Science.gov

Sample records for improved isolation protocol

  1. An improved protocol for intact chloroplasts and cpDNA isolation in conifers.

    PubMed

    Vieira, Leila do Nascimento; Faoro, Helisson; Fraga, Hugo Pacheco de Freitas; Rogalski, Marcelo; de Souza, Emanuel Maltempi; de Oliveira Pedrosa, Fábio; Nodari, Rubens Onofre; Guerra, Miguel Pedro

    2014-01-01

    Performing chloroplast DNA (cpDNA) isolation is considered a major challenge among different plant groups, especially conifers. Isolating chloroplasts in conifers by such conventional methods as sucrose gradient and high salt has not been successful. So far, plastid genome sequencing protocols for conifer species have been based mainly on long-range PCR, which is known to be time-consuming and difficult to implement. We developed a protocol for cpDNA isolation using three different conifer families: Araucaria angustifolia and Araucaria bidwilli (Araucariaceae), Podocarpus lambertii (Podocarpaceae) and Pinus patula (Pinaceae). The present protocol is based on high salt isolation buffer followed by saline Percoll gradient. Combining these two strategies allowed enhanced chloroplast isolation, along with decreased contamination caused by polysaccharides, polyphenols, proteins, and nuclear DNA in cpDNA. Microscopy images confirmed the presence of intact chloroplasts in high abundance. This method was applied to cpDNA isolation and subsequent sequencing by Illumina MiSeq (2×250 bp), using only 50 ng of cpDNA. Reference-guided chloroplast genome mapping showed that high average coverage was achieved for all evaluated species: 24.63 for A. angustifolia, 135.97 for A. bidwilli, 1196.10 for P. lambertii, and 64.68 for P. patula. Results show that this improved protocol is suitable for enhanced quality and yield of chloroplasts and cpDNA isolation from conifers, providing a useful tool for studies that require isolated chloroplasts and/or whole cpDNA sequences.

  2. An Improved Protocol for Intact Chloroplasts and cpDNA Isolation in Conifers

    PubMed Central

    Vieira, Leila do Nascimento; Faoro, Helisson; Fraga, Hugo Pacheco de Freitas; Rogalski, Marcelo; de Souza, Emanuel Maltempi; de Oliveira Pedrosa, Fábio; Nodari, Rubens Onofre; Guerra, Miguel Pedro

    2014-01-01

    Background Performing chloroplast DNA (cpDNA) isolation is considered a major challenge among different plant groups, especially conifers. Isolating chloroplasts in conifers by such conventional methods as sucrose gradient and high salt has not been successful. So far, plastid genome sequencing protocols for conifer species have been based mainly on long-range PCR, which is known to be time-consuming and difficult to implement. Methodology/Principal Findings We developed a protocol for cpDNA isolation using three different conifer families: Araucaria angustifolia and Araucaria bidwilli (Araucariaceae), Podocarpus lambertii (Podocarpaceae) and Pinus patula (Pinaceae). The present protocol is based on high salt isolation buffer followed by saline Percoll gradient. Combining these two strategies allowed enhanced chloroplast isolation, along with decreased contamination caused by polysaccharides, polyphenols, proteins, and nuclear DNA in cpDNA. Microscopy images confirmed the presence of intact chloroplasts in high abundance. This method was applied to cpDNA isolation and subsequent sequencing by Illumina MiSeq (2×250 bp), using only 50 ng of cpDNA. Reference-guided chloroplast genome mapping showed that high average coverage was achieved for all evaluated species: 24.63 for A. angustifolia, 135.97 for A. bidwilli, 1196.10 for P. lambertii, and 64.68 for P. patula. Conclusion Results show that this improved protocol is suitable for enhanced quality and yield of chloroplasts and cpDNA isolation from conifers, providing a useful tool for studies that require isolated chloroplasts and/or whole cpDNA sequences. PMID:24392157

  3. An improved protocol for isolation and culturing of mouse spermatogonial stem cells.

    PubMed

    Baazm, Maryam; Abolhassani, Farid; Abbasi, Mahdi; Habibi Roudkenar, Mehryar; Amidi, Fardin; Beyer, Cordian

    2013-08-01

    Spermatogonial stem cells (SSCs) represent a unique testicular cell type that has the capacity for proliferating, differentiating, and transmitting genetic information. This particular cell type is a strong focus of stem cell research, with isolation and maintenance of SSCs as an important issue. Therefore, we attempted to optimize SSCs handling and to analyze different media and feeder layers, such as adult and embryonic Sertoli cells. The expression patterns of SSC-specific proteins (α6 and β1 integrins, Stra8, and DAZL) and restoration of spermatogenesis were chosen as parameters to demonstrate the efficacy of the protocol. SSCs were isolated from testes of 3- to 6-day-old mice using a magnetic activated cell-sorting system and Thy-1 antibody. After enrichment, SSCs were cultured for 7 days with different media and feeder layers. Then, SSCs were transplanted to recipient mice. Culturing on adult and embryonic Sertoli cells and in the presence of different growth factors [glial cell line-derived neurotrophic factor (GDNF), GDNF family receptor α1 (GFR-α1), and basic fibroblast growth factor (bFGF) resulted in an undifferentiated SSC phenotype with typical stem cell characteristics observed in vivo. The established co-culture model could help to improve the recovery and quality of stem cell preparation of mammalian testis.

  4. An improved protocol for the isolation of total genomic DNA from Labyrinthulomycetes.

    PubMed

    Ranasinghe, Chaminda Padmashantha; Harding, Rob; Hargreaves, Megan

    2015-03-01

    Many protocols have been used for extraction of DNA from Thraustochytrids. These generally involve the use of CTAB, phenol/chloroform and ethanol. They also feature mechanical grinding, sonication, N2 freezing or bead beating. However, the resulting chemical and physical damage to extracted DNA reduces its quality. The methods are also unsuitable for large numbers of samples. Commercially-available DNA extraction kits give better quality and yields but are expensive. Therefore, an optimized DNA extraction protocol was developed which is suitable for Thraustochytrids to both minimise expensive and time-consuming steps prior to DNA extraction and also to improve the yield. The most effective method is a combination of single bead in TissueLyser (Qiagen) and Proteinase K. Results were conclusive: both the quality and the yield of extracted DNA were higher than with any other method giving an average yield of 8.5 µg/100 mg biomass.

  5. An improved protocol for the preparation and restriction enzyme digestion of pulsed-field gel electrophoresis agarose plugs for the analysis of Legionella isolates.

    PubMed

    Chang, Bin; Amemura-Maekawa, Junko; Watanabe, Haruo

    2009-01-01

    Pulsed-field gel electrophoresis (PFGE), which determines the genomic relatedness of isolates, is currently used for the epidemiological investigation of infectious agents such as bacteria. In particular, this method has been used for the epidemiological investigation of Legionella outbreaks. However, it takes 4 days to complete a Legionella-PFGE analysis. Due to partial digestion and DNA damage, the reproducibility of the obtained fragment digestion patterns is poor for this pathogen. In this study, we report an improved protocol that takes only 2 days to complete and that allows clear discrimination of the restriction profile with higher reproducibility than that previously achieved.

  6. Protocols for cofactor isolation of nitrogenase.

    PubMed

    Fay, Aaron W; Lee, Chi-Chung; Wiig, Jared A; Hu, Yilin; Ribbe, Markus W

    2011-01-01

    The iron-molybdenum cofactor (FeMoco) of the nitrogenase MoFe protein has remained a focal point in the field of bioinorganic chemistry for decades. This unique metal cluster has long been regarded as the actual site of dinitrogen reduction, and it is structurally complex and chemically unprecedented. A detailed characterization of the isolated FeMoco is crucial for elucidating the physiochemical properties of this biologically important cofactor. Such a study requires an effective technique to extract FeMoco intact, and in high yield, from the MoFe protein. A method involving the acid treatment of the MoFe protein and the subsequent extraction of FeMoco into an organic solvent was developed over 30 years ago and has been improved upon ever since. FeMoco isolated by this strategy is catalytically active and spectrally interesting, which provides a useful platform for future structure-function analyses of this unique cofactor. A general working protocol for FeMoco isolation is described in this chapter, along with some of the major modifications reported in the past years.

  7. An improved quantum key distribution protocol

    NASA Astrophysics Data System (ADS)

    Wu, Ting-wan; Wu, Guo-hua

    2008-08-01

    This paper presented an improved quantum key distribution protocol of the quantum cryptology. Using the same measure polarizer as BB84 protocol, the improved protocol we designed with not any classical channel, but a new looped quantum channel instead, so the job of sending and receiving can be finished only by one same person. It brings several good points: the utilization ratio of photons 100% in perfect condition, at least twice over other protocols, or even higher; the public channel easy to be attacked is avoided. Further, the improved protocol authenticates the legal communicators with pre-share information, so that no attacker can jump over the progress of authentication. Be alien from the protocol of BB84, the improved protocol uses message summary to detect whether messages intercepted by attacker. Because the message summary is encrypted by one-time-pad method using pre-share information, attacker could not alter the message summary and that not to be discovered. Moreover, some theoretical analysis to the improved protocol given with information theory: we used the measure channel concept for quantum detection, and calculated the information quantity obtained by attacker in the quantum secrecy communication. The analysis results provide the theory criterion for the legal communicators and the attackers.

  8. Improved RF Isolation Amplifier

    NASA Technical Reports Server (NTRS)

    Stevens, G. L.; Macconnell, J.

    1985-01-01

    Circuit has high reverse isolation and wide bandwidth. Wideband isolation amplifier has low intermodulation distortion and high reverse isolation. Circuit does not require selected or matched components or directional coupling device. Circuit used in applications requiring high reverse isolation such as receiver intermediate-frequency (IF) strips and frequency distribution systems. Also applicable in RF and video signaling.

  9. Protocol for miRNA isolation from biofluids.

    PubMed

    Lekchnov, Evgeny A; Zaporozhchenko, Ivan A; Morozkin, Evgeny S; Bryzgunova, Olga E; Vlassov, Valentin V; Laktionov, Pavel P

    2016-04-15

    MicroRNAs (miRNAs) have been identified as promising biomarkers in cancer and other diseases. Packaging of miRNAs into vesicles and complexes with proteins ensures their stability in biological fluids but also complicates their isolation. Conventional protocols used to isolate cell-free RNA are generally successful in overcoming these difficulties; however, they are costly, labor-intensive, or heavily reliant on the use of hazardous chemicals. Here we describe a protocol that is suitable for isolating miRNAs from biofluids, including blood plasma and urine. The protocol is based on precipitation of proteins, denaturation of miRNA-containing complexes with octanoic acid and guanidine isothiocyanate, and subsequent purification of miRNA on spin columns. The efficacy of miRNA extraction by phenol-chloroform extraction, miRCURY RNA isolation kit--biofluids (Exiqon), and the proposed protocol was compared by quantitative reverse-transcription PCR of miR-16 and miR-126. The proposed protocol was slightly more effective for isolating miRNA from plasma and significantly superior to the other two methods for miRNA isolation from urine. Spectrophotometry and SDS-PAGE data suggest that the disparity in performance between miRCURY Biofluids and the proposed protocol can be attributed to differences in precipitation mechanisms, as confirmed by the retention of different proteins in the supernatant.

  10. Bedside Reporting: Protocols for Improving Patient Care.

    PubMed

    Ferguson, Teresa D; Howell, Teresa L

    2015-12-01

    Bedside reporting continues to gain much attention and is being investigated to support the premise that "hand-off" communications enhance efficacy in delivery of patient care. Patient inclusion in shift reports enhances good patient outcomes, increased satisfaction with care delivery, enhanced accountability for nursing professionals, and improved communications between patients and their direct care providers. This article discusses the multiple benefits of dynamic dialogue between patients and the health care team, challenges often associated with bedside reporting, and protocols for managing bedside reporting with the major aim of improving patient care. Nursing research supporting the concept of bedside reporting is examined.

  11. Follicles of various maturation stages react differently to enzymatic isolation: a comparison of different isolation protocols.

    PubMed

    Lierman, Sylvie; Tilleman, Kelly; Cornelissen, Maria; De Vos, Winnok H; Weyers, Steven; T'Sjoen, Guy; Cuvelier, Claude A; De Sutter, Petra

    2015-02-01

    Isolation of human follicles is based on digestion of the tissue by combinations of enzymes. Follicle vitality and morphology are often based on the analysis of pooled follicles of different maturation stages. Information is therefore lacking on the effect of the isolation protocol to individual follicles of different maturation stages. A study was conducted using five protocols combining different enzymes and varying concentrations. Isolated follicles were classified according to their maturation stages, counted and characterized for vitality, morphology, early apoptosis and organization of transzonal projections. No statistical differences were found between the protocols when outcome parameters were analysed on a pool of follicles regardless of their maturation status. Differences were observed in quality when the follicles were analysed separately according to their maturation status. Combining morphologic characteristics and vitality, both Liberase DH and Liberase TM combined with collagenase IV were better at isolating high-quality primordial follicles, compared with collagenase IV. No statistical difference between the isolation protocols was found for primary follicles. If only high-quality isolated secondary follicles are needed, collagenase IV is found to be most advantageous. Follicles of different maturation stages react differently when enzymatic isolation protocols are compared. Copyright © 2014 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  12. REHABILITATION PROTOCOL AFTER ISOLATED POSTERIOR CRUCIATE LIGAMENT RECONSTRUCTION

    PubMed Central

    de Paula Leite Cury, Ricardo; Kiyomoto, Henry Dan; Rosal, Gustavo Fogolin; Bryk, Flávio Fernandes; de Oliveira, Victor Marques; de Camargo, Osmar Pedro Arbix

    2015-01-01

    To create a rehabilitation protocol following reconstruction of the posterior cruciate ligament (PCL), through a literature review. The literature review was conducted in the Medline and Embase databases, to search for data on biomechanical concepts and analyses relating to the posterior cruciate ligament of the knee. The search strategy was set up using the following rules: problem or injury in association with anatomical location terms; or surgical intervention procedure in association with rehabilitation terms. We began the process in this manner and subsequently introduced restrictions on certain terms to improve the search specificity. To design the protocol, a table was created for better data assessment, based on the time that elapsed between surgery and the start of physiotherapy. A rehabilitation protocol was created to improve weight-bearing control in the initial weeks after surgery, with the aid of a knee brace. Our aim was to achieve gains in total range of motion of the knee, which should be attained by the third month, thereby avoiding contractures resulting from the tissue healing process. Strengthening exercises and sensory-motor training were guided accordingly, thus avoiding overload on the graft and respecting the healing phases. The protocol proposed through this review was based on the current evidence relating to this subject. PMID:27047844

  13. Assessing an improved protocol for plasma microRNA extraction.

    PubMed

    Moret, Inés; Sánchez-Izquierdo, Dolors; Iborra, Marisa; Tortosa, Luis; Navarro-Puche, Ana; Nos, Pilar; Cervera, José; Beltrán, Belén

    2013-01-01

    The first step in biomarkers discovery is to identify the best protocols for their purification and analysis. This issue is critical when considering peripheral blood samples (plasma and serum) that are clinically interesting but meet several methodological problems, mainly complexity and low biomarker concentration. Analysis of small molecules, such as circulating microRNAs, should overcome these disadvantages. The present study describes an optimal RNA extraction method of microRNAs from human plasma samples. Different reagents and commercially available kits have been analyzed, identifying also the best pre-analytical conditions for plasma isolation. Between all of them, the column-based approaches were shown to be the most effective. In this context, miRNeasy Serum/Plasma Kit (from Qiagen) rendered more concentrated RNA, that was better suited for microarrays studies and did not require extra purification steps for sample concentration and purification than phenol based extraction methods. We also present evidences that the addition of low doses of an RNA carrier before starting the extraction process improves microRNA purification while an already published carrier dose can result in significant bias over microRNA profiles. Quality controls for best protocol selection were developed by spectrophotometry measurement of contaminants and microfluidics electrophoresis (Agilent 2100 Bioanalyzer) for RNA integrity. Selected donor and patient plasma samples and matched biopsies were tested by Affymetrix microarray technology to compare differentially expressed microRNAs. In summary, this study defines an optimized protocol for microRNA purification from human blood samples, increasing the performance of assays and shedding light over the best way to discover and use these biomarkers in clinical practice.

  14. Comparison between modified DNA extraction protocols and commercial isolation kits in grapevine (Vitis vinifera L.).

    PubMed

    Akkurt, M

    2012-08-13

    Various protocols have been developed and used for DNA extraction in grapevine. However, owing to the long duration of the isolation steps in previously developed protocols, researchers have preferred to use isolation kits for studies in recent years. In our study, the DNA yield and purity obtained using six methods--namely three DNA isolation protocols and three commercial DNA isolation kits--were compared. Modifications were made and the isolation steps were shortened in the previously developed DNA isolation protocols to achieve more rapid and practical protocols. The samples were taken from plants grown under vineyard and greenhouse conditions in two periods during spring and autumn. The best results among the six DNA isolation methods were discussed. The results were also supported with polymerase chain reaction analyses conducted with isolated DNAs.

  15. A quantitative telomeric chromatin isolation protocol identifies different telomeric states

    NASA Astrophysics Data System (ADS)

    Grolimund, Larissa; Aeby, Eric; Hamelin, Romain; Armand, Florence; Chiappe, Diego; Moniatte, Marc; Lingner, Joachim

    2013-11-01

    Telomere composition changes during tumourigenesis, aging and in telomere syndromes in a poorly defined manner. Here we develop a quantitative telomeric chromatin isolation protocol (QTIP) for human cells, in which chromatin is cross-linked, immunopurified and analysed by mass spectrometry. QTIP involves stable isotope labelling by amino acids in cell culture (SILAC) to compare and identify quantitative differences in telomere protein composition of cells from various states. With QTIP, we specifically enrich telomeric DNA and all shelterin components. We validate the method characterizing changes at dysfunctional telomeres, and identify and validate known, as well as novel telomere-associated polypeptides including all THO subunits, SMCHD1 and LRIF1. We apply QTIP to long and short telomeres and detect increased density of SMCHD1 and LRIF1 and increased association of the shelterins TRF1, TIN2, TPP1 and POT1 with long telomeres. Our results validate QTIP to study telomeric states during normal development and in disease.

  16. An improved key agreement protocol based on chaos

    NASA Astrophysics Data System (ADS)

    Wang, Xingyuan; Zhao, Jianfeng

    2010-12-01

    Cryptography based on chaos theory has developed fast in the past few years, but most of the researches focus on secret key cryptography. There are few public key encryption algorithms and cryptographic protocols based on chaos, which are also of great importance for network security. We introduce an enhanced key agreement protocol based on Chebyshev chaotic map. Utilizing the semi-group property of Chebyshev polynomials, the proposed key exchange algorithm works like Diffie-Hellman algorithm. The improved protocol overcomes the drawbacks of several previously proposed chaotic key agreement protocols. Both analytical and experimental results show that it is effective and secure.

  17. Improved Protocols of Secure Quantum Communication Using W States

    NASA Astrophysics Data System (ADS)

    Shukla, Chitra; Banerjee, Anindita; Pathak, Anirban

    2013-06-01

    Recently, Hwang et al. (Eur. Phys. J. D 61:785, 2011) and Yuan et al. (Int. J. Theor. Phys. 50:2403, 2011) have proposed two efficient protocols of secure quantum communication using 3-qubit and 4-qubit symmetric W state respectively. These two dense coding based protocols are generalized and their efficiencies are considerably improved. Simple bounds on the qubit efficiency of deterministic secure quantum communication (DSQC) and quantum secure direct communication (QSDC) protocols are obtained and it is shown that dense coding is not essential for designing of maximally efficient DSQC and QSDC protocols. This fact is used to design maximally efficient protocols of DSQC and QSDC using 3-qubit and 4-qubit W states.

  18. Protocol for simultaneous isolation of three important banana allergens.

    PubMed

    Nikolic, Jasna; Mrkic, Ivan; Grozdanovic, Milica; Popovic, Milica; Petersen, Arnd; Jappe, Uta; Gavrovic-Jankulovic, Marija

    2014-07-01

    Banana fruit (Musa acuminata) has become an important food allergen source in recent years. So far, 5 IgE reactive banana proteins have been identified, and the major allergens are: Mus a 2 (a class I chitinase, 31kDa), Mus a 4 (thaumatin-like protein, 21kDa), and Mus a 5 (β-1,3-glucanase, 33kDa). Due to variations in allergen expression levels, diagnostic reagents for food allergy can be improved by using individual allergen components instead of banana allergen extracts. The purpose of this study was to optimize the purification protocol of the three major allergens present in banana fruit: Mus a 2, Mus a 4 and Mus a 5. By employing a three-step purification protocol (a combination of anion-exchange, cation-exchange and reversed-phase chromatography) three important banana allergens were obtained in sufficient yield and high purity. Characterization of the purified proteins was performed by both biochemical (2-D PAGE, mass fingerprint and N-terminal sequencing) and immunochemical (immunoblot) methods. IgE reactivity to the purified allergens was tested by employing sera of five allergic patients. The purified allergens displayed higher sensitivity in IgE detection than the routinely used extracts. The three purified allergens are good candidates for reagents in component-based diagnosis of banana allergy.

  19. An Accurate Link Correlation Estimator for Improving Wireless Protocol Performance

    PubMed Central

    Zhao, Zhiwei; Xu, Xianghua; Dong, Wei; Bu, Jiajun

    2015-01-01

    Wireless link correlation has shown significant impact on the performance of various sensor network protocols. Many works have been devoted to exploiting link correlation for protocol improvements. However, the effectiveness of these designs heavily relies on the accuracy of link correlation measurement. In this paper, we investigate state-of-the-art link correlation measurement and analyze the limitations of existing works. We then propose a novel lightweight and accurate link correlation estimation (LACE) approach based on the reasoning of link correlation formation. LACE combines both long-term and short-term link behaviors for link correlation estimation. We implement LACE as a stand-alone interface in TinyOS and incorporate it into both routing and flooding protocols. Simulation and testbed results show that LACE: (1) achieves more accurate and lightweight link correlation measurements than the state-of-the-art work; and (2) greatly improves the performance of protocols exploiting link correlation. PMID:25686314

  20. IMPROVED ISOLATION & ELECTROSPINNING OF ZEIN

    USDA-ARS?s Scientific Manuscript database

    The major historical use of zein, the predominant corn protein, has been in the fibers market. In order to evaluate new formulations of zein in the fibers market a simple means of isolating and generating fibers is necessary. We have evaluated the ability to electrospin zein from numerous solvents...

  1. Algorithms for improved performance in cryptographic protocols.

    SciTech Connect

    Schroeppel, Richard Crabtree; Beaver, Cheryl Lynn

    2003-11-01

    Public key cryptographic algorithms provide data authentication and non-repudiation for electronic transmissions. The mathematical nature of the algorithms, however, means they require a significant amount of computation, and encrypted messages and digital signatures possess high bandwidth. Accordingly, there are many environments (e.g. wireless, ad-hoc, remote sensing networks) where public-key requirements are prohibitive and cannot be used. The use of elliptic curves in public-key computations has provided a means by which computations and bandwidth can be somewhat reduced. We report here on the research conducted in an LDRD aimed to find even more efficient algorithms and to make public-key cryptography available to a wider range of computing environments. We improved upon several algorithms, including one for which a patent has been applied. Further we discovered some new problems and relations on which future cryptographic algorithms may be based.

  2. Cryptanalysis and improvement of an improved two factor authentication protocol for telecare medical information systems.

    PubMed

    Chaudhry, Shehzad Ashraf; Naqvi, Husnain; Shon, Taeshik; Sher, Muhammad; Farash, Mohammad Sabzinejad

    2015-06-01

    Telecare medical information systems (TMIS) provides rapid and convenient health care services remotely. Efficient authentication is a prerequisite to guarantee the security and privacy of patients in TMIS. Authentication is used to verify the legality of the patients and TMIS server during remote access. Very recently Islam et al. (J. Med. Syst. 38(10):135, 2014) proposed a two factor authentication protocol for TMIS using elliptic curve cryptography (ECC) to improve Xu et al.'s (J. Med. Syst. 38(1):9994, 2014) protocol. They claimed their improved protocol to be efficient and provides all security requirements. However our analysis reveals that Islam et al.'s protocol suffers from user impersonation and server impersonation attacks. Furthermore we proposed an enhanced protocol. The proposed protocol while delivering all the virtues of Islam et al.'s protocol resists all known attacks.

  3. Adult Rhabdomyosarcoma Survival Improved With Treatment on Multimodality Protocols

    SciTech Connect

    Gerber, Naamit Kurshan; Wexler, Leonard H.; Singer, Samuel; Alektiar, Kaled M.; Keohan, Mary Louise; Shi, Weiji; Zhang, Zhigang; Wolden, Suzanne

    2013-05-01

    Purpose: Rhabdomyosarcoma (RMS) is a pediatric sarcoma rarely occurring in adults. For unknown reasons, adults with RMS have worse outcomes than do children. Methods and Materials: We analyzed data from all patients who presented to Memorial Sloan-Kettering Cancer Center between 1990 and 2011 with RMS diagnosed at age 16 or older. One hundred forty-eight patients met the study criteria. Ten were excluded for lack of adequate data. Results: The median age was 28 years. The histologic diagnoses were as follows: embryonal 54%, alveolar 33%, pleomorphic 12%, and not otherwise specified 2%. The tumor site was unfavorable in 67% of patients. Thirty-three patients (24%) were at low risk, 61 (44%) at intermediate risk, and 44 (32%) at high risk. Forty-six percent were treated on or according to a prospective RMS protocol. The 5-year rate of overall survival (OS) was 45% for patients with nonmetastatic disease. The failure rates at 5 years for patients with nonmetastatic disease were 34% for local failure and 42% for distant failure. Among patients with nonmetastatic disease (n=94), significant factors associated with OS were histologic diagnosis, site, risk group, age, and protocol treatment. On multivariate analysis, risk group and protocol treatment were significant after adjustment for age. The 5-year OS was 54% for protocol patients versus 36% for nonprotocol patients. Conclusions: Survival in adult patients with nonmetastatic disease was significantly improved for those treated on RMS protocols, most of which are now open to adults.

  4. The improvement and simulation for LEACH clustering routing protocol

    NASA Astrophysics Data System (ADS)

    Ji, Ai-guo; Zhao, Jun-xiang

    2017-01-01

    An energy-balanced unequal multi-hop clustering routing protocol LEACH-EUMC is proposed in this paper. The candidate cluster head nodes are elected firstly, then they compete to be formal cluster head nodes by adding energy and distance factors, finally the date are transferred to sink through multi-hop. The results of simulation show that the improved algorithm is better than LEACH in network lifetime, energy consumption and the amount of data transmission.

  5. Temporal arteritis: improving patient evaluation with a new protocol.

    PubMed

    Alberts, Michael

    2013-01-01

    The process of diagnosing temporal arteritis remains controversial. Although temporal artery biopsy has long been the standard tool of evaluation, its poor sensitivity has prompted investigation of other methods to aid in diagnosis. Improved clinical evaluation and various imaging techniques have been suggested as ways to establish the diagnosis through noninvasive means and to improve biopsy yield. To retrospectively report and evaluate the process and experience of the Kaiser Permanente Northwest Region in implementing a new protocol that includes an enhanced clinical evaluation as well as the incorporation of color duplex ultrasonography in addition to biopsy when appropriate for temporal arteritis evaluation. A 38% reduction in the number of temporal artery biopsies performed was achieved through the new protocol, which was created by a multidisciplinary process, including stakeholders from all departments involved. The percentage of abnormal biopsy results rose from 8.5% at baseline to 24%. No cases of the disease were missed after careful evaluation of clinical and medical-legal records. Adding specialist clinical evaluation and color duplex ultrasonography to the standard diagnostic workup for temporal arteritis creates a rapid, noninvasive, resource-sensible means to diagnose giant cell arteritis, to improve temporal artery biopsy yield, and to decrease the total number of biopsies done. The diagnosis can be made in some cases by clinical evaluation and color duplex ultrasonography alone, thereby saving the patient an unnecessary surgical procedure. Protocols such as this can be implemented by multidisciplinary cooperation in a patient-centered, integrated system.

  6. Temporal Arteritis: Improving Patient Evaluation with a New Protocol

    PubMed Central

    Alberts, Michael

    2013-01-01

    Context: The process of diagnosing temporal arteritis remains controversial. Although temporal artery biopsy has long been the standard tool of evaluation, its poor sensitivity has prompted investigation of other methods to aid in diagnosis. Improved clinical evaluation and various imaging techniques have been suggested as ways to establish the diagnosis through noninvasive means and to improve biopsy yield. Objective: To retrospectively report and evaluate the process and experience of the Kaiser Permanente Northwest Region in implementing a new protocol that includes an enhanced clinical evaluation as well as the incorporation of color duplex ultrasonography in addition to biopsy when appropriate for temporal arteritis evaluation. Results: A 38% reduction in the number of temporal artery biopsies performed was achieved through the new protocol, which was created by a multidisciplinary process, including stakeholders from all departments involved. The percentage of abnormal biopsy results rose from 8.5% at baseline to 24%. No cases of the disease were missed after careful evaluation of clinical and medical-legal records. Conclusions: Adding specialist clinical evaluation and color duplex ultrasonography to the standard diagnostic workup for temporal arteritis creates a rapid, noninvasive, resource-sensible means to diagnose giant cell arteritis, to improve temporal artery biopsy yield, and to decrease the total number of biopsies done. The diagnosis can be made in some cases by clinical evaluation and color duplex ultrasonography alone, thereby saving the patient an unnecessary surgical procedure. Protocols such as this can be implemented by multidisciplinary cooperation in a patient-centered, integrated system. PMID:23596371

  7. The evolution of improved and simplified superovulation protocols in cattle.

    PubMed

    Mapletoft, Reuben J; Bó, Gabriel A

    2011-01-01

    Superovulation protocols have improved greatly since the early days of bovine embryo transfer when purified gonadotrophins were not available, follicular wave dynamics were unknown physiological phenomena and prostaglandins were not available. Although superstimulatory protocols in cattle are normally initiated mid-cycle, elective control of follicular wave emergence and ovulation have had a great impact on the application of on-farm embryo transfer. However, the most common treatment for the synchronisation of follicular wave emergence involves the use of oestradiol, which cannot be used in many parts of the world. Therefore, the need for alternative treatments has driven recent research. An approach that has shown promise is to initiate follicle-stimulating hormone (FSH) treatments at the time of the emergence of the new follicular wave following ovulation induced by gonadotrophin-releasing hormone. Alternatively, it has been shown that it may be possible to ignore follicular wave status and, by extending the treatment protocol, induce subordinate follicles to superovulate. Finally, the short half-life of pituitary FSH necessitates twice-daily treatments, which are time-consuming, stressful and subject to error. Recent treatment protocols have permitted superstimulation with a single FSH treatment or two treatments 48h apart, reducing the need for animal handling during gonadotrophin treatments.

  8. Protocol guided bleeding management improves cardiac surgery patient outcomes.

    PubMed

    Pearse, B L; Smith, I; Faulke, D; Wall, D; Fraser, J F; Ryan, E G; Drake, L; Rapchuk, I L; Tesar, P; Ziegenfuss, M; Fung, Y L

    2015-10-01

    Excessive bleeding is a risk associated with cardiac surgery. Treatment invariably requires transfusion of blood products; however, the transfusion itself may contribute to postoperative sequelae. Our objective was to analyse a quality initiative designed to provide an evidenced-based approach to bleeding management. A retrospective analysis compared blood product transfusion and patient outcomes 15 months before and after implementation of a bleeding management protocol. The protocol incorporated point-of-care coagulation testing (POCCT) with ROTEM and Multiplate to diagnose the cause of bleeding and monitor treatment. Use of the protocol led to decreases in the incidence of transfusion of PRBCs (47·3% vs. 32·4%; P < 0·0001), FFP (26·9% vs. 7·3%; P < 0·0001) and platelets (36·1% vs. 13·5%; P < 0·0001). During the intra-operative period, the percentage of patients receiving cryoprecipitate increased (2·7% vs. 5·1%; P = 0·002), as did the number of units transfused (248 vs. 692; P < 0·0001). The proportion of patients who received tranexamic acid increased (13·7% to 68·2%; P < 0·0001). There were reductions in re-exploration for bleeding (5·6% vs. 3·4; P = 0·01), superficial chest wound (3·3% vs. 1·4%; P = 0·002), leg wound infection (4·6% vs. 2·0%; P < 0·0001) and a 12% reduction in mean length of stay from operation to discharge (95%: 9-16%, P < 0·0001). Acquisition cost of blood products decreased by $1 029 118 in the 15-month period with the protocol. The implementation of a bleeding management protocol supported by POCCT in a cardiac surgery programme was associated with significant reductions in the transfusion of allogeneic blood products, improved outcomes and reduced cost. © 2015 International Society of Blood Transfusion.

