Biochemical Characterization of the Flavivirus-resistance Mouse ABCF3 Protein and a Review of the Antibiotic Resistance and Dissemination Mechanisms in Bacteria

Mammalian ATP-binding cassette subfamily F member 3 (ABCF3) is a class 2 ABC protein that has previously been identified as a partner of the mouse flavivirus resistance protein 2′,5′-oligoadenylate synthetase 1B (OAS1B). The functions and natural substrates of ABCF3 are currently not known. In Chapter 1 of this study, we show that purified ABCF3 is an active ATPase. Binding analyses with a fluorescent ATP analog TNP-ATP suggested unequal contributions by the two nucleotide-binding domains. We further showed that ABCF3 activity is increased by lipids, including sphingosine, sphingomyelin, platelet-activating factor, and lysophosphatidylcholine. However, cholesterol inhibited ABCF3 activity, whereas alkyl ether lipids either inhibited or resulted in a biphasic response, suggesting small changes in lipid structure differentially affect ABCF3 activity. Point mutations in the two nucleotide-binding domains of ABCF3 affected basal and sphingosine-stimulated ATPase activity differently, further supporting different roles for the two catalytic pockets. We propose a model in which pocket 1 is the site of basal catalysis, whereas pocket 2 engages in ligand-stimulated ATP hydrolysis. Co-localization of the ABCF3–OAS1B complex to the virus-remodeled endoplasmic reticulum membrane has been shown before. We show that co-expression of ABCF3 and OAS1B in bacteria alleviated growth inhibition caused by expression of OAS1B alone, and significantly enhanced OAS1B levels, indirectly showing interaction between these two proteins in bacterial cells. As viral RNA synthesis requires large amounts of ATP, we conclude that lipid-stimulated ATP hydrolysis may contribute to the reduction in viral RNA production characteristic of the flavivirus resistance phenotype. Chapter 2 of this dissertation provides a comprehensive review of the major known antibiotic resistance mechanisms, including the function of ABC proteins, found in producer soil bacteria and discusses different horizontal gene transfer mechanisms that may play a role in the dissemination of resistance genes from producer and non-producer environmental bacteria to pathogenic bacteria in clinical settings. Many bacterial and eukaryotic ABC proteins are polyspecific in nature and are capable of transporting structurally diverse compounds, including drugs and lipids. These proteins are responsible for intrinsic or acquired multidrug resistance, which can also spread to pathogenic organisms through the horizontal transfer mechanisms discussed in this review

The Life Story of President Barack Obama: Using Critical Race Theory to Analyze Life Experience

Using Critical Race Theory (CRT) as a lens and interest convergence and color-blind ideology as foci to explore the life experience of President Barack Obama, this study provides a framework for using CRT as a lens for examining life experience

Race and Racism: A Critical Dialogue

This is a dialogue between Elizabeth Peterson, a black, female adult educator and Stephen Brookfield, a white male. The question was whether or not we could come together and engage in a critical dialogue about race and racism

Starting Well: Facilitating the Middle School Transition

School-based mental health programming is a viable intervention because it can provide evidenced-based-treatment (EBT) while avoiding the typical service barriers. In this study, 119 students (ages 10 to 12) were randomly assigned to participate in either a 24-session Coping Power Program (CPP) or a control group. Using the Behavior Assessment Scale for Children-2 (BASC-2), teachers reported significant improvements over time for all students (Attention Problems, Hyperactivity, Externalizing Problems, and Withdrawal, and Study Skills). However, teachers reported that students participating in the CPP showed significantly greater improvement than controls (Learning Problems, School Problems, Behavior Symptom Index, Social Skills, and Adaptive Skills). Effect sizes ranged from small (.19 for Withdrawal) to large (.76 for Adaptive Skills). Teacher reports showed greater improvement by the more experienced group leaders. Results validate the use of this EBT as a viable protocol. The improvement in both groups may suggest a spillover effect for untreated controls. The differential effect of group leader suggests that clinical experience may enhance EBT

Above- and below-ground Litter Manipulation: Effect on Retention and Release of DOC, DON and DIN in the Sikfokut Forest, Hungary

The above- and below-ground litter from fallen foliage and root exudates and their decomposition has an impact on forest soil. The objective of this research project was to determine the effect of above- and below-ground litter manipulation on the retention and release of dissolved organic carbon (DOC), dissolved organic nitrogen (DON), nitrate and ammonium in the soil profile at 0-5 and 5-15 cm depths. The soils were obtained from a Long Term Ecological Research site in the Sikfokut Forest in Hungary. The site is a mature oak forest and the soil has no organic layer; the leaf litter sits directly on the A horizon. There are six treatments applied to the soil: doubling of annual leaf litter (DL), doubling of annual wood litter (DW), removal of annual leaf litter (NL), removal of roots (NR), removal of leaf and roots (NI) and control (C). Each plot is 7 x 7 m, and each soil sample taken was 15 x 15 cm square. A stock solution of leaf litter was added in different concentrations to soils from each treatment in a 50 mL centrifuge tube. Because I was investigating retention or release of carbon and nitrogen, I inverted the tube gently over a period of 2 hours prior to centrifugation and removal of supernatant. The supernatant solution was analyzed for DOC, DON, ammonium and nitrate to determine retention and release values for each dependant upon the different treatments. The results of the study were interesting. Dissolved organic nitrogen was neither retained nor released in the 0-5 cm layer for any treatment yet nitrate, typically considered a mobile anion showed retention and release in this layer. In the 5-15 cm layer we had no retention or release of nitrate but dissolved organic nitrogen was retained and released between the solid and solution phases

