The development of mindfully-attentive nursing graduates is a heightened charge for pre-licensure nurse educators given an increasingly complex healthcare workplace in which nurse’s must detect patient risk among multiple stimuli, distractions, and interruption (Beyea, 2007; Cornell et al., 2010; Ebright, et al., 2006). Novice nurses frequently report symptoms of cognitive overload associated with error and other negative patient outcomes (Ebright, Urden, Patterson, & Chalko, 2004; McGillis Hall et al., 2010; Unver, Tastan, & Akbayrak, 2012) yet standard pre-licensure nursing curricula does not specifically prepare students for the needed attention regulation skills of safe nursing practice. Recent and accumulating neuroscientific research suggests a strong correlation between regular practice of mindfulness meditation (MM) (Eberth & Sedlmeier, 2012) and enhanced attentional capacity. This randomized controlled trial therefore investigated the effect of MM as compared to standard nursing education on the efficiency of attentional processes (alerting, orienting, and executive function) in pre-licensure registered nursing students (N=52) as well as on accuracy in performance of a nursing skill. It was framed by Posner and Gilbert’s (2002) neuropsychological Model of Attention. Main outcome data were collected using the Attention Network Test (ANT) (Fan, McCandliss, Sommer, Raz & Posner, 2002), and an investigator-developed Medication Administration Task (MAT). Examination of possible confounding influences of perceived stress using the Perceived Stress Scale - 10 (PSS-10) (Cohen, Kamarck, & Mermelstein, 1983) and mindfulness using the Five -Facet Mindfulness Questionnaire (FFMQ) (Baer, Smith, Hopkins, Krietemeyer, & Toney, 2006) were also included. Nursing students who participated in online training and four weeks of daily MM practice demonstrated improvement to executive attention efficiency as compared to a non-meditating control group F (1, 49) =4.26, p = .044, although interpretation was restricted by accompanied low power .53. MAT results on nursing skill accuracy were non-significant, but group differences on posttests of PSS-10 and FFMQ were significant F (2, 47) = 7.16, p = .002, power .92. After four weeks of meditation, participants in the MM group scored higher in mindfulness characteristics and lower in perceived stress than control group participants who did not meditate. / School of Nursing; / Nursing / PhD; / Dissertation;

Malaria is a debilitating and deadly disease that afflicts over 200 million people and kills over 600 thousand each year. Due to quickly evolving drug resistance and lack of an affordable vaccine, novel interventions are needed to fight the Plasmodium parasites that cause malaria. Targeting Plasmodium inside their mosquito hosts is one approach that could complement other preventative and medicinal interventions by reducing the ability of the mosquitoes to transmit the disease to humans. The research presented here uses paratransgenesis, the genetic modification of symbiotic bacteria within the mosquito midgut, to provide antimalarial protein to the mosquito and to interfere with the life cycle of Plasmodium within the insect host. <br>This research has produced three new antimalarial paratransgenic tools. The first tool is a set of new antimalarial effector proteins that were constructed by converting anti-Plasmodium mouse antibodies into single-chain variable fragment (scFv) versions for expression by bacteria. These antibodies bind to Plasmodium surface proteins and interfere with critical steps in the parasite life cycle. The second tool is a modified bacterial species, Pantoea agglomerans , which was engineered to secrete diverse antimalarial proteins via the hemolysin secretion pathway. Modified P. agglomerans were fed to mosquitoes and were capable of inhibiting the invasion of Plasmodium within the midgut. The third tool is another modified bacterial species, Asaia sp. SF2.1. Native Type II secretion signals were discovered that enable the creation of paratransgenic strains of these bacteria. Modified strains of Asaia sp. SF2.1 were also demonstrated to interfere with the invasion of Plasmodium within the mosquito. <br>These tools have laid the groundwork for the future use of paratransgenic bacteria to combat malaria in the wild. Asaia sp. SF2.1 bacteria, in particular, are capable of spreading throughout mosquito populations, so they provide their own drive mechanism to establish themselves within the mosquito vectors of malaria. While further modifications will be required to make these bacteria ready for field use, the findings of this research provide proof of concept that the bacteria are suitable for eventual use in malaria transmission-blocking interventions. / Bayer School of Natural and Environmental Sciences; / Biological Sciences / PhD; / Dissertation;