  9. Comparison between isolation protocols highlights intrinsic variability of human umbilical cord mesenchymal cells.

    PubMed

    Paladino, Fernanda Vieira; Peixoto-Cruz, Joana Silveira; Santacruz-Perez, Carolina; Goldberg, Anna Carla

    2016-03-01

    Mesenchymal stem cells (MSCs) though multipotent exhibit limited lifespan in vitro, with progressive reduction in capacity for self-renewal leading to irreversible arrest of cell division, which limits their use for therapeutic purposes. Human umbilical cord wall MSCs are easy to process and proliferate rapidly in culture, but variability of individual samples and impact upon in vitro expansion and aging processes is unknown. We compared isolation protocols to determine which one yields the highest number of viable cells with the best proliferation capacity. Three different protocols were tested: two were enzymatic procedures and one explant method. Isolated cells were evaluated in terms of proliferation, differentiation capacity, and phenotype. All samples were processed using one or more protocols. After passage 2 adherent cells displayed standard phenotypic and differentiation characteristics of MSCs, but our results show that isolating cells directly from Wharton's jelly is more advantageous. Cells obtained from explants presented similar characteristics to those from enzymatic protocols, but always reached proliferation arrest earlier, irrespective of initial population doubling times. From the same sample, cells obtained with enzymatic protocol ii reached later passages while exhibiting shorter doubling times in culture than cells from other protocols, that is, took longer to reach senescence. More important, each individual MSC sample exhibited different population doubling rates and reached senescence at different passages, irrespective of protocol. Thus, even when in strict conformity with procedures and quality control, each cord sample shows a unique behavior, a finding that should be taken into account when planning for therapeutic approaches.

  10. Broad application of a simple and affordable protocol for isolating plant RNA.

    PubMed

    Couto, Daniel; Stransfeld, Lena; Arruabarrena, Ana; Zipfel, Cyril; Lozano-Durán, Rosa

    2015-04-16

    Standard molecular biological methods involve the analysis of gene expression in living organisms under diverse environmental and developmental conditions. One of the most direct approaches to quantify gene expression is the isolation of RNA. Most techniques used to quantify gene expression require the isolation of RNA, usually from a large number of samples. While most published protocols, including those for commercial reagents, are either labour intensive, use hazardous chemicals and/or are costly, a previously published protocol for RNA isolation in Arabidopsis thaliana yields high amounts of good quality RNA in a simple, safe and inexpensive manner. We have tested this protocol in tomato and wheat leaves, as well as in Arabidopsis leaves, and compared the resulting RNA to that obtained using a commercial phenol-based reagent. Our results demonstrate that this protocol is applicable to other plant species, including monocots, and offers yield and purity at least comparable to those provided by commercial phenol-based reagents. Here, we show that this previously published RNA isolation protocol can be easily extended to other plant species without further modification. Due to its simplicity and the use of inexpensive reagents, this protocol is accessible and affordable and can be easily implemented to work on different plant species in laboratories worldwide.

  11. Protocol for Isolating the Mouse Circle of Willis.

    PubMed

    Hur, Justine Claire; Blaise, Régis; Limon, Isabelle

    2016-10-22

    The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and hypothalamus that supplies blood to the brain and surrounding structures. It has been implicated in several cerebrovascular disorders, including cerebral amyloid angiopathy (CAA)-associated vasculopathies, intracranial atherosclerosis and intracranial aneurysms. Studies of the molecular mechanisms underlying these diseases for the identification of novel drug targets for their prevention require animal models. Some of these models may be transgenic, whereas others will involve isolation of the cerebro-vasculature, including the CoW.The method described here is suitable for CoW isolation in any mouse lineage and has considerable potential for screening (expression of genes, protein production, posttranslational protein modifications, secretome analysis, etc.) studies on the large vessels of the mouse cerebro-vasculature. It can also be used for ex vivo studies, by adapting the organ bath system developed for isolated mouse olfactory arteries.

  12. Implementation of a Hydrotherapy Protocol to Improve Postpartum Pain Management.

    PubMed

    Batten, Meghann; Stevenson, Eleanor; Zimmermann, Deb; Isaacs, Christine

    2017-03-01

    A growing number of women are seeking alternatives to traditional pharmacologic pain management during birth. While there has been an extensive array of nonpharmacologic options developed for labor, there are limited offerings in the postpartum period. The purpose of this quality improvement project was to implement a hydrotherapy protocol in the early postpartum period to improve pain management for women choosing a nonmedicated birth. The postpartum hydrotherapy protocol was initiated in a certified nurse-midwife (CNM) practice in an urban academic medical center. All women who met criteria were offered a 30-minute warm water immersion bath at one hour postpartum. Pain scores were assessed prior to the bath, at 15 minutes after onset, and again at the conclusion (30 minutes). Women who completed the bath were also asked to complete a brief survey on their experience with postpartum hydrotherapy. In women who used the bath (N = 45), there was a significant reduction in pain scores (P < .001) between the onset of the bath and scores at both 15 minutes and 30 minutes. There was no significant difference between pain scores at 15 minutes and 30 minutes (P = .97). Of those women who completed a survey (n = 43), 97.7% reported both that the bath reduced their pain and improved their birth experience. One hundred percent reported they would use it again in another birth. This project demonstrated successful implementation of a hydrotherapy protocol as an alternative or adjunct to medication for early postpartum pain management that significantly reduced pain and improved the birth experience for those who used it. It offers a nonpharmacologic alternative where there have traditionally been limited options. © 2017 by the American College of Nurse-Midwives.

  13. Exosome-like vesicles in uterine aspirates: a comparison of ultracentrifugation-based isolation protocols.

    PubMed

    Campoy, Irene; Lanau, Lucia; Altadill, Tatiana; Sequeiros, Tamara; Cabrera, Silvia; Cubo-Abert, Montserrat; Pérez-Benavente, Assumpción; Garcia, Angel; Borrós, Salvador; Santamaria, Anna; Ponce, Jordi; Matias-Guiu, Xavier; Reventós, Jaume; Gil-Moreno, Antonio; Rigau, Marina; Colas, Eva

    2016-06-18

    Uterine aspirates are used in the diagnostic process of endometrial disorders, yet further applications could emerge if its complex milieu was simplified. Exosome-like vesicles isolated from uterine aspirates could become an attractive source of biomarkers, but there is a need to standardize isolation protocols. The objective of the study was to determine whether exosome-like vesicles exist in the fluid fraction of uterine aspirates and to compare protocols for their isolation, characterization, and analysis. We collected uterine aspirates from 39 pre-menopausal women suffering from benign gynecological diseases. The fluid fraction of 27 of those aspirates were pooled and split into equal volumes to evaluate three differential centrifugation-based procedures: (1) a standard protocol, (2) a filtration protocol, and (3) a sucrose cushion protocol. Characterization of isolated vesicles was assessed by electron microscopy, nanoparticle tracking analysis and immunoblot. Specifically for RNA material, we evaluate the effect of sonication and RNase A treatment at different steps of the protocol. We finally confirmed the efficiency of the selected methods in non-pooled samples. All protocols were useful to isolate exosome-like vesicles. However, the Standard procedure was the best performing protocol to isolate exosome-like vesicles from uterine aspirates: nanoparticle tracking analysis revealed a higher concentration of vesicles with a mode of 135 ± 5 nm, and immunoblot showed a higher expression of exosome-related markers (CD9, CD63, and CD81) thus verifying an enrichment in this type of vesicles. RNA contained in exosome-like vesicles was successfully extracted with no sonication treatment and exogenous nucleic acids digestion with RNaseA, allowing the analysis of the specific inner cargo by Real-Time qPCR. We confirmed the existence of exosome-like vesicles in the fluid fraction of uterine aspirates. They were successfully isolated by differential centrifugation

  14. Improvements to the NIST network time protocol servers

    NASA Astrophysics Data System (ADS)

    Levine, Judah

    2008-12-01

    The National Institute of Standards and Technology (NIST) operates 22 network time servers at various locations. These servers respond to requests for time in a number of different formats and provide time stamps that are directly traceable to the NIST atomic clock ensemble in Boulder. The link between the servers at locations outside of the NIST Boulder Laboratories and the atomic clock ensemble is provided by the Automated Computer Time Service (ACTS) system, which has a direct connection to the clock ensemble and which transmits time information over dial-up telephone lines with a two-way protocol to measure the transmission delay. I will discuss improvements to the ACTS servers and to the time servers themselves. These improvements have resulted in an improvement of almost an order of magnitude in the performance of the system.

  15. Quality: performance improvement, teamwork, information technology and protocols.

    PubMed

    Coleman, Nana E; Pon, Steven

    2013-04-01

    Using the Institute of Medicine framework that outlines the domains of quality, this article considers four key aspects of health care delivery which have the potential to significantly affect the quality of health care within the pediatric intensive care unit. The discussion covers: performance improvement and how existing methods for reporting, review, and analysis of medical error relate to patient care; team composition and workflow; and the impact of information technologies on clinical practice. Also considered is how protocol-driven and standardized practice affects both patients and the fiscal interests of the health care system.

  16. Improving security of the ping-pong protocol

    NASA Astrophysics Data System (ADS)

    Zawadzki, Piotr

    2013-01-01

    A security layer for the asymptotically secure ping-pong protocol is proposed and analyzed in the paper. The operation of the improvement exploits inevitable errors introduced by the eavesdropping in the control and message modes. Its role is similar to the privacy amplification algorithms known from the quantum key distribution schemes. Messages are processed in blocks which guarantees that an eavesdropper is faced with a computationally infeasible problem as long as the system parameters are within reasonable limits. The introduced additional information preprocessing does not require quantum memory registers and confidential communication is possible without prior key agreement or some shared secret.

  17. Isolation Improvement with Electromagnetic Band Gap Surfaces

    DTIC Science & Technology

    2012-01-01

    method ( FEM ) are shown to enable the design and opti- mization of the periodic unit cell geometry for particular frequency bands. Measured results show...conductive vias are “through holes” drilled and then plated through with nickel. As stated previously, the EBG ability to block elec- tromagnetic...electromagnetic iSolation improvement with eBg SurfaceS figure 7. Finite-element analysis ( FEM ) of a single EBG unit cell is conducted over the top and

  18. An improved protocol for DNA extraction from alkaline soil and sediment samples for constructing metagenomic libraries.

    PubMed

    Verma, Digvijay; Satyanarayana, T

    2011-09-01

    An improved single-step protocol has been developed for extracting pure community humic substance-free DNA from alkaline soils and sediments. The method is based on direct cell lysis in the presence of powdered activated charcoal and polyvinylpolypyrrolidone followed by precipitation with polyethyleneglycol and isopropanol. The strategy allows simultaneous isolation and purification of DNA while minimizing the loss of DNA with respect to other available protocols for metagenomic DNA extraction. Moreover, the purity levels are significant, which are difficult to attain with any of the methods reported in the literature for DNA extraction from soils. The DNA thus extracted was free from humic substances and, therefore, could be processed for restriction digestion, PCR amplification as well as for the construction of metagenomic libraries.

  19. An improved pyrite pretreatment protocol for kinetic and isotopic studies

    NASA Astrophysics Data System (ADS)

    Mirzoyan, Natella; Kamyshny, Alexey; Halevy, Itay

    2014-05-01

    An improved pyrite pretreatment protocol for kinetic and isotopic studies Natella Mirzoyan1, Alexey Kamyshny Jr.2, Itay Halevy1 1Earth and Planetary Sciences, Weizmann Institute of Science, Rehovot 76100, Israel 2Geological and Environmental Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, Israel Pyrite is one of the most abundant and widespread of the sulfide minerals with a central role in biogeochemical cycles of iron and sulfur. Due to its diverse roles in the natural and anthropogenic sulfur cycle, pyrite has been extensively studied in various experimental investigations of the kinetics of its dissolution and oxidation, the isotopic fractionations associated with these reactions, and the microbiological processes involved. Pretreatment of pyrite for removal of oxidation impurities to prevent experimental artifacts and inaccuracies is often practiced. While numerous pyrite-cleaning methods have been used in experiments, a common pyrite pretreatment method, often used to investigate pyrite chemistry by the isotopic fractionations associated with it, includes several rinses by HCl, acetone and deionized water. Elemental sulfur (S0) is a common product of incomplete pyrite oxidation. Removal of S0 is desirable to avoid experimental biases associated with its participation in pyrite transformations, but is more complicated than the removal of sulfate. Although rinsing with an organic solvent is in part aimed at removing S0, to the best of our knowledge, the extraction efficiency of S0 in existing protocols has not been assessed. We have developed and tested a new protocol for elemental sulfur removal from the surface of pyrite by ultrasonication with warm acetone. Our data demonstrate the presence of large fractions of S0 on untreated pyrite particle surfaces, of which only approximately 60% was removed by the commonly used pretreatment method. The new protocol described here was found to be more efficient at S0 removal than the commonly used method

  20. KeepEX, a simple dilution protocol for improving extracellular vesicle yields from urine.

    PubMed

    Puhka, M; Nordberg, M-E; Valkonen, S; Rannikko, A; Kallioniemi, O; Siljander, P; Af Hällström, T M

    2017-02-15

    Urinary extracellular vesicles (EVs) are a promising source of biomarkers, which can be obtained in a non-invasive manner. However, the yield of EVs from urine samples may be insufficient for various analyses due to the entrapment of EVs by the Tamm-Horsfall protein (THP) meshwork. Here, we developed a simple dilution protocol to increase the urinary EV yield by disrupting the interaction between THP filaments and EVs with the help of alkaline pH and lowered ionic concentration. The integrity of the EVs and THP was assessed by electron microscopy. The effect of the protocol on the EV yield was quantified against an undiluted control by western blotting of four EV markers, nanoparticle tracking analysis and measuring of the RNA/miRNA concentration of the EV samples. The average EV yield from the dilution protocol was 2-7 fold the yield from the undiluted control i.e. increased by 130-624% as measured by western blotting and NTA. The yield increased most from samples with a high THP to EV ratio. The morphology and size range of the EVs were unaltered by the protocol. However, RNA/miRNA yields were the same as from the undiluted control and THP filaments could still be detected in EV samples. The dilution protocol, that we named KeepEX, provides a simple and efficient way to prevent loss of EVs thus increasing their yield from urine. Since KeepEX does not require individual adjustment of sample pH nor extra centrifugation steps, it could be used on its own or in combination with other EV purification protocols to improve EV isolation particularly from small urine volumes. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Improved human mesenchymal stem cell isolation.

    PubMed

    Chan, Tzu-Min; Harn, Horng-Jyh; Lin, Hui-Ping; Chou, Pei-Wen; Chen, Julia Yi-Ru; Ho, Tsung-Jung; Chiou, Tzyy-Wen; Chuang, Hong-Meng; Chiu, Shao-Chih; Chen, Yen-Chung; Yen, Ssu-Yin; Huang, Mao-Hsuan; Liang, Bing-Chiang; Lin, Shinn-Zong

    2014-01-01

    Human mesenchymal stem cells (hMSCs) are currently available for a range of applications and benefits and have become a good material for regenerative medicine, tissue engineering, and disease therapy. Before ex vivo expansion, isolation and characterization of primary hMSCs from peripheral tissues are key steps for obtaining adequate materials for clinical application. The proportion of peripheral stem cells is very low in surrounding tissues and organs; thus the recovery ratio will be a limiting factor. In this review, we summarized current common methods used to isolate peripheral stem cells, as well as the new insights revealed to improve the quantity of stem cells and their stemness. These strategies offer alternative ways to acquire hMSCs in a convenient and/or effective manner, which is important for clinical treatments. Improved isolation and mass amplification of the hMSCs while ensuring their stemness and quantity will be an important step for clinical use. Enlarged suitable hMSCs are more clinically applicable for therapeutic transplants and may help people live longer and better.

  2. Transfer learning improves supervised image segmentation across imaging protocols.

    PubMed

    van Opbroek, Annegreet; Ikram, M Arfan; Vernooij, Meike W; de Bruijne, Marleen

    2015-05-01

    The variation between images obtained with different scanners or different imaging protocols presents a major challenge in automatic segmentation of biomedical images. This variation especially hampers the application of otherwise successful supervised-learning techniques which, in order to perform well, often require a large amount of labeled training data that is exactly representative of the target data. We therefore propose to use transfer learning for image segmentation. Transfer-learning techniques can cope with differences in distributions between training and target data, and therefore may improve performance over supervised learning for segmentation across scanners and scan protocols. We present four transfer classifiers that can train a classification scheme with only a small amount of representative training data, in addition to a larger amount of other training data with slightly different characteristics. The performance of the four transfer classifiers was compared to that of standard supervised classification on two magnetic resonance imaging brain-segmentation tasks with multi-site data: white matter, gray matter, and cerebrospinal fluid segmentation; and white-matter-/MS-lesion segmentation. The experiments showed that when there is only a small amount of representative training data available, transfer learning can greatly outperform common supervised-learning approaches, minimizing classification errors by up to 60%.

  3. An improved arbitrated quantum signature protocol based on the key-controlled chained CNOT encryption

    NASA Astrophysics Data System (ADS)

    Zhang, Long; Sun, Hong-Wei; Zhang, Ke-Jia; Jia, Heng-Yue

    2017-03-01

    In this paper, a new quantum encryption based on the key-controlled chained CNOT operations, which is named KCCC encryption, is proposed. With the KCCC encryption, an improved arbitrated quantum signature (AQS) protocol is presented. Compared with the existing protocols, our protocol can effectively prevent forgery attacks and disavowal attacks. Moreover, only single state is required in the protocol. We hope it is helpful to further research in the design of AQS protocols in future.

  4. An efficient protocol for isolation of inhibitor-free nucleic acids even from recalcitrant plants.

    PubMed

    Rezadoost, Mohammad Hossein; Kordrostami, Mojtaba; Kumleh, Hassan Hassani

    2016-06-01

    For fast and easy isolation of inhibitor-free genomic DNA even from the toughest plant leaf samples, including those high in polyphenols and polysaccharides, a protocol has been developed. To prevent the solubility of polysaccharides in the DNA extract, high salt concentration (1.4 M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) was used for the removal of polyphenols as polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by proteinase K and removed by centrifugation from plant extracts during the isolation process resulting in pure DNA and RNA ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA and RNA isolated from leaves and roots of recalcitrant plants which was free from contamination and color. The average yields of total RNA from roots and shoot of Betula and Grape ranged from 285 to 364 ng/µl with A260/A280 between 1.9 and 2.08. The RNA isolated with this protocol was verified to be suitable for PCR, quantitative real-time PCR, semi-quantitative reverse transcription polymerase chain reaction, cDNA synthesis and expression analysis. This protocol shown here is reproducible and can be used for a broad spectrum of plant species which have polyphenols and polysaccharide compounds.

  5. Isolation of Listeria monocytogenes from Food and Water: Official and Experimental Protocols.

    PubMed

    Azizoglu, Reha O; Gorski, Lisa; Kathariou, Sophia

    2014-05-01

    Listeria monocytogenes is frequently encountered in foods but often at low concentrations and typically in the presence of other microbiota, including nonpathogenic Listeria spp. The potential of L. monocytogenes to cause severe human disease mandates sensitive, accurate, and rapid detection in foods. Isolation of L. monocytogenes from foods is critical, not only for routine surveillance, but also for epidemiologic investigations. Isolation of the pathogen from water (especially surface water used for irrigation) is similarly important, as produce has been implicated in listeriosis outbreaks and contaminated water can be involved in contamination of produce. This unit provides basic protocols for the isolation of L. monocytogenes from foods and water.

  6. An improved method for isolation of epithelial and stromal cells from the human endometrium

    PubMed Central

    MASUDA, Ayako; KATOH, Noriko; NAKABAYASHI, Kazuhiko; KATO, Kiyoko; SONODA, Kenzo; KITADE, Mari; TAKEDA, Satoru; HATA, Kenichiro; TOMIKAWA, Junko

    2016-01-01

    We aimed to improve the efficiency of isolating endometrial epithelial and stromal cells (EMECs and EMSCs) from the human endometrium. We revealed by immunohistochemical staining that the large tissue fragments remaining after collagenase treatment, which are usually discarded after the first filtration in the conventional protocol, consisted of glandular epithelial and stromal cells. Therefore, we established protease treatment and cell suspension conditions to dissociate single cells from the tissue fragments and isolated epithelial (EPCAM-positive) and stromal (CD13-positive) cells by fluorescence-activated cell sorting. Four independent experiments showed that, on average, 1.2 × 106 of EMECs and 2.8 × 106 EMSCs were isolated from one hysterectomy specimen. We confirmed that the isolated cells presented transcriptomic features highly similar to those of epithelial and stromal cells obtained by the conventional method. Our improved protocol facilitates future studies to better understand the molecular mechanisms underlying the dynamic changes of the endometrium during the menstrual cycle. PMID:26853786

  7. Cryptanalysis and Improvement of the Semi-quantum Secret Sharing Protocol

    NASA Astrophysics Data System (ADS)

    Gao, Xiang; Zhang, Shibin; Chang, Yan

    2017-08-01

    Recently, Xie et al. Int. J. Theor. Phys. 54, 3819-3824, (2015) proposed a Semi-quantum secret sharing protocol (SQSS). Yin et al. Int. J. Theor. Phys. 55: 4027-4035, (2016) pointed out that this protocol suffers from the intercept-resend attack. Yin et al. also proposed an improved protocol. However, we find out that Yin et al.'s paper has some problems, we analyze Yin et al.'s paper, then proposed the improved semi-quantum secret sharing protocol. Our protocol is more secure and efficient, most importantly, our protocol satisfies the condition of semi-quantum.

  8. An alternative cetyltrimethylammonium bromide-based protocol for RNA isolation from blackberry (Rubus L.).

    PubMed

    Chen, Q; Yu, H W; Wang, X R; Xie, X L; Yue, X Y; Tang, H R

    2012-06-29

    Isolation of high-quality RNA free of contaminants, such as polyphenols, proteins, plant secondary metabolites, and genomic DNA from plant tissues, is usually a challenging but crucial step for molecular analysis. We developed a novel protocol based on the cetyltrimethylammonium bromide method to isolate high-quality RNA from blackberry plant tissues, especially fruits. Most DNA was removed when acetic acid was utilized, before RNA precipitation. Thus, lithium chloride, a reagent widely used for RNA purification, was not needed. The isolation time was shortened to less than 3 h. The RNA was quite pure, with little DNA contamination. The quality of the RNA was assessed by spectrophotometric readings and electrophoresis on agarose gels. It was good enough for downstream enzymatic reactions, such as reverse transcription-PCR, cloning and real-time PCR assay. The method yielded an amount of total RNA comparable to previously described protocols.

  9. A Hybrid Quorum Protocol for Improved Availability, Capacity, Load and Reduced Overhead

    NASA Astrophysics Data System (ADS)

    Pandey, Parul; Tripathi, Maheshwari

    2016-12-01

    Data replication is playing a vital role in the design of distributed information systems. This paper presents a novel and efficient distributed algorithm for managing replicated data and for better performance and availability. This paper presents an extension to existing wheel protocol for improved performance. Wheel protocol imposes a logical wheel structure on the set of copies of an object and gives smallest read quorum. In addition to small read quorum size for read intensive applications, it is necessary to have good write availability as well. This paper proposes two hybrid wheel protocols, which superimpose logarithmic and ring protocols on top of the wheel protocol. It shows that, both protocols help in improving write availability, read capacity, load and message overhead and also compare their performances with wheel and other protocols. Hybrid protocols expand usage of wheel protocol to different type of applications.

  10. A microscale protocol for the isolation of transferrin directly from serum.

    PubMed

    Penezić, Ana; Miljuš, Goran; Milutinović, Bojana; Nedić, Olgica

    2017-08-01

    A microscale procedure for the isolation of transferrin directly from human serum (hTf) is described in this study. The protocol is based on three precipitation steps without application of chromatography. It lasts 90min with the initial sample volume of 250μL. The yield of the isolated hTf is 58%, which is considerable in biochemical terms. The purity of the isolated hTf is 97%, as assessed by three methods: electrophoresis followed by protein staining, immunoblotting and HPLC. Immunoblotting with antibodies against other major serum proteins indicated that isolated hTf does not contain albumin, immunoglobulin G or alpha-2-macroglobulin. Lectin dot-blot demonstrated that isolated hTf preserved its glycan moieties. Fluorescent emission spectroscopy of the isolated hTf has shown no changes in tertiary structure. Isolated hTf was approximately 26% saturated with iron ion, which is comparable to physiological value (although a degree of saturation decreases to some extent during isolation procedure). Finally, co-immunoprecipitation experiment confirmed that isolated hTf retained its ligand characteristics crucial for the ligand-receptor type of interaction with the hTf receptor. To conclude, the procedure described in this work, is time and cost-effective, allows multiple sample handling and provides high-purity hTf isolate with preserved structural and functional properties. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Isolation and purification of rabbit mesenchymal stem cells using an optimized protocol.

    PubMed

    Lin, Chunbo; Shen, Maorong; Chen, Weiping; Li, Xiaofeng; Luo, Daoming; Cai, Jinhong; Yang, Yuan

    2015-11-01

    Mesenchymal stem cells were first isolated and grown in vitro by Friedenstein over 40 yr ago; however, their isolation remains challenging as they lack unique markers for identification and are present in very small quantities in mesenchymal tissues and bone marrow. Using whole marrow samples, common methods for mesenchymal stem cell isolation are the adhesion method and density gradient fractionation. The whole marrow sample adhesion method still results in the nonspecific isolation of mononuclear cells, and activation and/or potential loss of target cells. Density gradient fractionation methods are complicated, and may result in contamination with toxic substances that affect cell viability. In the present study, we developed an optimized protocol for the isolation and purification of mesenchymal stem cells based on the principles of hypotonic lysis and natural sedimentation.

  12. Mechanical ventilation weaning protocol improves medical adherence and results.

    PubMed

    Borges, Luís Guilherme Alegretti; Savi, Augusto; Teixeira, Cassiano; de Oliveira, Roselaine Pinheiro; De Camillis, Marcio Luiz Ferreira; Wickert, Ricardo; Brodt, Sérgio Fernando Monteiro; Tonietto, Túlio Frederico; Cremonese, Ricardo; da Silva, Leonardo Silveira; Gehm, Fernanda; Oliveira, Eubrando Silvestre; Barth, Jose Herve Diel; Macari, Juçara Gasparetto; de Barros, Cíntia Dias; Vieira, Sílvia Regina Rios

    2017-10-01

    Implementation of a weaning protocol is related to better patient prognosis. However, new approaches may take several years to become the standard of care in daily practice. We conducted a prospective cohort study to investigate the effectiveness of a multifaceted strategy to implement a protocol to wean patients from mechanical ventilation (MV) and to evaluate the weaning success rate as well as practitioner adherence to the protocol. We investigated all consecutive MV-dependent subjects admitted to a medical-surgical intensive care unit (ICU) for >24h over 7years. The multifaceted strategy consisted of continuing education of attending physicians and ICU staff and regular feedback regarding patient outcomes. The study was conducted in three phases: protocol development, protocol and multifaceted strategy implementation, and protocol monitoring. Data regarding weaning outcomes and physician adherence to the weaning protocol were collected during all phases. We enrolled 2469 subjects over 7years, with 1,943 subjects (78.7%) experiencing weaning success. Physician adherence to the protocol increased during the years of protocol and multifaceted strategy implementation (from 38% to 86%, p<0.01) and decreased in the protocol monitoring phase (from 73.9% to 50.0%, p<0.01). However, during the study years, the weaning success of all subjects increased (from 73.1% to 85.4%, p<0.001). When the weaning protocol was evaluated step-by-step, we found high adherence for noninvasive ventilation use (95%) and weaning predictor measurement (91%) and lower adherence for control of fluid balance (57%) and daily interruption of sedation (24%). Weaning success was higher in patients who had undergone the weaning protocol compared to those who had undergone weaning based in clinical practice (85.6% vs. 67.7%, p<0.001). A multifaceted strategy consisting of continuing education and regular feedback can increase physician adherence to a weaning protocol for mechanical ventilation

  13. The effect of different isolation protocols on detection and molecular characterization of Campylobacter from poultry.

    PubMed

    Ugarte-Ruiz, M; Wassenaar, T M; Gómez-Barrero, S; Porrero, M C; Navarro-Gonzalez, N; Domínguez, L

    2013-11-01

    We determined whether different methods to isolate Campylobacter (including the ISO standard 10272:2006-1) affected the genotypes detectable from poultry, at three points during slaughter: caecal content, neck skin and meat. Carcasses from 28 independent flocks were thus sampled (subset A). In addition, ten neck skin samples from four flocks, ten caecal samples from ten different flocks and ten unrelated meat samples obtained from local supermarkets were collected (subset B). Campylobacter was isolated using eight different protocols: with and without enrichment using Bolton broth, Preston broth or Campyfood broth (CFB), followed by culture on either modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA) or Campyfood agar (CFA). All obtained isolates were genotyped for flaA-SVR, and over half of the isolates were also typed by MLST. The strain richness, as a measure of number of detected fla-genotypes, obtained from subset A neck skin and caecal samples was higher than that of meat samples. In half of the cases, within a flock, at least one identical fla-genotype was obtained at all three slaughter stages, suggestive of autologous contamination of carcasses. Enrichment reduced the observed richness of isolates, while CFA plates increased richness compared to mCCDA plates, irrespective of inclusion of an enrichment step. Because the isolation protocol used influences both the yield and the fla-genotype richness obtained from poultry, this variable should be taken into account when different studies are being compared.

  14. Used protocols for isolation and propagation of ovarian stem cells, different cells with different traits.

    PubMed

    Yazdekhasti, Hossein; Rajabi, Zahra; Parvari, Soraya; Abbasi, Mehdi

    2016-10-20

    Although existence of ovarian stem cells (OSCs) in mammalian postnatal ovary is still under controversy, however, it has been almost accepted that OSCs are contributing actively to folliculogenesis and neo-oogenesis. Recently, various methods with different efficacies have been employed for OSCs isolation from ovarian tissue, which these methods could be chosen depends on aim of isolation and accessible equipments and materials in lab. Although isolated OSCs from different methods have various traits and characterizations, which might become from their different nature and origin, however these stem cells are promising source for woman infertility treatment or source of energy for women with a history of repeat IVF failure in near future. This review has brought together and summarized currently used protocols for isolation and propagation of OSCs in vitro.

  15. Improving outpatient safety through effective electronic communication: a study protocol.

    PubMed

    Hysong, Sylvia J; Sawhney, Mona K; Wilson, Lindsey; Sittig, Dean F; Esquivel, Adol; Watford, Monica; Davis, Traber; Espadas, Donna; Singh, Hardeep

    2009-09-25

    Health information technology and electronic medical records (EMRs) are potentially powerful systems-based interventions to facilitate diagnosis and treatment because they ensure the delivery of key new findings and other health related information to the practitioner. However, effective communication involves more than just information transfer; despite a state of the art EMR system, communication breakdowns can still occur. [1-3] In this project, we will adapt a model developed by the Systems Engineering Initiative for Patient Safety (SEIPS) to understand and improve the relationship between work systems and processes of care involved with electronic communication in EMRs. We plan to study three communication activities in the Veterans Health Administration's (VA) EMR: electronic communication of abnormal imaging and laboratory test results via automated notifications (i.e., alerts); electronic referral requests; and provider-to-pharmacy communication via computerized provider order entry (CPOE). Our specific aim is to propose a protocol to evaluate the systems and processes affecting outcomes of electronic communication in the computerized patient record system (related to diagnostic test results, electronic referral requests, and CPOE prescriptions) using a human factors engineering approach, and hence guide the development of interventions for work system redesign. This research will consist of multiple qualitative methods of task analysis to identify potential sources of error related to diagnostic test result alerts, electronic referral requests, and CPOE; this will be followed by a series of focus groups to identify barriers, facilitators, and suggestions for improving the electronic communication system. Transcripts from all task analyses and focus groups will be analyzed using methods adapted from grounded theory and content analysis.

