bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Ovo Live Imaging of Avian Embryos: Figure 1.
Authors: Kulesa Paul M. , Bailey Caleb M. , Cooper Cameron , Fraser Scott E. 
Published: June 2010

INTRODUCTIONVertebrate development is best studied in an intact embryo model, but a robust interface between time-lapse microscopy and in vivo embryo health and maintenance can be difficult to achieve in model systems that rely on external factors for life support. This protocol presents a system for in ovo culture and time-lapse imaging of fluorescently labeled cells within living avian embryos, using a Teflon membrane that is oxygen-permeable and liquid-impermeable. The protocol describes the Teflon membrane assembly (the assembly size can be changed to fit smaller eggs, such as those of the quail), its interface with the egg window, and the use of an upright microscope and heated chamber. The use of the system is demonstrated in chick embryos by following individual fluorescently labeled neural crest cells, a multipotent stem cell-like population that differentiates into a wide range of derivatives and travels extensively throughout the embryo. By combining in ovo culture with confocal or two-photon four-dimensional time-lapse imaging, embryo health can be maintained for up to 5 d, and neural crest cell behaviors can be visualized for long periods of time (~36 h). This technique has been adapted to study somitogenesis.
Multi-Position Photoactivation and Multi-Time Acquisition for Large-Scale Cell Tracing in Avian Embryos
Authors: Steen Joseph , Morrison Jason A. , Kulesa Paul M. 
Published: June 2010

INTRODUCTIONVertebrate development is a complex orchestration of cell and tissue movements. Tracing individual cell positions can rapidly become a large-scale problem because cell numbers often grow exponentially in the early embryo. A typical approach consists of fluorescently marking small numbers of cells within a large number of embryos, followed by comprehensive three-dimensional static or time-lapse imaging to map cell positions. However, for large-scale cell tracing, such as during organogenesis, the time, effort, and expense of this approach can be limiting. The multi-position acquisition method can be used to capture more than one location on a microscope stage and allow for multi-specimen imaging. When combined with photoactivation cell labeling, a tool for selective cell marking using laser excitation, multi-position imaging offers a powerful tool for rapid data acquisition. This protocol describes the technique and demonstrates its use to map cell movements in the chick spinal cord, using slice culture explants. The details of multiple slice culture preparation, multi-position photoactivation, and multi-time acquisition are described. These events are coordinated by setting up blocks of microscope instructions that execute sequentially. This method significantly decreases the time, effort, microscopy, and embryo costs by a factor of the number of specimens imaged per session, typically six.
Purification of the Mitotic Checkpoint Complex (MCC) and the Anaphase Promoting Complex/Cyclosome (APC/C) from HeLa Cells
Authors: Sudakin Valery 
Published: June 2010

INTRODUCTIONIn this protocol, active anaphase promoting complex/cyclosome (APC/C) and its inhibitor, mitotic checkpoint complex (MCC), are purified from HeLa cell extracts. The two complexes are first separated by gel filtration and ion exchange chromatography. The MCC is further purified by an additional gel filtration step. The APC/C activity and the ability of MCC to inhibit the APC/C are assayed at every stage of the purification procedure by in vitro ubiquitination assays. The composition and distribution of the complexes are monitored by Western blot analysis.
Barriers to research utilisation by staff nurses in a university hospital
Authors: Kocaman Gulseren , Seren Seyda , Lash Ayhan A , Kurt Seyyare , Bengu Nergiz , Yurumezoglu Havva A 
Published: June 2010