A role for protein kinase C (PKC) in the denervation-induced supersensitivity of the rat vas deferens was investigated. Chronic, surgical denervation of the rat vas deferens (up to 8 days) resulted in tissues that produced enhanced contractile responses to norepinephrine (NE) in isolated organ baths. Single challenges of NE (10 $\mu$M) produced 0.6 $\pm$ 0.1 g of maximal tension in the control vas whereas in the paired, denervated tissue 2.2 $\pm$ 0.3 g of tension was recorded (n = 6). Cumulative concentration-effect curves to NE produced in the denervated vas deferens were shifted 18-fold to the left of the control response. Neurokinin A (NKA) responses after denervation of the tissue were not significantly different from the control. Denervation did not alter the contractile response to phorbol diacetate (PDA), a PKC activator. Pretreatment of denervated and control vas deferens with 100 $\mu$M nifedipine (a calcium channel blocker), significantly attenuated the contractile response to NE. The responses in the control tissues were depressed by 88%, those in the denervated vasa were only antagonized by 65% after nifedipine treatment. Exposure of denervated and control vas deferens to 100 $\mu$M NE resulted in no significant accumulation of diacylglycerol (DAG) from basal values. The molecular species of DAG produced after receptor stimulation, in either tissue group, were not different from those found in resting tissues. Denervation also had no effect on the binding characteristics of membrane-associated PKC when assayed using the specific ligand, ($\sp3$H) phorbol dibutyrate. The PKC activity of resting vas deferens was not altered by chronic surgical denervation. Denervated and control vas deferens that were stimulated with 100 $\mu$M NE showed a time-dependent translocation of PKC from the cytosolic to the membrane fraction of the tissue. In both tissue groups exposure to NE resulted in a 3-4 fold increase in the membrane-bound PKC activity, which remained elevated above basal values for up to 20 min. The rate of translocation of PKC was faster in denervated vasa (maximal at 5 min after NE) when compared to the control (maximal at 20 min), but the maximal amount of the enzyme activated was the same for the two tissue groups. The ability of NKA, 60 mM K$\sp+$-depolarization and PDB (PKC activator) to produce translocation of the PKC was not altered by denervation of the vas deferens. (Abstract shortened by UMI.)

To test whether SKF 38393 could ontogenetically sensitize dopamine (DA) D$\sb1$ receptors and whether this sensitization would be associated with biochemical changes, intact and neonatal 6-hydroxydopamine (6-OHDA)-lesioned rats (200 $\mu$g i.c.v.) were treated daily from birth with SKF 38393 (3.0 mg/kg i.p. x 28 days) or its vehicle. In DA D$\sb1$ neonatally sensitized 6-OHDA rats, enhanced locomotor responses were observed with the first SKF 38393 challenge dose (3.0 mg/kg i.p.) at 6 weeks. This response increased further with weekly SKF 38393 treatments. Enhanced stereotyped behaviors were seen in both lesioned and sensitized rats at 8 weeks. There was no change in the percentage of high affinity D$\sb1$ sites in these groups of rats. Striatal mRNA levels for D$\sb1$ receptors were reduced in the lesioned rats, but restored to control level after treatments with SKF 38393 in adulthood. Basal, DA-, NaF- and forskolin-stimulated adenylate cyclase activities were similar among treatment groups. Striatal DA content was reduced ($>$99%), whereas serotonin (5-HT) content was elevated ($>$50%) in the 6-OHDA groups. To study possible interaction between DA and 5-HT systems, the effects of a series of 5-HT agents on the induction of oral activity were determined. The 5-HT$\sb{\rm 1C}$ receptor agonist, m-chlorophenylpiperazine (m-CPP), produced a marked increase in oral activity in 6-OHDA-lesioned rats. The respective 5-HT$\sb{\rm 1A}$ and 5-HT$\sb{\rm 1B}$ agonists, 8-OH-DPAT and CGS-12066B did not increase oral activity. The m-CPP-induced oral response in the lesioned rats was attenuated by mianserin, a 5-HT$\sb{\rm 1C}$ antagonist, but not by ketanserin or MDL-72222, 5-HT$\sb2$ and 5-HT$\sb3$ antagonists, respectively. Although the supersensitized oral response of lesioned rats to m-CPP was not attenuated by SCH 23390, the enhanced response of SKF 38393 was attenuated by mianserin. Additionally, mRNA levels for 5-HT$\sb{\rm 1C}$ receptor were not altered in both intact and lesioned rats. These findings demonstrate that ontogenetic treatments of neonatal 6-OHDA-lesioned rats with a D$\sb1$ agonist produce partial sensitization of DA D$\sb1$ receptors in adulthood without altered biochemical markers, and that this neonatal lesion is associated with both supersensitized DA D$\sb1$ and 5-HT$\sb{\rm 1C}$ receptors. Moreover, induction of oral activity by DA agonists is mediated via a serotonergic system.