  16. Improving outpatient safety through effective electronic communication: a study protocol

    PubMed Central

    Hysong, Sylvia J; Sawhney, Mona K; Wilson, Lindsey; Sittig, Dean F; Esquivel, Adol; Watford, Monica; Davis, Traber; Espadas, Donna; Singh, Hardeep

    2009-01-01

    Background Health information technology and electronic medical records (EMRs) are potentially powerful systems-based interventions to facilitate diagnosis and treatment because they ensure the delivery of key new findings and other health related information to the practitioner. However, effective communication involves more than just information transfer; despite a state of the art EMR system, communication breakdowns can still occur. [1-3] In this project, we will adapt a model developed by the Systems Engineering Initiative for Patient Safety (SEIPS) to understand and improve the relationship between work systems and processes of care involved with electronic communication in EMRs. We plan to study three communication activities in the Veterans Health Administration's (VA) EMR: electronic communication of abnormal imaging and laboratory test results via automated notifications (i.e., alerts); electronic referral requests; and provider-to-pharmacy communication via computerized provider order entry (CPOE). Aim Our specific aim is to propose a protocol to evaluate the systems and processes affecting outcomes of electronic communication in the computerized patient record system (related to diagnostic test results, electronic referral requests, and CPOE prescriptions) using a human factors engineering approach, and hence guide the development of interventions for work system redesign. Design This research will consist of multiple qualitative methods of task analysis to identify potential sources of error related to diagnostic test result alerts, electronic referral requests, and CPOE; this will be followed by a series of focus groups to identify barriers, facilitators, and suggestions for improving the electronic communication system. Transcripts from all task analyses and focus groups will be analyzed using methods adapted from grounded theory and content analysis. PMID:19781075

  17. Improvement on "Quantum Key Agreement Protocol with Maximally Entangled States"

    NASA Astrophysics Data System (ADS)

    Chong, Song-Kong; Tsai, Chia-Wei; Hwang, Tzonelih

    2011-06-01

    Recently, Hsueh and Chen [in Proceedings of the 14th Information Security Conference, National Taiwan University of Science and Technology, Taipei, pp. 236-242, 2004] proposed a quantum key agreement (QKA) protocol with maximally entangled states. Their protocol allows two users to negotiate a secret key in such a way that no one can predetermine the shared key alone. This study points out two security flaws in their protocol: (1) a legitimate but malicious user can fully control the shared key alone; (2) an eavesdropper can obtain the shared key without being detected. A possible solution is presented to avoid these attacks and also Tsai et al.'s CNOT attack [in Proceedings of the 20th Cryptology and Information Security Conference, National Chiao Tung University, Hsinchu, pp. 210-213, 2010] on Hsueh and Chen protocol to obtain the shared key without being detected.

  18. Hypertension Improvement Project (HIP): study protocol and implementation challenges

    PubMed Central

    Dolor, Rowena J; Yancy, William S; Owen, William F; Matchar, David B; Samsa, Gregory P; Pollak, Kathryn I; Lin, Pao-Hwa; Ard, Jamy D; Prempeh, Maxwell; McGuire, Heather L; Batch, Bryan C; Fan, William; Svetkey, Laura P

    2009-01-01

    Background Hypertension affects 29% of the adult U.S. population and is a leading cause of heart disease, stroke, and kidney failure. Despite numerous effective treatments, only 53% of people with hypertension are at goal blood pressure. The chronic care model suggests that blood pressure control can be achieved by improving how patients and physicians address patient self-care. Methods and design This paper describes the protocol of a nested 2 × 2 randomized controlled trial to test the separate and combined effects on systolic blood pressure of a behavioral intervention for patients and a quality improvement-type intervention for physicians. Primary care practices were randomly assigned to the physician intervention or to the physician control condition. Physician randomization occurred at the clinic level. The physician intervention included training and performance monitoring. The training comprised 2 internet-based modules detailing both the JNC-7 hypertension guidelines and lifestyle modifications for hypertension. Performance data were collected for 18 months, and feedback was provided to physicians every 3 months. Patient participants in both intervention and control clinics were individually randomized to the patient intervention or to usual care. The patient intervention consisted of a 6-month behavioral intervention conducted by trained interventionists in 20 group sessions, followed by 12 monthly phone contacts by community health advisors. Follow-up measurements were performed at 6 and 18 months. The primary outcome was the mean change in systolic blood pressure at 6 months. Secondary outcomes were diastolic blood pressure and the proportion of patients with adequate blood pressure control at 6 and 18 months. Discussion Overall, 8 practices (4 per treatment group), 32 physicians (4 per practice; 16 per treatment group), and 574 patients (289 control and 285 intervention) were enrolled. Baseline characteristics of patients and providers and the

  19. DNA isolation protocol for the medicinal plant lemon balm (Melissa officinalis, Lamiaceae).

    PubMed

    Ghaffariyan, S; Mohammadi, S A; Aharizad, S

    2012-04-27

    Lemon balm (Melissa officinalis) is a medicinal plant that is widely used as a sedative or calmant, spasmolytic and antibacterial agent and sleep aid. This has led to a high demand for lemon balm products, resulting in the extinction of this species in some of its natural habitats. Molecular techniques have increasingly been used in plant diversity conservation and isolation of PCR amplifiable genomic DNA is an important pre-requisite. Lemon balm contains high levels of polyphenols and polysaccharides, which pose a major challenge for the isolation of high-quality DNA. We compared different genomic DNA extraction protocols, including traditional phenol-chloroform DNA extraction protocols and two commercial kits for DNA purification for their ability to produce good-quality DNA from fresh leaves of five lemon balm genotypes. Quality and quantity of the DNA samples were determined using 0.8% agarose gel electrophoresis and a spectrophotometer. The DNA purity was further confirmed by PCR amplification using barley retrotransposon LTR base primers. The spectral quality of DNA as measured by the A(260)/A(280) ratio ranged from 1.46 to 2.37. The Fermentase genomic DNA purification kit and the CTAB extraction protocol using PVP and ammonium acetate to overcome the high levels of polyphenols and polysaccharides yielded high-quality DNA with a mean A(260)/A(280) ratio of 1.87. The quantity of DNA and its PCR purity were similar with all the protocols, but considering the time and cost required for extraction of DNA from a large number of samples, the CTAB protocol using PVP and ammonium acetate is suitable for lemon balm.

  20. A protocol for the culture and isolation of murine synovial fibroblasts

    PubMed Central

    Zhao, Jinjun; Ouyang, Qingqing; Hu, Ziyou; Huang, Qin; Wu, Jing; Wang, Ran; Yang, Min

    2016-01-01

    The culture of synovial fibroblasts (SFs) is one of the most effective tools for investigating the pathology and physiology of synovial tissues and should prove useful for identifying the importance of SFs in disease as well as for the development of novel therapeutic approaches for several chronic joint diseases, such as rheumatoid arthritis. However, thus far, a detailed protocol for the primary culture and isolation of murine SFs has not been established. Therefore, the present study describes an easy and convenient method for isolating and culturing SFs from C57BL/6 mice. This protocol can be divided into 4 stages: Isolation of synovial tissues, isolation of SFs, seeding of SFs for growth in culture and purity analysis of SFs using the four cell markers, vimentin, cluster of differentiation 90.2 (CD90.2; Thy-1.2), intracellular adhesion molecule 1 (CD54) and vascular cell adhesion molecule 1 (CD106). This method is efficient and a purified population of SFs can be obtained 10 days after the initiation of culture. PMID:27446536

  1. Comparative evaluation of three lignin isolation protocols for various wood species.

    PubMed

    Guerra, Anderson; Filpponen, Ilari; Lucia, Lucian A; Argyropoulos, Dimitris S

    2006-12-27

    Milled wood lignin (MWL), cellulolytic enzyme lignin (CEL), and enzymatic mild acidolysis lignin (EMAL) were isolated from different wood species and characterized by various techniques. The EMAL protocol offered gravimetric lignin yields 2-5 times greater than those of the corresponding MWL and CEL. The purities of the EMALs were 3.75-10.6% higher than those of their corresponding CELs, depending upon the wood species from which they were isolated. Molecular weight analyses showed that the EMAL protocol isolates lignin fractions that are not accessed by the other procedures evaluated, while 31P NMR spectroscopy revealed that MWL is more condensed and bears more phenolic hydroxyl groups than EMAL and CEL. The yields and purities of EMAL, MWL, and CEL from hardwood were greater than those obtained for the examined softwoods. Structural details obtained by DFRC (derivatization followed by reductive cleavage)/31P NMR revealed different contents of condensed and uncondensed beta-O-aryl ether structures, dibenzodioxocins, and condensed and uncondensed phenolic hydroxyl and carboxylic acid groups within lignins isolated from different wood species.

  2. An improved ATAC-seq protocol reduces background and enables interrogation of frozen tissues.

    PubMed

    Corces, M Ryan; Trevino, Alexandro E; Hamilton, Emily G; Greenside, Peyton G; Sinnott-Armstrong, Nicholas A; Vesuna, Sam; Satpathy, Ansuman T; Rubin, Adam J; Montine, Kathleen S; Wu, Beijing; Kathiria, Arwa; Cho, Seung Woo; Mumbach, Maxwell R; Carter, Ava C; Kasowski, Maya; Orloff, Lisa A; Risca, Viviana I; Kundaje, Anshul; Khavari, Paul A; Montine, Thomas J; Greenleaf, William J; Chang, Howard Y

    2017-10-01

    We present Omni-ATAC, an improved ATAC-seq protocol for chromatin accessibility profiling that works across multiple applications with substantial improvement of signal-to-background ratio and information content. The Omni-ATAC protocol generates chromatin accessibility profiles from archival frozen tissue samples and 50-μm sections, revealing the activities of disease-associated DNA elements in distinct human brain structures. The Omni-ATAC protocol enables the interrogation of personal regulomes in tissue context and translational studies.

  3. An improved protocol for primary culture of cardiomyocyte from neonatal mice.

    PubMed

    Sreejit, P; Kumar, Suresh; Verma, Rama S

    2008-01-01

    The primary culture of neonatal mice cardiomyocyte model enables researchers to study and understand the morphological, biochemical, and electrophysiological characteristics of the heart, besides being a valuable tool for pharmacological and toxicological studies. Because cardiomyocytes do not proliferate after birth, primary myocardial culture is recalcitrant. The present study describes an improved method for rapid isolation of cardiomyocytes from neonatal mice, as well as the maintenance and propagation of such cultures for the long term. Immunocytochemical and gene expression data also confirmed the presence of several cardiac markers in the beating cells during the long-term culture condition used in this protocol. The whole culture process can be effectively shortened by reducing the enzyme digestion period and the cardiomyocyte enrichment step.

  4. An Improved RSA Based User Authentication and Session Key Agreement Protocol Usable in TMIS.

    PubMed

    Amin, Ruhul; Biswas, G P

    2015-08-01

    Recently, Giri et al.'s proposed a RSA cryptosystem based remote user authentication scheme for telecare medical information system and claimed that the protocol is secure against all the relevant security attacks. However, we have scrutinized the Giri et al.'s protocol and pointed out that the protocol is not secure against off-line password guessing attack, privileged insider attack and also suffers from anonymity problem. Moreover, the extension of password guessing attack leads to more security weaknesses. Therefore, this protocol needs improvement in terms of security before implementing in real-life application. To fix the mentioned security pitfalls, this paper proposes an improved scheme over Giri et al.'s scheme, which preserves user anonymity property. We have then simulated the proposed protocol using widely-accepted AVISPA tool which ensures that the protocol is SAFE under OFMC and CL-AtSe models, that means the same protocol is secure against active and passive attacks including replay and man-in-the-middle attacks. The informal cryptanalysis has been also presented, which confirmed that the proposed protocol provides well security protection on the relevant security attacks. The performance analysis section compares the proposed protocol with other existing protocols in terms of security and it has been observed that the protocol provides more security and achieves additional functionalities such as user anonymity and session key verification.

  5. A Xenogeneic-Free Protocol for Isolation and Expansion of Human Adipose Stem Cells for Clinical Uses

    PubMed Central

    Escobedo-Lucea, Carmen; Bellver, Carmen; Gandia, Carolina; Sanz-Garcia, Andres; Esteban, Francisco J.; Mirabet, Vicente; Forte, Giancarlo; Moreno, Isabel; Lezameta, Melissa; Ayuso-Sacido, Angel; Garcia-Verdugo, José M.

    2013-01-01

    Human adipose stem cells (hASCs) play a crucial role in the fields of regenerative medicine and tissue engineering for different reasons: the abundance of adipose tissue, their easy harvesting, the ability to multipotent differentiation and the fact that they do not trigger allogeneic blood response or secrete cytokines that act as immunosuppressants. The vast majority of protocols use animal origin reagents, with the underlying risk of transmitting infections by non-human pathogens. We have designed a protocol to isolate and maintain the properties of hASCs avoiding xenogeneic reagents. These changes not only preserve hASCs morphology, but also increase cell proliferation and maintain their stem cell marker profile. On the other hand, human serum albumin (HSA), Tryple® and human Serum (HS), do not affect hASCs multipotent differentiation ability. The amendments introduced do not trigger modifications in the transcriptional profile of hASCs, alterations in key biochemical pathways or malignization. Thus, we have proven that it is possible to isolate and maintain hASCs avoiding animal reagents and, at the same time, preserving crucial culture parameters during long term culture. Thereby we have revealed a novel and effective tool for the improvement of clinical, cell-based therapies. PMID:23874459

  6. Modified Test Protocol Improves Sensitivity of the Stereo Fly Test.

    PubMed

    De La Cruz, Angie; Morale, Sarah E; Jost, Reed M; Kelly, Krista R; Birch, Eileen E

    2016-01-01

    Stereoacuity measurement is a common element of pediatric ophthalmic examinations. Although the Stereo Fly Test is routinely used to establish the presence of coarse stereopsis (3000 arcsecs), it often yields a false negative "pass" due to learned responses and non-stereoscopic cues. We developed and evaluated a modified Stereo Fly Test protocol aimed at increasing sensitivity, thus reducing false negatives. The Stereo Fly Test was administered according to manufacturer instructions to 321 children aged 3-12 years. Children with a "pass" outcome (n = 147) were re-tested wearing glasses fitted with polarizers of matching orientation for both eyes to verify that they were responding to stereoscopic cues (modified protocol). The response to the standard Stereo Fly Test was considered a false negative (pass) if the child still pinched above the plate after disparity cues were eliminated. Randot(®) Preschool Stereoacuity and Butterfly Tests were used as gold standards. Sensitivity was 81% (95% CI: 0.75 - 0.86) for standard administration of the Stereo Fly Test (19% false negative "pass"). The modified protocol increased sensitivity to 90% (95% CI: 0.85 - 0.94). The modified two-step protocol is a simple and convenient way to administer the Stereo Fly Test with increased sensitivity in a clinical setting. © 2016 Board of regents of the University of Wisconsin System, American Orthoptic Journal, Volume 66, 2016, ISSN 0065-955X, E-ISSN 1553-4448.

  7. An improved protocol to study the plant cell wall proteome

    PubMed Central

    Printz, Bruno; Dos Santos Morais, Raphaël; Wienkoop, Stefanie; Sergeant, Kjell; Lutts, Stanley; Hausman, Jean-Francois; Renaut, Jenny

    2015-01-01

    Cell wall proteins were extracted from alfalfa stems according to a three-steps extraction procedure using sequentially CaCl2, EGTA, and LiCl-complemented buffers. The efficiency of this protocol for extracting cell wall proteins was compared with the two previously published methods optimized for alfalfa stem cell wall protein analysis. Following LC-MS/MS analysis the three-steps extraction procedure resulted in the identification of the highest number of cell wall proteins (242 NCBInr identifiers) and gave the lowest percentage of non-cell wall proteins (about 30%). However, the three protocols are rather complementary than substitutive since 43% of the identified proteins were specific to one protocol. This three-step protocol was therefore selected for a more detailed proteomic characterization using 2D-gel electrophoresis. With this technique, 75% of the identified proteins were shown to be fraction-specific and 72.7% were predicted as belonging to the cell wall compartment. Although, being less sensitive than LC-MS/MS approaches in detecting and identifying low-abundant proteins, gel-based approaches are valuable tools for the differentiation and relative quantification of protein isoforms and/or modified proteins. In particular isoforms, having variations in their amino-acid sequence and/or carrying different N-linked glycan chains were detected and characterized. This study highlights how the extracting protocols as well as the analytical techniques devoted to the study of the plant cell wall proteome are complementary and how they may be combined to elucidate the dynamism of the plant cell wall proteome in biological studies. Data are available via ProteomeXchange with identifier PXD001927. PMID:25914713

  8. DNA isolation protocols affect the detection limit of PCR approaches of bacteria in samples from the human gastrointestinal tract.

    PubMed

    Zoetendal, E G; Ben-Amor, K; Akkermans, A D; Abee, T; de Vos, W M

    2001-11-01

    A major concern in molecular ecological studies is the lysis efficiency of different bacteria in a complex ecosystem. We used a PCR-based 16S rDNA approach to determine the effect of two DNA isolation protocols (i.e. the bead beating and Triton-X100 method) on the detection limit of seven feces-associated bacterial species of different genera. Glycogen was used in these protocols to improve the precipitation of small concentrations of DNA in ethanol without affecting the sequential procedures. The PCR detection limit of 16S rDNA amplicons on agarose gel from the seven strains tested varied between 8.0 (+/- 1.3) x 10(4) and 4.3 (+/- 1.6) x 10(6) cells for the bead beating method, and between 8.0 (+/- 1.3) x 10(4) and 5.4 (+/- 0.7) x 10(8) cells for the Triton X-100 method. These large differences are most like due to the difference in cell lysis efficiency, since a competitive PCR experiment did not indicate any preference for gram negative, low G+C gram positive or high G+C gram positive bacteria. Denaturing gradient gel electrophoresis (DGGE) analysis was performed to investigate the effect of both DNA isolation protocols on the lysis efficiency of bacteria in fecal samples. A higher diversity in fecal samples was observed with the bead beating method than with the Triton-X100 method. Bands in the bead beating method-derived DGGE profiles corresponding to bands of cloned sequences of the Clostridium coccoides-Eubacterium rectale group and uncultured Fusobacterium prausnitzii were absent or had low intensity in the Triton X-100 method-derived profiles. The applicability of the bead beating method was further investigated by analyzing biopsy samples from the human colon which contain approximately 10(6) cells.

  9. Cryptanalysis and improvement of quantum secure communication network protocol with entangled photons for mobile communications

    NASA Astrophysics Data System (ADS)

    Gao, Gan

    2014-12-01

    Recently, a communication protocol called controlled bidirectional quantum secret direct communication for mobile networks was proposed by Chou et al (2014 Mobile Netw. Appl. 19 121). We study the security of the proposed communication protocol and find that it is not secure. The controller, Telecom Company, may eavesdrop secret messages from mobile devices without being detected. Finally, we give a possible improvement of the communication protocol.

  10. Security improvement on an anonymous key agreement protocol based on chaotic maps

    NASA Astrophysics Data System (ADS)

    Xue, Kaiping; Hong, Peilin

    2012-07-01

    In 2009, Tseng et al. proposed a password sharing and chaotic map based key agreement protocol (Tseng et al.'s protocol). They claimed that the protocol provided mutual authentication between a server and a user, and allowed the user to anonymously interact with the server to establish a shared session key. However, in 2011, Niu et al. have proved that Tseng et al.'s protocol cannot guarantee user anonymity and protocol security when there is an internal adversary who is a legitimate user. Also it cannot provide perfect forward secrecy. Then Niu et al. introduced a trust third party (TTP) into their protocol designing (Niu et al.'s protocol). But according to our research, Niu et al.'s protocol is found to have several unsatisfactory drawbacks. Based on reconsidering Tseng et al.'s protocol without introducing TTP, we give some improvements to meet the original security and performance requirements. Meanwhile our proposed protocol overcomes the security flaws of Tseng et al.'s protocol.

  11. An improved coding method of quantum key distribution protocols based on Fibonacci-valued OAM entangled states

    NASA Astrophysics Data System (ADS)

    Lai, Hong; Luo, Ming-Xing; Zhan, Cheng; Pieprzyk, Josef; Orgun, Mehmet A.

    2017-09-01

    We propose an improved coding method of quantum key distribution protocols based on a recently proposed (QKD) protocol using Fibonacci-valued OAM entangled states. To be exact, we define a new class of Fibonacci-matrix coding and Fibonacci-matrix representation and show how they can be used to extend and improve the original protocols. Compared with the original protocols, our protocol not only greatly improves the encoding efficiency but also has verifiability.

  12. Cryptanalysis and improvement of a quantum private set intersection protocol

    NASA Astrophysics Data System (ADS)

    Cheng, Xiaogang; Guo, Ren; Chen, Yonghong

    2017-02-01

    A recent Quantum Private Set Intersection (QPSI) scheme is crypt-analyzed. The original claimed communication overhead is shown to be not accurate. And the original security definition is passive and not fair. To ensure fairness, a passive third party is introduced. It is also shown that unconditional fairness of QPSI protocol is impossible. Since otherwise, it would violate a well-known impossible quantum cryptography result.

  13. Development of an efficient protocol of RNA isolation from recalcitrant tree tissues.

    PubMed

    Wang, Xuchu; Tian, Weimin; Li, Yinxin

    2008-01-01

    Isolation of RNA from recalcitrant tree tissues has been problematic due to large amounts of secondary metabolites and interfering compounds in their cells. We have developed an efficient RNA extraction method, which yielded high-quality RNA preparations from tissues of the lychee tree. The method reported here utilized EDTA, LSS, and CTAB to successfully inhibit RNase activities. It was found that a high ionic strength brought about by 2 M NaCl was necessary. In addition, secondary metabolites and other interfering compounds were effectively removed using sodium borate and PVPP under a deoxidized condition. The quality of purified RNA was tested by both RACE and Northern blotting analysis, ensuring that the RNA could be used for subsequent gene expression analysis. This method has been successfully applied to purify RNA from 15 other plant species. In conclusion, the protocol reported here is expected to have excellent applications for RNA isolation from recalcitrant plant tissues.

  14. Optimized Protocol for Isolation of Multipotent Mesenchymal Stromal Cells from Human Umbilical Cord.

    PubMed

    Romanov, Yu A; Balashova, E E; Volgina, N E; Kabaeva, N V; Dugina, T N; Sukhikh, G T

    2015-11-01

    Extraembryonic tissues, in particular, umbilical cord stroma are promising sources of multipotent mesenchymal stromal cells for regenerative medicine. In recent years, methods for isolation of mesenchymal stromal cells from different compartments of the umbilical cords based on enzymatic disaggregation of the tissue or on tissue explants have been proposed. Here we propose a protocol of isolation of multipotent mesenchymal stromal cells from the whole umbilical cord that combines the advantages of each approach and ensures sufficient cell yield for further experimental and clinical applications. A combination of short-term incubation of tissue fragments on cold collagenase solution followed by their culturing in the form of explants significantly increased the yield of cells with high proliferative activity, typical pluripotent mesenchymal stromal cell phenotype, and preserved differentiation capacity.

  15. Protocols for nuclei isolation and nuclear protein extraction from the resurrection plant Xerophyta viscosa for proteomic studies.

    PubMed

    Abdalla, Kamal Omer; Thomson, Jennifer Ann; Rafudeen, Muhammad Suhail

    2009-01-15

    The plant nucleus is an important subcellular organelle but the isolation of pure and enriched nuclei from plants and subsequent extraction of nuclear proteins for proteomic studies is challenging. Here, we present protocols for nuclei isolation and nuclear protein extraction from the resurrection plant, Xerophyta viscosa, and show optimization and modification of the most critical steps.

  16. Isolation of osteogenic progenitors from human amniotic fluid using a single step culture protocol

    PubMed Central

    Antonucci, Ivana; Iezzi, Irene; Morizio, Elisena; Mastrangelo, Filiberto; Pantalone, Andrea; Mattioli-Belmonte, Monica; Gigante, Antonio; Salini, Vincenzo; Calabrese, Giuseppe; Tetè, Stefano; Palka, Giandomenico; Stuppia, Liborio

    2009-01-01

    Background Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases. In the present study, we report a novel single step protocol for the osteoblastic differentiation of human amniotic fluid cells. Results The described protocol is able to provide osteoblastic cells producing nodules of calcium mineralization within 18 days from withdrawal of amniotic fluid samples. These cells display a complete expression of osteogenic markers (COL1, ONC, OPN, OCN, OPG, BSP, Runx2) within 30 days from withdrawal. In order to test the ability of these cells to proliferate on surfaces commonly used in oral osteointegrated implantology, we carried out cultures onto different test disks, namely smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium). Electron microscopy analysis evidenced the best cell growth on this latter surface. Conclusion The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology. PMID:19220883

  17. High Hydrostatic Pressure Pretreatment of Whey Protein Isolates Improves Their Digestibility and Antioxidant Capacity

    PubMed Central

    Iskandar, Michèle M.; Lands, Larry C.; Sabally, Kebba; Azadi, Behnam; Meehan, Brian; Mawji, Nadir; Skinner, Cameron D.; Kubow, Stan

    2015-01-01

    Whey proteins have well-established antioxidant and anti-inflammatory bioactivities. High hydrostatic pressure processing of whey protein isolates increases their in vitro digestibility resulting in enhanced antioxidant and anti-inflammatory effects. This study compared the effects of different digestion protocols on the digestibility of pressurized (pWPI) and native (nWPI) whey protein isolates and the antioxidant and anti-inflammatory properties of the hydrolysates. The pepsin-pancreatin digestion protocol was modified to better simulate human digestion by adjusting temperature and pH conditions, incubation times, enzymes utilized, enzyme-to-substrate ratio and ultrafiltration membrane molecular weight cut-off. pWPI showed a significantly greater proteolysis rate and rate of peptide appearance regardless of digestion protocol. Both digestion methods generated a greater relative abundance of eluting peptides and the appearance of new peptide peaks in association with pWPI digestion in comparison to nWPI hydrolysates. Hydrolysates of pWPI from both digestion conditions showed enhanced ferric-reducing antioxidant power relative to nWPI hydrolysates. Likewise, pWPI hydrolysates from both digestion protocols showed similar enhanced antioxidant and anti-inflammatory effects in a respiratory epithelial cell line as compared to nWPI hydrolysates. These findings indicate that regardless of considerable variations of in vitro digestion protocols, pressurization of WPI leads to more efficient digestion that improves its antioxidant and anti-inflammatory properties. PMID:28231198

  18. Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol

    NASA Astrophysics Data System (ADS)

    Livshts, Mikhail A.; Khomyakova, Elena; Evtushenko, Evgeniy G.; Lazarev, Vassili N.; Kulemin, Nikolay A.; Semina, Svetlana E.; Generozov, Edward V.; Govorun, Vadim M.

    2015-11-01

    Exosomes, small (40-100 nm) extracellular membranous vesicles, attract enormous research interest because they are carriers of disease markers and a prospective delivery system for therapeutic agents. Differential centrifugation, the prevalent method of exosome isolation, frequently produces dissimilar and improper results because of the faulty practice of using a common centrifugation protocol with different rotors. Moreover, as recommended by suppliers, adjusting the centrifugation duration according to rotor K-factors does not work for “fixed-angle” rotors. For both types of rotors - “swinging bucket” and “fixed-angle” - we express the theoretically expected proportion of pelleted vesicles of a given size and the “cut-off” size of completely sedimented vesicles as dependent on the centrifugation force and duration and the sedimentation path-lengths. The proper centrifugation conditions can be selected using relatively simple theoretical estimates of the “cut-off” sizes of vesicles. Experimental verification on exosomes isolated from HT29 cell culture supernatant confirmed the main theoretical statements. Measured by the nanoparticle tracking analysis (NTA) technique, the concentration and size distribution of the vesicles after centrifugation agree with those theoretically expected. To simplify this “cut-off”-size-based adjustment of centrifugation protocol for any rotor, we developed a web-calculator.

  19. Laboratory diagnosis of contagious ecthyma: comparison of different PCR protocols with virus isolation in cell culture.

    PubMed

    Kottaridi, Christine; Nomikou, Kiki; Lelli, Rossella; Markoulatos, Panayotis; Mangana, Olga

    2006-06-01

    A new polymerase chain reaction (PCR) assay for rapid diagnosis of contagious ecthyma was designed and applied to 21 clinical samples from Greece. This assay, which detects a highly conserved gene from the parapox genome, was evaluated for its sensitivity and specificity in order to be considered as a useful diagnostic tool. A comparative study with two published PCR protocols one using primers PPP1-PPP3, PPP1-PPP4 which targets putative virion envelope gene B2L and the other using VIR1-VIR2 primers which amplifies ORF virus interferon resistant (VIR) gene, as well as cell culture virus neutralization assay was carried out. All samples tested were amplified successfully with the PCR protocol established in the laboratory. The combination of primers PPP1-PPP3 and PPP1-PPP4 in a semi-nested PCR gave a positive result in 20 of 21 samples while primers VIR1-VIR2 failed to amplify successfully 7 of 21 samples. The diagnostic value of parapox viral DNA amplification was also compared with the results of virus isolation by cell culture and was positive in three samples that the virus isolation was obtained.

  20. Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol

    PubMed Central

    Livshts, Mikhail A.; Khomyakova, Elena; Evtushenko, Evgeniy G.; Lazarev, Vassili N.; Kulemin, Nikolay A.; Semina, Svetlana E.; Generozov, Edward V.; Govorun, Vadim M.

    2015-01-01

    Exosomes, small (40–100 nm) extracellular membranous vesicles, attract enormous research interest because they are carriers of disease markers and a prospective delivery system for therapeutic agents. Differential centrifugation, the prevalent method of exosome isolation, frequently produces dissimilar and improper results because of the faulty practice of using a common centrifugation protocol with different rotors. Moreover, as recommended by suppliers, adjusting the centrifugation duration according to rotor K-factors does not work for “fixed-angle” rotors. For both types of rotors – “swinging bucket” and “fixed-angle” – we express the theoretically expected proportion of pelleted vesicles of a given size and the “cut-off” size of completely sedimented vesicles as dependent on the centrifugation force and duration and the sedimentation path-lengths. The proper centrifugation conditions can be selected using relatively simple theoretical estimates of the “cut-off” sizes of vesicles. Experimental verification on exosomes isolated from HT29 cell culture supernatant confirmed the main theoretical statements. Measured by the nanoparticle tracking analysis (NTA) technique, the concentration and size distribution of the vesicles after centrifugation agree with those theoretically expected. To simplify this “cut-off”-size-based adjustment of centrifugation protocol for any rotor, we developed a web-calculator. PMID:26616523

  1. Inter-comparison of NIOSH and IMPROVE protocols for OC and EC determination: implications for inter-protocol data conversion

    NASA Astrophysics Data System (ADS)

    Wu, Cheng; Huang, X. H. Hilda; Ng, Wai Man; Griffith, Stephen M.; Zhen Yu, Jian

    2016-09-01

    Organic carbon (OC) and elemental carbon (EC) are operationally defined by analytical methods. As a result, OC and EC measurements are protocol dependent, leading to uncertainties in their quantification. In this study, more than 1300 Hong Kong samples were analyzed using both National Institute for Occupational Safety and Health (NIOSH) thermal optical transmittance (TOT) and Interagency Monitoring of Protected Visual Environment (IMPROVE) thermal optical reflectance (TOR) protocols to explore the cause of EC disagreement between the two protocols. EC discrepancy mainly (83 %) arises from a difference in peak inert mode temperature, which determines the allocation of OC4NSH, while the rest (17 %) is attributed to a difference in the optical method (transmittance vs. reflectance) applied for the charring correction. Evidence shows that the magnitude of the EC discrepancy is positively correlated with the intensity of the biomass burning signal, whereby biomass burning increases the fraction of OC4NSH and widens the disagreement in the inter-protocol EC determination. It is also found that the EC discrepancy is positively correlated with the abundance of metal oxide in the samples. Two approaches (M1 and M2) that translate NIOSH TOT OC and EC data into IMPROVE TOR OC and EC data are proposed. M1 uses direct relationship between ECNSH_TOT and ECIMP_TOR for reconstruction: M1 : ECIMP_TOR = a × ECNSH_TOT + b; while M2 deconstructs ECIMP_TOR into several terms based on analysis principles and applies regression only on the unknown terms: M2 : ECIMP_TOR = AECNSH + OC4NSH - (a × PCNSH_TOR + b), where AECNSH, apparent EC by the NIOSH protocol, is the carbon that evolves in the He-O2 analysis stage, OC4NSH is the carbon that evolves at the fourth temperature step of the pure helium analysis stage of NIOSH, and PCNSH_TOR is the pyrolyzed carbon as determined by the NIOSH protocol. The implementation of M1 to all urban site data (without considering seasonal specificity

  2. Improving the security of protocols of quantum key agreement solely using Bell states and Bell measurement

    NASA Astrophysics Data System (ADS)

    Zhu, Zhen-Chao; Hu, Ai-Qun; Fu, An-Min

    2015-11-01

    In a recent study, Shukla et al. (Quantum Inf Process 13:2391-2405, 2014) proposed two quantum key agreement protocols based on Bell state and Bell measurement, and they claimed that their two protocols were secure. However, in this study, we will show that the three-party protocol they proposed is not secure. Any participant in the protocol can directly obtain other two participants' secret keys. More seriously, two dishonest participants in the protocol can conclude to determine the shared key alone. Furthermore, we will show that there is another minor flaw in their two protocols; that is, eavesdroppers can flip any bit of the final key without introducing any error. In the end, some possible improvements are proposed to avoid these flaws.

