grupos focales y entrevistas de padres étnicamente diversos y de bajos ingresos y entrevistas semiestructuradas del personal de atención primaria de la salud obtuvieron perspectivas sobre el contenido y la estructura de una intervención de eSalud destinada a mejorar los hábitos de estilo de vida para reducir la obesidade infantil. Os membros da equipe de pesquisa analisaram os dados para identificar necessidades específicas e estratégias de comunicação para a população-alvo. Resultados. Os pais aprovaram aprender mais sobre alimentação saudável, atividade física e os efeitos das bebidas açucaradas no peso de seus filhos. Os pais relataram preferência por cores vivas, textos curtos, vídeos, questionários curtos e recursos adicionais para explorar sozinhos e com seus filhos. A equipe de saúde relatou limitações de tempo durante as visitas à clínica e interesse em incluir o progresso da família em seu EHR. Com uma intervenção de eSaúde, os pais esperam um melhor relacionamento e maior confiança com o pediatra de seus filhos. Conclusão. Os pais e funcionários viram valor nesta ferramenta e forneceram recomendações úteis para o seu design. Essa intervenção de eSaúde pode criar uma parceria de saúde entre o profissional de saúde e a família, fornecer conteúdo personalizado e ser ampliada para abordar amplamente as disparidades de saúde

The MALDI TOF E2/E3 ligase assay as universal tool for drug discovery in the ubiquitin pathway

AbstractIn many diseases, components of the ubiquitin system - such as E2/E3 ligases and deubiquitylases - are dysregulated. The ubiquitin system has therefore become an emergent target for the treatment of a number of diseases, including cancer, neurodegeneration and autoimmunity. Despite of the efforts in this field, primary screenings of compound libraries to individuate new potential therapeutic molecules targeting the ubiquitin pathway have been strongly limited by the lack of robust and fast high-throughput assays. Here we report the first label-free high-throughput screening (HTS) assay for ubiquitin E2 conjugating enzymes and E3 ligases based on Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight (MALDI TOF) mass spectrometry. The MALDI TOF E2/E3 assay allows us to test E2 conjugating enzymes and E3 ligases for their ubiquitin transfer activity, to identify E2/E3 active pairs, inhibitor potency and specificity and to screen compound librariesin vitrowithout synthesis of chemical or fluorescent probes. We demonstrate that the MALDI TOF E2/E3 assay is a universal tool for drug discovery screening in the ubiquitin pathway as it is suitable for working with all E3 ligase families and requires a reduced amount of reagents, compared to standard biochemical assays.</jats:p

Acetylation at lysine 86 of escherichia coli HUβ modulates the DNA-binding capability of the protein

DNA-binding protein HU is highly conserved in bacteria and has been implicated in a range of cellular processes and phenotypes. Like eukaryotic histones, HU is subjected to post-translational modifications. Specifically, acetylation of several lysine residues have been reported in both homologs of Escherichia coli HU. Here, we investigated the effect of acetylation at Lys67 and Lys86, located in the DNA binding-loop and interface of E. coli HUβ, respectively. Using the technique of genetic code expansion, homogeneous HUβ(K67ac) and HUβ(K86ac) protein units were obtained. Acetylation at Lys86 seemed to have negligible effects on protein secondary structure and thermal stability. Nevertheless, we found that this site-specific acetylation can regulate DNA binding by the HU homodimer but not the heterodimer. Intriguingly, while Lys86 acetylation reduced the interaction of the HU homodimer with short double-stranded DNA containing a 2-nucleotide gap or nick, it enhanced the interaction with longer DNA fragments and had minimal effect on a short, fully complementary DNA fragment. These results demonstrate the complexity of post-translational modifications in functional regulation, as well as indicating the role of lysine acetylation in tuning bacterial gene transcription and epigenetic regulation

A Radio Pulsar/X-ray Binary Link

Radio pulsars with millisecond spin periods are thought to have been spun up by transfer of matter and angular momentum from a low-mass companion star during an X-ray-emitting phase. The spin periods of the neutron stars in several such low-mass X-ray binary (LMXB) systems have been shown to be in the millisecond regime, but no radio pulsations have been detected. Here we report on detection and follow-up observations of a nearby radio millisecond pulsar (MSP) in a circular binary orbit with an optically identified companion star. Optical observations indicate that an accretion disk was present in this system within the last decade. Our optical data show no evidence that one exists today, suggesting that the radio MSP has turned on after a recent LMXB phase.Comment: published in Scienc

Development of mammalian cell logic gates controlled by unnatural amino acids

Genetic code expansion employing an orthogonal aminoacyl-tRNA synthetase/tRNA pair for site-specific unnatural amino acid incorporation has a wide range of applications. However, a blank (orthogonal) codon is needed for each orthogonal tRNA. Although the amber stop codon is commonly used as the blank codon, the use of stop codons as the blank codons inevitably limits the number of possible different unnatural amino acids to be incorporated into proteins in a single cell. To overcome this limitation, we evaluated the capability of 11 Pyl tRNA variants for decoding quadruplet codons in mammalian cells. With efficient quadruplet-decoding orthogonal tRNA variants available, it is possible for simultaneous incorporation of more than three different unnatural amino acids in mammalian cells