Functional Studies of Novel Bioactives From Complex Host-Microbiomes as Drug Leads for Cancer, Infectious Disease, Depression and Pain

The drug discovery process has become increasingly complex in comparison to early efforts particularly those involving diseases or conditions that affect large numbers of the human population. It is difficult to point directly to one factor leading evolution of the process but the definition of a mechanism of pharmacological action for a drug lead has noticeably become a higher priority. This evolution has helped diminish the stigma of natural products and compound supply issues. The issue of compound supply has also been addressed to some extent with the emergence of recombinant technologies and new synthetic methodology. With approximately 60 years of research performed after the development of SCUBA, the drug discovery opportunities in the sea are still too numerous to count. Since the FDA approval of the direct-from-the-sea calcium channel blocker Prialt (ziconotide), marine natural products has been validated as a source for new medicines. However, the demand for natural products is extremely high due to the development of high-throughput assays and this bottleneck has created the need for an intense focus on increasing the rate of isolating and elucidating new bioactive secondary metabolites. A cystine-rich peptide asteropusin A (ASPA) was isolated from the marine sponge Asteropus sp. and its structure determined by X-ray crystallography and NMR spectroscopy. Administration of ASPA to veratridine-stimulated cerebrocortical neuron cells enhances calcium influx but does not modify the oscillation frequency or amplitude of the neuronal calcium alone. Ion channel modulation is an emerging target for drug therapy and the discovery of an ion channel interacting knottin from Porifera accompanied by such high quality spatial details is an uncomcombination. Improvements made to the isolation of peptide metabolites from marine sponges of the Family Theonellidae enables the isolation of significant amounts of potential angiogenesis inhibitors like theopapuamide with high purity. The improvements are particularly relevant in future cases where nuisance components like the aurantoside dyes can lead to false indications of cytotoxicity or antifungal activity. The elucidation of a new theopapuamide analog by CID-MS from Theonella invaginata indicates the high level of peptide diversity found within this Family. Two functional studies of marine natural products contribute to the understanding of their mechanisms of biological activity which can provide insight into their future development as drugs. In the first study, aaptamine was found to possess anxiolytic effects in vivo using a chick anxiety model. A large number of neurological receptors and enzymes were screenedin vitro. In vivo functional challenges were then performed to validate the anxiolytic targetamong several putative candidates previously identified by the in vitro studies. The results of those challenges eliminated several anxiety linked receptor targets and indicated aaptamine as a modulator of monoamine oxidase inhibition activity in vivo as an alternative explanation. The second functional study evaluates new latrunculin B analogs and their correlation of predicted binding with G-actin to the inhibition of polymerization. The latrunculins are well-studied sponge derived inhibitors of actin polymerization and the results validate a method for in silico prediction activity this cancer drug target. Natural products which are isolated in high yields can imply the absence of a utility for humans considering that discovery efforts areprimarily focused on producing drug and agrochemical agents. However, compounds which are abundant in one organism imply an ecological impact that can be studied for the development of an alternative use. A significant quantity of fragrant oil was obtained from a Jamaican Plakortis sp. by cryo-trap. The oil was determined to be exclusively n-decan-2-one. The antifouling character of the oil was evaluated by its effects on surface attachment of a Gram negative bacterial model using confocal fluorescence microscopy as well as its effects on the attachment of Dreissena polymorpha (zebra mussel). The ketone ( n-decan-2-one) inhibited attachment of the bacteria and zebra mussels. Although the aliphatic ketone alone is not a potential commercial alternative for antifouling coatings, incorporating the functionality into coating design is a feasible alternative. The unusual amount of oil extracted from the imported fire ant ( Solenopsis sp.) may be an indication of the presence of oleaginous microorganisms or enzymes supporting the digestion of raw sugars. Heat of combustion of the ant oil was 133,000 BTU/gal, an amount within the range of reported values for vegetable oil and biodiesel. This investigation offers a unique perspective of a potentially new source of microorganisms or enzymes useful for reducing the cost of producing an alternative fuel

Is there a correlation between infection control performance and other hospital quality measures?