  3. Improvement of "Novel Multiparty Quantum Key Agreement Protocol with GHZ States"

    NASA Astrophysics Data System (ADS)

    Gu, Jun; Hwang, Tzonelih

    2017-10-01

    Quantum key agreement (QKA) protocol is a method for negotiating a fair and secure key among mutually untrusted participants. Recently, Xu et al. (Quantum Inf. Process. 13:2587-2594, 2014) proposed a multi-party QKA protocol based on Greenberger-Horne-Zeilinger (GHZ) states. However, this study points out that Xu et al.'s protocol cannot provide the fairness property. That is, the last involved participant in the protocol can manipulate the final shared secret key without being detected by the other participants. Moreover, according to Yu et al.'s research (2015), Xu et al.'s protocol cannot avoid the public discussion attack too. To avoid these weaknesses, an improved QKA protocol is proposed.

  4. Improvement of "Novel Multiparty Quantum Key Agreement Protocol with GHZ States"

    NASA Astrophysics Data System (ADS)

    Gu, Jun; Hwang, Tzonelih

    2017-08-01

    Quantum key agreement (QKA) protocol is a method for negotiating a fair and secure key among mutually untrusted participants. Recently, Xu et al. (Quantum Inf. Process. 13:2587-2594, 2014) proposed a multi-party QKA protocol based on Greenberger-Horne-Zeilinger (GHZ) states. However, this study points out that Xu et al.'s protocol cannot provide the fairness property. That is, the last involved participant in the protocol can manipulate the final shared secret key without being detected by the other participants. Moreover, according to Yu et al.'s research (2015), Xu et al.'s protocol cannot avoid the public discussion attack too. To avoid these weaknesses, an improved QKA protocol is proposed.

  5. An optimized protocol for isolating primary epithelial cell chromatin for ChIP.

    PubMed

    Browne, James A; Harris, Ann; Leir, Shih-Hsing

    2014-01-01

    A critical part of generating robust chromatin immunoprecipitation (ChIP) data is the optimization of chromatin purification and size selection. This is particularly important when ChIP is combined with next-generation sequencing (ChIP-seq) to identify targets of DNA-binding proteins, genome-wide. Current protocols refined by the ENCODE consortium generally use a two-step cell lysis procedure that is applicable to a wide variety of cell types. However, the isolation and size selection of chromatin from primary human epithelial cells may often be particularly challenging. These cells tend to form sheets of formaldehyde cross-linked material in which cells are resistant to membrane lysis, nuclei are not released and subsequent sonication produces extensive high molecular weight contamination. Here we describe an optimized protocol to prepare high quality ChIP-grade chromatin from primary human bronchial epithelial cells. The ENCODE protocol was used as a starting point to which we added the following key steps to separate the sheets of formaldehyde-fixed cells prior to lysis. (1) Incubation of the formaldehyde-fixed adherent cells in Trypsin-EDTA (0.25% room temperature) for no longer than 5 min. (2) Equilibration of the fixed cells in detergent-free lysis buffers prior to each lysis step. (3) The addition of 0.5% Triton X-100 to the complete cell membrane lysis buffer. (4) Passing the cell suspension (in complete cell membrane lysis buffer) through a 25-gauge needle followed by continuous agitation on ice for 35 min. Each step of the modified protocol was documented by light microscopy using the Methyl Green-Pyronin dual dye, which stains cytoplasm red (Pyronin) and the nuclei grey-blue (Methyl green). This modified method is reproducibly effective at producing high quality sheared chromatin for ChIP and is equally applicable to other epithelial cell types.

  6. Improving security in the Fiber Distributed Data Interface (FDDI) protocol

    NASA Astrophysics Data System (ADS)

    Jones, Benjamin E.

    1992-09-01

    The arrival of high speed packet switched fiber optic LAN's has allowed local area design architectures to be used for larger metropolitan area network (MAN) implementations. The current LAN security mechanisms used in larger and faster fiber optic LAN's and MAN's are often inappropriate or unacceptable for use with emerging applications. The protocol of the Fiber Distributed Data Interface (FDDI) standard provides a natural means for message integrity and availability verification. However, privacy in FDDI is facilitated at higher layers through a generic LAN standard. This thesis proposes a modification to the FDDI protocol implemented at the medium access control (MAC) sublayer, which integrates a confidentiality mechanism for data transfer. The modification provides a simple comprehensive security package to meet the high performance needs of current and emerging applications. In the proposed modification, the inherent properties of the ring are exploited using a unique Central Key Translator to distribute initial session keys. A symmetric bit stream cipher based on modulo2 addition is used for encryption/decryption by the transmitting and receiving stations. Part of the plain text from transmitted message frames is used as feedback to generate new session keys.

  7. Protocol for Maximizing Energy Savings and Indoor Environmental Quality Improvements when Retrofiting Apartments

    SciTech Connect

    Noris, Federico; Delp, William W.; Vermeer, Kimberly; Adamkiewicz, Gary; Singer, Brett C.; Fisk, William J.

    2012-06-18

    The current focus on building energy retrofit provides an opportunity to simultaneously improve indoor environmental quality (IEQ). Toward this end, we developed a protocol for selecting packages of retrofits that both save energy and improve IEQ in apartments. The protocol specifies the methodology for selecting retrofits from a candidate list while addressing expected energy savings, IEQ impacts, and costs in an integrated manner. Interviews, inspections and measurements are specified to collect the needed input information. The protocol was applied to 17 apartments in three buildings in two different climates within California. Diagnostic measurements and surveys conducted before and after retrofit implementation indicate enhanced apartment performance.

  8. Protocol for maximizing energy savings and indoor environmental quality improvements when retrofitting apartments

    SciTech Connect

    Noris, Federico; Delp, William W.; Vermeer, Kimberly; Adamkiewicz, Gary; Singer, Brett C.; Fisk, William J.

    2013-06-01

    The current focus on building energy retrofit provides an opportunity to simultaneously improve indoor environmental quality (IEQ). Toward this end, we developed a protocol for selecting packages of retrofits that both save energy and improve IEQ in apartments. The protocol specifies the methodology for selecting retrofits from a candidate list while addressing expected energy savings, IEQ impacts, and costs in an integrated manner. Interviews, inspections and measurements are specified to collect the needed input information. The protocol was applied to 17 apartments in three buildings in two different climates within California. Diagnostic measurements and surveys conducted before and after retrofit implementation indicate enhanced apartment performance.

  9. An improved lookup protocol model for peer-to-peer networks

    NASA Astrophysics Data System (ADS)

    Fan, Wei; Ye, Dongfen

    2011-12-01

    With the development of the peer-to-peer (P2P) technology, file sharing is becoming the hottest, fastest growing application on the Internet. Although we can benefit from different protocols separately, our research shows that if there exists a proper model, most of the seemingly different protocols can be classified to a same framework. In this paper, we propose an improved Chord arithmetic based on the binary tree for P2P networks. We perform extensive simulations to study our proposed protocol. The results show that the improved Chord reduces the average lookup path length without increasing the joining and departing complexity.

  10. Cryptanalysis on the improved multiparty quantum secret sharing protocol based on the GHZ state

    NASA Astrophysics Data System (ADS)

    Chen, Xiu-Bo; Yang, Shuai; Su, Yuan; Yang, Yi-Xian

    2012-11-01

    Recently, Liu et al (2011 Phys. Scr. 84045015) pointed out that the multiparty quantum secret sharing (MQSS) protocol based on the GHZ state (Hwang et al 2011 Phys. Scr. 83045004) is insecure. They found that an inside participant can deduce half of the sender's secret information directly just by his piece of the secret. In order to resist this attack, an improvement was put forward. However, in this paper, we find that Liu et al's improved protocol is still insecure. We give details of three attack strategies to steal the secret information. It is shown that the eavesdropper can steal half or all of the secret information. Furthermore, a simple and ingenious MQSS protocol is proposed. We perform explicit cryptanalysis to prove that our improved protocol can resist the attacks from both the outside attackers and the inside participants, even the collusion attack.

  11. Optimisation of the protocol for detection of Aeromonas species in ready-to-eat salads, and its use to speciate isolates and establish their prevalence.

    PubMed

    Mattick, K L; Donovan, T J

    1998-12-01

    Aeromonas spp. are detected in more than 500 cases of gastrointestinal infection each year in England and Wales. This study aimed to identify their prevalence in ready-to-eat salads, which are a potential source of aeromonas infection. The protocol for isolation of mesophilic Aeromonas spp. from salads was optimised. Using the improved method, Aeromonas spp were isolated from 19 of 25 samples (25 g) of ready-to-eat salad products. Aeromonas organisms were counted, isolates were identified to species level, and the effect of pH on colonisation of salads was assessed. Aeromonas was present at high levels in six salads (> or = 100 cfu/g). The major species present in salads was Aeromonas caviae, but A.hydrophila and A.sobria, which have more pathogenic potential, were also isolated. It is hoped that this study will help to assess the risk to public health of aeromonas in salads.

  12. Intelligent QoS routing algorithm based on improved AODV protocol for Ad Hoc networks

    NASA Astrophysics Data System (ADS)

    Huibin, Liu; Jun, Zhang

    2016-04-01

    Mobile Ad Hoc Networks were playing an increasingly important part in disaster reliefs, military battlefields and scientific explorations. However, networks routing difficulties are more and more outstanding due to inherent structures. This paper proposed an improved cuckoo searching-based Ad hoc On-Demand Distance Vector Routing protocol (CSAODV). It elaborately designs the calculation methods of optimal routing algorithm used by protocol and transmission mechanism of communication-package. In calculation of optimal routing algorithm by CS Algorithm, by increasing QoS constraint, the found optimal routing algorithm can conform to the requirements of specified bandwidth and time delay, and a certain balance can be obtained among computation spending, bandwidth and time delay. Take advantage of NS2 simulation software to take performance test on protocol in three circumstances and validate the feasibility and validity of CSAODV protocol. In results, CSAODV routing protocol is more adapt to the change of network topological structure than AODV protocol, which improves package delivery fraction of protocol effectively, reduce the transmission time delay of network, reduce the extra burden to network brought by controlling information, and improve the routing efficiency of network.

  13. Improved CMOS field isolation using Germaniun/Boron implantation

    SciTech Connect

    Pfiester, J.R.; Alvis, J.R. )

    1988-08-01

    A novel germanium/boron implantation technique for improving the electrical field isolation for high-density CMOS circuits is demonstrated. Germanium implantation causes a reduction in dopant diffusion and segregation during field oxidation and is shown to increase the p-well field threshold voltage by as much as 40 percent with no significant degradation to junction or device performance. Selective germanium implantation with a blanket boron field implant can also improve the electrical field isolation behavior for CMOS circuits.

  14. A DNA extraction protocol for improved DNA yield from individual mosquitoes

    PubMed Central

    Nieman, Catelyn C.; Yamasaki, Youki; Collier, Travis C.; Lee, Yoosook

    2015-01-01

    Typical DNA extraction protocols from commercially available kits provide an adequate amount of DNA from a single individual mosquito sufficient for PCR-based assays. However, next-generation sequencing applications and high-throughput SNP genotyping assays exposed the limitation of DNA quantity one usually gets from a single individual mosquito. Whole genome amplification could alleviate the issue but it also creates bias in genome representation. While trying to find alternative DNA extraction protocols for improved DNA yield, we found that a combination of the tissue lysis protocol from Life Technologies and the DNA extraction protocol from Qiagen yielded a higher DNA amount than the protocol using the Qiagen or Life Technologies kit only. We have not rigorously tested all the possible combinations of extraction protocols; we also only tested this on mosquito samples. Therefore, our finding should be noted as a suggestion for improving people’s own DNA extraction protocols and not as an advertisement of a commercially available product. PMID:26937269

  15. New research and tools lead to improved earthquake alerting protocols

    USGS Publications Warehouse

    Wald, David J.

    2009-01-01

    What’s the best way to get alerted about the occurrence and potential impact of an earthquake? The answer to that question has changed dramatically of late, in part due to improvements in earthquake science, and in part by the implementation of new research in the delivery of earthquake information

  16. An efficient protocol for tissue sampling and DNA isolation from the stem bark of Leguminosae trees.

    PubMed

    Novaes, R M L; Rodrigues, J G; Lovato, M B

    2009-02-03

    Traditionally, molecular studies of plant species have used leaves as the source of DNA. However, sampling leaves from tall tree species can be quite difficult and expensive. We developed a sequence of procedures for using stem bark as a source of DNA from Leguminosae trees of the Atlantic Forest and the Cerrado. Leguminosae is an important species-rich family in these two highly diverse and endangered biomes. A modified CTAB protocol for DNA isolation is described, and details of the procedures for sampling and storage of the bark are given. The procedures were initially developed for three species, and then their applicability for 15 other species was evaluated. DNA of satisfactory quality was obtained from the bark of all species. The amounts of DNA obtained from leaves were slightly higher than from bark samples, while its purity was the same. Storing the bark frozen or by drying in silica gel yielded similar results. Polymerase chain reaction amplification worked for both plastid and nuclear genomes. This alternative for isolating DNA from bark samples of trees facilitates field work with these tree species.

  17. Improved protocol for chromatofocusing on the ProteomeLab PF2D.

    PubMed

    Barré, Olivier; Solioz, Marc

    2006-10-01

    Beckman-Coulter has recently introduced the ProteomeLab PF2D for 2-D liquid separation of protein samples. The system features separation in the first dimension by chromatofocusing, followed by RP chromatography in the second dimension, allowing the analysis of complex proteomics samples. When used by the standard protocol, reproducibility and column life times are limited, making the use of the instrument very costly. We here present an improved protocol for chromatofocusing, which enhances column life by at least fivefold.

  18. Isolated Teres Major Rupture: A case report with a suggested dedicated imaging protocol and review of the literature

    PubMed Central

    Fitzpatrick, Darren; Cagle, Paul; Flatow, Evan

    2016-01-01

    Isolated injuries to the teres major muscle occur in competitive sporting activities such as baseball pitching, hockey and tennis. We report a similar event of a physically fit man sustaining an isolated teres major rupture while waterskiing. Non-operative management was chosen, with pain resolution and no appreciable functional limitations at follow up. Because teres major muscle injury was suspected at the time of imaging, we present a dedicated imaging protocol to optimize assessment for teres major injury. PMID:27200170

  19. A standardized protocol to reduce pediatric spine surgery infection: a quality improvement initiative.

    PubMed

    Ryan, Sheila L; Sen, Anish; Staggers, Kristen; Luerssen, Thomas G; Jea, Andrew

    2014-09-01

    Quality improvement methods are being implemented in various areas of medicine. In an effort to reduce the complex (instrumented) spine infection rate in pediatric patients, a standardized protocol was developed and implemented at an institution with a high case volume of instrumented spine fusion procedures in the pediatric age group. Members of the Texas Children's Hospital Spine Study Group developed the protocol incrementally by using the current literature and prior institutional experience until consensus was obtained. The protocol was prospectively applied to all children undergoing complex spine surgery starting August 21, 2012. Acute infections were defined as positive wound cultures within 12 weeks of surgery, defined in alignment with current hospital infection control criteria. Procedures and infections were measured before and after protocol implementation. This protocol received full review and approval of the Baylor College of Medicine institutional review board. Nine spine surgeons performed 267 procedures between August 21, 2012, and September 30, 2013. The minimum follow-up was 12 weeks. The annual institutional infection rate prior to the protocol (2007-2011) ranged from 3.4% to 8.9%, with an average of 5.8%. After introducing the protocol, the infection rate decreased to 2.2% (6 infections of 267 cases) (p = 0.0362; absolute risk reduction 3.6%; relative risk 0.41 [95% CI 0.18-0.94]). Overall compliance with data form completion was 63.7%. In 4 of the 6 cases of infection, noncompliance with completion of the data collection form was documented; moreover, 2 of the 4 spine surgeons whose patients experienced infections had the lowest compliance rates in the study group. The standardized protocol for complex spine surgery significantly reduced surgical site infection at the authors' institution. The overall compliance with entry into the protocol was good. Identification of factors associated with post-spine surgery wound infection will allow

  20. Compliance with an empirical antimicrobial protocol improves the outcome of complicated intra-abdominal infections: a prospective observational study

    PubMed Central

    Guilbart, M.; Zogheib, E.; Ntouba, A.; Rebibo, L.; Régimbeau, J. M.; Mahjoub, Y.; Dupont, H.

    2016-01-01

    Background Despite improvements in medical and surgical care, mortality attributed to complicated intra-abdominal infections (cIAI) remains high. Appropriate initial antimicrobial therapy (ABT) is key to successful management. The main causes of non-compliance with empirical protocols have not been clearly described. Methods An empirical ABT protocol was designed according to guidelines, validated in the institution and widely disseminated. All patients with cIAI (2009–2011) were then prospectively studied to evaluate compliance with this protocol and its impact on outcome. Patients were classified into two groups according to whether or not they received ABT in compliance with the protocol. Results 310 patients were included: 223 (71.9%) with community-acquired and 87 (28.1%) with healthcare-associated cIAI [mean age 60(17–97) yr, mean SAPS II score 24(16)]. Empirical ABT complied with the protocol in 52.3% of patients. The appropriateness of empirical ABT to target the bacteria isolated was 80%. Independent factors associated with non-compliance with the protocol were the anaesthetist's age ≥36 yr [OR 2.1; 95%CI (1.3–3.4)] and the presence of risk factors for multidrug-resistant bacteria (MDRB) [OR 5.4; 95%CI (3.0–9.5)]. Non-compliance with the protocol was associated with higher mortality (14.9 vs 5.6%, P=0.011) and morbidity: relaparotomy (P=0.047), haemodynamic failure (P=0.001), postoperative pneumonia (P=0.025), longer duration of mechanical ventilation (P<0.001), longer ICU stay (P<0.001) and longer hospital stay (P=0.002). On multivariate logistic regression analysis, non-compliance with the ABT protocol was independently associated with mortality [OR 2.4; 95% CI (1.1–5.7), P=0.04]. Conclusions Non-compliance with empirical ABT guidelines in cIAI is associated with increased morbidity and mortality. Information campaigns should target older anaesthetists and risk factors for MDRB. PMID:27317705

  1. Ischemic postconditioning of the isolated human myocardium: Role of the applied protocol.

    PubMed

    Casós, Kelly; Pérez, María-Llanos; Blasco-Lucas, Arnau; Ferrer-Curriu, Gemma; Gracia-Baena, Juan Manuel; Sureda, Carlos; Permanyer, Eduard; Igual, Alberto; Galiñanes, Manuel

    2015-09-01

    Ischemic postconditioning (IPostC), has been proposed as a useful approach to reduce infarct size in all species, but its clinical utility remains unclear. To investigate the role played by the protocol used on the efficacy of IPostC in protecting the diseased human myocardium. Myocardial atrial samples from patients were subjected to a 90 min ischemia/120 min reoxygenation followed by different IPostC protocols to investigate the role of the time of ischemia (30, 60, 90 and 120 s) and the number of cycles (1, 2, 3 and 4) with 60 and 120 s of total ischemic time. Muscles were also subjected to ischemic preconditioning (IPreC). The release of lactate dehydrogenase (LDH) and the measurement of tetrazolium bromide (MTT) were determined. IPostC increased the LDH and decreased the MTT values from those of control, independently of the duration of the conditioning ischemia. LDH and MTT values also worsened by augmenting the number of IPostC cycles whereas they were significantly improved by IPreC. However, analysis of individual results indicated that in approximately 1/3 of the cases IPostC exhibited some degree of protection especially in the presence of increased ischemic injury. The present findings show that IPostC of the human myocardium may be influenced by the protocol used and also by the degree of the preceding ischemic injury. IPostC was beneficial in approximately 1/3 of the cases; however in the remaining cases it increased ischemic damage and, therefore, these results raise a word of caution on its broad clinical use.

  2. Isolating Spermathecae and Determining Mating Status of Drosophila suzukii: A Protocol for Tissue Dissection and Its Applications

    PubMed Central

    Avanesyan, Alina; Jaffe, Benjamin D.; Guédot, Christelle

    2017-01-01

    The spotted wing drosophila, Drosophila suzukii (Diptera: Drosophilidae), is an emerging invasive pest, which attacks a wide variety of fruits and berries. Although previous studies have focused on different aspects of D. suzukii reproductive biology, there are no protocols available for determining the mating status of D. suzukii females and drosophilids in general. In this study, a step-by-step protocol for tissue dissection, isolating spermathecae, and determining the mating status of females was developed specifically for D. suzukii. This protocol is an effective and relatively quick method for determining female mating status. It has important applications from exploring reproductive output of D. suzukii females to understanding the biology of D. suzukii winter morph, which presumably plays the main role in the overwintering of this invasive species. We demonstrated applicability of this protocol for both field collected flies and flies reared in the lab, including fly specimens stored on a long-term basis. PMID:28287438

  3. Kyushu-TCP: Improving Fairness of High-Speed Transport Protocols

    NASA Astrophysics Data System (ADS)

    Yoshimizu, Suguru; Koga, Hiroyuki; Kouyama, Katsushi; Shimamura, Masayoshi; Kumazoe, Kazumi; Tsuru, Masato

    With the emergence of bandwidth-greedy application services, high-speed transport protocols are expected to effectively and aggressively use large amounts of bandwidth in current broadband and multimedia networks. However, when high-speed transport protocols compete with other standard TCP flows, they can occupy most of the available bandwidth leading to disruption of service. To deploy high-speed transport protocols on the Internet, such unfair situations must be improved. In this paper, therefore, we propose a method to improve fairness, called Kyushu-TCP (KTCP), which introduces a non-aggressive period in the congestion avoidance phase to give other standard TCP flows more chances of increasing their transmission rates. This method improves fairness in terms of the throughput by estimating the stably available bandwidth-delay product and adjusting its transmission rate based on this estimation. We show the effectiveness of the proposed method through simulations.

  4. An improved and general streamlined phylogenetic protocol applied to the fatty acid desaturase family.

    PubMed

    Wilding, Matthew; Nachtschatt, Matthias; Speight, Robert; Scott, Colin

    2017-10-01

    Numerous tools to generate phylogenetic estimates are available, but there is no single protocol that will produce an accurate phylogenetic tree for any dataset. Here, we investigated some of those tools, paying particular attention to different alignment algorithms, in order to produce a phylogeny for the integral membrane fatty acid desaturase (FAD) family. Herein, we report a novel streamlined protocol which utilises peptide pattern recognition (PPR). This protocol can theoretically be applied universally to generate accurate multiple sequence alignments and improve downstream phylogenetic analyses. Applied to the desaturases, the protocol generated the first detailed phylogenetic estimates for the family since 2003, which suggested they may have evolved from three functionally distinct desaturases and further, that desaturases evolved first in cyanobacteria. In addition to the phylogenetic outputs, we mapped PPR sequence motifs onto an X-ray protein structure to provide insights into biochemical function and demonstrate the complementarity of PPR and phylogenetics. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Improved regeneration and transformation protocols for three strawberry cultivars

    PubMed Central

    Zakaria, Hossam; Hussein, Gihan M; Abdel-Hadi, Abdel-Hadi A; Abdallah, Naglaa A

    2014-01-01

    Strawberry (Fragaria × ananassa) is an economically important soft fruit crop with polyploid genome which makes the breeding of new cultivars difficult. Simple and efficient method for transformation and regeneration is required for cultivars improvement in strawberry. In the present study, adventitious shoot regeneration has been investigated in three cultivated strawberry plants, i.e., Festival, Sweet Charly and Florida via direct organogenesis using the in vitro juvenile leaves as explants. Explants were collected after sub-culturing on a propagation medium composed of MS supplemented with 0.5 mg/l BA; 0.1 mg/l GA3 and 0.1 mg/l IBA. To select the suitable organogenesis, the explants of the three cultivars were cultured on MS medium supplemented with different concentrations of TDZ (1, 2, 3, and 4 mg/l), then incubated at a temperature of 22 °C ± 2. Medium containing 2 mg/l TDZ revealed the best regeneration efficiency with the three cultivars (72% for Festival, and 73% for Sweet Charly and Florida). After 4 weeks, the produced shoots were cultured on MS medium with different concentrations of BA and Kin to enhance shoot elongation. Results showed that the medium containing 1.5 mg/l BA and 0.5 mg/l Kin revealed highest elongation efficiency (88% and 94%) for Festival and Sweet Charly, respectively. On the other hand, medium containing 1.5 mg/l BA and 0.1 mg/l Kin showed highest elongation efficiency (90%) in Florida. Elongated shoots were successfully rooted on MS medium containing 1.5 mg/l NAA. Furthermore, transformation of the two cultivars, Festival and Sweet Charly, has been established via Agrobacterium strain LBA44404 containing the plasmid pISV2678 with gus-intron and bar genes. Three days post co-cultivation, GUS activity was screening using the histochemical assay. The results showed 16% and 18% of the tested plant materials has changed into blue color for Festival and Sweet Charly, respectively. Out of 120 explants only 13 shoots were developed on

  6. Improved regeneration and transformation protocols for three strawberry cultivars.

    PubMed

    Zakaria, Hossam; Hussein, Gihan M; Abdel-Hadi, Abdel-Hadi A; Abdallah, Naglaa A

    2014-01-01

    Strawberry (Fragaria × ananassa) is an economically important soft fruit crop with polyploid genome which makes the breeding of new cultivars difficult. Simple and efficient method for transformation and regeneration is required for cultivars improvement in strawberry. In the present study, adventitious shoot regeneration has been investigated in three cultivated strawberry plants, i.e., Festival, Sweet Charly and Florida via direct organogenesis using the in vitro juvenile leaves as explants. Explants were collected after sub-culturing on a propagation medium composed of MS supplemented with 0.5 mg/l BA; 0.1 mg/l GA3 and 0.1 mg/l IBA. To select the suitable organogenesis, the explants of the three cultivars were cultured on MS medium supplemented with different concentrations of TDZ (1, 2, 3, and 4 mg/l), then incubated at a temperature of 22 °C ± 2. Medium containing 2 mg/l TDZ revealed the best regeneration efficiency with the three cultivars (72% for Festival, and 73% for Sweet Charly and Florida). After 4 weeks, the produced shoots were cultured on MS medium with different concentrations of BA and Kin to enhance shoot elongation. Results showed that the medium containing 1.5 mg/l BA and 0.5 mg/l Kin revealed highest elongation efficiency (88% and 94%) for Festival and Sweet Charly, respectively. On the other hand, medium containing 1.5 mg/l BA and 0.1 mg/l Kin showed highest elongation efficiency (90%) in Florida. Elongated shoots were successfully rooted on MS medium containing 1.5 mg/l NAA. Furthermore, transformation of the two cultivars, Festival and Sweet Charly, has been established via Agrobacterium strain LBA44404 containing the plasmid pISV2678 with gus-intron and bar genes. Three days post co-cultivation, GUS activity was screening using the histochemical assay. The results showed 16% and 18% of the tested plant materials has changed into blue color for Festival and Sweet Charly, respectively. Out of 120 explants only 13 shoots were developed on

  7. Protocol to cryopreserve and isolate nuclei from adipose tissue without dimethyl sulfoxide.

    PubMed

    Almeida, M M; Caires, L C J; Musso, C M; Campos, J M S; Maranduba, C M C; Macedo, G C; Mendonça, J P R F; Garcia, R M G

    2014-12-19

    Cryopreservation injuries involve nuclear DNA damage. A protocol for cryopreserving and isolating adipocyte nuclei is proposed. Adipose tissue samples were directly analyzed (NoCRYO-0h), or stored at -196°C for 7 days without 10% dimethyl sulfoxide (DMSO) (CRYO-WO-DMSO) or with DMSO (CRYO-W-DMSO). To determine the effect of DMSO on cryopreservation treatment, adipose tissue samples were stored at 4°C for 24 h with 10% DMSO (NoCRYO-W-DMSO-24h) and without (NoCRYO-WO-DMSO-24h). Samples were processed in isolation buffer, and nuclear integrity was measured by flow cytometry. The coefficient of variation, forward scatter, side scatter, and number of nuclei analyzed were evaluated. Pea (Pisum sativum) was used to measure the amount of DNA. All groups contained similar amounts of DNA to previously reported values and a satisfactory number of nuclei were analyzed. CRYO-W-DMSO presented a higher coefficient of variation (3.19 ± 0.09) compared to NoCRYO-0h (1.85 ± 0.09) and CRYO-WO-DMSO (2.02 ± 0.02). The coefficient of variation was increased in NoCRYO-W-DMSO-24h (3.80 ± 0.01) compared to NoCRYO-WO-DMSO-24h (2.46 ± 0.03). These results relate DMSO presence to DNA damage independently of the cryopreservation process. CRYO-W-DMSO showed increased side scatter (93.46 ± 5.03) compared to NoCRYO-0h (41.13 ± 3.19) and CRYO-WO-DMSO (48.01 ± 2.28), indicating that cryopreservation with DMSO caused chromatin condensation and/or nuclear fragmentation. CRYO-W-DMSO and CRYO-WO-DMSO presented lower forward scatter (186.33 ± 9.33 and 196.89 ± 26.86, respectively) compared to NoCRYO-0h (322.80 ± 3.36), indicating that cryopreservation reduced nuclei size. Thus, a simple method for cryopreservation and isolation of adipocyte nuclei causing less damage to DNA integrity was proposed.

  8. Improving International-Level Chess Players' Performance with an Acceptance-Based Protocol: Preliminary Findings

    ERIC Educational Resources Information Center

    Ruiz, Francisco J.; Luciano, Carmen

    2012-01-01

    This study compared an individual, 4-hr intervention based on acceptance and commitment therapy (ACT) versus a no-contact control condition in improving the performance of international-level chess players. Five participants received the brief ACT protocol, with each matched to another chess player with similar characteristics in the control…

  9. Substance Abuse Treatment And Family Therapy. A Treatment Improvement Protocol (TIP) Series 39

    ERIC Educational Resources Information Center

    Gilbert, Jonathan Max; Oliff, Helen; Sutton, David; Bartlett, Catalina; Henderson, Randi

    2004-01-01

    This Treatment Improvement Protocol (TIP) addresses substance abuse treatment in the context of family therapy. TIPs are best-practice guidelines for the treatment of substance use disorders that make the latest research in substance abuse treatment available to counselors and educators. The content was generated by a panel of experts in the…

  10. Substance Abuse Treatment: Group Therapy. Treatment Improvement Protocol (TIP) Series 41

    ERIC Educational Resources Information Center

    Gilbert, Jonathan Max; Hills, Susan; Rife, Mary Lou

    2005-01-01

    This Treatment Improvement Protocol (TIP) presents an overview of the role and efficacy of group therapy in substance abuse treatment. TIPs are best-practice guidelines for the treatment of substance use disorders that make the latest research in substance abuse treatment available to counselors and educators. The content was generated by a panel…

  11. Substance Abuse Treatment for Persons With Co-Occurring Disorders. Treatment Improvement Protocol (TIP) Series 42

    ERIC Educational Resources Information Center

    Sacks, Stanley; Ries, Richard K.

    2005-01-01

    This Treatment Improvement Protocol (TIP) provides guidelines for counselors and others working in the field of co-occurring substance use and mental disorders. TIPs are best-practice guidelines for the treatment of substance use disorders that make the latest research in substance abuse treatment available to counselors and educators. The content…

  12. Improving International-Level Chess Players' Performance with an Acceptance-Based Protocol: Preliminary Findings

    ERIC Educational Resources Information Center

    Ruiz, Francisco J.; Luciano, Carmen

    2012-01-01

    This study compared an individual, 4-hr intervention based on acceptance and commitment therapy (ACT) versus a no-contact control condition in improving the performance of international-level chess players. Five participants received the brief ACT protocol, with each matched to another chess player with similar characteristics in the control…

  13. Addressing Viral Hepatitis in People with Substance Use Disorders. Treatment Improvement Protocol (TIP) Series 53

    ERIC Educational Resources Information Center

    Substance Abuse and Mental Health Services Administration, 2011

    2011-01-01

    Treatment Improvement Protocols (TIPs) are developed by the Center for Substance Abuse Treatment (CSAT), part of the Substance Abuse and Mental Health Services Administration (SAMHSA) within the U.S. Department of Health and Human Services (HHS). Each TIP involves the development of topic-specific best-practice guidelines for the prevention and…

  14. Substance Abuse Treatment for Persons with Co-Occurring Disorders. Treatment Improvement Protocol (TIP) Series 42

    ERIC Educational Resources Information Center

    Substance Abuse and Mental Health Services Administration, 2005

    2005-01-01

    Treatment Improvement Protocols (TIPs), developed by the Center for Substance Abuse Treatment (CSAT), part of the Substance Abuse and Mental Health Services Administration (SAMHSA) within the U.S. Department of Health and Human Services (DHHS), are best-practice guidelines for the treatment of substance use disorders. CSAT draws on the experience…

  15. An improved protocol for the Pd-catalyzed alpha-arylation of aldehydes with aryl halides.

    PubMed

    Martín, Rubén; Buchwald, Stephen L

    2008-10-16

    An improved protocol for the Pd-catalyzed alpha-arylation of aldehydes with aryl halides has been developed. The new catalytic system allows for the coupling of an array of substrates including challenging electron-rich aryl bromides and less reactive aryl chlorides. The utility of this method has been demonstrated in a new total synthesis of (+/-)-sporochnol.