Trehalose Is A Chemical Attractant In The Establishment Of Coral Symbiosis

Coral reefs have evolved with a crucial symbiosis between photosynthetic dinoflagellates (genus Symbiodinium) and their cnidarian hosts (Scleractinians). Most coral larvae take up Symbiodinium from their environment; however, the earliest steps in this process have been elusive. Here we demonstrate that the disaccharide trehalose may be an important signal from the symbiont to potential larval hosts. Symbiodinium freshly isolated from Fungia scutaria corals constantly released trehalose (but not sucrose, maltose or glucose) into seawater, and released glycerol only in the presence of coral tissue. Spawning Fungia adults increased symbiont number in their immediate area by excreting pellets of Symbiodinium, and when these naturally discharged Symbiodinium were cultured, they also released trehalose. In Y-maze experiments, coral larvae demonstrated chemoattractant and feeding behaviors only towards a chamber with trehalose or glycerol. Concomitantly, coral larvae and adult tissue, but not symbionts, had significant trehalase enzymatic activities, suggesting the capacity to utilize trehalose. Trehalase activity was developmentally regulated in F. scutaria larvae, rising as the time for symbiont uptake occurs. Consistent with the enzymatic assays, gene finding demonstrated the presence of a trehalase enzyme in the genome of a related coral, Acropora digitifera, and a likely trehalase in the transcriptome of F. scutaria. Taken together, these data suggest that adult F. scutaria seed the reef with Symbiodinium during spawning and the exuded Symbiodinium release trehalose into the environment, which acts as a chemoattractant for F. scutaria larvae and as an initiator of feeding behavior- the first stages toward establishing the coral-Symbiodinium relationship. Because trehalose is a fixed carbon compound, this cue would accurately demonstrate to the cnidarian larvae the photosynthetic ability of the potential symbiont in the ambient environment. To our knowledge, this is the first report of a chemical cue attracting the motile coral larvae to the symbiont

The effects of environmental enrichment on nicotine sensitization in a rodent model of schizophrenia

Environmental enrichment, for more than fifty years, has shown to increase learning in behaviors and to alter some brain structures (Renner and Rosenzweig). Some brain changes that occur when environmental enrichment is implemented include the following: increases in cortical thickness, especially the occipital cortex, increases in size of neuronal cell bodies, number of dendrites and dendritic spines, increases in astrocyte branching, increases in the number of brain blood capillaries, and increases in mitochondria (an indication of higher metabolic activity) (Stairs and Bard). It has been shown in research studies that rats in the environmental enrichment group are less sensitive to nicotine effects, both repeated and acute, than rats in isolated situations (Green et al). This is so because enrichment changes the intensity of the acute administration of drugs of abuse. Rats are stimulated by the environment, rather than a particular stimulant

Effects of a DVD-delivered randomized controlled physical activity intervention on functional health in cancer survivors

BACKGROUND: Supervised physical activity interventions improve functional health during cancer survivorship, but remain costly and inaccessible for many. We previously reported on the benefits of a DVD-delivered physical activity program (FlexToBa™) in older adults. This is a secondary analysis of the intervention effects among cancer survivors in the original sample. METHODS: Low active, older adults who self-reported a history of cancer (N = 46; M time since diagnosis = 10.7 ± 9.4 years) participated in a 6-month, home-based physical activity intervention. Participants were randomized to either the DVD-delivered physical activity program focused on flexibility, toning, and balance (FlexToBa™; n = 22) or an attentional control condition (n = 24). Physical function was assessed by the Short Physical Performance Battery (SPPB) at baseline, end of intervention, and at 12 and 24 months after baseline. RESULTS: Repeated measures linear mixed models indicated a significant group*time interaction for the SPPB total score (β = - 1.14, p = 0.048), driven by improved function from baseline to six months in the FlexToBa™ group. The intervention group also had improved balance (β = - 0.56, p = 0.041) compared with controls. Similar trends emerged for the SPPB total score during follow-up; the group*time interaction from 0 to 12 months approached significance (β = - 0.97, p = 0.089) and was significant from 0 to 24 months (β = - 1.84, p = 0.012). No significant interactions emerged for other outcomes (ps \u3e 0.11). CONCLUSIONS: A DVD-delivered physical activity intervention designed for cancer-free older adults was capable of eliciting and maintaining clinically meaningful functional improvements in a subgroup of cancer survivors, with similar effects to the original full sample. These findings inform the dissemination of evidence-based physical activity programs during survivorship. TRIAL REGISTRATION: ClinicalTrials.gov NCT01030419 . Registered 11 December 2009