Quality measures are increasingly reported by hospitals to the Centers for Medicare and Medicaid Services (CMS), yet there may be tradeoffs in performance between infection control (IC) and other quality measures. Hospitals that performed best on IC measures did not perform well on most CMS non–IC quality measures

Dynamic DNA methylation across diverse human cell lines and tissues

As studies of DNA methylation increase in scope, it has become evident that methylation has a complex relationship with gene expression, plays an important role in defining cell types, and is disrupted in many diseases. We describe large-scale single-base resolution DNA methylation profiling on a diverse collection of 82 human cell lines and tissues using reduced representation bisulfite sequencing (RRBS). Analysis integrating RNA-seq and ChIP-seq data illuminates the functional role of this dynamic mark. Loci that are hypermethylated across cancer types are enriched for sites bound by NANOG in embryonic stem cells, which supports and expands the model of a stem/progenitor cell signature in cancer. CpGs that are hypomethylated across cancer types are concentrated in megabase-scale domains that occur near the telomeres and centromeres of chromosomes, are depleted of genes, and are enriched for cancer-specific EZH2 binding and H3K27me3 (repressive chromatin). In noncancer samples, there are cell-type specific methylation signatures preserved in primary cell lines and tissues as well as methylation differences induced by cell culture. The relationship between methylation and expression is context-dependent, and we find that CpG-rich enhancers bound by EP300 in the bodies of expressed genes are unmethylated despite the dense gene-body methylation surrounding them. Non-CpG cytosine methylation occurs in human somatic tissue, is particularly prevalent in brain tissue, and is reproducible across many individuals. This study provides an atlas of DNA methylation across diverse and well-characterized samples and enables new discoveries about DNA methylation and its role in gene regulation and disease

Looking Back, Looking Forward: A New Look at the Historic Resources of the Maryland Port Towns

This document has had referenced material removed in respect for the owner's copyright. A complete version of this document, which includes said referenced material, resides in the University of Maryland, College Park's library collection.During the fall of 2008, the historic preservation studio of the University of Maryland’s Graduate Program in Historic Preservation developed a heritage resource study for the Maryland Port Towns, a group of four individual municipalities located on the Anacostia River in Prince George’s County, Maryland. The client, the Port Towns Community Development Corporation, made it clear from the beginning that the study was to dovetail with their already extensive efforts for social and economic development in the Port Towns. The study that follows is the culmination of the efforts of the nine-member studio team. Titled Looking Back, Looking Forward: A New Look at the Heritage Resources of the Maryland Port Towns, the study initially developed from two principal questions: • What existing historic resources are located in the Port Towns? • What can be done to preserve, enhance, and highlight the existing historic resources located in the Port Towns to meet the socioeconomic goals set by the Port Towns Community Development Corporation

Whole-Genome SNP Association in the Horse: Identification of a Deletion in Myosin Va Responsible for Lavender Foal Syndrome

Lavender Foal Syndrome (LFS) is a lethal inherited disease of horses with a suspected autosomal recessive mode of inheritance. LFS has been primarily diagnosed in a subgroup of the Arabian breed, the Egyptian Arabian horse. The condition is characterized by multiple neurological abnormalities and a dilute coat color. Candidate genes based on comparative phenotypes in mice and humans include the ras-associated protein RAB27a (RAB27A) and myosin Va (MYO5A). Here we report mapping of the locus responsible for LFS using a small set of 36 horses segregating for LFS. These horses were genotyped using a newly available single nucleotide polymorphism (SNP) chip containing 56,402 discriminatory elements. The whole genome scan identified an associated region containing these two functional candidate genes. Exon sequencing of the MYO5A gene from an affected foal revealed a single base deletion in exon 30 that changes the reading frame and introduces a premature stop codon. A PCR–based Restriction Fragment Length Polymorphism (PCR–RFLP) assay was designed and used to investigate the frequency of the mutant gene. All affected horses tested were homozygous for this mutation. Heterozygous carriers were detected in high frequency in families segregating for this trait, and the frequency of carriers in unrelated Egyptian Arabians was 10.3%. The mapping and discovery of the LFS mutation represents the first successful use of whole-genome SNP scanning in the horse for any trait. The RFLP assay can be used to assist breeders in avoiding carrier-to-carrier matings and thus in preventing the birth of affected foals

Association of objective sedentary behaviour and self-rated health in English older adults