  16. Substance Abuse Treatment For Adults in the Criminal Justice System. Treatment Improvement Protocol (TIP) Series 44

    ERIC Educational Resources Information Center

    Bartlett, Catalina; Dinsmore, Janet; Gilbert, J. Max; Kornblum, Annette; Latham, Joyce; Oliff, Helen; Paisner, Susan; Sutton, David

    2005-01-01

    This Treatment Improvement Protocol (TIP) provides guidelines for counselors and criminal justice personnel who treat offenders with substance use disorders. TIPs are best-practice guidelines that make the latest research in substance abuse treatment available to counselors and educators. The content was generated by a panel of experts in the…

  17. Structured forensic interview protocols improve the quality and informativeness of investigative interviews with children: A review of research using the NICHD Investigative Interview Protocol

    PubMed Central

    Lamb, Michael E.; Orbach, Yael; Hershkowitz, Irit; Esplin, Phillip W.; Horowitz, Dvora

    2007-01-01

    Objective To show how the results of research on children’s memory, communicative skills, social knowledge, and social tendencies can be translated into guidelines that improve the quality of forensic interviews of children. Method We review studies designed to evaluate children’s capacities as witnesses, explain the development of the structured NICHD Investigative Interview Protocol, and discuss studies designed to assess whether use of the Protocol enhances the quality of investigative interviews. Results Controlled studies have repeatedly shown that the quality of interviewing reliably and dramatically improves when interviewers employ the NICHD Protocol. No other technique has been proven to be similarly effective. Conclusions Use of the structured NICHD Protocol improves the quality of information obtained from alleged victims by investigators, thereby increasing the likelihood that interventions will be appropriate. PMID:18023872

  18. A Post-operative Feeding Protocol to Improve Outcomes for Neonates With Critical Congenital Heart Disease.

    PubMed

    Newcombe, Jennifer; Fry-Bowers, Eileen

    2017-01-04

    Neonates with critical congenital heart disease (CCHD) are vulnerable to malnutrition during the post-operative period due to hypermetabolism and hypercatabolism. To improve nutritional outcomes during hospitalization, a nurse led post-operative enteral feeding protocol was implemented at a large U.S. children's hospital. During an eight-month implementation period, twenty-one neonates met protocol inclusion criteria. Days for neonates to achieve goal caloric feedings (120kcal/kg/day) were decreased. A one-way repeated measures analysis of variance showed serum albumin levels and serial anthropometric measurements improved significantly throughout hospitalization (p<0.005). Results from this quality improvement project show standardizing nutritional care for neonates with CCHD during the post-operative period is an effective way to improve nutritional outcomes and shorten length of hospital stay.

  19. Quality improvement initiative: Preventative Surgical Site Infection Protocol in Vascular Surgery.

    PubMed

    Parizh, David; Ascher, Enrico; Raza Rizvi, Syed Ali; Hingorani, Anil; Amaturo, Michael; Johnson, Eric

    2017-01-01

    Objective A quality improvement initiative was employed to decrease single institution surgical site infection rate in open lower extremity revascularization procedures. In an attempt to lower patient morbidity, we developed and implemented the Preventative Surgical Site Infection Protocol in Vascular Surgery. Surgical site infections lead to prolonged hospital stays, adjunctive procedure, and additive costs. We employed targeted interventions to address the common risk factors that predispose patients to post-operative complications. Methods Retrospective review was performed between 2012 and 2016 for all surgical site infections after revascularization procedures of the lower extremity. A quality improvement protocol was initiated in January 2015. Primary outcome was the assessment of surgical site infection rate reduction in the pre-protocol vs. post-protocol era. Secondary outcomes evaluated patient demographics, closure method, perioperative antibiotic coverage, and management outcomes. Results Implementation of the protocol decreased the surgical site infection rate from 6.4% to 1.6% p = 0.0137). Patient demographics and comorbidities were assessed and failed to demonstrate a statistically significant difference among the infection and no-infection groups. Wound closure with monocryl suture vs. staple proved to be associated with decreased surgical site infection rate ( p < 0.005). Conclusions Preventative measures, in the form of a standardized protocol, to decrease surgical site infections in the vascular surgery population are effective and necessary. Our data suggest that there may be benefit in the incorporation of MRSA and Gram-negative coverage as part of the Surgical Care Improvement Project perioperative guidelines.

  20. Change to a standardized airway clearance protocol for children with bronchiolitis leads to improved care.

    PubMed

    Jarvis, Kristen; Pirvu, Daniela; Barbee, Kim; Berg, Nancy; Meyer, Michelle; Gaulke, Lenny; Pate, Brian M; Roberts, Cristine

    2014-01-01

    Nurses at a pediatric hospital identified the liberal utilization of deep nasopharyngeal (NP) suction as the primary airway clearance modality in young children with bronchiolitis. This invasive practice lacked supporting evidence and a standardized approach. Nurses created an interdisciplinary team to develop a less invasive airway clearance protocol. Data from 2years, both pre and post protocol implementation, were analyzed. An 11% decrease in deep NP suctioning resulted in improved or unchanged balancing measures and perceptions of quality of care. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Abbreviated breast magnetic resonance protocol: Value of high-resolution temporal dynamic sequence to improve lesion characterization.

    PubMed

    Oldrini, Guillaume; Fedida, Benjamin; Poujol, Julie; Felblinger, Jacques; Trop, Isabelle; Henrot, Philippe; Darai, Emile; Thomassin-Naggara, Isabelle

    2017-10-01

    To evaluate the added value of ULTRAFAST-MR sequence to an abbreviated FAST protocol in comparison with FULL protocol to distinguish benign from malignant lesions in a population of women, regardless of breast MR imaging indication. From March 10th to September 22th, 2014, we retrospectively included a total of 70 consecutive patients with 106 histologically proven lesions (58 malignant and 48 benign) who underwent breast MR imaging for preoperative breast staging (n=38), high-risk screening (n=7), problem solving (n=18), and nipple discharge (n=4) with 12 time resolved imaging of contrast kinetics (TRICKS) acquisitions during contrast inflow interleaved in a regular high-resolution dynamic MRI protocol (FULL protocol). Two readers scored MR exams as either positive or negative and described significant lesions according to Bi-RADS lexicon with a TRICKS images (ULTRAFAST), an abbreviated protocol (FAST) and all images (FULL protocol). Sensitivity, specificity, positive and negative predictive values, and accuracy were calculated for each protocol and compared with McNemar's test. For all readers, the combined FAST-ULTRAFAST protocol significantly improved the reading with a specificity of 83.3% and 70.8% in comparison with FAST protocol or FULL protocol, respectively, without change in sensitivity. By adding ULTRAFAST protocol to FAST protocol, readers 1 and 2 were able to correctly change the diagnosis in 22.9% (11/48) and 10.4% (5/48) of benign lesions, without missing any malignancy, respectively. Both interpretation and image acquisition times for combined FAST-ULTRAFAST protocol and FAST protocol were shorter compared to FULL protocol (p<0.001). Compared to FULL protocol, adding ULTRAFAST to FAST protocol improves specificity, mainly in correctly reclassifying benign masses and reducing interpretation and acquisition time, without decreasing sensitivity. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Protocols for Improvement of Black Pepper (Piper nigrum L.) Utilizing Biotechnological Tools.

    PubMed

    Nirmal Babu, K; Divakaran, Minoo; Yamuna, G; Ravindran, P N; Peter, K V

    2016-01-01

    Black pepper, Piper nigrum L., the "King of spices" is the most widely used spice growing in the South-Western region of India. The humid tropical evergreen forest bordering the Malabar Coast (Western Ghats is one of the hot spot areas of plant bio-diversity on earth) is its center of origin and diversity. However, the crop faces constraints like rampant fungal and viral diseases, lack of disease free planting material, hence biotechnological tools can be utilized to address these problems and strides have been made successfully. The standardization of micropropagation, somatic embryogenesis, in vitro conservation, protoplast isolation, and genetic transformation protocols are described here. The protocols could be utilized to achieve similar goals in the related species of Piper too.

  3. Evaluation of four protocols for the detection and isolation of thermophilic Campylobacter from different matrices.

    PubMed

    Ugarte-Ruiz, M; Gómez-Barrero, S; Porrero, M C; Alvarez, J; García, M; Comerón, M C; Wassenaar, T M; Domínguez, L

    2012-07-01

      To identify the optimal method for detection of thermophilic Campylobacter at various stages in the food chain, three culture-dependent (direct plating, Bolton and Preston enrichment) and one molecular method (qPCR) were compared for three matrices: poultry faeces (n = 38), neck skin (n = 38) and packed fresh meat (n = 38).   Direct plating was compared to enrichment with either Bolton broth (ISO 10272:2006-1) or Preston broth, followed by culture on two selective agars: modified charcoal cefoperazone desoxycholate agar (mCCDA) and Campyfood agar (CFA). Direct plating on CFA provided the highest number of positive samples for faeces and neck skin samples. Enrichment of meat samples in Preston followed by plating on mCCDA gave significantly higher number of positives than the recommended ISO method. Real-time qPCR yielded the highest number of positive samples.   Direct plating on CFA is optimal for Campylobacter isolation from highly contaminated samples such as faeces or neck skin. When enrichment is required for less-contaminated samples such as poultry meat, Preston broth is the best choice. The maximum of detectable cells predicted by qPCR is a sensitive and powerful evaluation tool.   The recommended ISO protocol had the least sensitivity, and application of this method could result in underreporting. We detected a high prevalence of Campylobacter on packed meat to be distributed, which suggests this is still a significant risk for consumers. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  4. Efficient protocol for isolation and purification of different soyasaponins from soy hypocotyls.

    PubMed

    Zhao, Dayun; Yan, Mingxia; Huang, Yuai; Sun, Xiangjun

    2012-12-01

    Soyasaponins are naturally occurring triterpenoid glycosides associated with many biological activities. The aim of the present study was to develop an effective method for isolation and purification of differently glycosylated, acetylated, and 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP)-conjugated soyasaponins from soy hypocotyls. Both gel filtration using Sephadex LH-20 chromatography (Amersham Pharmacia Biotech AB; elution phase: methanol, flow rate: 3.0 mL/min, sample loading: 60 mg) and high-speed countercurrent chromatography (stationary phase: n-butanol-acetic acid (5.0%, v/v), mobile phase: water flow rate: 3.0 mL/min, sample loading: 100 mg) could effectively fractionate isoflavones and soyasaponins from the crude extract with yield of soyasaponin complexes 20.5 mg and 22.3 mg, respectively. After fractionation, the soyasaponin complexes could be purified further using preparative HPLC to separate individuals. A total of nine soyasaponins, triacetyl soyasaponin Ab (yield 1.55%, HPLC purity >98%), Aa (2.68%, >99%), Ab (18.53%, >98%), Ae (0.85%, >98%), Ba (0.63%, >91%), Af (1.12%, >85%), Bb (3.45%, >98%) and Be (0.59%, >76.8%) were obtained. DDMP-conjugated groups, αg (2.06%, >85%), βg (7.59%, >85%), and γg (0.29%, >85%) that were very labile even in mild conditions, were also collected. The method described here can be used as an effective protocol to separate different soyasaponins occurring in the original sample.

  5. A reliable protocol for the isolation of viable, chondrogenically differentiated human mesenchymal stem cells from high-density pellet cultures.

    PubMed

    Ullah, Mujib; Hamouda, Houda; Stich, Stefan; Sittinger, Michael; Ringe, Jochen

    2012-12-01

    Administration of chondrogenically differentiated mesenchymal stem cells (MSC) is discussed as a promising approach for the regenerative treatment of injured or diseased cartilage. The high-density pellet culture is the standard culture for chondrogenic differentiation, but cells in pellets secrete extracellular matrix (ECM) that they become entrapped in. Protocols for cell isolation from pellets often result in cell damage and dedifferentiation towards less differentiated MSC. Therefore, our aim was to develop a reliable protocol for the isolation of viable, chondrogenically differentiated MSC from high-density pellet cultures. Human bone marrow MSC were chondrogenically stimulated with transforming growth factor-β3, and the cartilaginous structure of the pellets was verified by alcian blue staining of cartilage proteoglycans, antibody staining of cartilage collagen type II, and quantitative real-time reverse-transcription polymerase chain reaction of the marker genes COL2A1 and SOX9. Trypsin and collagenases II and P were tested alone or in combination, and for different concentrations and times, to find a protocol for optimized pellet digestion. Whereas trypsin was not able to release viable cells, 90-min digestion with 300 U of collagenase II, 20 U of collagenase P, and 2 mM CaCl2 worked quite well and resulted in about 2.5×10(5) cells/pellet. The protocol was further optimized for the separation of released cells and ECM from each other. Cells were alcian blue and collagen type II positive and expressed COL2A1 and SOX9, verifying a chondrogenic character. However, they had different morphological shapes. The ECM was also uniformly alcian blue and collagen type II positive but showed different organizational and structural forms. To conclude, our protocol allows the reliable isolation of a defined number of viable, chondrogenically differentiated MSC from high-density pellet cultures. Such cells, as well as the ECM components, are of interest as

  6. A Reliable Protocol for the Isolation of Viable, Chondrogenically Differentiated Human Mesenchymal Stem Cells from High-Density Pellet Cultures

    PubMed Central

    Ullah, Mujib; Hamouda, Houda; Stich, Stefan; Sittinger, Michael

    2012-01-01

    Abstract Administration of chondrogenically differentiated mesenchymal stem cells (MSC) is discussed as a promising approach for the regenerative treatment of injured or diseased cartilage. The high-density pellet culture is the standard culture for chondrogenic differentiation, but cells in pellets secrete extracellular matrix (ECM) that they become entrapped in. Protocols for cell isolation from pellets often result in cell damage and dedifferentiation towards less differentiated MSC. Therefore, our aim was to develop a reliable protocol for the isolation of viable, chondrogenically differentiated MSC from high-density pellet cultures. Human bone marrow MSC were chondrogenically stimulated with transforming growth factor-β3, and the cartilaginous structure of the pellets was verified by alcian blue staining of cartilage proteoglycans, antibody staining of cartilage collagen type II, and quantitative real-time reverse-transcription polymerase chain reaction of the marker genes COL2A1 and SOX9. Trypsin and collagenases II and P were tested alone or in combination, and for different concentrations and times, to find a protocol for optimized pellet digestion. Whereas trypsin was not able to release viable cells, 90-min digestion with 300 U of collagenase II, 20 U of collagenase P, and 2 mM CaCl2 worked quite well and resulted in about 2.5×105 cells/pellet. The protocol was further optimized for the separation of released cells and ECM from each other. Cells were alcian blue and collagen type II positive and expressed COL2A1 and SOX9, verifying a chondrogenic character. However, they had different morphological shapes. The ECM was also uniformly alcian blue and collagen type II positive but showed different organizational and structural forms. To conclude, our protocol allows the reliable isolation of a defined number of viable, chondrogenically differentiated MSC from high-density pellet cultures. Such cells, as well as the ECM components, are of interest as

  7. Protocol: an improved and universal procedure for whole-mount immunolocalization in plants.

    PubMed

    Pasternak, Taras; Tietz, Olaf; Rapp, Katja; Begheldo, Maura; Nitschke, Roland; Ruperti, Benedetto; Palme, Klaus

    2015-01-01

    Rapid advances in microscopy have boosted research on cell biology. However sample preparation enabling excellent reproducible tissue preservation and cell labeling for in depth microscopic analysis of inner cell layers, tissues and organs still represents a major challenge for immunolocalization studies. Here we describe a protocol for whole-mount immunolocalization of proteins which is applicable to a wide range of plant species. The protocol is improved and robust for optimal sample fixation, tissue clearing and multi-protein staining procedures and can be used in combination with simultaneous detection of specific sequences of nucleic acids. In addition, cell wall and nucleus labelling can be implemented in the protocol, thereby allowing a detailed analysis of morphology and gene expression patterns with single-cell resolution. Besides enabling accurate, high resolution and reproducible protein detection in expression and localization studies, the procedure takes a single working day to complete without the need for robotic equipment.

  8. Improving shift report focus and consistency with the situation, background, assessment, recommendation protocol.

    PubMed

    Cornell, Paul; Gervis, Mary Townsend; Yates, Lauren; Vardaman, James M

    2013-01-01

    The Situation, Background, Assessment, Recommendation (SBAR) protocol was used to improve shift reports in 4 medical-surgical units. The SBAR protocol is increasingly advocated for use during shift reports, but data on the efficacy are limited. Nurses were trained on SBAR in 4 medical-surgical units in a tertiary care hospital. Nurse tasks, tools, and locations were recorded during observation audits. The average time for shift reports did not decrease using SBAR. Nurses spent significantly more time on tasks specific to report. There was significantly more dialogue and less writing with SBAR. The introduction of SBAR made reports more focused, with more time spent discussing the patient and less on transcribing information. The SBAR protocol provides a concise and prioritized structure that enables consistent, comprehensive, and patient-centric reports.

  9. Improved yield of canine islet isolation from deceased donors.

    PubMed

    Harrington, Stephen; Williams, S Janette; Otte, Vern; Barchman, Sally; Jones, Cheryl; Ramachandran, Karthik; Stehno-Bittel, Lisa

    2017-08-22

    Canine diabetes is a strikingly prevalent and growing disease, and yet the standard treatment of a twice-daily insulin injection is both cumbersome to pet owners and only moderately effective. Islet transplantation has been performed with repeated success in canine research models, but has unfortunately not been made available to companion animals. Standard protocols for islet isolation, developed primarily for human islet transplantation, include beating-heart organ donation, vascular perfusion of preservation solutions, specialized equipment. Unfortunately, these processes are prohibitively complex and expensive for veterinary use. The aim of the study was to develop a simplified approach for isolating canine islets that is compatible with the financial and logistical restrictions inherent to veterinary medicine for the purpose of translating islet transplantation to a clinical treatment for canine diabetes. Here, we describe simplified strategies for isolating quality islets from deceased canine donors without vascular preservation and with up to 90 min of cold ischemia time. An average of more than 1500 islet equivalents per kg of donor bodyweight was obtained with a purity of 70% (N = 6 animals). Islets were 95% viable and responsive to glucose stimulation for a week. We found that processing only the body and tail of the pancreas increased isolation efficiency without sacrificing islet total yield. Islet yield per gram of tissue increased from 773 to 1868 islet equivalents when the head of the pancreas was discarded (N = 3/group). In summary, this study resulted in the development of an efficient and readily accessible method for obtaining viable and functional canine islets from deceased donors. These strategies provide an ethical means for obtaining donor islets.

  10. Utilization of a standardized tracheostomy capping and decannulation protocol to improve patient safety.

    PubMed

    Pandian, Vinciya; Miller, Christina R; Schiavi, Adam J; Yarmus, Lonny; Contractor, Anisha; Haut, Elliott R; Feller-Kopman, David J; Mirski, Marek A; Morad, Athir H; Carey, John P; Hillel, Alexander T; Maragos, Carol S; Bhatti, Nasir I

    2014-08-01

    To develop and assess the feasibility of a new standardized protocol to guide tracheostomy decannulation. Descriptive review of quality improvement project. A quality improvement project was conducted in the inpatient setting of a tertiary urban academic hospital. Adult patients who had received a tracheostomy and for whom the indication for tracheostomy had resolved were included. A multidisciplinary task force reviewed input from clinicians caring for tracheostomy patients and developed a protocol for screening, capping, and decannulation. The primary outcome measured was successful decannulation. Fifty-seven patients were screened for a capping trial over a 12-month period; 54 were capped. Six patients were lost to follow-up. Fifty patients passed the capping trial, and all 50 were decannulated successfully. When decannulation was pursued in one patient who had twice failed the screening criteria and subsequent capping trials, the patient failed decannulation and ultimately required reintubation for the management of secretions. The screening tool had high sensitivity (90%) and positive predictive value (100%) for successful decannulation. Additionally, the number of reported patient safety concerns decreased from seven in the 6 months preceding implementation of the program to one report in the 6 months after implementation. The new tracheostomy capping and decannulation protocol assisted in predicting both successful and failed decannulation. Although several patients failed certain capping criteria initially, the protocol stipulated modifications of care that enabled successful decannulation. The screening tool had high sensitivity and promoted communication, standardization of practice, and patient safety. © 2014 The American Laryngological, Rhinological and Otological Society, Inc.

  11. An improved authenticated key agreement protocol for telecare medicine information system.

    PubMed

    Liu, Wenhao; Xie, Qi; Wang, Shengbao; Hu, Bin

    2016-01-01

    In telecare medicine information systems (TMIS), identity authentication of patients plays an important role and has been widely studied in the research field. Generally, it is realized by an authenticated key agreement protocol, and many such protocols were proposed in the literature. Recently, Zhang et al. pointed out that Islam et al.'s protocol suffers from the following security weaknesses: (1) Any legal but malicious patient can reveal other user's identity; (2) An attacker can launch off-line password guessing attack and the impersonation attack if the patient's identity is compromised. Zhang et al. also proposed an improved authenticated key agreement scheme with privacy protection for TMIS. However, in this paper, we point out that Zhang et al.'s scheme cannot resist off-line password guessing attack, and it fails to provide the revocation of lost/stolen smartcard. In order to overcome these weaknesses, we propose an improved protocol, the security and authentication of which can be proven using applied pi calculus based formal verification tool ProVerif.

  12. Improved patient survival using a modified resuscitation protocol for out-of-hospital cardiac arrest.

    PubMed

    Garza, Alex G; Gratton, Matthew C; Salomone, Joseph A; Lindholm, Daniel; McElroy, James; Archer, Rex

    2009-05-19

    Cardiac arrest continues to have poor survival in the United States. Recent studies have questioned current practice in resuscitation. Our emergency medical services system made significant changes to the adult cardiac arrest resuscitation protocol, including minimizing chest compression interruptions, increasing the ratio of compressions to ventilation, deemphasizing or delaying intubation, and advocating chest compressions before initial countershock. This retrospective observational cohort study reviewed all adult primary ventricular fibrillation and pulseless ventricular tachycardia cardiac arrests 36 months before and 12 months after the protocol change. Primary outcome was survival to discharge; secondary outcomes were return of spontaneous circulation and cerebral performance category. Survival of out-of-hospital arrest of presumed primary cardiac origin improved from 7.5% (82 of 1097) in the historical cohort to 13.9% (47 of 339) in the revised protocol cohort (odds ratio, 1.80; 95% confidence interval, 1.19 to 2.70). Similar increases in return of spontaneous circulation were achieved for the subset of witnessed cardiac arrest patients with initial rhythm of ventricular fibrillation from 37.8% (54 of 143) to 59.6% (34 of 57) (odds ratio, 2.44; 95% confidence interval, 1.24 to 4.80). Survival to hospital discharge also improved from an unadjusted survival rate of 22.4% (32 of 143) to 43.9% (25 of 57) (odds ratio, 2.71; 95% confidence interval, 1.34 to 1.59) with the protocol. Of the 25 survivors, 88% (n=22) had favorable cerebral performance categories on discharge. The changes to our prehospital protocol for adult cardiac arrest that optimized chest compressions and reduced disruptions increased the return of spontaneous circulation and survival to discharge in our patient population. These changes should be further evaluated for improving survival of out-of-hospital cardiac arrest patients.

  13. Improvement of pancreatic islet cell isolation for transplantation

    PubMed Central

    Noguchi, Hirofumi; Naziruddin, Bashoo; Onaca, Nicolas; Jackson, Andrew; Nobuyo, Hatanaka; Teru, Okitsu; Naoya, Kobayashi; Klintmalm, Göran; Levy, Marlon

    2007-01-01

    Pancreatic islet transplantation is a promising treatment for diabetes but still faces several challenges. Poor islet isolation efficiency and poor long-term insulin independence are currently two major issues, although donor shortage and the need for immunosuppressants also need to be addressed. We established the Kyoto islet isolation method (KIIM), which has enabled us to isolate and transplant islets even from non–heart-beating donors. KIIM involves 1) cooling the donor pancreas in situ, 2) preserving the ducts with modified Kyoto solution, 3) using a modified two-layer pancreas preservation method, and 4) adjusting the density of the density gradient centrifugation and using an iodixanol-based solution for purification. KIIM has enabled us to transplant 17 islet preparations out of 21 isolations (an 81% success rate). All transplanted islets functioned, and all transplanted patients had improved glycemic control without hypoglycemic unawareness. Recently, we used KIIM for islet isolation from a brain-dead donor at Baylor, which resulted in a very high islet yield (789,984 IE) with high viability (100% by fluorescein diacetate/propidium iodide staining and a stimulation index of 4.7). This preliminary evidence suggests that KIIM may also be promising for islet isolation from brain-dead donors. In addition, to assess engrafted islet mass, we developed a secretory unit of islet transplant objects (SUITO) index: fasting C-peptide (ng/dL) / [fasting blood glucose (mg/dL) – 63] × 1500. This simple index has enabled us to monitor the engrafted islet mass. This index should be useful when deciding whether to perform additional islet transplantations to maintain insulin independence. Poor islet isolation efficacy and poor long-term results could be resolved with ongoing research. PMID:17948109

  14. A Depression Screening Protocol for Patients With Acute Stroke: A Quality Improvement Project.

    PubMed

    McIntosh, Celia

    2017-02-01

    Depression after stroke is common and is associated with poor functional recovery, suicidal ideation, decreased quality of life, and increased mortality. Despite this knowledge, poststroke depression (PSD) is often underdetected and thus undertreated. PSD is clinically important for the caregiver, the family, and the stroke survivor. Inconsistencies in screening and treatment practices may further contribute to these negative outcomes. The purposes of this evidence-based clinical scholarship project were to (1) determine the efficacy of an evidence-based depression screening protocol in improving early detection and treatment of PSD and (2) identify if there were any relationships between the protocol interventions, depression scores, and diagnosis. A retrospective chart review was conducted in a convenience sample of 79 hospitalized patients with acute stroke. Depression was assessed using the Patient Health Questionnaire-9. Demographic data and medical and protocol variables were also collected. Descriptive statistics, chi-square test, and Pearson correlation test were used for data analysis. Of the 79 participants, 56% were men, 65% were White, 77% were admitted with ischemic stroke, and 48% were identified as being depressed (Patient Health Questionnaire-9 depression scale > 4). Individuals with a history of depression (χ = 17.09, p = .002) were also more likely to have higher levels of depression severity as compared with patients who did not have a history of depression. After the intervention, patients screening positive were more likely to receive an educational booklet on stroke and depression (χ = 30.0, p = .000) and be medically treated for PSD before discharge (χ = 5.57, p = .018). Nurses' documentation of screening results also improved (χ = 9.19, p = .002). Implementation of the Evidence Based Depression Screening and Treatment (EBDST) protocol improved early detection and treatment of PSD in the hospitalized patients with acute stroke before

  15. Protocol to Isolate a Large Amount of Functional Oligodendrocyte Precursor Cells from the Cerebral Cortex of Adult Mice and Humans

    PubMed Central

    Medina-Rodríguez, Eva María; Arenzana, Francisco Javier; Bribián, Ana; de Castro, Fernando

    2013-01-01

    During development, oligodendrocytes are generated from oligodendrocyte precursor cells (OPCs), a cell type that is a significant proportion of the total cells (3-8%) in the adult central nervous system (CNS) of both rodents and humans. Adult OPCs are responsible for the spontaneous remyelination that occurs in demyelinating diseases like Multiple Sclerosis (MS) and they constitute an interesting source of cells for regenerative therapy in such conditions. However, there is little data regarding the neurobiology of adult OPCs isolated from mice since an efficient method to isolate them has yet to be established. We have designed a protocol to obtain viable adult OPCs from the cerebral cortex of different mouse strains and we have compared its efficiency with other well-known methods. In addition, we show that this protocol is also useful to isolate functional OPCs from human brain biopsies. Using this method we can isolate primary cortical OPCs in sufficient quantities so as to be able to study their survival, maturation and function, and to facilitate an evaluation of their utility in myelin repair. PMID:24303061

  16. Improved Coomassie Blue Dye-Based Fast Staining Protocol for Proteins Separated by SDS-PAGE

    PubMed Central

    Májek, Pavel; Riedelová-Reicheltová, Zuzana; Pecánková, Klára; Dyr, Jan E.

    2013-01-01

    The time required to visualize proteins using Coomassie Blue dye has been significantly reduced with the introduction of fast staining protocols based on staining with a Coomassie Blue dye solution at boiling temperatures. However, fast stainings suffer from high gel backgrounds, reducing the signal-to-noise ratio and limiting the number of detectable spots in the case of 2D SDS-PAGE. The aim of this work was to eliminate the high gel background, and thus improve fast staining protocols based on Coomassie Blue dye. We show that merely replacing water with a 4 mM EDTA washing solution at boiling temperatures, results in a transparent gel background within 50 to 60 minutes of destaining. Moreover, when a combination of imidazole-zinc reverse staining and Coomassie Blue-based fast staining is used the sensitivity is improved significantly; nanogram amounts of proteins can be detected using 1D SDS-PAGE, and about 30% to 60% more spots can be detected with 2D SDS-PAGE in plasma, platelet, and rat brain tissue samples. This work represents an optimized fast staining protocol with improved sensitivity, requiring between 60 to 75 minutes to complete protein visualization. PMID:24278455

  17. Improved coomassie blue dye-based fast staining protocol for proteins separated by SDS-PAGE.

    PubMed

    Májek, Pavel; Riedelová-Reicheltová, Zuzana; Pecánková, Klára; Dyr, Jan E

    2013-01-01

    The time required to visualize proteins using Coomassie Blue dye has been significantly reduced with the introduction of fast staining protocols based on staining with a Coomassie Blue dye solution at boiling temperatures. However, fast stainings suffer from high gel backgrounds, reducing the signal-to-noise ratio and limiting the number of detectable spots in the case of 2D SDS-PAGE. The aim of this work was to eliminate the high gel background, and thus improve fast staining protocols based on Coomassie Blue dye. We show that merely replacing water with a 4 mM EDTA washing solution at boiling temperatures, results in a transparent gel background within 50 to 60 minutes of destaining. Moreover, when a combination of imidazole-zinc reverse staining and Coomassie Blue-based fast staining is used the sensitivity is improved significantly; nanogram amounts of proteins can be detected using 1D SDS-PAGE, and about 30% to 60% more spots can be detected with 2D SDS-PAGE in plasma, platelet, and rat brain tissue samples. This work represents an optimized fast staining protocol with improved sensitivity, requiring between 60 to 75 minutes to complete protein visualization.

  18. An Improved Protocol for Quantification of Freshwater Actinobacteria by Fluorescence In Situ Hybridization

    PubMed Central

    Sekar, Raju; Pernthaler, Annelie; Pernthaler, Jakob; Warnecke, Falk; Posch, Thomas; Amann, Rudolf

    2003-01-01

    We tested a previously described protocol for fluorescence in situ hybridization of marine bacterioplankton with horseradish peroxidase-labeled rRNA-targeted oligonucleotide probes and catalyzed reporter deposition (CARD-FISH) in plankton samples from different lakes. The fraction of Bacteria detected by CARD-FISH was significantly lower than after FISH with fluorescently monolabeled probes. In particular, the abundances of aquatic Actinobacteria were significantly underestimated. We thus developed a combined fixation and permeabilization protocol for CARD-FISH of freshwater samples. Enzymatic pretreatment of fixed cells was optimized for the controlled digestion of gram-positive cell walls without causing overall cell loss. Incubations with high concentrations of lysozyme (10 mg ml−1) followed by achromopeptidase (60 U ml−1) successfully permeabilized cell walls of Actinobacteria for subsequent CARD-FISH both in enrichment cultures and environmental samples. Between 72 and >99% (mean, 86%) of all Bacteria could be visualized with the improved assay in surface waters of four lakes. For freshwater samples, our method is thus superior to the CARD-FISH protocol for marine Bacteria (mean, 55%) and to FISH with directly fluorochrome labeled probes (mean, 67%). Actinobacterial abundances in the studied systems, as detected by the optimized protocol, ranged from 32 to >55% (mean, 45%). Our findings confirm that members of this lineage are among the numerically most important Bacteria of freshwater picoplankton. PMID:12732568

  19. Different exercise protocols improve metabolic syndrome markers, tissue triglycerides content and antioxidant status in rats

    PubMed Central

    2011-01-01

    Background An increase in the prevalence of obesity entails great expenditure for governments. Physical exercise is a powerful tool in the combat against obesity and obesity-associated diseases. This study sought to determine the effect of three different exercise protocols on metabolic syndrome and lipid peroxidation markers and the activity of antioxidant enzymes in adult Wistar rats (120 days old). Methods Animals were randomly divided into four groups: the control (C) group was kept sedentary throughout the study; the aerobic group (A) swam1 h per day, 5 days per week, at 80% lactate threshold intensity; the strength group (S) performed strength training with four series of 10 jumps, 5 days per week; and the Concurrent group (AS) was trained using the aerobic protocol three days per week and the strength protocol two days per week. Results Groups A and S exhibited a reduction in body weight compared to group C. All exercised animals showed a reduction in triglyceride concentrations in fatty tissues and the liver. Exercised animals also exhibited a reduction in lipid peroxidation markers (TBARS) and an increase in serum superoxide dismutase activity. Animals in group A had increased levels of liver catalase and superoxide dismutase activities. Conclusions We concluded that all physical activity protocols improved the antioxidant systems of the animals and decreased the storage of triglycerides in the investigated tissues. PMID:22182600

  20. Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells.