The hypothesis that antioxidant vitamins (ascorbate and tocopherols) along with urate protect blood plasma lipids from oxidation was tested. Dietary fat is also an important factor influencing plasma lipid peroxidation. The purpose of this study was to investigate the role of plasma antioxidants and dietary fat on low density lipoprotein (LDL) and plasma lipid oxidation. In the first part of this study, we compared the ability of urate and ascorbate to protect human LDL from in vitro oxidation. LDL oxidation was initiated by 15 mM of a water soluble azo-initiator in the presence or absence of ascorbate or urate. The rate of lipid hydroperoxide (LOOH) formation was increased after the LDL tocopherols were totally consumed, i.e., after the lag phase. Urate (50 $\mu$M) was more effective than ascorbate (50 $\mu$M) in extending the lag phase. Moreover, urate was consumed more slowly than ascorbate under identical oxidation conditions. The combination af 25 $\mu$M ascorbate and 25 $\mu$M urate was more effective in extending the lag phase than ascorbate alone but less effective than urate alone. An empirical mathematical model was developed to describe the oxidation kinetics of LDL tocopherols. In the second part of this study, we studied the role of dietary fat and dietary $\alpha$-tocopherol ($\alpha$-toc) levels on rat plasma oxidation. The fatty acid composition of plasma was found to be modulated by the type of dietary fat. Neither dietary fat nor $\alpha$-toc influenced the plasma levels of water soluble antioxidants (ascorbate, urate and sulfhydryl content). Rat plasma was oxidized either by a water soluble azo-initiator (25 mM) or a lipid soluble azo-initiator (10 mM). In both cases, the rate of LOOH formation in plasma from rats fed butter oil diets was markedly suppressed compared to the plasma from rats fed corn oil diets. When oxidation was initiated by a lipid soluble azo-initiator, plasma from rats fed $\alpha$-toc supplemented diets showed higher LOOH levels than plasma from rats fed $\alpha$-toc deficient diets. Surprisingly, when oxidation was initiated by water soluble azo-initiator, tocopherol appeared to act as a pro-oxidant. The results suggest that urate may be more significant than ascorbate in delaying the consumption of tocopherols in human LDL and that low dietary PUFAs levels are more important in preventing the in vitro oxidation of plasma lipids than high dietary levels of $\alpha$-tocopherol.

The primary purpose of this study was to investigate the admission/selection process of the Radiologic Technology Program at East Tennessee State University to ascertain predictive validity of the admission process and to identify specific indicators leading to program completion. The population for this investigation consisted of 510 students who had applied to East Tennessee State University and the Radiologic Technology Program from 1991 through 1993. Data were collected from institutional and program academic records. Correlation research was chosen to establish validity. One-way Analysis of Variance and t-test were applied to investigate different admission parameters and discriminate analysis was completed due to prior groupings in the initial academic analysis. An alpha level of.05 was selected for this study. The population (N = 510) consisted of four groups of students: Program completers (graduates), program non-completers, students interviewed/not admitted, and students not interviewed nor admitted. One way analysis of variance (ANOVA) determined significant differences in all admission parameters between the four groups. Discriminate analysis of program completers and non-completers found that curriculum GPA ranking was significant explaining 47% of the shared variance. The population of students admitted to the program (N = 110) was further statistically analyzed by t-Tests and no significant differences were identified between program completers and non completers. Based on the findings, the following conclusions were drawn: (1) The admission parameters utilized by the Radiologic Technology Program at East Tennessee State University were statistically significant in identifying differences among the four groups of students; (2) No significant differences were identified between program completers and non-completers; (3) The admission model utilized by the Radiologic Technology Program is equitable. The following recommendations were suggested: (1) Further studies should be conducted incorporating demographic factors; (2) Studies addressing selection parameter reliability should be conducted; (3) Reliability of each interview question should be ascertained; (4) Studies involving common admission parameters at different sponsoring institutions should be investigated, and (5) Incorporation of qualitative methodology regarding program completion versus non-completion could be beneficial.

