Development of a Single-Step Subtraction Method for Eukaryotic 18S and 28S Ribonucleic Acids

The abundance of mammalian 18S and 28S ribosomal RNA can decrease the detection sensitivity of bacterial or viral targets in complex host-pathogen mixtures. A method to capture human RNA in a single step was developed and characterized to address this issue. For this purpose, capture probes were covalently attached to magnetic microbeads using a dendrimer linker and the solid phase was tested using rat thymus RNA (mammalian components) with Escherichia coli RNA (bacterial target) as a model system. Our results indicated that random capture probes demonstrated better performance than specific ones presumably by increasing the number of possible binding sites, and the use of a tetrame-thylammonium-chloride (TMA-Cl-) based buffer for the hybridization showed a beneficial effect in the selectivity. The subtraction efficiency determined through real-time RT-PCR revealed capture-efficiencies comparable with commercially available enrichment kits. The performance of the solid phase can be further fine tuned by modifying the annealing time and temperature

PENGARUH KNOWLEDGE MANAGEMENT DAN PENGEMBANGAN KARIR TERHADAP KINERJA KARYAWAN KLINIK UTAMA SPESIALIS MATA SMEC MANADO

Suatu organisasi dibentuk untuk mencaspai tujuan bersama, namun untuk mencapai tujuan secara efektif diperlukan manajemen yang baik dan benar. Pengembangan karir merupakan salah satu cara yang ditempuh organisasi dalam meningkatkan kinerja stafnya. knowledge management dalam organisasi meliputi proses dari knowledge management yang mempengaruhi proses operasional, proses operasional mempengaruhi keluaran langsung dan keluaran langsung mempengaruhi kinerja organisasi.Tujuan penelitian ini untuk menganalisis pengaruh knowledge management dan pengembangan karir terhadap kinerja karyawan Klinik Utama Spesialis Mata SMEC Manado. Penelitian ini merupakan penelitian kuantitatif dengan desain pengumpulan data secara cross sectional. Penelitian ini dilaksanakan di Klinik Utama Spesialis Mata SMEC Manado. Penelitian ini telah dilaksanakan pada bulan Oktober 2022 sampai Januari 2023. Populasi yakni karyawan Klinik Utama Spesialis Mata SMEC Manado sebanyak 67 karyawan. Sampel penelitian sejumlah 67 karyawan. Pada penelitian ini, penulis menyebarkan kuesioner. Didapatkan hasil bahwa secara simultan (bersamaan) variabel knowledge management dan pengembangan karir berpengaruh signifikan terhadap variabel kinerja karyawan. Diharapkan kepada Manajemen Klinik Utama Spesialis Mata SMEC Manado kiranya dapat mendengarkan dan menanggapi keluhan yang datang dari karyawan serta diharapkan juga lebih memberikan perhatian berupa penghargaan yang dapat diwujudkan dalam bentuk pujian atas prestasi atau bonus

Understanding how appraisal of doctors produces its effects: a realist review protocol

INTRODUCTION: UK doctors are now required to participate in revalidation to maintain their licence to practise. Appraisal is a fundamental component of revalidation. However, objective evidence of appraisal changing doctors' behaviour and directly resulting in improved patient care is limited. In particular, it is not clear how the process of appraisal is supposed to change doctors' behaviour and improve clinical performance. The aim of this research is to understand how and why appraisal of doctors is supposed to produce its effect. METHODS AND ANALYSIS: Realist review is a theory-driven interpretive approach to evidence synthesis. It applies realist logic of inquiry to produce an explanatory analysis of an intervention that is, what works, for whom, in what circumstances, in what respects. Using a realist review approach, an initial programme theory of appraisal will be developed by consulting with key stakeholders in doctors' appraisal in expert panels (ethical approval is not required), and by searching the literature to identify relevant existing theories. The search strategy will have a number of phases including a combination of: (1) electronic database searching, for example, EMBASE, MEDLINE, the Cochrane Library, ASSIA, (2) 'cited by' articles search, (3) citation searching, (4) contacting authors and (5) grey literature searching. The search for evidence will be iteratively extended and refocused as the review progresses. Studies will be included based on their ability to provide data that enable testing of the programme theory. Data extraction will be conducted, for example, by note taking and annotation at different review stages as is consistent with the realist approach. The evidence will be synthesised using realist logic to interrogate the final programme theory of the impact of appraisal on doctors' performance. The synthesis results will be written up according to RAMESES guidelines and disseminated through peer-reviewed publication and presentations. TRIAL REGISTRATION NUMBER: The protocol is registered with PROSPERO 2014:CRD42014007092.UK Department of HealthNational Institute for Health Research (NIHR