    PubMed

    Kegel, Victoria; Deharde, Daniela; Pfeiffer, Elisa; Zeilinger, Katrin; Seehofer, Daniel; Damm, Georg

    2016-03-30

    Beside parenchymal hepatocytes, the liver consists of non-parenchymal cells (NPC) namely Kupffer cells (KC), liver endothelial cells (LEC) and hepatic Stellate cells (HSC). Two-dimensional (2D) culture of primary human hepatocyte (PHH) is still considered as the "gold standard" for in vitro testing of drug metabolism and hepatotoxicity. It is well-known that the 2D monoculture of PHH suffers from dedifferentiation and loss of function. Recently it was shown that hepatic NPC play a central role in liver (patho-) physiology and the maintenance of PHH functions. Current research focuses on the reconstruction of in vivo tissue architecture by 3D- and co-culture models to overcome the limitations of 2D monocultures. Previously we published a method to isolate human liver cells and investigated the suitability of these cells for their use in cell cultures in Experimental Biology and Medicine(1). Based on the broad interest in this technique the aim of this article was to provide a more detailed protocol for the liver cell isolation process including a video, which will allow an easy reproduction of this technique. Human liver cells were isolated from human liver tissue samples of surgical interventions by a two-step EGTA/collagenase P perfusion technique. PHH were separated from the NPC by an initial centrifugation at 50 x g. Density gradient centrifugation steps were used for removal of dead cells. Individual liver cell populations were isolated from the enriched NPC fraction using specific cell properties and cell sorting procedures. Beside the PHH isolation we were able to separate KC, LEC and HSC for further cultivation. Taken together, the presented protocol allows the isolation of PHH and NPC in high quality and quantity from one donor tissue sample. The access to purified liver cell populations could allow the creation of in vivo like human liver models.

  1. A protocol for exosome isolation and characterization: evaluation of ultracentrifugation, density-gradient separation, and immunoaffinity capture methods.

    PubMed

    Greening, David W; Xu, Rong; Ji, Hong; Tauro, Bow J; Simpson, Richard J

    2015-01-01

    Exosomes are 40-150 nm extracellular vesicles that are released from a multitude of cell types, and perform diverse cellular functions including intercellular communication, antigen presentation, and transfer of tumorigenic proteins, mRNA and miRNA. Exosomes are important regulators of the cellular niche, and their altered characteristics in many diseases, such as cancer, suggest their importance for diagnostic and therapeutic applications, and as drug delivery vehicles. Exosomes have been purified from biological fluids and in vitro cell cultures using a variety of strategies and techniques. In this chapter, we reveal the protocol and key insights into the isolation, purification and characterization of exosomes, distinct from shed microvesicles and apoptotic blebs. Using the colorectal cancer cell line LIM1863 as a cell model, a comprehensive evaluation of exosome isolation methods including ultracentrifugation (UC-Exos), OptiPrep™ density-based separation (DG-Exos), and immunoaffinity capture using anti-EpCAM-coated magnetic beads (IAC-Exos) were examined. All exosome isolation methodologies contained 40-150 nm vesicles based on electron microscopy, and positive for exosome markers (Alix, TSG101, HSP70) based on immunoblotting. This protocol employed a proteomic profiling approach to characterize the protein composition of exosomes, and label-free spectral counting to evaluate the effectiveness of each method in exosome isolation. Based on the number of MS/MS spectra identified for exosome markers and proteins associated with their biogenesis, trafficking, and release, IAC-Exos was shown to be the most effective method to isolate exosomes. However, the use of density-based separation (DG-Exos) provides significant advantages for exosome isolation when the use of immunoaffinity capture is limited (due to antibody availability and suitability of exosome markers).

  2. Impact of cell lines included in enterovirus isolation protocol on perception of nonpolio enterovirus species C diversity.

    PubMed

    Adeniji, Johnson Adekunle; Faleye, Temitope Oluwasegun Cephas

    2014-10-01

    There has been under-reporting of nonpolio enterovirus species Cs (NPESCs) in Nigeria despite the fact that most isolates recovered from the Nigerian vaccine derived poliovirus serotype 2 (VDPV2) outbreak were recombinants with nonstructural region of NPESC origin. It has been suggested that cell lines included in enterovirus isolation protocols might account for this phenomenon and this study examined this suggestion. Fifteen environmental samples concentrated previously and analysed using L20B and RD cell lines as part of the poliovirus environmental surveillance (ES) program in Nigeria were randomly selected and inoculated into two cell lines (MCF-7 and LLC-MK2). Isolates were identified as enteroviruses and species C members using different RT-PCR assays, culture in L20B cell line and sequencing of partial VP1. Forty-eight (48) isolates were recovered from the 15 samples, 47 (97.9%) of which were enteroviruses. Of the enteroviruses, 32 (68.1%) belonged to enterovirus species C (EC) of which 19 (40.4%) were polioviruses and 13 (27.7%) were NPESC members. All 13 NPESC isolates were recovered on MCF-7. Results of the study show that NPESCs are circulating in Nigeria and their under-reporting was due to the combination of cell lines used for enterovirus isolation in previous reports.

  3. A modified protocol to maximize differentiation of human preadipocytes and improve metabolic phenotypes

    PubMed Central

    Lee, Mi-Jeong; Wu, Yuanyuan; Fried, Susan K.

    2012-01-01

    Adipose stromal cells proliferate and differentiate into adipocytes, providing a valuable model system for studies of adipocyte biology. We compared differentiation protocols for human preadipocytes and report on their metabolic phenotypes. By simply prolonging the adipogenic induction period from the first 3 days to 7 days, the proportion of cells (passage 5–6) acquiring adipocyte morphology increased from 30–70% to over 80% in human subcutaneous preadipocytes. These morphological changes were accompanied by increases in the adipogenic marker expression and improved adipocyte metabolic phenotypes: enhanced responses to beta-adrenergically-stimulated lipolysis and to insulin-stimulated glucose metabolism into triglyceride. Confirming previous studies, FBS dose-dependently inhibited adipogenesis. However, in subcutaneous preadipocytes that differentiate well (donor-dependant high capacity and subcultured fewer than 2 times), the use of 7d-induction protocols in both 3% FBS and serum-free conditions allowed >80% differentiation. Responsiveness to β-adrenergically stimulated lipolysis was lower in 3% FBS. Rates of insulin-stimulated glucose uptake were higher in adipocytes differentiated with 3% FBS, while the sensitivity to insulin was almost identical between the two groups. In summary, extending the length of the induction period in adipogenic cocktail improves the degree of differentiation and responses to key metabolic hormones. This protocol permits functional analysis of metabolic phenotypes in valuable primary human adipocyte cultures through multiple passages. PMID:22627913

  4. Improved single-cell protocol for preimplantation genetic diagnosis of spinal muscular atrophy.

    PubMed

    Burlet, Philippe; Frydman, Nelly; Gigarel, Nadine; Bonnefont, Jean Paul; Kerbrat, Violaine; Tachdjian, Gérard; Frydman, René; Munnich, Arnold; Steffann, Julie; Ray, Pierre F

    2005-09-01

    To develop and validate a simple and reliable single-cell analysis protocol for the preimplantation genetic diagnosis (PGD) of spinal muscular atrophy (SMA). Molecular tests based on specific enzymatic digestion have already been described for SMA diagnosis. We modified the amplified DNA fragments so as to introduce a novel restriction site that provides an internal control for the completeness of the digestion. The genetics and reproduction departments of two teaching hospitals. Six informed couples at risk of transmitting SMA. All patients underwent standard procedures associated with intracytoplasmic sperm injection. Improvement of SMA diagnostic efficiency and accuracy on single cell. One hundred fifty lymphocytes were analyzed with our protocol. One hundred percent diagnostic accuracy was achieved from both homozygous normal and SMN1-deleted leukocytes. Successful molecular analysis was achieved for 36 of 42 biopsied embryos (86%). Twenty-five normal embryos were transferred, but no pregnancy was achieved. We developed an improved protocol for PGD of SMA that is simple, robust, and accurate; unfortunately, no pregnancies were achieved for any of the six patients who have undergone PGD in the program thus far.

  5. Using shadow page cache to improve isolated drivers performance.

    PubMed

    Zheng, Hao; Dong, Xiaoshe; Wang, Endong; Chen, Baoke; Zhu, Zhengdong; Liu, Chengzhe

    2015-01-01

    With the advantage of the reusability property of the virtualization technology, users can reuse various types and versions of existing operating systems and drivers in a virtual machine, so as to customize their application environment. In order to prevent users' virtualization environments being impacted by driver faults in virtual machine, Chariot examines the correctness of driver's write operations by the method of combining a driver's write operation capture and a driver's private access control table. However, this method needs to keep the write permission of shadow page table as read-only, so as to capture isolated driver's write operations through page faults, which adversely affect the performance of the driver. Based on delaying setting frequently used shadow pages' write permissions to read-only, this paper proposes an algorithm using shadow page cache to improve the performance of isolated drivers and carefully study the relationship between the performance of drivers and the size of shadow page cache. Experimental results show that, through the shadow page cache, the performance of isolated drivers can be greatly improved without impacting Chariot's reliability too much.

  6. Using Shadow Page Cache to Improve Isolated Drivers Performance

    PubMed Central

    Dong, Xiaoshe; Wang, Endong; Chen, Baoke; Zhu, Zhengdong; Liu, Chengzhe

    2015-01-01

    With the advantage of the reusability property of the virtualization technology, users can reuse various types and versions of existing operating systems and drivers in a virtual machine, so as to customize their application environment. In order to prevent users' virtualization environments being impacted by driver faults in virtual machine, Chariot examines the correctness of driver's write operations by the method of combining a driver's write operation capture and a driver's private access control table. However, this method needs to keep the write permission of shadow page table as read-only, so as to capture isolated driver's write operations through page faults, which adversely affect the performance of the driver. Based on delaying setting frequently used shadow pages' write permissions to read-only, this paper proposes an algorithm using shadow page cache to improve the performance of isolated drivers and carefully study the relationship between the performance of drivers and the size of shadow page cache. Experimental results show that, through the shadow page cache, the performance of isolated drivers can be greatly improved without impacting Chariot's reliability too much. PMID:25815373

  7. Antibiotic prophylaxis audit and questionnaire study: Traffic Light Poster improves adherence to protocol in gastrointestinal surgery.

    PubMed

    Cameron, Michaella; Jones, Stacey; Adedeji, Olufunso

    2015-07-01

    To measure adherence to antibiotic prophylaxis (AP) protocol amongst surgeons and anesthetists and explore their understanding of AP prescribing in practice. A prospective audit of AP in gastrointestinal surgery and re-audit after intervention. A questionnaire survey of practice. 58 (38%- clean; 62%- clean contaminated) operations were audited and 73 (48%-clean; 51%-clean contaminated) operations were re-audited after intervention with "Traffic Light Poster" (TFP) .55 colleagues (32 consultants and 23 trainees) were recruited for questionnaire survey in three West Midlands hospitals. Audit and Re-Audits. Only 31% of procedures followed the protocol correctly in the initial audit and this increased to 73% in the re-audit. 73% of patients undergoing clean procedures received AP inappropriately in the initial audit but reduced significantly to 20% (p < 0.002) in the re-audit. In the initial audit, 62% of clean contaminated procedures did not receive the appropriate first line AP but this fell to 35% (p < 0.05) in the re-audit. Questionnaire Survey- Only 30% of respondents would not give AP in clean surgery as recommended. 45% would use appropriate AP for clean-contaminated wounds. 73% of respondents will give AP at induction, 20% 1 h pre op and 7% just before incision. There is poor compliance with AP protocols in gastrointestinal surgery in part due to general lack of awareness. An educational intervention in the form of a 'Traffic Light Poster' improved adherence to AP protocol two fold. There was improved rationalizing of AP. Clean procedures, in particular, had less inappropriate prescribing. Copyright © 2015 IJS Publishing Group Limited. Published by Elsevier Ltd. All rights reserved.

  8. Improvement in 2-year survival for ventricular assist device patients after implementation of an intensive surveillance protocol.

    PubMed

    Pamboukian, Salpy V; Tallaj, José A; Brown, Robert N; Holman, William L; Blood, Margaret; George, James F; Costanzo, Maria Rosa; Cadeiras, Martin; Smallfield, Melissa C; McGiffin, David C; Kirklin, James K

    2011-08-01

    This study was conducted to determine the effect of a disease-management model termed an "intensive surveillance protocol" (ISP) on survival in ventricular assist device (VAD) patients. This intervention consisted of a formalized, protocol-driven, multi-disciplinary team approach to VAD patient follow-up initiated August 1, 2006. The goal was to attain an internal program benchmark of 70% survival at 2 years. Historically, 2-year survival after VAD implant has been sub-optimal, and no patient management algorithms have been formally tested to determine their effect on 2-year survival. The study comprised 76 patients, of whom 26 had a VAD as destination therapy (DT) and 50 as a bridge to transplant (BTT), from July 1, 2003, to June 30, 2008. Survival before and after initiation of ISP was compared. A parametric hazard multivariable analysis, with a time-varying covariable for implementation of ISP, was used to evaluate of other factors affecting survival. Survival at 16 months was 100% for DT patients who received a VAD after August 1, 2006 vs 64% for the earlier era (p = 0.06). For BTT, 16- month survival was 71% vs 43% (p = 0.03). Predicted 2-year survival before and after implementation of the ISP improved from 30% to 87% for DT (p = 0.02) and from 20% to 61% for BTT patients (p = 0.01). Predictors of midterm survival by multivariable analysis included ISP (p = 0.004), younger age (p = 0.03), non-emergent implant (p < 0.0001), and isolated left ventricular VAD (p < 0.0001). After adjustment for covariables, the ISP was associated with a 70% reduction in the hazard for death for the entire cohort (p = 0.004). The effect of ISP was also significant in the patients who received the HeartMate XVE (Thoratec, Pleasanton, CA), which spanned both eras of the study. Survival improved for DT and BTT VAD patients after implementation of the ISP, with a dramatic decrease in hazard for death. Although the transition from pulsatile to axial flow technology occurred during the

  9. Improved High-Voltage Gas Isolator for Ion Thruster

    NASA Technical Reports Server (NTRS)

    Banks, Bruce

    2007-01-01

    A report describes an improved high-voltage isolator for preventing electrical discharge along the flow path of a propellant gas being fed from a supply at a spacecraft chassis electrical potential to an ion thruster at a potential as high as multiple kilovolts. The isolator must survive launch vibration and must remain electrically nonconductive for thousands of hours under conditions that, in the absence of proper design, would cause formation of electrically conductive sputtered metal, carbon, and/or decomposed hydrocarbons on its surfaces. The isolator includes an alumina cylinder containing a spiral channel filled with a porous medium made from alumina microbeads fired together with an alumina slurry. Connections to gas-transport tubes are made at both ends of the alumina cylinder by means of metal caps containing fine-mesh screens to prevent passage of loose alumina particles. The outer surface of the alumina cylinder is convoluted to lengthen the electrical path between the metal caps and to afford shadow shielding to minimize the probability of formation of a continuous deposit that would electrically connect the ends. A flanged cylindrical metal cap that surrounds the alumina cylinder without touching one of the ends provides additional shadow shielding.

  10. Improved selective medium for the isolation of Treponema hyodysenteriae.

    PubMed Central

    Kunkle, R A; Kinyon, J M

    1988-01-01

    An agar medium with improved selection for Treponema hyodysenteriae was developed. Cultures of T. hyodysenteriae and T. innocens, feces from 11 clinically normal pigs, and colonic contents from 6 pigs with gross lesions consistent with swine dysentery were diluted in phosphate-buffered saline and plated on Trypticase soy agar (BBL Microbiology Systems, Cockeysville, Md.) with 5% citrated bovine blood (TSA), TSA with 400 micrograms of spectinomycin per ml (TSA-S400), TSA-S400 with 25 micrograms each of colistin and vancomycin per ml, and TSA with 5% pig feces extract and five antimicrobial agents (spiramycin, rifampin, vancomycin, colistin, and spectinomycin) (BJ). Viable numbers of T. hydodysenteriae grown on BJ were virtually identical to those for TSA, TSA-S400, and TSA-S400 with colistin and vancomycin. Pure cultures of four isolates of T. hyodysenteriae and three isolates of T. innocens were sustained through six subcultures on BJ. Fecal floras were completely inhibited on BJ for 14 of 17 fecal samples from both groups of pigs. A total of 461 colonic specimens from naturally occurring cases of porcine intestinal disease were plated on TSA-S400 and BJ. T. hyodysenteriae was isolated on both TSA-S400 and BJ for 69 specimens and on BJ alone for an additional 19 specimens. PMID:3235663

  11. Simulation improves procedural protocol adherence during central venous catheter placement: a randomized-controlled trial

    PubMed Central

    Peltan, Ithan D.; Shiga, Takashi; Gordon, James A.; Currier, Paul F.

    2015-01-01

    Background Simulation training may improve proficiency at and reduces complications from central venous catheter (CVC) placement, but the scope of simulation’s effect remains unclear. This randomized controlled trial evaluated the effects of a pragmatic CVC simulation program on procedural protocol adherence, technical skill, and patient outcomes. Methods Internal medicine interns were randomized to standard training for CVC insertion or standard training plus simulation-based mastery training. Standard training involved a lecture, a video-based online module, and instruction by the supervising physician during actual CVC insertions. Intervention-group subjects additionally underwent supervised training on a venous access simulator until they demonstrated procedural competence. Raters evaluated interns’ performance during internal jugular CVC placement on actual patients in the medical intensive care unit. Generalized estimating equations were used to account for outcome clustering within trainees. Results We observed 52 interns place 87 CVCs. Simulation-trained interns exhibited better adherence to prescribed procedural technique than interns who received only standard training (p=0.024). There were no significant differences detected in first-attempt or overall cannulation success rates, mean needle passes, global assessment scores or complication rates. Conclusions Simulation training added to standard training improved protocol adherence during CVC insertion by novice practitioners. This study may have been too small to detect meaningful differences in venous cannulation proficiency and other clinical outcomes, highlighting the difficulty of patient-centered simulation research in settings where poor outcomes are rare. For high-performing systems, where protocol deviations may provide an important proxy for rare procedural complications, simulation may improve CVC insertion quality and safety. PMID:26154250

  12. Improvement in β-cell secretory capacity after human islet transplantation according to the CIT07 protocol.

    PubMed

    Rickels, Michael R; Liu, Chengyang; Shlansky-Goldberg, Richard D; Soleimanpour, Scott A; Vivek, Kumar; Kamoun, Malek; Min, Zaw; Markmann, Eileen; Palangian, Maral; Dalton-Bakes, Cornelia; Fuller, Carissa; Chiou, Allen J; Barker, Clyde F; Luning Prak, Eline T; Naji, Ali

    2013-08-01

    The Clinical Islet Transplantation 07 (CIT07) protocol uses antithymocyte globulin and etanercept induction, islet culture, heparinization, and intensive insulin therapy with the same low-dose tacrolimus and sirolimus maintenance immunosuppression as in the Edmonton protocol. To determine whether CIT07 improves engrafted islet β-cell mass, our center measured β-cell secretory capacity from glucose-potentiated arginine tests at days 75 and 365 after transplantation and compared those results with the results previously achieved by our group using the Edmonton protocol and normal subjects. All subjects were insulin free, with CIT07 subjects receiving fewer islet equivalents from a median of one donor compared with two donors for Edmonton protocol subjects. The acute insulin response to glucose-potentiated arginine (AIRpot) was greater in the CIT07 protocol than in the Edmonton protocol and was less in both cohorts than in normal subjects, with similar findings for C-peptide. The CIT07 subjects who completed reassessment at day 365 exhibited increasing AIRpot by trend relative to that of day 75. These data indicate that engrafted islet β-cell mass is markedly improved with the CIT07 protocol, especially given more frequent use of single islet donors. Although several peritransplant differences may have each contributed to this improvement, the lack of deterioration in β-cell secretory capacity over time in the CIT07 protocol suggests that low-dose tacrolimus and sirolimus are not toxic to islets.

  13. An Improved Quantum Information Hiding Protocol Based on Entanglement Swapping of χ-type Quantum States

    NASA Astrophysics Data System (ADS)

    Xu, Shu-Jiang; Chen, Xiu-Bo; Wang, Lian-Hai; Ding, Qing-Yan; Zhang, Shu-Hui

    2016-06-01

    In 2011, Qu et al. proposed a quantum information hiding protocol based on the entanglement swapping of χ-type quantum states. Because a χ-type state can be described by the 4-particle cat states which have good symmetry, the possible output results of the entanglement swapping between a given χ-type state and all of the 16 χ-type states are divided into 8 groups instead of 16 groups of different results when the global phase is not considered. So it is difficult to read out the secret messages since each result occurs twice in each line (column) of the secret messages encoding rule for the original protocol. In fact, a 3-bit instead of a 4-bit secret message can be encoded by performing two unitary transformations on 2 particles of a χ-type quantum state in the original protocol. To overcome this defect, we propose an improved quantum information hiding protocol based on the general term formulas of the entanglement swapping among χ-type states. Supported by the National Natural Science Foundation of China under Grant Nos. 61572297, 61303199, 61272514, and 61373131, the Shandong Provincial Natural Science Foundation of China under Grant Nos. ZR2013FM025, ZR2013FQ001, ZR2014FM003, and ZY2015YL018, the Shandong Provincial Outstanding Research Award Fund for Young Scientists of China under Grant Nos. BS2015DX006 and BS2014DX007, the National Development Foundation for Cryptological Research, China under Grant No. MMJJ201401012, the Priority Academic Program Development of Jiangsu Higher Education Institutions and Jiangsu Collaborative Innovation Center on Atmospheric Environment and Equipment Technology Funds, and the Shandong Academy of Sciences Youth Fund Project, China under Grant Nos. 2015QN003 and 2013QN007

  14. An occupation-based video feedback intervention for improving self-awareness: protocol and rationale.

    PubMed

    Schmidt, Julia; Fleming, Jennifer; Ownsworth, Tamara; Lannin, Natasha A

    2015-02-01

    Impaired self-awareness can limit rehabilitation outcomes for people with traumatic brain injury (TBI). Video feedback on occupational performance has been found to improve self-awareness after TBI when delivered according to specific principles. The purpose of this article is to describe an occupation-based video feedback intervention found to be effective in a randomized controlled trial to assist with translation into clinical practice. The intervention uses therapist-mediated video feedback on clients' occupational performance, aiming to facilitate self-reflection on performance and improve self-awareness. This paper describes the theoretical background, intervention principles, and protocol of the intervention. Therapists can use video feedback intervention, incorporating the principles in this article, to improve people's intellectual awareness and ability to recognize and correct errors during task performance after TBI without a negative impact on emotional status.

  15. Treatment Protocols to Reduce Injury and Improve Stone Breakage in SWL

    NASA Astrophysics Data System (ADS)

    McAteer, James A.; Evan, Andrew P.; Connors, Bret A.; Pishchalnikov, Yuri A.; Williams, James C.; Lingeman, James E.

    2008-09-01

    Here we provide a capsule summary of key observations showing that adverse effects can be reduced and stone breakage outcomes can be improved by the choice of the treatment protocol used in SWL. The take home message is—technique in lithotripsy can be used to significant advantage. SW-rate is key, and so is the sequence of SW delivery. Patient studies have shown that stone breakage is significantly improved at 60SW/min compared to a rate of 120SW/min, and laboratory experiments with pigs show that acute SWL injury to the kidney can be reduced dramatically by further slowing the SW firing rate to 30SW/min. The sequence of SW administration has a profound effect on the kidney, and renal injury is significantly reduced when the treatment protocol incorporates a priming dose of SW's followed by a brief pause before treatment is resumed. Continued developments in lithotripsy technology are welcome and will hopefully lead to improved SWL systems. Current experience suggests, however, that technology is not a substitute for expert technique, and attention to the fundamentals of SW delivery is essential to achieve the best possible outcomes regardless of the lithotripter at hand.

  16. Rate allocation protocol using competitive pricing for improving performance of multicast sessions

    NASA Astrophysics Data System (ADS)

    Levy, Zohar; Dolev, Danny

    1998-10-01

    Rate allocation using the Max-Min fairness criterion may highly discriminate against multicast and long unicast sessions and may lead to sever network underutilization. In this paper, we present a solution for rate allocation that is based on competitive pricing. The resultant allocation increases fairness towards multicast sessions and improves network utilization considerably. The solution requires no re-routing of sessions. The economy on which we base our solution is simple enough, enabling its implementation for practical use. We present a distributed asynchronous protocol suitable for the ATM ABR service, which achieves the economy's allocation efficiently and with short convergence time.

  17. Development and validation of a PulseNet standardized protocol for subtyping isolates of Cronobacter species.

    PubMed

    Brengi, Silvina P; O'Brien, Stephen B; Pichel, Mariana; Iversen, Carol; Arduino, Matthew; Binsztein, Norma; Jensen, Bette; Pagotto, Franco; Ribot, Efrain M; Stephan, Roger; Cernela, Nicole; Cooper, Kara; Fanning, Séamus

    2012-09-01

    Cronobacter (formerly known as Enterobacter sakazakii) is a genus comprising seven species regarded as opportunistic pathogens that can be found in a wide variety of environments and foods, including powdered infant formula (PIF). Cronobacter sakazakii, the major species of this genus, has been epidemiologically linked to cases of bacteremia, meningitis in neonates, and necrotizing enterocolitis, and contaminated PIF has been identified as an important source of infection. Robust and reproducible subtyping methods are required to aid in the detection and investigation, of foodborne outbreaks. In this study, a pulsed-field gel electrophoresis (PFGE) protocol was developed and validated for subtyping Cronobacter species. It was derived from an existing modified PulseNet protocol, wherein XbaI and SpeI were the primary and secondary restriction enzymes used, generating an average of 14.7 and 20.3 bands, respectively. The PFGE method developed was both reproducible and discriminatory for subtyping Cronobacter species.

  18. Three-year outcome of isolated glomerulitis on 3-month protocol biopsies of donor HLA antibody negative patients.

    PubMed

    Buob, David; Grimbert, Philippe; Glowacki, François; Labalette, Myriam; Dufossé, Françoise; Nochy, Dominique; Copin, Marie-Christine; Boleslawski, Emmanuel; Noël, Christian; Hazzan, Marc

    2012-06-01

    Transplant glomerulitis (TG) can lead to the diagnosis of acute humoral rejection when associated with C4d. Recent data have shown that, in patients with donor-specific antibodies, TG is a sign of humoral rejection, even in the absence of C4d. However, the clinical significance of isolated TG, i.e. TG without C4d deposition or morphological evidence of rejection, has not been specifically studied in protocol biopsies of recipients without donor-specific antibodies. We compared 20 isolated TG-patients with 44 selected recipients without TG or any rejection-associated change. The two groups had similar baseline characteristics. After a 3 year follow-up, renal function, acute rejection rate, and development of HLA antibodies were not significantly different between the two groups. Isolated TG had no deleterious consequences on the 3 year graft outcome. Eleven patients of the glomerulitis-group had another allograft biopsy during follow-up: glomerular lesions returned to normal in six patients whereas the persistence of glomerulitis or features consistent with chronic transplant glomerulopathy were noticed in the remaining five patients. Four of these five patients had pretransplant non-donor specific HLA antibodies. In conclusion, although isolated TG had no impact on allograft function at 3 year, histological outcome could be related to patient sensitization. © 2012 The Authors. Transplant International © 2012 European Society for Organ Transplantation.

  19. New ruthenium-based protocol for cleavage of terminal olefins to primary alcohols: improved synthesis of a bicyclic nucleoside.

    PubMed

    Sharma, Pawan K; Nielsen, Poul

    2004-08-20

    A new protocol for the oxidative cleavage of terminal alkenes to give exclusively primary alcohols in high yields is introduced. The protocol is based on RuO4-mediated dihydroxylation, NaIO4-mediated diol cleavage, and NaBH4-mediated reduction, but the introduction of a reducing step before the diol cleavage removes the formation of byproducts and improves the yield significantly. The new protocol has been developed and used for the improved preparation of a [3.2.0]bicycloarabinonucleoside with important potential in antisense and antigene technology.

  20. U22 protocol as measure of symptomatic improvement after catheter ablation of atrial fibrillation

    PubMed Central

    Höglund, Niklas; Rönn, Folke; Tollefsen, Titti; Jensen, Steen m.

    2013-01-01

    Introduction. Left atrial catheter ablation is useful as symptomatic treatment in selected patients with atrial fibrillation (AF). Evaluation requires measurement of arrhythmia-related symptoms. Many of the published protocols have drawbacks and have been used in AF only, with no possible comparison to other ablations that compete for the same resources. U22 is a published protocol that quantifies paroxysmal tachycardia symptoms through scales with 11 answer alternatives, translated into discrete numerical scales 0–10. It has been shown to reflect the clinical improvement after ablation of supraventricular tachycardia. Here we report the use of U22 in measuring improvement after catheter ablation for AF. Material and methods. A total of 105 patients underwent first-time ablation for AF and answered U22 and SF-36 forms at baseline and follow-up 304 (SD 121) days after ablation. Independently, the patients underwent a clinical follow-up. All decisions regarding medication and reablation were taken without knowledge of the symptom scores. Results. The U22 scores for well-being, arrhythmia as cause for impaired well-being, derived time-aspect score for arrhythmia, and discomfort during attack detected relevant improvements of symptoms after the ablation. U22 showed larger improvement in patients undergoing only one procedure than in patients who later underwent repeated interventions, thus reflecting the independent clinical decision for reablation. Conclusion.U22 quantifies the symptomatic improvement after AF ablation with adequate internal consistency and construct validity. U22 mirrors aspects of the arrhythmia symptomatology other than SF-36. PMID:24102147

  1. Does the presence of a condition-specific obstetric protocol lead to detectable improvements in pregnancy outcomes?

    PubMed Central

    Bailit, Jennifer L.; Grobman, William; McGee, Paula; Reddy, Uma M.; Wapner, Ronald J.; Varner, Michael W.; Thorp, John M.; Leveno, Kenneth J.; Iams, Jay D.; Tita, Alan T. N.; Saade, George; Sorokin, Yoram; Rouse, Dwight J.; Blackwell, Sean C.

    2015-01-01

    Objective To evaluate whether the presence of condition-specific obstetric protocols within a hospital was associated with better maternal and neonatal outcomes. Study Design Cohort study of a random sample of deliveries performed at 25 hospitals over three years. Condition-specific protocols were collected from all hospitals and categorized independently by two authors. Data on maternal and neonatal outcomes, as well as data necessary for risk adjustment were collected. Risk-adjusted outcomes were compared according to whether the patient delivered in a hospital with condition-specific obstetric protocols at the time of delivery. Results Hemorrhage-specific protocols were not associated with a lower rate of postpartum hemorrhage or with fewer cases of EBL >1000cc. Similarly, in the presence of a shoulder dystocia protocol, there were no differences in the frequency of shoulder dystocia or number of shoulder dystocia maneuvers used. Conversely, preeclampsia-specific protocols were associated with fewer ICU admissions (OR 0.28, 95% CI 0.18–0.44) and fewer cases of severe maternal hypertension (OR 0.86, 95% CI 0.77–0.96). Conclusion The presence of condition-specific obstetric protocols was not consistently shown to be associated with improved risk-adjusted outcomes. Our study would suggest that the presence or absence of a protocol does not matter and regulations to require protocols are not fruitful. PMID:25659468

  2. Development of an improved vaccine evaluation protocol to compare the efficacy of Newcastle disease vaccines.

    PubMed

    Cardenas-Garcia, Stivalis; Diel, Diego G; Susta, Leonardo; Lucio-Decanini, Eduardo; Yu, Qingzhong; Brown, Corrie C; Miller, Patti J; Afonso, Claudio L

    2015-03-01

    While there is typically 100% survivability in birds challenged with vNDV under experimental conditions, either with vaccines formulated with a strain homologous or heterologous (different genotype) to the challenge virus, vaccine deficiencies are often noted in the field. We have developed an improved and more stringent protocol to experimentally evaluate live NDV vaccines, and showed for the first time under experimental conditions that a statistically significant reduction in mortality can be detected with genotype matched vaccines. Using both vaccine evaluation protocols (traditional and improved), birds were challenged with a vNDV of genotype XIII and the efficacy of live heterologous (genotype II) and homologous (genotype XIII) NDV vaccines was compared. Under traditional vaccination conditions there were no differences in survival upon challenge, but the homologous vaccine induced significantly higher levels of antibodies specific to the challenge virus. With the more stringent challenge system (multiple vaccine doses and early challenge with high titers of vNDV), the birds administered the homologous vaccine had superior humoral responses, reduced clinical signs, and reduced mortality levels than those vaccinated with the heterologous vaccine. These results provide basis for the implementation of more sensitive methods to evaluate vaccine efficacy. Published by Elsevier Ltd.