The vascular endothelial cell (EC) plays an important role in regulating vascular tone and local blood flow by sensing chemical and mechanical stimuli on the vascular wall and releasing a host of vasoactive substances upon activations of endogenous or exogenous vasoactive substances. The central hypothesis is that local control of blood now and autoregulatory behavior in the microcirculation is distinctive at different levels of the vasculature and is dependent on the cellular activities of the EC and its interaction with the local environment. The in vivo as well as the ex vivo, flow-controlled preparations of the hamster cheek pouch were utilized. Inhibition of Endothelium-Derive Relaxing Factor (EDRF) synthesis and the functional impairment by light-dye (L-D) treatment were used to remove functional characteristics of the EC. It is found that the EC played differential roles in modulating vascular tone and blood flow in distinct segments of arterioles. Impairment of the EC by L-D treatment significantly reduced both acetylcholine (Ach)-induced dilation and the local angiotensin conversion in small (4th order) arterioles (A$\sb4$). Whereas, data obtained after inhibition of EDRF synthesis indicated that EDRF pathway appeared to be the dominant regulatory mechanism mediating agonists (e.g. Ach)-induced responses in these small vessels. In large (2nd order) arterioles (A$\sb2$), on the other hand, neither L-D treatment nor EDRF inhibition affected Ach-induced dilation or local angiotensin conversion; therefore, these responses seemed to be independent of the EC or EDRF pathway. Autoregulation was observed in both A$\sb2$ and A$\sb4$ when perfusion flow (shear stress) and perfusion pressure (stretch) were elevated. Nevertheless, the underlying regulatory mechanisms in response to mechanical stimuli differed in these series-arranged arterioles. The EC/EDRF-dependent, flow-induced dilation was dominant in A$\sb2$; whereas, the myogenic autoregulation (which appears to be independent of the EC) played major role in A$\sb4$. Therefore, the function of the EC does not appear homogenous throughout the arteriolar portion of the microcirculation. Thus, the local control of blood flow and autoregulatory behavior in the microcirculation is distinctive at different levels of the vasculature; whereas, the differential role of the EC in discrete segments of series-arranged arterioles seems to be the determinant for these differences. These differential modulations of vascular tone and blood flow by the EC at discrete levels of the microcirculation may have important implications in pathological conditions, such as hypertension, diabetes, and atherosclerosis.

Chlamydia trachomatis (CT) is the most prevalent sexually-transmitted infection in the United States. It has been suggested that CT infections can become latent. This has not been substantiated. CT persistence was examined at the molecular and cellular level in vitro and in vivo. Penicillin treatment of CT in vitro results in abnormal inclusions and reduced recovery of infectious CT. Penicillin did not inhibit initial stages of infection, but did downregulate CT rRNA levels after 25 hours post-inoculation (p.i.). DNA amplification was employed to differentiate between a resolved infection and a persistent one. Utilizing a primer pair that amplified a 144 bp fragment in the CT MOMP gene, CT-persistently-infected McCoy cells maintained in penicillin medium were examined. Though undetectable by other assay methods, these cells harbored the CT genome for 18 passages. Removal of penicillin 1, 3 or 6 passages p.i. and subsequent cultivation in permissive medium resulted in "recovery" to productive infection. Removal of penicillin at later passages resulted in low level inclusion formation but no infectious progeny. Penicillin treatment in vitro resulted in a persistent infection undetectable by most methods. Female C$\sb3$H/HeNCRL mice were inoculated with CT intrauterinely and intravaginally in two separate experiments. In one, CT infection was established in untreated and Depo-Provera (DP)-pretreated mice. DP pretreatment enhanced vaginal shedding of infectious CT. A negative vaginal culture did not correlate with elimination of CT from the upper tract. In the second, penicillin therapy did not halt vaginal CT shedding, however, it reduced frequency of recurrent vaginal CT shedding. To examine reactivation, culture-negative mice ($\geq$2 successive vaginal cultures) were injected with cortisone-acetate (CA) or DP; mice from same subpopulation injected with saline served as controls. Transient vaginal CT shedding was reactivated in penicillin-treated mice (14% CA-injected), and in unmedicated mice (28% CA-injected, 33% DP-injected). Saline injection did not reactivate vaginal CT shedding. At time of sacrifice (16 or 22 weeks p.i.) no infectious CT was detected in upper tract tissues, although tissue damage was observed in most mice (70-71%). It is unknown if these mice harbored a persistent infection undetectable by culture. Further work utilizing molecular techniques is needed to resolve this question.