Aim.  To identify self‐reported perceptions of the barriers to research utilisation by Turkish staff nurses working in a university hospital and to ascertain whether involvement in research‐related activities influenced their perception of the barriers.Background.  Research utilisation in nursing has been a prominent concern for the last 30 years. While investigators in several countries have identified nurses’ perception of barriers to research utilisation, this is the first study to investigate such barriers in Turkey.Design.  This is a descriptive, cross sectional study with a sample of 329 staff nurses working in a university hospital. Two instruments were used; a demographics questionnaire that investigated research activities and the Barriers Scale to measure perceptions. An additional item, ‘studies are published in English’ was added to the scale.Results.  The lack of time to implement new ideas was the most frequently, 84·5%, perceived barrier. The second most frequently perceived barrier was difficulty in reading English. The top six barriers were all related to the setting. Additionally, research‐active nurses found insufficient time less of a barrier but unawareness of research findings and being isolated from knowledgeable colleagues as greater barriers to research utilisation.Conclusions.  Turkish nurses’ perception of barriers is similar to the findings in other countries, i.e., that the top six barriers were related to setting. Language barrier compares similarly with studies in other countries that investigated this variable.Relevance to clinical practice.  The gap between theory and practice in nursing can be minimised by identifying strategies to increase research utilisation in clinical settings. Such strategies can also improve patient care outcomes.
Directed mutagenesis of Mycobacterium smegmatis 16S rRNA to reconstruct the in vivo evolution of aminoglycoside resistance in Mycobacterium tuberculosis
Authors: Shcherbakov Dmitri , Akbergenov Rashid , Matt Tanja , Sander Peter , Andersson Dan I. , Böttger Erik C. 
Published: June 2010

SummaryDrug resistance in Mycobacterium tuberculosis is a global problem, with major consequences for treatment and public health systems. As the emergence and spread of drug‐resistant tuberculosis epidemics is largely influenced by the impact of the resistance mechanism on bacterial fitness, we wished to investigate whether compensatory evolution occurs in drug‐resistant clinical isolates of M. tuberculosis. By combining information from molecular epidemiology studies of drug‐resistant clinical M. tuberculosis isolates with genetic reconstructions and measurements of aminoglycoside susceptibility and fitness in Mycobacterium smegmatis, we have reconstructed a plausible pathway for how aminoglycoside resistance develops in clinical isolates of M. tuberculosis. Thus, we show by reconstruction experiments that base changes in the highly conserved A‐site of 16S rRNA that: (i) cause aminoglycoside resistance, (ii) confer a high fitness cost and (iii) destabilize a stem‐loop structure, are associated with a particular compensatory point mutation that restores rRNA secondary structure and bacterial fitness, while maintaining to a large extent the drug‐resistant phenotype. The same types of resistance and associated mutations can be found in M. tuberculosis in clinical isolates, suggesting that compensatory evolution contributes to the spread of drug‐resistant tuberculosis disease.
Gain‐of‐function mutations cluster in distinct regions associated with the signalling pathway in the PAS domain of the aerotaxis receptor, Aer
Authors: Campbell Asharie J. , Watts Kylie J. , Johnson Mark S. , Taylor Barry L. 
Published: June 2010

SummaryThe Aer receptor monitors internal energy (redox) levels in Escherichia coli with an FAD‐containing PAS domain. Here, we randomly mutagenized the region encoding residues 14–119 of the PAS domain and found 72 aerotaxis‐defective mutants, 24 of which were gain‐of‐function, signal‐on mutants. The mutations were mapped onto an Aer homology model based on the structure of the PAS–FAD domain in NifL from Azotobacter vinlandii. Signal‐on lesions clustered in the FAD binding pocket, the β‐scaffolding and in the N‐cap loop. We suggest that the signal‐on lesions mimic the ‘signal‐on’ state of the PAS domain, and therefore may be markers for the signal‐in and signal‐out regions of this domain. We propose that the reduction of FAD rearranges the FAD binding pocket in a way that repositions the β‐scaffolding and the N‐cap loop. The resulting conformational changes are likely to be conveyed directly to the HAMP domain, and on to the kinase control module. In support of this hypothesis, we demonstrated disulphide band formation between cysteines substituted at residues N98C or I114C in the PAS β‐scaffold and residue Q248C in the HAMP AS‐2 helix.
Metabolic shutdown in Escherichia coli cells lacking the outer membrane channel TolC
Authors: Dhamdhere Girija , Zgurskaya Helen I. 
Published: June 2010