Phylogeny of Celastraceae (spindle-tree family) subfamilies Hippocrateoideae and Salacioideae inferred from chloroplast and nuclear genes

The phylogeny of Celastraceae (the spindle-tree family) subfamilies Hippocrateoideae and Salacioideae, which include about 360 species of shrubs, trees, and vines in 25 genera, was inferred using plastid (matK, trnL-F) and nuclear (ITS and 26S rDNA) genes. Together, subfamilies Hippocrateoideae and Salacioideae contain all members of the former Hippocrateaceae, which are now recognized as a derived group within Celastraceae sensu stricto. Based on our results, Brassiantha, a monotypic genus endemic to New Guinea, is more closely related to the clade of Dicarpellum (New Caledonia) and Hypsophila (Queensland, Australia) than it is to the former Hippocrateaceae, in contrast to previous studies. This well supported resolution indicates that having a nectary disk positioned outside the stamens has been convergently derived in these two separate lineages. The clade of Kokoona and Lophopetalum was resolved as sister to the clade of Hippocrateoideae, Sarawakodendron, and Salacioideae. This resolution of Kokoona and Lophopetalum supports previous assertions that they are a "transitional link" between Celastraceae sensu stricto and the former Hippocrateaceae. Sarawakodendron, a monotypic genus endemic to Borneo, was resolved as sister to the clade of Salacioideae, which supports earlier assertions that Sarawakodendron is "transitional" between Kokoona, Lophopetalum, and Salacioideae. Based on our inferred phylogeny, arils as mucilaginous pulp are derived within Salacioideae and winged arils may be primitive within the former Hippocrateaceae as a whole. Finally, the former Hippocrateaceae had an Old World, rather than a New World, origin.College Honors

Effect of probe characteristics on the subtractive hybridization efficiency of human genomic DNA

<p>Abstract</p> <p>Background</p> <p>The detection sensitivity of low abundance pathogenic species by polymerase chain reaction (PCR) can be significantly enhanced by removing host nucleic acids. This selective removal can be performed using a magnetic bead-based solid phase with covalently immobilized capture probes. One of the requirements to attain efficient host background nucleic acids subtraction is the capture probe characteristics.</p> <p>Findings</p> <p>In this study we investigate how various capture probe characteristics influence the subtraction efficiency. While the primary focus of this report is the impact of probe length, we also studied the impact of probe conformation as well as the amount of capture probe attached to the solid phase. The probes were immobilized on magnetic microbeads functionalized with a phosphorous dendrimer. The subtraction efficiency was assessed by quantitative real time PCR using a single-step capture protocol and genomic DNA as target. Our results indicate that short probes (100 to 200 bp) exhibit the best subtraction efficiency. Additionally, higher subtraction efficiencies with these probes were obtained as the amount of probe immobilized on the solid phase decreased. Under optimal probes condition, our protocol showed a 90 - 95% subtraction efficiency of human genomic DNA.</p> <p>Conclusions</p> <p>The characteristics of the capture probe are important for the design of efficient solid phases. The length, conformation and abundance of the probes determine the capture efficiency of the solid phase.</p

Nursing home administrators’ perspectives on a study feedback report : a cross sectional survey