  3. Improving detection of avian malaria from host blood: a step towards a standardised protocol for diagnostics.

    PubMed

    Niebuhr, Chris N; Blasco-Costa, Isabel

    2016-10-01

    Avian malaria, caused by Plasmodium spp., has been linked to the mortality and population-level declines in native birds in some regions. While molecular diagnostic methods have greatly improved our ability to detect infections of both human and bird malaria, failing to identify false negatives remains an important handicap, particularly for avian malaria due to host DNA presence in the bird blood cells. In an attempt to improve the accuracy of diagnostics by PCR, we evaluated the performance of a commercial silica-membrane-based DNA extraction kit by modifying the protocol with four unpooled elution volume alternatives. Our results suggest that the best template is the DNA extract obtained from the second eluate of a first 50 μL elution step. In one case, the only band visible was from this second eluate and, thus, may not have been identified as positive for Plasmodium spp. if a different elution protocol had been followed. Our results are likely explained by the concept of size exclusion chromatography by which particles of different sizes will elute at different rates. Overall, first elution templates may consist of a lower ratio of parasite to host DNA, while second eluates may contain a higher parasite to host DNA ratio. A low ratio of parasite to host DNA is a concern in detecting chronic infections, in which birds typically carry low levels of parasitemia, making accurate diagnostics imperative when identifying reservoirs of disease that could lead to spillback events.

  4. A Standardized Shift Handover Protocol: Improving Nurses’ Safe Practice in Intensive Care Units

    PubMed Central

    Malekzadeh, Javad; Mazluom, Seyed Reza; Etezadi, Toktam; Tasseri, Alireza

    2013-01-01

    Introduction: For maintaining the continuity of care and improving the quality of care, effective inter-shift information communication is necessary. Any handover error can endanger patient safety. Despite the importance of shift handover, there is no standard handover protocol in our healthcare settings. Methods: In this one-group pretest-posttest quasi-experimental study conducted in spring and summer of 2011, we recruited a convenience sample of 56 ICU nurses. The Nurses’ Safe Practice Evaluation Checklist was used for data collection. The Content Validity Index and the inter-rater correlation coefficient of the checklist was 0.92 and 89, respectively. We employed the SPSS 11.5 software and the Mc Nemar and paired-samples t test for data analysis. Results: Study findings revealed that nurses’ mean score on the Safe Practice Evaluation Checklist increased significantly from 11.6 (2.7) to 17.0 (1.8) (P < 0.001). Conclusion: using a standard handover protocol for communicating patient’s needs and information improves nurses’ safe practice in the area of basic nursing care. PMID:25276725

  5. A Structured Rehabilitation Protocol for Improved Multifunctional Prosthetic Control: A Case Study.

    PubMed

    Roche, Aidan Dominic; Vujaklija, Ivan; Amsüss, Sebastian; Sturma, Agnes; Göbel, Peter; Farina, Dario; Aszmann, Oskar C

    2015-11-06

    Advances in robotic systems have resulted in prostheses for the upper limb that can produce multifunctional movements. However, these sophisticated systems require upper limb amputees to learn complex control schemes. Humans have the ability to learn new movements through imitation and other learning strategies. This protocol describes a structured rehabilitation method, which includes imitation, repetition, and reinforcement learning, and aims to assess if this method can improve multifunctional prosthetic control. A left below elbow amputee, with 4 years of experience in prosthetic use, took part in this case study. The prosthesis used was a Michelangelo hand with wrist rotation, and the added features of wrist flexion and extension, which allowed more combinations of hand movements. The participant's Southampton Hand Assessment Procedure score improved from 58 to 71 following structured training. This suggests that a structured training protocol of imitation, repetition and reinforcement may have a role in learning to control a new prosthetic hand. A larger clinical study is however required to support these findings.

  6. The protocol for the isolation and cryopreservation of osteoclast precursors from mouse bone marrow and spleen.

    PubMed

    Boraschi-Diaz, Iris; Komarova, Svetlana V

    2016-01-01

    Osteoclasts are responsible for physiological bone remodeling as well as pathological bone destruction in osteoporosis, periodontitis and rheumatoid arthritis, and thus represent a pharmacological target for drug development. We aimed to characterize and compare the cytokine-induced osteoclastogenesis of bone marrow and spleen precursors. Established protocols used to generate osteoclasts from bone marrow were modified to examine osteoclastogenesis of the spleen cells of healthy mice. Osteoclast formation was successfully induced from spleen precursors using receptor activator of nuclear factor κB ligand (50 ng/ml) and macrophage colony stimulating factor (50 ng/ml). Compared to bone marrow cultures, differentiation from spleen required a longer cultivation time (9 days for spleen, as compared to 5 days for marrow cultures) and a higher plating density of non-adherent cells (75,000/cm(2) for spleen, as compared to 50,000/cm(2) for bone marrow). Osteoclasts generated from spleen precursors expressed osteoclast marker genes calcitonin receptor, cathepsin K and matrix metalloproteinase 9 and were capable of resorbing hydroxyapatite. The differentiation capacity of spleen and bone marrow precursors was comparable for BALB/c, C57BL/6 and FVB mice. We also developed and tested a cryopreservation protocol for the osteoclast precursors. While 70-80 % of cells were lost during the first week of freezing, during the subsequent 5 weeks the losses were within 2-5 % per week. Osteoclastogenesis from the recovered bone marrow precursors was successful up to 5 weeks after freezing. Spleen precursors retained their osteoclastogenic capacity for 1 week after freezing, but not thereafter. The described protocol is useful for the studies of genetically modified animals as well as for screening new osteoclast-targeting therapeutics.

  7. Improved hybrid isolator with maglev actuator integrated in air spring for active-passive isolation of ship machinery vibration

    NASA Astrophysics Data System (ADS)

    Li, Yan; He, Lin; Shuai, Chang-geng; Wang, Chun-yu

    2017-10-01

    A hybrid isolator consisting of maglev actuator and air spring is proposed and developed for application in active-passive vibration isolation system of ship machinery. The dynamic characteristics of this hybrid isolator are analyzed and tested. The stability and adaptability of this hybrid isolator to shock and swing in the marine environment are improved by a compliant gap protection technique and a disengageable suspended structure. The functions of these new engineering designs are proved by analytical verification and experimental validation of the designed stiffness of such a hybrid isolator, and also by shock adaptability testing of the hybrid isolator. Finally, such hybrid isolators are installed in an engineering mounting loaded with a 200-kW ship diesel generator, and the broadband and low-frequency sinusoidal isolation performance is tested.

  8. A pilot study of an acupuncture protocol to improve visual function in retinitis pigmentosa patients

    PubMed Central

    Bittner, Ava K; Gould, Jeffrey M; Rosenfarb, Andy; Rozanski, Collin; Dagnelie, Gislin

    2014-01-01

    Background Patients with retinitis pigmentosa are motivated to try complementary or integrative therapies to slow disease progression. Basic science, clinical research and retinitis pigmentosa patients' self-reports support the hypothesis that acupuncture may improve visual function. Methods A prospective, case series, pilot study enrolled 12 adult patients with RP treated at an academic medical centre with a standardised protocol that combined electroacupuncture to the forehead and below the eyes and acupuncture to the body, at 10 half-hour sessions over two weeks. Pre- and post-treatment tests included Early Treatment Diabetic Retinopathy Study visual acuity (VA), Pelli-Robson contrast sensitivity (CS), Goldmann visual fields, and dark-adapted full-field stimulus threshold (FST)(n = 9). Scotopic Sensitivity Tester-1 (SST-1) dark-adaptometry was performed on the last two subjects. Results Six of 12 subjects had measurable, significant visual function improvements after treatment. Three of nine subjects tested with the FST had a significant 10.3 to 17.5 dB (that is, 13- to 53-fold) improvement in both eyes at one week after acupuncture, maintained for at least 10 to 12 months, which was well outside typical test-retest variability (95% CI: 3–3.5 dB) previously found in retinitis pigmentosa. SST-1 dark-adaptation was shortened in both subjects tested on average by 48.5 per cent at one week (range 36 to 62 per cent across 10 to 30 dB), which was outside typical coefficients of variation of less than 30 per cent previously determined in patients with retinitis pigmentosa and normals. Four of the five subjects with psychophysically measured scotopic sensitivity improvements reported subjective improvements in vision at night or in dark environments. One subject had 0.2 logMAR improvement in VA; another had 0.55 logCS improvement. Another subject developed more than 20 per cent improvement in the area of the Goldmann visual fields. The acupuncture protocol was

  9. A pilot study of an acupuncture protocol to improve visual function in retinitis pigmentosa patients.

    PubMed

    Bittner, Ava K; Gould, Jeffrey M; Rosenfarb, Andy; Rozanski, Collin; Dagnelie, Gislin

    2014-05-01

    Patients with retinitis pigmentosa are motivated to try complementary or integrative therapies to slow disease progression. Basic science, clinical research and retinitis pigmentosa patients' self-reports support the hypothesis that acupuncture may improve visual function. A prospective, case series, pilot study enrolled 12 adult patients with RP treated at an academic medical centre with a standardised protocol that combined electroacupuncture to the forehead and below the eyes and acupuncture to the body, at 10 half-hour sessions over two weeks. Pre- and post-treatment tests included Early Treatment Diabetic Retinopathy Study visual acuity (VA), Pelli-Robson contrast sensitivity (CS), Goldmann visual fields, and dark-adapted full-field stimulus threshold (FST)(n = 9). Scotopic Sensitivity Tester-1 (SST-1) dark-adaptometry was performed on the last two subjects. Six of 12 subjects had measurable, significant visual function improvements after treatment. Three of nine subjects tested with the FST had a significant 10.3 to 17.5 dB (that is, 13- to 53-fold) improvement in both eyes at one week after acupuncture, maintained for at least 10 to 12 months, which was well outside typical test-retest variability (95% CI: 3-3.5 dB) previously found in retinitis pigmentosa. SST-1 dark-adaptation was shortened in both subjects tested on average by 48.5 per cent at one week (range 36 to 62 per cent across 10 to 30 dB), which was outside typical coefficients of variation of less than 30 per cent previously determined in patients with retinitis pigmentosa and normals. Four of the five subjects with psychophysically measured scotopic sensitivity improvements reported subjective improvements in vision at night or in dark environments. One subject had 0.2 logMAR improvement in VA; another had 0.55 logCS improvement. Another subject developed more than 20 per cent improvement in the area of the Goldmann visual fields. The acupuncture protocol was completed and well tolerated

  10. Phenotypic and functional consequences of different isolation protocols on skin mononuclear phagocytes.

    PubMed

    Botting, Rachel A; Bertram, Kirstie M; Baharlou, Heeva; Sandgren, Kerrie J; Fletcher, James; Rhodes, Jake W; Rana, Hafsa; Plasto, Toby M; Wang, Xin Maggie; Lim, Jake J K; Barnouti, Laith; Kohout, Mark P; Papadopoulos, Tim; Merten, Steve; Olbourne, Norman; Cunningham, Anthony L; Haniffa, Muzlifah; Harman, Andrew N

    2017-06-01

    Mononuclear phagocytes are present in skin and mucosa and represent one of the first lines of defense against invading pathogens, which they detect via an array of pathogen-binding receptors expressed on their surface. However, their extraction from tissue is difficult, and the isolation technique used has functional consequences on the cells obtained. Here, we compare mononuclear phagocytes isolated from human skin using either enzymatic digestion or spontaneous migration. Cells isolated via enzymatic digestion are in an immature state, and all subsets are easily defined. However, cells isolated by spontaneous migration are in a mature state, and CD141 cross-presenting DCs (cDC1) are more difficult to define. Different pathogen-binding receptors are susceptible to cleavage by blends of collagenase, demonstrating that great care must be taken in choosing the correct enzyme blend to digest tissue if carrying out pathogen-interaction assays. Finally, we have optimized mononuclear phagocyte culture conditions to enhance their survival after liberation from the tissue. © The Author(s).

  11. Improving anticoagulation therapy using point-of-care testing and a standardized protocol.

    PubMed

    Franke, Curtis A; Dickerson, Lori M; Carek, Peter J

    2008-01-01

    Many patients in primary care require anticoagulation with warfarin for the prevention of venous and systemic embolism. Achieving the goal international normalized ratio (INR) with warfarin is challenging. The purpose of this quality improvement initiative was to increase the proportion of patients taking warfarin with an INR value within the goal range. We included all patients identified on an anticoagulation log in the family medicine residency practice during 3 time periods: baseline, after point-of-care (POC) testing was initiated (intervention period 1), and after a standardized warfarin-dosing protocol was implemented (intervention period 2). Educational sessions were conducted during each intervention period. Measures included the frequency of INR monitoring and the percentage of office visits in which patients' values were within the goal INR range. Data were analyzed using descriptive statistics, the Student t test, and the chi2 test. At baseline, patients had an average of 2.6 INR tests performed, and 30.8% were within the INR goal range. Using POC testing, the frequency of monitoring increased to 4.3 INR tests per patient (P = .04), but the percentage of patients within the INR goal remained low at 32.1% (P=.88). When physicians implemented the standardized protocol to guide warfarin dosing, the frequency of testing was similar (3.8 tests per patient), but the percentage of patients within the INR goal increased to 45.9% (P<.04). POC testing increased the frequency of INR testing, and additional use of a standardized protocol for warfarin dosing increased the percentage of patients within the INR goal range. This model of anticoagulation management could be easily implemented in any family medicine office.

  12. Gene fishing: the use of a simple protocol to isolate multiple homeodomain classes from diverse invertebrate taxa.

    PubMed

    Lee, Shannon E; Gates, Ruth D; Jacobs, David K

    2003-04-01

    Comparison of relevant gene sequence and functional data is central to understanding the evolution of metazoan development. The conservation of portions of regulatory genes, such as homeoboxes, allows for the design of PCR-based sequence isolation and amplification strategies. Here we describe a simple protocol that uses a degenerate primer pair to isolate a variety of homeobox-containing genes from diverse metazoan taxa. In a nonexhaustive survey, we have isolated 28 gene sequence fragments from 15 taxa, representing eight invertebrate phyla (Mollusca, Echiura, Annelida, Platyhelminth, Acoela, Ctenophora, Cnidaria, and Porifera). Based on BLAST and parsimony analyses, these gene fragments affiliate with several gene groups (PAIRED-like, HOX, and ParaHOX) and several single genes, including pancreas/duodenum homeoboxes (Pdx), empty spiracles (ems/Emx), gastrulation brain homeoboxes (Gbx), hematopoietically expressed homeoboxes (HEX), brain specific homeobox (bsh/BarH1/BarH2), NK-1 (NK-1/s59/slouch), and ladybird (Lbl/Lbe/Lbx). In several cases, these fragments represent the first reported orthologue for the phylum or superphyletic group (i.e., Lophotrochozoa).

  13. Improved protocol for the simultaneous extraction and column-based separation of DNA and RNA from different soils.

    PubMed

    Töwe, Stefanie; Wallisch, Stefanie; Bannert, Andrea; Fischer, Doreen; Hai, Brigitte; Haesler, Felix; Kleineidam, Kristina; Schloter, Michael

    2011-03-01

    We developed an improved protocol, allowing the simultaneous extraction of DNA and RNA from soil using phenol-chloroform with subsequent column-based separation of DNA and RNA (PCS). We compared this new approach with the well established protocol published by Griffiths et al. (2000), where DNA and RNA are separated by selective enzymatic digestions and two commercial kits used for DNA or RNA extraction, respectively, using four different agricultural soils. We compared yield and purity of the nucleic acids as well as abundance and diversity profiles of the soil bacterial communities targeting the nosZ gene via quantitative real-time PCR and terminal restriction fragment length polymorphism on DNA and RNA level. The newly developed protocol provided purer nucleic acid extracts compared to the used kit-based protocols. All protocols were suitable for DNA- and RNA-based gene quantification, however high variations between replicates were obtained for RNA samples using the original Griffiths protocol. Diversity patterns of nosZ were highly influenced by the extraction protocol used both on the DNA and RNA level. Finally, our data showed that the new protocol allows a simultaneous and reproducible extraction and separation of DNA and RNA, which were suitable for reliable analyses of gene and transcript copy numbers and diversity pattern. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. A Mirror Therapy-Based Action Observation Protocol to Improve Motor Learning After Stroke.

    PubMed

    Harmsen, Wouter J; Bussmann, Johannes B J; Selles, Ruud W; Hurkmans, Henri L P; Ribbers, Gerard M

    2015-07-01

    Mirror therapy is a priming technique to improve motor function of the affected arm after stroke. To investigate whether a mirror therapy-based action observation (AO) protocol contributes to motor learning of the affected arm after stroke. A total of 37 participants in the chronic stage after stroke were randomly allocated to the AO or control observation (CO) group. Participants were instructed to perform an upper-arm reaching task as fast and as fluently as possible. All participants trained the upper-arm reaching task with their affected arm alternated with either AO or CO. Participants in the AO group observed mirrored video tapes of reaching movements performed by their unaffected arm, whereas participants in the CO group observed static photographs of landscapes. The experimental condition effect was investigated by evaluating the primary outcome measure: movement time (in seconds) of the reaching movement, measured by accelerometry. Movement time decreased significantly in both groups: 18.3% in the AO and 9.1% in the CO group. Decrease in movement time was significantly more in the AO compared with the CO group (mean difference = 0.14 s; 95% confidence interval = 0.02, 0.26; P = .026). The present study showed that a mirror therapy-based AO protocol contributes to motor learning after stroke. © The Author(s) 2014.

  15. Fluorescence-Activated Cell Sorting (FACS) Protocol for Podocyte Isolation in Adult Zebrafish.

    PubMed

    Bates, Thomas J D; Naumann, Uta; Englert, Christoph

    2016-01-01

    Zebrafish is becoming a very important model for studying human diseases. The conserved structure of the nephrons in the kidney allows the user to answer questions relating to study human kidney disorders. Wt1a-expressing podocytes are the most important cells within the glomeruli of adult zebrafish. In order to understand the molecular characteristics of these cells, within damage models, we have established a method for isolating them.

  16. Improvement of halophilic cellulase production from locally isolated fungal strain

    PubMed Central

    Gunny, Ahmad Anas Nagoor; Arbain, Dachyar; Jamal, Parveen; Gumba, Rizo Edwin

    2014-01-01

    Halophilic cellulases from the newly isolated fungus, Aspergillus terreus UniMAP AA-6 were found to be useful for in situ saccharification of ionic liquids treated lignocelluloses. Efforts have been taken to improve the enzyme production through statistical optimization approach namely Plackett–Burman design and the Face Centered Central Composite Design (FCCCD). Plackett–Burman experimental design was used to screen the medium components and process conditions. It was found that carboxymethylcellulose (CMC), FeSO4·7H2O, NaCl, MgSO4·7H2O, peptone, agitation speed and inoculum size significantly influence the production of halophilic cellulase. On the other hand, KH2PO4, KOH, yeast extract and temperature had a negative effect on enzyme production. Further optimization through FCCCD revealed that the optimization approach improved halophilic cellulase production from 0.029 U/ml to 0.0625 U/ml, which was approximately 2.2-times greater than before optimization. PMID:26150755

  17. Improvement of halophilic cellulase production from locally isolated fungal strain.

    PubMed

    Gunny, Ahmad Anas Nagoor; Arbain, Dachyar; Jamal, Parveen; Gumba, Rizo Edwin

    2015-07-01

    Halophilic cellulases from the newly isolated fungus, Aspergillus terreus UniMAP AA-6 were found to be useful for in situ saccharification of ionic liquids treated lignocelluloses. Efforts have been taken to improve the enzyme production through statistical optimization approach namely Plackett-Burman design and the Face Centered Central Composite Design (FCCCD). Plackett-Burman experimental design was used to screen the medium components and process conditions. It was found that carboxymethylcellulose (CMC), FeSO4·7H2O, NaCl, MgSO4·7H2O, peptone, agitation speed and inoculum size significantly influence the production of halophilic cellulase. On the other hand, KH2PO4, KOH, yeast extract and temperature had a negative effect on enzyme production. Further optimization through FCCCD revealed that the optimization approach improved halophilic cellulase production from 0.029 U/ml to 0.0625 U/ml, which was approximately 2.2-times greater than before optimization.

  18. Application of a simple and affordable protocol for isolating plant total nucleic acids for RNA and DNA virus detection.

    PubMed

    Arruabarrena, Ana; Benítez-Galeano, María José; Giambiasi, Mario; Bertalmío, Ana; Colina, Rodney; Hernández-Rodríguez, Lester

    2016-11-01

    Standard molecular methods for plant virus diagnosis require the purification of RNA or DNA extracts from a large number of samples, with sufficient concentration and quality for their use in PCR, RT-PCR, or qPCR analysis. Most methods are laborious and use either hazardous and/or costly chemicals. A previously published protocol for RNA isolation from several plant species yields high amounts of good quality RNA-DNA mixture in a simple, safe and inexpensive manner. In the present work, this method was tested to obtain RNA-DNA extracts from leaves of tomato, potato and three species of citrus, and was compared with two commercial kits. The results demonstrated that this protocol offers at least comparable nucleic acid quality, quantity and purity to those provided by commercial phenol-based or spin column systems and that are suitable to be used in PCR, RT-PCR and qPCR for virus and viroid detection. Because of its easy implementation and the use of safe and inexpensive reagents, it can be easily implemented to work in plant virus and viroid detection in different plant species. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Cryptanalysis and Improvement for the Quantum Private Comparison Protocol Based on Triplet Entangled State and Single-Particle Measurement

    NASA Astrophysics Data System (ADS)

    Ting, Xu; Tian-Yu, Ye

    2017-03-01

    Quantum private comparison (QPC) aims to accomplish the equality comparison of secret inputs from two users on the basis of not leaking their contents out. Recently, Chen et al. proposed the QPC protocol based on triplet GHZ state and single-particle measurement (Optics Communications 283, 1561-1565 (2010)). In this paper, they suggested the standard model of a semi-honest third party (TP) for the first time, and declared that their protocol is secure. Subsequently, Lin et al. pointed out that in Chen et al.'s protocol, one user can extract the other user's secret without being discovered by performing the intercept-resend attack, and suggested two corresponding improvements (Optics Communications 284, 2412-2414 (2011)). However, Yang et al. first pointed out that the model of TP adopted by both Chen et al.'s protocol and Lin et al.'s improved protocols is unreasonable, and thought that a practical TP may also try any possible means to steal the users' secrets except being corrupted by the adversary including the dishonest user (Quantum Inf Process 12, 877-885 (2013). In this paper, after taking the possible attacks from TP into account, we propose the eavesdropping strategy of TP toward Lin et al.'s improved protocols and suggest two feasible solutions accordingly.

  20. A protocol to improve genotyping of problematic microsatellite loci of Trypanosoma brucei gambiense from body fluids.

    PubMed

    Kaboré, Jacques; De Meeûs, Thierry; Macleod, Annette; Ilboudo, Hamidou; Capewell, Paul; Camara, Mamadou; Gaston Belem, Adrien Marie; Bucheton, Bruno; Jamonneau, Vincent

    2013-12-01

    Microsatellite genotyping of Trypanosoma brucei gambiense, the causative agent of human African trypanosomiasis or sleeping sickness, and population genetics tools, are useful for inferring population parameters such as population size and dispersal. Amplifying parasite DNA directly from body fluids (i.e., blood, lymph or cerebrospinal fluid) allows avoiding costly and tedious isolation phases. It is however associated to increased frequencies of amplification failures (allelic dropouts and/or null alleles) at some loci. In this paper, we present a study focused on three T. brucei gambiense microsatellite loci suspected to present amplification problems when amplified from body fluids sampled in Guinean sleeping sickness foci. We checked for the real nature of blank and apparent homozygous genotypes of parasite DNA directly amplified from body fluids and tested the effect of three different DNA quantities of trypanosomes. Our results show that some initially blank and homozygous genotypes happen to be actual heterozygous genotypes. In Guinea, lymph from the cervical nymph nodes, known to contain the highest concentrations of parasites, appeared to provide the best amplification results. Simply repeating the PCR may be enough to retrieve the correct genotype, but we also show that increasing initial DNA content provides better results while undertaking first amplification. We finally propose an optimal protocol for amplifying trypanosome's DNA directly from body fluids that should be adapted to local characteristics and/or constraints. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. New protocol for the isolation of nitrocellulose from gunpowders: utility in their identification.

    PubMed

    López-López, María; de la Ossa, María Angeles Fernández; Galindo, Jorge Sáiz; Ferrando, Jose Luis; Vega, Alfonso; Torre, Mercedes; García-Ruiz, Carmen

    2010-06-15

    In this work, a new approach for the isolation of nitrocellulose from smokeless gunpowders has been developed. A multistep solvent extraction method was needed to purify nitrocellulose contained in gunpowders. For single-base or double-base gunpowders six consecutive solvent extractions were selected: three extractions with methanol (to remove nitroglycerin, 2,4-dinitrotoluene, ethyl-centralite, diphenylamine, and diphenylamine derivatives); one extraction with dichloromethane (to remove colorants and plasticizers of organic nature); one extraction with methanol (to facilitate a final polar extraction); and one extraction with water (to remove ionic components) were necessary at 35 degrees C. For the triple-base gunpowder studied, eight solvent extractions were needed due to a high concentration of the water-soluble nitroguanidine was present. In addition to the same five initial phases used for the single-base and double-base gunpowders, three water extraction phases at a higher temperature (75 degrees C instead of 35 degrees C) were also needed. A final step to solubilize nitrocellulose in methyl ethyl ketone was used to remove inert components (mainly graphite). Nitrocellulose isolated from these propellants was characterized by Fourier-Transformed Infrared Spectroscopy (FTIR spectroscopy). The same FTIR spectra were observed for nitrocelluloses isolated from different types of gunpowders. A comparison of FTIR spectra of nitrocellulose samples of different nitration degree evidenced that the bands regions most affected by this factor were: 3600-3400cm(-1), corresponding to the stretching vibrations of residual hydroxyl groups; 1200-1000cm(-1), attributed to the valence vibrations nuCO of the glucopyranose cycle; and 750-690cm(-1), assigned to vibrations of the nitrate group. In both cases, the bands appearing in these regions were more pronounced in the spectra of nitrocellulose samples of low nitration degree.

  2. Isolating the Unique Effects of the Unified Protocol Treatment Modules Using Single Case Experimental Design.

    PubMed

    Sauer-Zavala, Shannon; Cassiello-Robbins, Clair; Conklin, Laren R; Bullis, Jacqueline R; Thompson-Hollands, Johanna; Kennedy, Katherine A

    2017-03-01

    The Unified Protocol (UP) for the Transdiagnostic Treatment of Emotional Disorders is a cognitive-behavioral intervention designed to treat the range of anxiety, depressive, and related disorders. Thus far, the UP treatment modules have only been studied when they are delivered in their entirety and presented in a standard sequence. To personalize the presentation of the UP modules for a given patient's presentation (e.g., providing the modules in a varied order, dropping irrelevant modules), it is first necessary to establish that each module leads to change in the skill it is designed to promote, and that these changes can occur in the absence of the other modules. Using a multiple baseline design in accordance with the single-case reporting guidelines in behavioral interventions (SCRIBE), eight patients with heterogeneous emotional disorders were randomly assigned to a 1- or 3-week baseline assessment phase followed by four sessions of one of four UP modules (psychoeducation, emotional awareness, cognitive flexibility, and countering emotional behaviors). Results provide preliminary support for the notion that each UP module under study leads to change in its associated skill in the absence of the other modules (five of eight patients demonstrated reliable change in the module-specific skill). In addition, exploratory analyses suggest that the emotion awareness training and cognitive flexibility modules appeared to exhibit change specific to their associated skills, psychoeducation, and countering emotional behaviors demonstrated somewhat more broad-based change across skills.

  3. Systematic study of cell isolation from bovine nucleus pulposus: Improving cell yield and experiment reliability.

    PubMed

    Lee, Juliana T Y; Cheung, Kenneth M C; Leung, Victor Y L

    2015-12-01

    Differences in matrix compositions in human nucleus pulposus (NP) clinical samples demand different cell isolation protocols for optimal results but there is no clear guide about this to date. Sub-optimal protocols may result in low cell yield, limited reliability of results or even failure of experiments. Cell yield, viability and attachment of cells isolated from bovine NP tissue with different protocols were estimated by cell counting, Trypan blue staining and cell culturing respectively. RNA was extracted from isolated cells and quantified by Nanodrop spectrometry and RT-qPCR. Higher collagenase concentration, longer digestion duration and pronase pre-treatment increased the cell yield. Cell viability remained high (<5% dead cells) even after 0.2% collagenase treatment for overnight. NP cells remained to have high ACAN, COL2A1, CDH2, KRT18, and KRT19 expression compared to muscle cells for different cell isolation conditions tested. Digestion by collagenase alone without the use of pronase could isolate cells from human degenerated NP tissue but clusters of cells were observed. We suggest the use of the disappearance of tissue as an indirect measure of cells released. This study provides a guide for researchers to decide the parameters involved in NP cell isolation for optimal outcome.

  4. Improving the efficiency of single and multiple teleportation protocols based on the direct use of partially entangled states

    SciTech Connect

    Fortes, Raphael; Rigolin, Gustavo

    2013-09-15

    We push the limits of the direct use of partially pure entangled states to perform quantum teleportation by presenting several protocols in many different scenarios that achieve the optimal efficiency possible. We review and put in a single formalism the three major strategies known to date that allow one to use partially entangled states for direct quantum teleportation (no distillation strategies permitted) and compare their efficiencies in real world implementations. We show how one can improve the efficiency of many direct teleportation protocols by combining these techniques. We then develop new teleportation protocols employing multipartite partially entangled states. The three techniques are also used here in order to achieve the highest efficiency possible. Finally, we prove the upper bound for the optimal success rate for protocols based on partially entangled Bell states and show that some of the protocols here developed achieve such a bound. -- Highlights: •Optimal direct teleportation protocols using directly partially entangled states. •We put in a single formalism all strategies of direct teleportation. •We extend these techniques for multipartite partially entangle states. •We give upper bounds for the optimal efficiency of these protocols.

  5. Optimized dissociation protocol for isolating human glioma stem cells from tumorspheres via fluorescence-activated cell sorting.

    PubMed

    Lv, Donglai; Ma, Qing-Hua; Duan, Jiang-Jie; Wu, Hai-Bo; Zhao, Xi-Long; Yu, Shi-Cang; Bian, Xiu-Wu

    2016-07-10

    Fluorescence-activated cell sorting (FACS) based on the surface marker CD133 is the most common method for isolating glioma stem cells (GSCs) from heterogeneous glioma cell populations. Optimization of this method will have profound implications for the future of GSC research. Five commonly used digestion reagents, Liberase-TL, trypsin, TrypLE, Accutase, and non-enzymatic cell dissociation solution (NECDS), were used to dissociate glioma tumorspheres derived from two primary glioma specimens (091214 and 090116) and the cell lines U87 and T98G. The dissociation time, cell viability, retention of CD133, and stemness capacity were assessed. The results showed that single cells derived from the Liberase-TL (200 µg/ml) group exhibited high viability and less damage to the antigen CD133. However, the efficiency of NECDS for dissociating the tumorspheres into single cells was fairly low. Meanwhile, the use of this digestion reagent resulted in obvious cellular and antigenic impairments. Taken together, Liberase-TL (200 µg/ml) is an ideal reagent for isolating GSCs from tumorspheres. In contrast, the use of NECDS for such a protocol should be carefully considered. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  6. Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass

    PubMed Central

    O'Neil, Gregory W.; Williams, John R.; Wilson-Peltier, Julia; Knothe, Gerhard; Reddy, Christopher M.