SummaryThe outer membrane channel TolC is a key component of multidrug efflux and type I secretion transporters in Escherichia coli. Mutational inactivation of TolC renders cells highly susceptible to antibiotics and leads to defects in secretion of protein toxins. Despite impairment of various transport functions, no growth defects were reported in cells lacking TolC. Unexpectedly, we found that the loss of TolC notably impairs cell division and growth in minimal glucose medium. The TolC‐dependent phenotype was further exacerbated by the loss of ygiB and ygiC genes expressed in the same operon as tolC and their homologues yjfM and yjfC located elsewhere on the chromosome. Our results show that this growth deficiency is caused by depletion of the critical metabolite NAD+ and high NADH/NAD+ ratios. The increased amounts of PspA and decreased rates of NADH oxidation in ΔtolC membranes indicated stress on the membrane and dissipation of a proton motive force. We conclude that inactivation of TolC triggers metabolic shutdown in E. coli cells grown in minimal glucose medium. The ΔtolC phenotype is partially rescued by YgiBC and YjfMC, which have parallel functions independent from TolC.
UVA1 phototherapy: a review of mechanism and therapeutic application
Authors: York Nathan R. , Jacobe Heidi T. 
Published: June 2010

AbstractUltraviolet radiation (UVR) phototherapy has been associated with both deleterious and beneficial effects to patients with both localized and systemic skin disorders. Phototherapy is advantageous in diseases of the epidermis and dermis, as it provides the most direct approach minimizing systemic side effects. Most recently, ultraviolet A1 (UVA1) phototherapy has emerged as a specific UVR phototherapeutic mechanism. It has shown to be therapeutic in a number of sclerosing skin conditions and other dermatitides, in many cases proving to be more effective than other phototherapy modalities. Treatment advantages of UVA1 phototherapy include the ability to penetrate into the deep layers of the skin to affect changes on disease‐causing T cells, as well as activation of endothelial cells to promote neovascularization. UVA1 therapy also has been shown to be relatively free of side effects associated with other phototherapy regimens, including erythema and cellular transformation. These properties make UVA1 phototherapy an important treatment option for many debilitating skin conditions.
Nonmutilating palmoplantar and periorificial kertoderma: a variant of Olmsted syndrome or a distinct entity?
Authors: Nofal Ahmad , Assaf Magda , Nassar Amani , Nofal Eman , Shehab Marwa , El‐Kabany Moustafa 
Published: June 2010

AbstractBackground  Olmsted syndrome is a rare keratinization disorder characterized by mutilating palmoplantar and periorificial keratoderma as the two major diagnostic features. Some authors believe that atypical cases without this standard combination may not really belong to Olmsted syndrome. Herein, we describe two familial cases with congenital nonmutilating palmoplantar and periorificial keratoderma, and discuss their similarities and differences with Olmsted syndrome.Patients  The study included two sisters who presented with focal and punctate nonmutilating palmoplantar keratoderma (PPK), periorificial hyperkeratotic plaques, and widely distributed keratotic lesions. Fragile denuded areas of the skin were found in sites exposed to trauma. Fingernails showed a characteristic form of leukonychia.Results  Histopathology of plantar keratoderma showed psoriasiform hyperplasia with marked compact hyperkeratosis, while vicinity of denuded skin revealed thin parakeratotic zone and dissolution of the granular cell layer. Immunohistochemistry demonstrated suprabasal staining pattern for acidic keratin (AE1) and uniform positivity, starting four to six layers above the basal layer, for cytokeratin 10. Electron microscopy showed defective keratinization. Cytogenetic studies revealed normal karyotype and no chromosomal breakage.Conclusion  Our cases share Olmsted syndrome in the early onset, and the presence of symmetrical PPK, periorificial keratoderma and keratotic lesions. However, the striking nonmutilating nature of PPK and the presence of unique features in our patients suggest a newly described keratinization disorder.
Progressive multifocal leukoencephalopathy and antipsoriatic drugs
Authors: Di Lernia Vito 
Published: June 2010