Abstract Objective Reducing sedentary behaviour (SB) might improve the health of older adults. However, we know little about how objectively measured SB impacts on self-rated health in older adults. We aimed to explore the associations between objectively measured SB and self-rated health in English older adults. Results A random sub-sample of older adults (≥ 65 years old) from the 2008 Health Survey for England wore an ActiGraph GT1M accelerometer for 7 days. Self-rated health was measured using an item from the General Health Questionnaire. Linear regression and analysis of covariance were used to test the associations between percentage time spent in SB and mean daily minutes in SB and self-rated health (very good/good; fair; bad/very bad), adjusting for covariates. Valid accelerometry datasets were returned by 578 individuals. Significant negative associations between percentage time and mean daily minutes in SB and self-rated health were found. In particular, individuals spending reduced percentages of time being sedentary had higher self-rated health. In conclusion, SB appears to be associated with self-rated health in older people independently from MVPA. If longitudinal research could determine how changes in SB influence self-rated health as individuals’ age, this might be an important lifestyle variable to target for health improvement

The FLUXNET2015 dataset and the ONEFlux processing pipeline for eddy covariance data

The FLUXNET2015 dataset provides ecosystem-scale data on CO2, water, and energy exchange between the biosphere and the atmosphere, and other meteorological and biological measurements, from 212 sites around the globe (over 1500 site-years, up to and including year 2014). These sites, independently managed and operated, voluntarily contributed their data to create global datasets. Data were quality controlled and processed using uniform methods, to improve consistency and intercomparability across sites. The dataset is already being used in a number of applications, including ecophysiology studies, remote sensing studies, and development of ecosystem and Earth system models. FLUXNET2015 includes derived-data products, such as gap-filled time series, ecosystem respiration and photosynthetic uptake estimates, estimation of uncertainties, and metadata about the measurements, presented for the first time in this paper. In addition, 206 of these sites are for the first time distributed under a Creative Commons (CC-BY 4.0) license. This paper details this enhanced dataset and the processing methods, now made available as open-source codes, making the dataset more accessible, transparent, and reproducible.Peer reviewe

Clinical Sequencing Exploratory Research Consortium: Accelerating Evidence-Based Practice of Genomic Medicine

Despite rapid technical progress and demonstrable effectiveness for some types of diagnosis and therapy, much remains to be learned about clinical genome and exome sequencing (CGES) and its role within the practice of medicine. The Clinical Sequencing Exploratory Research (CSER) consortium includes 18 extramural research projects, one National Human Genome Research Institute (NHGRI) intramural project, and a coordinating center funded by the NHGRI and National Cancer Institute. The consortium is exploring analytic and clinical validity and utility, as well as the ethical, legal, and social implications of sequencing via multidisciplinary approaches; it has thus far recruited 5,577 participants across a spectrum of symptomatic and healthy children and adults by utilizing both germline and cancer sequencing. The CSER consortium is analyzing data and creating publically available procedures and tools related to participant preferences and consent, variant classification, disclosure and management of primary and secondary findings, health outcomes, and integration with electronic health records. Future research directions will refine measures of clinical utility of CGES in both germline and somatic testing, evaluate the use of CGES for screening in healthy individuals, explore the penetrance of pathogenic variants through extensive phenotyping, reduce discordances in public databases of genes and variants, examine social and ethnic disparities in the provision of genomics services, explore regulatory issues, and estimate the value and downstream costs of sequencing. The CSER consortium has established a shared community of research sites by using diverse approaches to pursue the evidence-based development of best practices in genomic medicine

Imaging the cytoplasmic domain of the rosette cellulose-synthesizing terminal complex

textA new technique for imaging terminal cellulose-synthesizing complexes in isolated plasma membrane sheets was developed. Protoplasts bound to poly-L-lysine-coated substrates were burst in a sonicator bath. This process served to partially extract the plasma membrane, allowing the imaging of cellulose microfibrils through holes in the plasma membrane and the tracing of these cellulose microfibrils back to rosette terminal complexes. Using this new technique, the cytoplasmic domain of the rosette terminal complex was imaged. This domain appears to be hexagonal. A new model of the complete rosette cellulose-synthesizing terminal complex includes the hexagonal geometry of the cytoplasmic domain. This new technique effectively augments freeze fracture data in providing new data about the structure-function of the cellulose synthesizing terminal complex.Biological Sciences, School o