A Playground Plan for South Shore Prek-8 School

The purpose of this project was to create a comprehensive playground safety manual of procedures for South Shore PreK-8 School, that can possibly be implemented at other K-5 schools. The manual will make it clear to both students and staff playground safety procedures for students and instructions for staff in supervision. The information gathered finds that when students play in safe manners with specific rules and expectations, that the number of incidents from injury and conflict are reduced. Children need to be taught rules for games, instead of expected to figure out rules for games they do not know how to play. Safe play includes students who have learned how to take turns, and handle conflict or ask for help when needed. Lessons need to be taught to achieve the above objectives, as well as timelines to have in place for lessons or trainings to occur. Statistics and effective studies are included in this project

Identification of transposon insertion mutants of Francisella tularensis tularensis strain Schu S4 deficient in intracellular replication in the hepatic cell line HepG2

BACKGROUND: Francisella tularensis is a zoonotic intracellular bacterial pathogen that causes tularemia. The subspecies tularensis is highly virulent and is classified as a category A agent of biological warfare because of its low infectious dose by an aerosol route, and its ability to cause severe disease. In macrophages F. tularensis exhibits a rather novel intracellular lifestyle; after invasion it remains in a phagosome for three to six hours before escaping to, and replicating in the cytoplasm. The molecular mechanisms that allow F. tularensis to invade and replicate within a host cell have not been well defined. METHODS: We constructed a stable transposon mutagenesis library of virulent strain Schu S4 using a derivative of the EZ::TN transposon system(®). Approximately 2000 mutants were screened for the inability to invade, and replicate in the hepatic carcinoma cell line HepG2. These mutants were also tested for replication within the J774.1 macrophage-like cell line. RESULTS: Eighteen mutants defective in intracellular replication in HepG2 cells were identified. Eight of these mutants were auxotrophs; seven had mutations in nucleotide biosynthesis pathways. The remaining mutants had insertions in genes that were predicted to encode putative transporters, enzymes involved in protein modification and turnover, and hypothetical proteins. A time course of the intracellular growth of a pyrB mutant revealed that this mutant was only able to grow at low levels within HepG2 cells but grew like wild-type bacteria in J774.1 cells. This pyrB mutant was also attenuated in mice. CONCLUSION: This is the first reported large-scale mutagenesis of a type A strain of F. tularensis and the first identification of mutants specifically defective in intracellular growth in a hepatic cell line. We have identified several genes and pathways that are key for the survival and growth of F. tularensis in a hepatic cell line, and a number of novel intracellular growth-defective mutants that have not been previously characterized in other pathogens. Further characterization of these mutants will help provide a better understanding of the pathogenicity of F. tularensis, and may have practical applications as targets for drugs or attenuated vaccines

Azithromycin effectiveness against intracellular infections of Francisella

<p>Abstract</p> <p>Background</p> <p>Macrolide antibiotics are commonly administered for bacterial respiratory illnesses. Azithromycin (Az) is especially noted for extremely high intracellular concentrations achieved within macrophages which is far greater than the serum concentration. Clinical strains of Type B <it>Francisella </it>(<it>F.</it>) <it>tularensis </it>have been reported to be resistant to Az, however our laboratory <it>Francisella </it>strains were found to be sensitive. We hypothesized that different strains/species of <it>Francisella </it>(including Type A) may have different susceptibilities to Az, a widely used and well-tolerated antibiotic.</p> <p>Results</p> <p><it>In vitro </it>susceptibility testing of Az confirmed that <it>F. tularensis subsp. holarctica </it>Live Vaccine Strain (LVS) (Type B) was not sensitive while <it>F. philomiragia, F. novicida</it>, and Type A <it>F. tularensis </it>(NIH B38 and Schu S4 strain) were susceptible. In J774A.1 mouse macrophage cells infected with <it>F. philomiragia, F. novicida</it>, and <it>F. tularensis </it>LVS, 5 μg/ml Az applied extracellularly eliminated intracellular <it>Francisella </it>infections. A concentration of 25 μg/ml Az was required for <it>Francisella-</it>infected A549 human lung epithelial cells, suggesting that macrophages are more effective at concentrating Az than epithelial cells. Mutants of RND efflux components (<it>tolC </it>and <it>ftlC</it>) in <it>F. novicida </it>demonstrated less sensitivity to Az by MIC than the parental strain, but the <it>tolC </it>disc-inhibition assay demonstrated increased sensitivity, indicating a complex role for the outer-membrane transporter. Mutants of <it>acrA </it>and <it>acrB </it>mutants were less sensitive to Az than the parental strain, suggesting that AcrAB is not critical for the efflux of Az in <it>F. novicida</it>. In contrast, <it>F. tularensis </it>Schu S4 mutants Δ<it>acrB </it>and Δ<it>acrA </it>were more sensitive than the parental strain, indicating that the AcrAB may be important for Az efflux in <it>F. tularensis </it>Schu S4. <it>F. novicida </it>LPS O-antigen mutants (<it>wbtN, wbtE, wbtQ </it>and <it>wbtA</it>) were found to be less sensitive <it>in vitro </it>to Az compared to the wild-type. Az treatment prolonged the survival of <it>Galleria </it>(<it>G</it>.) <it>mellonella </it>infected with <it>Francisella</it>.</p> <p>Conclusion</p> <p>These studies demonstrate that Type A <it>Francisella </it>strains, as well as <it>F. novicida </it>and <it>F. philomiragia</it>, are sensitive to Az <it>in vitro. Francisella </it>LPS and the RND efflux pump may play a role in Az sensitivity. Az also has antimicrobial activity against intracellular <it>Francisella</it>, suggesting that the intracellular concentration of Az is high enough to be effective against multiple strains/species of <it>Francisella</it>, especially in macrophages. Az treatment prolonged survival an <it>in vivo </it>model of <it>Francisella-</it>infection.</p