AbstractProgressive Multifocal Leukoencephalopathy (PML) is a rare and often fatal opportunistic demyelinating disorder secondary to central nervous system infection by the polyomavirus John Cunningham (JC), a common infecting agent in human populations, with 50–70% of healthy individuals having antibodies to the virus. Although human immunodeficiency virus (HIV) infection remains the most common predisposing factor for PML, which can occur also in association with hematologic malignant neoplasms or iatrogenic immunosuppression in the setting of organ transplantations, recently JC virus has been recognized as an important pathogen in patients with autoimmune and rheumatic diseases receiving immunosuppressive treatments. Following the availability of new biologic drugs, additional cases of PML have been reported in dermatologic patients as well. In particular, the occurrence of PML in psoriatic patients who had been taking efalizumab in the absence of any other concurrent immunosuppressive agents resulted in the decision of European Medicine Agency (EMEA) in February 2009 to recommend suspension of the marketing authorization for this drug in Europe. Some months later the manufacturer of the drug decided to voluntarily withdraw the product from the U.S. market. Since PML can occur as a potential side effect of different immunosuppressive drugs, including conventional treatments for psoriasis, and in consideration of the future development of new immunosuppressive biological agents, dermatologists need to become familiar with opportunistic infections including PML.
Feasibility of antegrade radical prostatectomy for clinically locally advanced prostate cancer: a comparative study with clinically localized disease
Authors: Yamamoto Shinya , Kawakami Satoru , Yonese Junji , Fujii Yasuhisa , Tsukamoto Tetsuro , Okubo Yuhei , Kijima Toshiki , Ishikawa Yuichi , Fukui Iwao 
Published: June 2010

Objectives:  To investigate intraoperative and early postoperative complications of antegrade radical prostatectomy with intended wide resection (aRP) for clinically locally advanced prostate cancer (cLAD) and to compare with those of aRP for clinically localized prostate cancer (cLD).Methods:  Between March 1994 and June 2007, 800 consecutive Japanese patients including 625 with cLD and 175 with cLAD underwent aRP and bilateral limited lymphadenectomy. Clinicopathological data including intraoperative and early postoperative complications (within 30 days after operation) were compared between cLD and cLAD groups.Results:  No deaths occurred. Operative time and blood loss did not differ significantly between the groups. Intraoperative and early postoperative complications were observed in 11 (1.4%) and 123 (15.4%) of the entire cohort, respectively. Prevalent early postoperative complications were pelvic hematoma, wound infection, urinary retention and lymphocele or prolonged lymph drainage. There were no significant differences in the entire intraoperative and early postoperative complications between the groups. The majority of the early postoperative complications were minor.Conclusions:  aRP for cLAD is technically feasible and a safe surgical procedure. If radical prostatectomy could be established as a standard treatment for cLAD in the future, aRP might be valuable as the first step of multimodal treatments.
Evolution of pancreas transplant surgery
Authors: Lam Vincent W. T. , Pleass Henry C. C. , Hawthorne Wayne , Allen Richard D. M. 
Published: June 2010

AbstractBackground: Type 1 diabetes mellitus is a chronic condition often leading to disabling complications including retinopathy, neuropathy and cardiovascular disease which can be modified by intensive treatment with insulin. Such treatment, however, is associated with a restrictive lifestyle and risk of hypoglycaemic morbidity and mortality.Methods: This review examines the role of pancreas transplantation in patients with Type 1 diabetes mellitus.Results: Pancreas transplantation is currently the only proven option to achieve long‐term insulin independence, resulting in an improvement or stabilization of those diabetic related complications. The hazards of pancreas transplantation as a major operation are well known.Balancing the risks of a surgical procedure, with the benefits of restoring normoglycaemia remains an important task for the pancreas transplant surgeon. Pancreas transplantation is not an emergency operation to treat poorly managed and non‐compliant patients with debilitating complications. It is a highly specialized procedure which has evolved both in terms of the surgical technique, patient selection and assessment.Conclusion: Pancreas transplantation has emerged as the single most effective way to achieve normal glucose homeostasis in patients with Type 1 diabetes mellitus.
Role of an acute assessment and review area for general surgical patients
Authors: Perry Ellen , Mackintosh Stuart , Connor Saxon 
Published: June 2010