BackgroundThis project is part of the Translating Research in Elder Care (TREC) program of research, a multi-level and longitudinal research program being conducted in 36 nursing homes in three Canadian Prairie Provinces. The overall goal of TREC is to improve the quality of care for older persons living in nursing homes and the quality of work life for care providers. The purpose of this paper is to report on development and evaluation of facility annual reports (FARs) from facility administrators&rsquo; perspectives on the usefulness, meaningfulness, and understandability of selected data from the TREC survey. MethodsA cross sectional survey design was used in this study. The feedback reports were developed in collaboration with participating facility administrators. FARs presented results in four contextual areas: workplace culture, feedback processes, job satisfaction, and staff burnout. Six weeks after FARs were mailed to each administrator, we conducted structured telephone interviews with administrators to elicit their evaluation of the FARs. Administrators were also asked if they had taken any actions as a result of the FAR. Descriptive and inferential statistics, as well as content analysis for open-ended questions, were used to summarize findings. ResultsThirty-one facility administrators (representing thirty-two facilities) participated in the interviews. Six administrators had taken action and 18 were planning on taking action as a result of FARs. The majority found the four contextual areas addressed in FAR to be useful, meaningful, and understandable. They liked the comparisons made between data from years one and two and between their facility and other TREC study sites in their province. Twenty-two indicated that they would like to receive information on additional areas such as aggressive behaviours of residents and information sharing. Twenty-four administrators indicated that FARs contained enough information, while eight found FARs &lsquo;too short&rsquo;. Administrators who reported that the FAR contained enough information were more likely to take action within their facilities than administrators who reported that they needed more information. ConclusionsAlthough the FAR was brief, the presentation of the four contextual areas was relevant to the majority of administrators and prompted them to plan or to take action within their facility. <br /

Genistein Improves Neuropathology and Corrects Behaviour in a Mouse Model of Neurodegenerative Metabolic Disease

BACKGROUND: Neurodegenerative metabolic disorders such as mucopolysaccharidosis IIIB (MPSIIIB or Sanfilippo disease) accumulate undegraded substrates in the brain and are often unresponsive to enzyme replacement treatments due to the impermeability of the blood brain barrier to enzyme. MPSIIIB is characterised by behavioural difficulties, cognitive and later motor decline, with death in the second decade of life. Most of these neurodegenerative lysosomal storage diseases lack effective treatments. We recently described significant reductions of accumulated heparan sulphate substrate in liver of a mouse model of MPSIIIB using the tyrosine kinase inhibitor genistein. METHODOLOGY/PRINCIPAL FINDINGS: We report here that high doses of genistein aglycone, given continuously over a 9 month period to MPSIIIB mice, significantly reduce lysosomal storage, heparan sulphate substrate and neuroinflammation in the cerebral cortex and hippocampus, resulting in correction of the behavioural defects observed. Improvements in synaptic vesicle protein expression and secondary storage in the cerebral cortex were also observed. CONCLUSIONS/SIGNIFICANCE: Genistein may prove useful as a substrate reduction agent to delay clinical onset of MPSIIIB and, due to its multimodal action, may provide a treatment adjunct for several other neurodegenerative metabolic diseases

Patterns of ash (Fraxinus excelsior L.) colonization in mountain grasslands: the importance of management practices

International audienceWoody colonization of grasslands is often associated with changes in abiotic or biotic conditions or a combination of both. Widely used as fodder and litter in the past traditional agro-pastoral system, ash (Fraxinus excelsior L.) has now become a colonizing species of mountain grasslands in the French Pyrenees. Its present distribution is dependent on past human activities and it is locally controlled by propagule pressure and abiotic conditions. However, even when all favourable conditions are met, all the potentially colonizable grasslands are not invaded. We hypothesize that management practices should play a crucial role in the control of ash colonization. From empirical field surveys we have compared the botanical composition of a set of grasslands (present and former) differing in management practices and level of ash colonization. We have displayed a kind of successional gradient positively linked to both ash cover and height but not to the age of trees. We have tested the relationships between ash presence in grassland and management types i.e. cutting and/or grazing, management intensity and some grassland communities' features i.e. total and local specific richness and species heterogeneity. Mixed use (cutting and grazing) is negatively linked to ash presence in grassland whereas grazing alone positively. Mixed use and high grazing intensity are directly preventing ash seedlings establishment, when low grazing intensity is allowing ash seedlings establishment indirectly through herbaceous vegetation neglected by livestock. Our results show the existence of a limit between grasslands with and without established ashes corresponding to a threshold in the intensity of use. Under this threshold, when ash is established, the colonization process seems to become irreversible. Ash possesses the ability of compensatory growth and therefore under a high grazing intensity develops a subterranean vegetative reproduction. However the question remains at which stage of seedling development and grazing intensity these strategies could occur