Regulation of Virulence of Entamoeba histolytica by the URE3-BP Transcription Factor

It is not understood why only some infections with Entamoeba histolytica result in disease. The calcium-regulated transcription factor upstream regulatory element 3-binding protein (URE3-BP) was initially identified by virtue of its role in regulating the expression of two amebic virulence genes, the Gal/GalNac lectin and ferredoxin. Here we tested whether this transcription factor has a broader role in regulating virulence. A comparison of in vivo to in vitro parasite gene expression demonstrated that 39% of in vivo regulated transcripts contained the URE3 motif recognized by URE3-BP, compared to 23% of all promoters (P < 0.0001). Amebae induced to express a dominant positive mutant form of URE3-BP had an increase in an elongated morphology (30% ± 6% versus 14% ± 5%; P = 0.001), a 2-fold competitive advantage at invading the intestinal epithelium (P = 0.017), and a 3-fold increase in liver abscess size (0.1 ± 0.1 g versus 0.036 ± 0.1 g; P = 0.03). These results support a role for URE3-BP in virulence regulation

Propagation of local decohering action in distributed quantum systems

We study propagation of the decohering influence caused by a local measurement performed on a distributed quantum system. As an example, the gas of bosons forming a Bose-Einstein condensate is considered. We demonstrate that the local decohering perturbation exerted on the measured region propagates over the system in the form of a decoherence wave, whose dynamics is governed by elementary excitations of the system. We argue that the post-measurement evolution of the system (determined by elementary excitations) is of importance for transfer of decoherence, while the initial collapse of the wave function has negligible impact on the regions which are not directly affected by the measurement.Comment: 6 REVTeX pages, no figures. Introduction and discussion sections are extende

Relationship between quantum decoherence times and solvation dynamics in condensed phase chemical systems

A relationship between the time scales of quantum coherence loss and short-time solvent response for a solute/bath system is derived for a Gaussian wave packet approximation for the bath. Decoherence and solvent response times are shown to be directly proportional to each other, with the proportionality coefficient given by the ratio of the thermal energy fluctuations to the fluctuations in the system-bath coupling. The relationship allows the prediction of decoherence times for condensed phase chemical systems from well developed experimental methods.Comment: 10 pages, no figures, late

Affect in response to stressors and coping strategies: an ecological momentary assessment study of borderline personality disorder

Background: Affect instability is a core symptom of borderline personality disorder (BPD). Ecological momentary assessment allows for an understanding of real-time changes in affect in response to various daily stressors. The purpose of this study was to explore changes in affect in response to specific stressors and coping strategies in subjects with BPD utilizing ecological momentary assessment (EMA) methodology. Methods: Subjects (n = 50) with BPD were asked to complete real-time assessments about stressors experienced, affect felt, and coping strategies employed six times per day for a 1-week period. Mixed effect regression models were used to measure the effect of stressors and coping strategies on affect change. Results: While most stressors led to experiencing more negative affect, only being in a disagreement was independently associated with increased negative affect. Among coping strategies, only doing something good for oneself independently reduced negative affect, controlling for all other coping strategies used. Conclusions: These findings provide valuable insights into affective instability in BPD and can help inform treatment with individuals with the disorder

Switchgrass (\u3ci\u3ePanicum virgatum\u3c/i\u3e L.) polyubiquitin gene (\u3ci\u3ePvUbi1\u3c/i\u3e and \u3ci\u3ePvUbi2\u3c/i\u3e) promoters for use in plant transformation

Abstract Background The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (Panicum virgatum L.) ubiquitin genes (PvUbi1 and PvUbi2) were cloned and characterized. Reporter constructs were produced containing the isolated 5\u27 upstream regulatory regions of the coding sequences (i.e. PvUbi1 and PvUbi2 promoters) fused to the uidA coding region (GUS) and tested for transient and stable expression in a variety of plant species and tissues. Results PvUbi1 consists of 607 bp containing cis-acting regulatory elements, a 5\u27 untranslated region (UTR) containing a 93 bp non-coding exon and a 1291 bp intron, and a 918 bp open reading frame (ORF) that encodes four tandem, head -to-tail ubiquitin monomer repeats followed by a 191 bp 3\u27 UTR. PvUbi2 consists of 692 bp containing cis-acting regulatory elements, a 5\u27 UTR containing a 97 bp non-coding exon and a 1072 bp intron, a 1146 bp ORF that encodes five tandem ubiquitin monomer repeats and a 183 bp 3\u27 UTR. PvUbi1 and PvUbi2 were expressed in all examined switchgrass tissues as measured by qRT-PCR. Using biolistic bombardment, PvUbi1 and PvUbi2 promoters showed strong expression in switchgrass and rice callus, equaling or surpassing the expression levels of the CaMV 35S, 2x35S, ZmUbi1, and OsAct1 promoters. GUS staining following stable transformation in rice demonstrated that the PvUbi1 and PvUbi2 promoters drove expression in all examined tissues. When stably transformed into tobacco (Nicotiana tabacum), the PvUbi2+3 and PvUbi2+9 promoter fusion variants showed expression in vascular and reproductive tissues. Conclusions The PvUbi1 and PvUbi2 promoters drive expression in switchgrass, rice and tobacco and are strong constitutive promoter candidates that will be useful in genetic transformation of monocots and dicots