AbstractBackground:  Increasing demand for acute surgical services is one of the major challenges facing modern health systems. The aim of this study was to assess the impact of implementing a dedicated surgical assessment and review area (SARA) on patient flow for acute general surgical patients at a major teaching hospital in New Zealand.Methods:  A specific area within inpatient surgical beds was redeveloped and staffed by a dedicated surgical team. Prospective data pertaining to patient flow and outcomes were collected (10‐month period) and compared with historical controls (preceding 14 months prior to implementation).Results:  During the 24‐month period 9182 acute general surgical patients were admitted (3836 [41.1%] post implementation of SARA). Subsequent to the introduction of SARA, 865 (22.5%) patients were referred directly from their general practitioner. Length of stay was reduced in all patients from 2.58 to 2.04 days (P < 0.001) and in those who did not require surgery from 2.56 to 1.96 days (P < 0.001). However, the number of days in which the department of surgery had outlying patients increased significantly from 76.7% to 86.3%, P < 0.001.Conclusions:  The introduction of a dedicated SARA significantly reduced hospital stay and improved efficiencies of the emergency department.
Physician assistants: trialling a new surgical health professional in Australia
Authors: Ho Phyllis , Pesicka Dawanda , Schafer Amy , Maddern Guy 
Published: June 2010

AbstractBackground:  The Australia health workforce productivity Commission Research Report in 2005 identified workforce shortages. One of the recommendations is that new models of health care be established. As a result South Australia is trialling United States trained physician assistants in a pilot program. This paper summaries the review of literature of the physician assistant role and safety in the surgical setting.Methods:  A literature search using Medline and Pubmed from 1966 until 2009 with key words: physician assistants, midlevel providers, surgery. The references of the results were also searched for suitable articles. The Google search engine was also used with the above keywords to search for latest developments from nontraditional sources.Results:  There were over 200 suitable articles relating to the quality and safety of physician assistants. The overwhelming majority of the articles originate from the United States and these vary in quality. There were 13 published studies identified that documented physician assistants in the surgical setting.Conclusion:  From the published data physician assistants have been shown to provide safe and provide high quality care in surgical units. It is important that prior to their commencement their role is defined to alleviate conflict and confusion in the team. Continued auditing should be conducted to monitor progress and impact.
Guidelines for patient selection and performance of carotid artery stenting
Authors:   The Carotid Stenting Guidelines Committee
Published: June 2010

AbstractBackground:  The endovascular treatment of carotid atherosclerosis with carotid artery stenting (CAS) remains controversial. Carotid endarterectomy remains the benchmark in terms of procedural mortality and morbidity. At present, there are no consensus Australasian guidelines for the safe performance of CAS.Methods:  We applied a modified Delphi consensus method of iterative consultation between the College representatives on the Carotid Stenting Guidelines Committee (CSGC).Results:  Selection of patients suitable for CAS needs careful consideration of clinical and patho‐anatomical criteria and cannot be directly extrapolated from clinical indicators for carotid endarterectomy (CEA). Randomized controlled trials (including pooled analyses of results) comparing CAS with CEA for treatment of symptomatic stenosis have demonstrated that CAS is more hazardous than CEA. On current evidence, the CGSC therefore recommends that CAS should not be performed in the majority of patients requiring carotid revascularisation. The evidence for CAS in patients with symptomatic severe carotid stenosis who are considered medically high risk is weak, and there is currently no evidence to support CAS as a treatment for asymptomatic carotid stenosis. The use of distal protection devices during CAS remains controversial with increased risk of clinically silent stroke. The knowledge requirements for the safe performance of CAS include an understanding of the evidence base from randomized controlled trials, carotid and aortic arch anatomy and pathology, clinical stroke syndromes, the differing treatment options for stroke and carotid atherosclerosis, and recognition and management of periprocedural complications. It is critical that all patients being considered for a carotid intervention have adequate pre‐procedural neuro‐imaging and an independent, standardized neurological assessment before and after the procedure. Maintenance of proficiency in CAS requires active involvement in surgical/endovascular audit and continuing medical education programs. These standards should apply in the public and private health care settings.Conclusion:  These guidelines represent the consensus of an inter‐collegiate committee in order to direct appropriate patient selection and the range of cognitive and technical requirements to perform CAS. Advances in endovascular technologies and the results of randomized controlled trials will guide future revisions of these guidelines.