In an effort to elucidate the mechanism by which indomethacin (IN) attenuates the stimulatory effect of estradiol (E$\sb2$) on rabbit splenic red pulp macrophages (RPM), thirty-nine female New Zealand White rabbits were divided into 10 groups: ovariectomized (OVX), OVX/IN at 0.1 and 5.0 mg/kg body weight (bw)/day; sham OVX (SOVX), SOVX/IN at 0.1 and 5.0 mg/kg bw/day; OVX/25 mg E2, OVX/25 mg E$\sb2$/IN at 0.1 and 5.0 mg/kg bw/day; intact Control. Quantitative changes in RPM population in response to the treatments were measured using a 0 to 4 histologic grading scale. Estradiol treatment resulted in increased RPM grade when compared to the OVX non-E$\sb2$ groups. Indomethacin addition decreased mean RPM grade in the SOVX/IN 5.0 group when compared to its E$\sb2$ control group. Indomethacin administration had no significant effect on levels of PGE$\sb2$ in the spleen, blood or urine (p $>$.05). Hematocrits were reduced in both OVX and OVX/E$\sb2$ groups and this decrease was exacerbated by the high IN dose. The results from this study suggest that the effect of IN on E$\sb2$-induced RPM activation may be mediated through a non-prostaglandin pathway. The observed hematocrit changes are possibly the result of direct action of IN and E$\sb2$ on erythrocytes. To further investigate whether a direct interaction of IN and E$\sb2$ with rabbit erythrocytes may be responsible for the decreases in hematocrit observed in vivo, an in vitro study was conducted to determine the effect of these drugs on erythrocyte fragility characteristics. Two ml aliquots of treated New Zealand White rabbit whole blood were assayed as; Control, IN (9.6 $\mu$g/ml), E$\sb2$ (500 pg/ml) and IN plus E$\sb2$, for changes in erythrocyte fragility. Osmotic (OF) and mechanical (MF) fragility were evaluated under approximate physiological conditions by measurement of hemoglobin release at 545 nm. Blood samples at 39.5$\sp\circ$C were assayed immediately after drug addition (initial) and again 4 hours after incubation (final). Eight replicates of each experiment were run. Results of the OF assays showed a significant increase (p $<$.05) in mean 50% hemolysis point between IN (final) and IN plus E$\sb2$ (final) when compared to their mean initial values and to the mean final Control value. The OF hemolysis dispersion was increased by IN and IN plus E$\sb2$ treatment when final values were compared to initial values. The mean final values for MF increased with IN, E$\sb2$ and IN plus E$\sb2$ treatment versus the mean final Control value (p $<$.05). While the increase in MF from IN was greater than that from E$\sb2$, the MF from the combination (IN plus E$\sb2$) was not greater than from IN alone (p $>$.05). The IN-induced increases in both OF and MF indicate a difference in degrees of interaction with the erythrocyte from that of E$\sb2$, which only affected MF and whose effect was not additive or synergistic with that of IN. The in vitro experimental results demonstrate that the increased fragility produced by IN and E$\sb2$ on rabbit erythrocytes may account for the observed in vivo reduction in hematocrit. Increased erythrocyte fragility would also lead to their accelerated clearance from the circulation by splenic RPM and subsequent increases in activity of these macrophages. This elevation in splenic RPM population may also be enhanced by direct E$\sb2$ stimulation of macrophage proliferation.

Enoyl-acyl carrier protein (ACP) reductase from chloroplast nonaggregated fatty acid synthetase (FAS) of Euglena gracilis variety bacillaris was purified to a single band on a denaturing polyacrylamide gel. The enzyme was partially characterized with respect to substrate specificity, reduced nucleotide requirement, and the effect of ACP and Ca$\sp{++}$ on enzyme activity. Antibodies against the purified protein were raised in hens and isolated from eggs. ACP was purified from Euglena in yields of about 1mg/100g (wet weight) of cells. Antibodies were raised against the purified protein. ACP antibodies inhibited the Euglena chloroplast FAS using Euglena or E. coli ACP as a substrate. Comparisons with other ACPs included the following items: biological activity, pI, behavior in size exclusion media, and amino acid sequence of the N-terminal portion of the molecule. ACPs from E. coli and Euglena have been shown to interact with melittin, a cationic peptide from bee venom. E. coli ACP is a small (Mr, 8847), acidic, Ca$\sp{++}$-binding protein which possesses some characteristics resembling those of regulatory Ca$\sp{++}$-binding proteins including interaction with melittin. Melittin inhibited activity of the nonaggregated FAS from Euglena using either E. coli or Euglena ACP as a substrate. The peptide also inhibited activity of the aggregated FAS from Euglena. Antibodies against melittin were raised. Anti-melittin inhibited activity of both the nonaggregated and aggregated FAS enzyme systems from Euglena relative to nonimmune antibody. Investigation of inhibition of the nonaggregated FAS enzyme system demonstrated that acetyl-CoA-ACP transacylase, malonyl-CoA-ACP transacylase, and keto-acyl-ACP synthetase activities were inhibited to different degrees by anti-melittin antibodies, while keto-acyl-ACP reductase and enoyl-ACP reductase enzyme activities were not inhibited.