    2016-01-01

    The need to replace petroleum fuels with alternatives from renewable and more environmentally sustainable sources is of growing importance. Biomass-derived biofuels have gained considerable attention in this regard, however first generation biofuels from edible crops like corn ethanol or soybean biodiesel have generally fallen out of favor. There is thus great interest in the development of methods for the production of liquid fuels from domestic and superior non-edible sources. Here we describe a detailed procedure for the production of a purified biodiesel from the marine microalgae Isochrysis. Additionally, a unique suite of lipids known as polyunsaturated long-chain alkenones are isolated in parallel as potentially valuable coproducts to offset the cost of biodiesel production. Multi-kilogram quantities of Isochrysis are purchased from two commercial sources, one as a wet paste (80% water) that is first dried prior to processing, and the other a dry milled powder (95% dry). Lipids are extracted with hexanes in a Soxhlet apparatus to produce an algal oil ("hexane algal oil") containing both traditional fats (i.e., triglycerides, 46-60% w/w) and alkenones (16-25% w/w). Saponification of the triglycerides in the algal oil allows for separation of the resulting free fatty acids (FFAs) from alkenone-containing neutral lipids. FFAs are then converted to biodiesel (i.e., fatty acid methyl esters, FAMEs) by acid-catalyzed esterification while alkenones are isolated and purified from the neutral lipids by crystallization. We demonstrate that biodiesel from both commercial Isochrysis biomasses have similar but not identical FAME profiles, characterized by elevated polyunsaturated fatty acid contents (approximately 40% w/w). Yields of biodiesel were consistently higher when starting from the Isochrysis wet paste (12% w/w vs. 7% w/w), which can be traced to lower amounts of hexane algal oil obtained from the powdered Isochrysis product. PMID:27404113

  7. Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass.

    PubMed

    O'Neil, Gregory W; Williams, John R; Wilson-Peltier, Julia; Knothe, Gerhard; Reddy, Christopher M

    2016-06-24

    The need to replace petroleum fuels with alternatives from renewable and more environmentally sustainable sources is of growing importance. Biomass-derived biofuels have gained considerable attention in this regard, however first generation biofuels from edible crops like corn ethanol or soybean biodiesel have generally fallen out of favor. There is thus great interest in the development of methods for the production of liquid fuels from domestic and superior non-edible sources. Here we describe a detailed procedure for the production of a purified biodiesel from the marine microalgae Isochrysis. Additionally, a unique suite of lipids known as polyunsaturated long-chain alkenones are isolated in parallel as potentially valuable coproducts to offset the cost of biodiesel production. Multi-kilogram quantities of Isochrysis are purchased from two commercial sources, one as a wet paste (80% water) that is first dried prior to processing, and the other a dry milled powder (95% dry). Lipids are extracted with hexanes in a Soxhlet apparatus to produce an algal oil ("hexane algal oil") containing both traditional fats (i.e., triglycerides, 46-60% w/w) and alkenones (16-25% w/w). Saponification of the triglycerides in the algal oil allows for separation of the resulting free fatty acids (FFAs) from alkenone-containing neutral lipids. FFAs are then converted to biodiesel (i.e., fatty acid methyl esters, FAMEs) by acid-catalyzed esterification while alkenones are isolated and purified from the neutral lipids by crystallization. We demonstrate that biodiesel from both commercial Isochrysis biomasses have similar but not identical FAME profiles, characterized by elevated polyunsaturated fatty acid contents (approximately 40% w/w). Yields of biodiesel were consistently higher when starting from the Isochrysis wet paste (12% w/w vs. 7% w/w), which can be traced to lower amounts of hexane algal oil obtained from the powdered Isochrysis product.

  8. Use of a protocolized approach to the management of sepsis can improve time to first dose of antibiotics.

    PubMed

    Tipler, Pamela S; Pamplin, Jeremy; Mysliwiec, Vincent; Anderson, David; Mount, Cristin A

    2013-04-01

    The Surviving Sepsis Guidelines established recommendations for early recognition and rapid treatment of patients with sepsis. Recognizing systemic difficulties that delayed the application of early goal-directed therapy, the Emergency Department and Critical Care leadership instituted a sepsis protocol to identify patients with sepsis and expedite antibiotic delivery. We aimed to determine if the sepsis protocol improved the time to first dose of antibiotics in patients diagnosed with sepsis. We performed a retrospective chart review of patients with sepsis comparing the time from antibiotic order placement to the first dose of antibiotic therapy over a 3-year period. Patients who received vancomycin and ciprofloxacin underwent additional subgroup analysis, as these antibiotics were made available by protocol for use without infectious disease consultation. The average time to first dose of antibiotics for the presepsis protocol group was 160 minutes, and the average time for the sepsis protocol group was 99 minutes. Fifty-eight patients received vancomycin, and 30 received ciprofloxacin, with a decrease in time of 65 minutes and 41 minutes, respectively. Initiation of a sepsis protocol, which emphasizes early goal-directed therapy, can improve time to administration of first dose of antibiotics. Published by Elsevier Inc.

  9. Early gonadotropin-releasing hormone antagonist start improves follicular synchronization and pregnancy outcome as compared to the conventional antagonist protocol.

    PubMed

    Park, Chan Woo; Hwang, Yu Im; Koo, Hwa Seon; Kang, Inn Soo; Yang, Kwang Moon; Song, In Ok

    2014-12-01

    To assess whether an early GnRH antagonist start leads to better follicular synchronization and an improved clinical pregnancy rate (CPR). A retrospective cohort study. A total of 218 infertile women who underwent IVF between January 2011 and February 2013. The initial cohort (Cohort I) that underwent IVF between January 2011 and March 2012 included a total of 68 attempted IVF cycles. Thirty-four cycles were treated with the conventional GnRH antagonist protocol, and 34 cycles with an early GnRH antagonist start protocol. The second cohort (Cohort II) that underwent IVF between June 2012 and February 2013 included a total of 150 embryo-transfer (ET) cycles. Forty-three cycles were treated with the conventional GnRH antagonist protocol, 34 cycles with the modified early GnRH antagonist start protocol using highly purified human menopause gonadotropin and an addition of GnRH agonist to the luteal phase support, and 73 cycles with the GnRH agonist long protocol. The analysis of Cohort I showed that the number of mature oocytes retrieved was significantly higher in the early GnRH antagonist start cycles than in the conventional antagonist cycles (11.9 vs. 8.2, p=0.04). The analysis of Cohort II revealed higher but non-significant CPR/ET in the modified early GnRH antagonist start cycles (41.2%) than in the conventional antagonist cycles (30.2%), which was comparable to that of the GnRH agonist long protocol cycles (39.7%). The modified early antagonist start protocol may improve the mature oocyte yield, possibly via enhanced follicular synchronization, while resulting in superior CPR as compared to the conventional antagonist protocol, which needs to be studied further in prospective randomized controlled trials.

  10. Dispatcher-assisted cardiopulmonary resuscitation protocol improves diagnosis and resuscitation recommendations for out-of-hospital cardiac arrest.

    PubMed

    Besnier, Emmanuel; Damm, Cedric; Jardel, Benoit; Veber, Benoit; Compere, Vincent; Dureuil, Bertrand

    2015-12-01

    Despite recent efforts, most people are not trained in cardiopulmonary resuscitation (CPR), which has a major impact on survival following cardiac arrest (CA). We have set up a dispatcher-assisted CPR protocol at our call centre, based on international guidelines issued in 2010. The aim of our study was to evaluate the impact of this protocol on CA diagnosis and quantity of recommendations given by telephone dispatchers to untrained witnesses. We performed a 'before and after' monocentric observational study. Data were compared before and a short time after (2 months) implementation of the protocol. We included patients presenting as an out-of-hospital CA in the presence of a witness untrained in CPR. Fisher's test was used to compare periods. P < 0.05 was considered significant. During the 8 month period before the protocol, 115 victims were potentially eligible for CPR. Diagnosis was achieved in 63.5% of cases and CPR recommendations given in 6.1%. After implementation of the protocol, 130 victims were potentially eligible for CPR. Frequency of CA diagnosis was significantly higher after the protocol with 76% of cases (P = 0.0359). Frequency of CPR recommendations given to witnesses was also significantly higher after the protocol, with a fivefold increase up to 29.2% (P < 0.0001). Implementation of a dispatcher-assisted CPR protocol was efficient in improving both CA diagnosis and CPR recommendations given to untrained witnesses for out-of-hospital CA with a very short time of dispatcher training. It is a simple and efficacious measure, at no additional cost and with the promises of improving prognosis following cardiac arrest in a centre not equipped with computerised dispatcher support programmes. © 2015 Australasian College for Emergency Medicine and Australasian Society for Emergency Medicine.

  11. Multisession Anodal tDCS Protocol Improves Motor System Function in an Aging Population

    PubMed Central

    Dumel, G.; Bourassa, M.-E.; Desjardins, M.; Voarino, N.; Charlebois-Plante, C.; Doyon, J.; De Beaumont, Louis

    2016-01-01

    Objectives. The primary objective of this study was to investigate the effects of five consecutive, daily 20-minute sessions of M1 a-tDCS on motor learning in healthy, cognitively intact, aging adults. Design. A total of 23 participants (51 to 69 years old) performed five consecutive, daily 20-minute sessions of a serial reaction time task (SRT task) concomitant with either anodal (n = 12) or sham (n = 11) M1 a-tDCS. Results. We found a significant group × training sessions interaction, indicating that whereas aging adults in the sham group exhibited little-to-no sequence-specific learning improvements beyond the first day of training, reproducible improvements in the ability to learn new motor sequences over 5 consecutive sessions were the net result in age-equivalent participants from the M1 a-tDCS group. A significant main effect of group on sequence-specific learning revealed greater motor learning for the M1 a-tDCS group when the five learning sessions were averaged. Conclusion. These findings raise into prominence the utility of multisession anodal TDCS protocols in combination with motor training to help prevent/alleviate age-associated motor function decline. PMID:26881118

  12. Optimized protocols for improving the likelihood of cloning recombinant progeny from Plasmodium yoelii genetic crosses

    PubMed Central

    Qi, Yanwei; Zhu, Feng; Li, Jian; Fu, Yong; Pattaradilokrat, Sittiporn; Hong, Linxian; Liu, Shengfa; Huang, Fusheng; Xu, Wenyue; Su, Xin-zhuan

    2012-01-01

    Genetic cross is a powerful tool for studying malaria genes contributing to drug resistance, parasite development, and pathogenesis. Cloning and identification of recombinant progeny (RP) is laborious and expensive, especially when a large proportion of progeny derived from self-fertilization are present in the uncloned progeny of a genetic cross. Since the frequency of cross-fertilization affects the number of recombinant progeny in a genetic cross, it is important to optimize the procedure of a genetic cross to maximize the cross-fertilization. Here we investigated the factors that might influence the chances of obtaining RP from a genetic cross and showed that different Plasmodium yoelii strains/subspecies/clones had unique abilities in producing oocysts in a mosquito midgut. When a genetic cross is performed between two parents producing different numbers of functional gametocytes, the ratio of parental parasites must be adjusted to improve the chance of obtaining RP. An optimized parental ratio could be established based on oocyst counts from single infection of each parent before crossing experiments, which may reflect the efficiency of gametocyte production and/or fertilization. The timing of progeny cloning is also important; cloning of genetic cross progeny from mice directly infected with sporozoites (vs. frozen blood after needle passage) at a time when parasitemia is low (usually <1%) could improve the chance of obtaining RP. This study provides an optimized protocol for efficiently cloning RPs from a genetic cross of malaria parasites. PMID:23116600

  13. Improving safety in care homes: protocol for evaluation of the Walsall and Wolverhampton care home improvement programme.

    PubMed

    Damery, Sarah; Flanagan, Sarah; Rai, Kiran; Combes, Gill

    2017-01-26

    Improving safety in care homes is becoming increasingly important. Care home residents typically have multiple physical and/or cognitive impairments, and adverse events like falls often lead to hospital attendance or admission. Developing a safety culture is associated with beneficial impacts on safety outcomes, but the complex needs of care home residents, coupled with staffing pressures in the sector, pose challenges for positive safety practices to become embedded at the individual and organisational levels. Staff training and education can positively enforce safety culture and reduce the incidence of harms, but improvement initiatives are often short lived and thorough evaluation is uncommon. This protocol outlines an evaluation of a large-scale care home improvement programme in the West Midlands. The programme will run in 35 care homes across Walsall and Wolverhampton over 24 months, and we anticipate that 30 care homes will participate in the evaluation (n = 1500 staff). The programme will train staff and managers in service improvement techniques, with the aim of strengthening safety culture and reducing adverse safety event rates. The evaluation will use a pre-post design with mixed methods. Quantitative data will focus on: care home manager and staff surveys administered at several time points and analysis of adverse event rates. Data on hospital activity by residents at participating care homes will be compared to matched controls. Qualitative data on experience of training and the application of learning to practice will be collected via semi-structured interviews with staff (n = 48 to 64) and programme facilitators (n = 6), and staff focus groups (n = 36 to 48 staff). The primary outcome measure is the change in mean score on the safety climate domain of the Safety Attitudes Questionnaire between baseline and programme end. This mixed methods evaluation of a large-scale care home improvement programme will allow a substantial amount of

  14. Single prokaryotic cell isolation and total transcript amplification protocol for transcriptomic analysis

    PubMed Central

    Kang, Yun; McMillan, Ian; Norris, Michael H; Hoang, Tung T.

    2015-01-01

    Until recently, transcriptome analyses of single cells have been confined to eukaryotes. The information obtained from single cell transcripts can provide detailed insight into spatiotemporal gene-expression, and could be even more valuable if expanded to prokaryotic cells. Transcriptome analysis of single prokaryotic cells is a recently developed and powerful tool. Here, we describe a procedure that allows amplification of the total transcript of a single prokaryotic cell for in-depth analysis. This is performed by utilizing a laser capture microdissection instrument for single cell isolation, followed by reverse transcription via Moloney Murine Leukemia virus, degradation of chromosomal DNA with McrBC and DpnI restriction enzymes, ss-cDNA ligation using T4 polynucleotide kinase and CircLigase, and polymerization of ss-cDNA to ds-cDNA by ϕ 29 polymerase. This procedure takes ~5 days, and sufficient amounts of ds-cDNA can be obtained from single cell RNA template for further microarray analysis. PMID:26042386

  15. Single prokaryotic cell isolation and total transcript amplification protocol for transcriptomic analysis.

    PubMed

    Kang, Yun; McMillan, Ian; Norris, Michael H; Hoang, Tung T

    2015-07-01

    Until recently, transcriptome analyses of single cells have been confined to eukaryotes. The information obtained from single-cell transcripts can provide detailed insight into spatiotemporal gene expression, and it could be even more valuable if expanded to prokaryotic cells. Transcriptome analysis of single prokaryotic cells is a recently developed and powerful tool. Here we describe a procedure that allows amplification of the total transcript of a single prokaryotic cell for in-depth analysis. This is performed by using a laser-capture microdissection instrument for single-cell isolation, followed by reverse transcription via Moloney murine leukemia virus, degradation of chromosomal DNA with McrBC and DpnI restriction enzymes, single-stranded cDNA (ss-cDNA) ligation using T4 polynucleotide kinase and CircLigase, and polymerization of ss-cDNA to double-stranded cDNA (ds-cDNA) by Φ29 polymerase. This procedure takes ∼5 d, and sufficient amounts of ds-cDNA can be obtained from single-cell RNA template for further microarray analysis.

  16. Simple and rapid protocol for the isolation of PCR-amplifiable DNA from medicinal plants.

    PubMed

    Alatar, A A; Mahmoud, M A; Al-Sohaibani, S A; Abd-Elsalam, K A

    2012-02-13

    Medicinal plant species has a valuable economic importance because of its usage as pharmaceuticals, nutritional, as well as its use in popular medication. For DNA-based techniques, nanogram quantities of the purified DNA are requisite to amplify and yield sufficient amounts of PCR products. SDS-based DNA isolation method was used to extract DNA from 11 species of different aromatic and medicinal plants collected from Saudi Arabia. Three hundred milligrams of fresh shredded plant material was necessary. The DNA purity was further confirmed by agarose gel, restriction endonuclease digestion and microsatellite primed-polymerase chain reaction (MP-PCR). DNA yields ranged from 10-20 μg (in 100-μL elution volumes) from all plant material evaluated. The DNA obtained was free of any contaminating proteins, polysaccharides and colored pigments. The extracted genomic DNA was found suitable for restriction digestion and PCR amplification. Our experimental procedure provides an easy, suitable, non-toxic, cheap, and quick process for the amplification of DNA from medical plant tissue.

  17. An improved method to isolate lichen algae by gel filtration.

    PubMed

    Pérez, M J; Vicente, C; Legaz, M E

    1985-08-01

    Photobiont cells of the lichen Evernia prunastri have completely been separated from their fungal partner by filtration through a bed of Sepharose 2B. Both mannitol and ribitol have been quantified by gas-liquid chromatography in the different steps of the isolation procedure. Absence of mannitol, which is exclusively produced by the mycobiont, has been used as the best probe to monitor isolation.

  18. Improving credibility assessment in child sexual abuse allegations: the role of the NICHD investigative interview protocol.

    PubMed

    Hershkowitz, Irit; Fisher, Sara; Lamb, Michael E; Horowitz, Dvora

    2007-02-01

    The study was designed to explore whether the credibility of children's statements regarding their alleged experiences of child sexual abuse could be assessed in a more valid and reliable way when investigative interviews were conducted using the NICHD protocol rather than in an unstructured manner. Forty-two experienced Israeli youth investigators each assessed the credibility of allegations of sexual abuse made by alleged victims of sexual abuse when interviewed either with or without the protocol. Half of the alleged incidents were judged likely to have happened ("plausible") on the basis of independent evidence, while half were deemed unlikely to have happened ("implausible"). More non-protocol than protocol interviews were rated as "No judgment possible" rather than either credible or incredible. Allegations made in protocol interviews were more accurately rated as credible or incredible when they were either plausible or implausible, respectively, than those made in non-protocol statements. Levels of inter-rater reliability were also higher when protocol interviews were rated. The differences were significant only for plausible cases, however. The use of the NICHD protocol facilitated the assessment of credibility by child investigators although incredible allegations (those describing incidents that were unlikely to have happened) remained difficult to detect, even when the protocol was used.

  19. Learning from positively deviant wards to improve patient safety: an observational study protocol.

    PubMed

    Baxter, Ruth; Taylor, Natalie; Kellar, Ian; Lawton, Rebecca

    2015-12-11

    Positive deviance is an asset-based approach to improvement which has recently been adopted to improve quality and safety within healthcare. The approach assumes that solutions to problems already exist within communities. Certain groups or individuals identify these solutions and succeed despite having the same resources as others. Within healthcare, positive deviance has previously been applied at individual or organisational levels to improve specific clinical outcomes or processes of care. This study explores whether the positive deviance approach can be applied to multidisciplinary ward teams to address the broad issue of patient safety among elderly patients. Preliminary work analysed National Health Service (NHS) Safety Thermometer data from 34 elderly medical wards to identify 5 'positively deviant' and 5 matched 'comparison' wards. Researchers are blinded to ward status. This protocol describes a multimethod, observational study which will (1) assess the concurrent validity of identifying positively deviant elderly medical wards using NHS Safety Thermometer data and (2) generate hypotheses about how positively deviant wards succeed. Patient and staff perceptions of safety will be assessed on each ward using validated surveys. Correlation and ranking analyses will explore whether this survey data aligns with the routinely collected NHS Safety Thermometer data. Staff focus groups and researcher fieldwork diaries will be completed and qualitative thematic content analysis will be used to generate hypotheses about the strategies, behaviours, team cultures and dynamics that facilitate the delivery of safe patient care. The acceptability and sustainability of strategies identified will also be explored. The South East Scotland Research Ethics Committee 01 approved this study (reference: 14/SS/1085) and NHS Permissions were granted from all trusts. Findings will be published in peer-reviewed, scientific journals, and presented at academic conferences. This study

  20. Learning from positively deviant wards to improve patient safety: an observational study protocol

    PubMed Central

    Baxter, Ruth; Taylor, Natalie; Kellar, Ian; Lawton, Rebecca

    2015-01-01

    Introduction Positive deviance is an asset-based approach to improvement which has recently been adopted to improve quality and safety within healthcare. The approach assumes that solutions to problems already exist within communities. Certain groups or individuals identify these solutions and succeed despite having the same resources as others. Within healthcare, positive deviance has previously been applied at individual or organisational levels to improve specific clinical outcomes or processes of care. This study explores whether the positive deviance approach can be applied to multidisciplinary ward teams to address the broad issue of patient safety among elderly patients. Methods and analysis Preliminary work analysed National Health Service (NHS) Safety Thermometer data from 34 elderly medical wards to identify 5 ‘positively deviant’ and 5 matched ‘comparison’ wards. Researchers are blinded to ward status. This protocol describes a multimethod, observational study which will (1) assess the concurrent validity of identifying positively deviant elderly medical wards using NHS Safety Thermometer data and (2) generate hypotheses about how positively deviant wards succeed. Patient and staff perceptions of safety will be assessed on each ward using validated surveys. Correlation and ranking analyses will explore whether this survey data aligns with the routinely collected NHS Safety Thermometer data. Staff focus groups and researcher fieldwork diaries will be completed and qualitative thematic content analysis will be used to generate hypotheses about the strategies, behaviours, team cultures and dynamics that facilitate the delivery of safe patient care. The acceptability and sustainability of strategies identified will also be explored. Ethics and dissemination The South East Scotland Research Ethics Committee 01 approved this study (reference: 14/SS/1085) and NHS Permissions were granted from all trusts. Findings will be published in peer

  1. A History of the Improvement of Internet Protocols Over Satellites Using ACTS

    NASA Technical Reports Server (NTRS)

    Allman, Mark; Kruse, Hans; Ostermann, Shawn

    2000-01-01

    This paper outlines the main results of a number of ACTS experiments on the efficacy of using standard Internet protocols over long-delay satellite channels. These experiments have been jointly conducted by NASAs Glenn Research Center and Ohio University over the last six years. The focus of our investigations has been the impact of long-delay networks with non-zero bit-error rates on the performance of the suite of Internet protocols. In particular, we have focused on the most widely used transport protocol, the Transmission Control Protocol (TCP), as well as several application layer protocols. This paper presents our main results, as well as references to more verbose discussions of our experiments.

  2. Study protocol: mobile improvement of self-management ability through rural technology (mI SMART).

    PubMed

    Mallow, Jennifer A; Theeke, Laurie A; Long, Dustin M; Whetsel, Tara; Theeke, Elliott; Mallow, Brian K

    2015-01-01

    There are 62 million Americans currently residing in rural areas who are more likely to have multiple chronic conditions and be economically disadvantaged, and in poor health, receive less recommended preventive services and attend fewer visits to health care providers. Recent advances in mobile healthcare (mHealth) offer a promising new approach to solving health disparities and improving chronic illness care. It is now possible and affordable to transmit health information, including values from glucometers, automated blood pressure monitors, and scales, through Bluetooth-enabled devices. Additionally, audio and video communications technologies can allow healthcare providers to conduct many parts of a physical exam remotely from varied settings. These technologies could remove geographical distance as a barrier to care and diminish the access to care issues faced by patients who live rurally. However, currently there is lack of studies that provide evidence of feasibility, acceptability, and effectiveness of mHealth initiatives on improved outcomes of care, a needed step to make the translation to implementation studies in healthcare systems. The purpose of this paper is to present the protocol for the first study of mI SMART (mobile Improvement of Self-Management Ability through Rural Technology), a new integrated mHealth intervention. Our objective is to provide evidence of feasibility and acceptability for the use of mI SMART in an underserved population and establish evidence for the refinement of mI SMART. The proposed study will take place at Milan Puskar Health Right, a free primary care clinic in the state of West Virginia. The clinic provides health care at no cost to uninsured, low income; adults aged 18-64 living in West Virginia. We will enroll 30 participants into this feasibility study with plans of implementing a longitudinal randomized, comparative effectiveness design in the future. Data collection will include tracking of barriers and

  3. Vibration isolation for line of sight performance improvement

    NASA Technical Reports Server (NTRS)

    Rodden, J. J.; Dougherty, H. J.; Haile, W. B.

    1987-01-01

    Diagrams of the Reaction Wheel Assembly (RWA) are presented along with charts and graphs illustrating jitter error model, induced vibration tests, radial displacement transfer function, and axial displacement power spectra density. The RWA isolator specification requirements are listed.

  4. From Human Monocytes to Genome-Wide Binding Sites - A Protocol for Small Amounts of Blood: Monocyte Isolation/ChIP-Protocol/Library Amplification/Genome Wide Computational Data Analysis

    PubMed Central

    Weiterer, Sebastian; Uhle, Florian; Bhuju, Sabin; Jarek, Michael; Weigand, Markus A.; Bartkuhn, Marek

    2014-01-01

    Chromatin immunoprecipitation in combination with a genome-wide analysis via high-throughput sequencing is the state of the art method to gain genome-wide representation of histone modification or transcription factor binding profiles. However, chromatin immunoprecipitation analysis in the context of human experimental samples is limited, especially in the case of blood cells. The typically extremely low yields of precipitated DNA are usually not compatible with library amplification for next generation sequencing. We developed a highly reproducible protocol to present a guideline from the first step of isolating monocytes from a blood sample to analyse the distribution of histone modifications in a genome-wide manner. Conclusion: The protocol describes the whole work flow from isolating monocytes from human blood samples followed by a high-sensitivity and small-scale chromatin immunoprecipitation assay with guidance for generating libraries compatible with next generation sequencing from small amounts of immunoprecipitated DNA. PMID:24732314

  5. A childhood chemotherapy protocol improves overall survival among adults with T-lymphoblastic lymphoma

    PubMed Central

    Xia, Zhong-jun; Chen, Xiao-qin; Geng, Qi-rong; Wang, Wei-da; Wang, Liang; Lu, Yue

    2016-01-01

    A broadly accepted standard treatment for adult T-lymphoblastic lymphoma (T-LBL) has not yet been defined. To address that issue, we retrospectively compared three chemotherapy regimens used to treat 110 adult patients with newly diagnosed T-LBL. These included two adult regimens (ECOG2993 and hyper-CVAD) and a childhood regimen (BFM-90). These intensive drug regimens are mainly used to treat childhood and adult acute lymphoblastic leukemia. They included induction, consolidation, and maintenance chemotherapy protocols and were administered over the course of 2 years. Seventy-five patients (80%) achieved a complete remission (CR). Within a median follow-up time of 31 months (range: 5–152 months), the 5-year overall survival (OS) and progression-free survival (PFS) rates were 47.7% (95% CI, 35.0–69.8%) and 45.7% (95% CI, 27.6–56.6%), respectively. Shorter survival was associated with age > 40 years, poor ECOG PS and bone marrow involvement. Elevated lactic dehydrogenase (LDH) level, Ann Arbor stage and International Prognostic Index (IPI) score had no prognostic value. The childhood chemotherapy regimen improved CR and the overall survival rate more than the adult regimen in patients aged < 40 years. PMID:27150061

  6. An improved control mode for the ping-pong protocol operation in imperfect quantum channels

    NASA Astrophysics Data System (ADS)

    Zawadzki, Piotr

    2015-07-01

    Quantum direct communication (QDC) can bring confidentiality of sensitive information without any encryption. A ping-pong protocol, a well-known example of entanglement-based QDC, offers asymptotic security in a perfect quantum channel. However, it has been shown (Wójcik in Phys Rev Lett 90(15):157901, 2003. doi:10.1103/PhysRevLett.90.157901) that it is not secure in the presence of losses. Moreover, legitimate parities cannot rely on dense information coding due to possible undetectable eavesdropping even in the perfect setting (Pavičić in Phys Rev A 87(4):042326, 2013. doi:10.1103/PhysRevA.87.042326). We have identified the source of the above-mentioned weaknesses in the incomplete check of the EPR pair coherence. We propose an improved version of the control mode, and we discuss its relation to the already-known attacks that undermine the QDC security. It follows that the new control mode detects these attacks with high probability and independently on a quantum channel type. As a result, an asymptotic security of the QDC communication can be maintained for imperfect quantum channels, also in the regime of dense information coding.

  7. On the Improvement of Free-Energy Calculation from Steered Molecular Dynamics Simulations Using Adaptive Stochastic Perturbation Protocols

    PubMed Central

    Perišić, Ognjen; Lu, Hui

    2014-01-01

    The potential of mean force (PMF) calculation in single molecule manipulation experiments performed via the steered molecular dynamics (SMD) technique is a computationally very demanding task because the analyzed system has to be perturbed very slowly to be kept close to equilibrium. Faster perturbations, far from equilibrium, increase dissipation and move the average work away from the underlying free energy profile, and thus introduce a bias into the PMF estimate. The Jarzynski equality offers a way to overcome the bias problem by being able to produce an exact estimate of the free energy difference, regardless of the perturbation regime. However, with a limited number of samples and high dissipation the Jarzynski equality also introduces a bias. In our previous work, based on the Brownian motion formalism, we introduced three stochastic perturbation protocols aimed at improving the PMF calculation with the Jarzynski equality in single molecule manipulation experiments and analogous computer simulations. This paper describes the PMF reconstruction results based on full-atom molecular dynamics simulations, obtained with those three protocols. We also want to show that the protocols are applicable with the second-order cumulant expansion formula. Our protocols offer a very noticeable improvement over the simple constant velocity pulling. They are able to produce an acceptable estimate of PMF with a significantly reduced bias, even with very fast perturbation regimes. Therefore, the protocols can be adopted as practical and efficient tools for the analysis of mechanical properties of biological molecules. PMID:25232859

  8. Clinical Guidelines for the Use of Buprenorphine in the Treatment of Opioid Addiction. Treatment Improvement Protocol (TIP) Series 40

    ERIC Educational Resources Information Center

    Boone, Margaret; Brown, Nancy J.; Moon, Mary A.; Schuman, Deborah J.; Thomas, Josephine; Wright, Denise L.

    2004-01-01

    This Treatment Improvement Protocol (TIP) addresses the clinical use of buprenorphine in the treatment of opioid addiction. TIPs are best-practice guidelines for the treatment of substance use disorders that make the latest research in substance abuse treatment available to counselors and educators. The content was generated by a panel of experts…

  9. Improvement of parameters of freezing medium and freezing protocol for bull sperm using two osmotic supports.

    PubMed

    Chaveiro, A; Machado, L; Frijters, A; Engel, B; Woelders, H

    2006-06-01

    The aim of this study was to improve the freezing protocol of bull sperm, by investigating the influence on sperm viability after freeze/thawing of different freezing medium components, as well as the effect of cooling rates in the different stages of the cooling protocol, in single factor experiments. The experimental variables were: (1) salt-based versus a sugar-based medium (Tris versus sucrose); (2) glycerol concentration; (3) detergent (Equex) concentration; (4) presence of bicarbonate; (5) rate of cooling from 22 degrees C to holding temperature (CR1); (6) holding temperature (HT); (7) rate of cooling from holding temperature to -6 degrees C (CR2); (8) rate of cooling from -10 to -100 degrees C (CR3). All experiments were performed using five bulls per experiment (three ejaculates per bull). Sperm motility after freezing and thawing was assessed by CASA system, and sperm membrane integrity was assessed by flow cytometry. Sucrose-based medium did not offer a clear significant benefit compared to Tris medium. The concentration of Equex that gave the best results in Tris-based media group and sucrose-based media group was in a range between 2-7 and 4-7 g/l, respectively. In both media groups, a glycerol concentration of 800 mM was the best in any post-thaw viability parameters. In the Tris media group, the presence of bicarbonate had a negative effect on sperm viability. CR1 and CR2 had no significant effect on any of the post-thaw sperm viability parameters, but a CR1=0.2 degrees C/min and CR2=4 degrees C/min appeared to give better results in both media. The holding temperature (HT) that gave the best results was found to be in the range of 5-9 degrees C. There was a significant disadvantage of using a low CR3 of 10 degrees C/min, while 150 degrees C/min appeared to be the best cooling rate for either medium.

  10. The Agricultural Model Intercomparison and Improvement Project (AgMIP): Protocols and Pilot Studies

    NASA Technical Reports Server (NTRS)

    Rosenzweig, C.; Jones, J. W.; Hatfield, J. L.; Ruane, A. C.; Boote, K. J.; Thorburn, P.; Antle, J. M.; Nelson, G. C.; Porter, C.; Janssen, S.; hide

    2012-01-01

    The Agricultural Model Intercomparison and Improvement Project (AgMIP) is a major international effort linking the climate, crop, and economic modeling communities with cutting-edge information technology to produce improved crop and economic models and the next generation of climate impact projections for the agricultural sector. The goals of AgMIP are to improve substantially the characterization of world food security due to climate change and to enhance adaptation capacity in both developing and developed countries. Analyses of the agricultural impacts of climate variability and change require a transdisciplinary effort to consistently link state-of-the-art climate scenarios to crop and economic models. Crop model outputs are aggregated as inputs to regional and global economic models to determine regional vulnerabilities, changes in comparative advantage, price effects, and potential adaptation strategies in the agricultural sector. Climate, Crop Modeling, Economics, and Information Technology Team Protocols are presented to guide coordinated climate, crop modeling, economics, and information technology research activities around the world, along with AgMIP Cross-Cutting Themes that address uncertainty, aggregation and scaling, and the development of Representative Agricultural Pathways (RAPs) to enable testing of climate change adaptations in the context of other regional and global trends. The organization of research activities by geographic region and specific crops is described, along with project milestones. Pilot results demonstrate AgMIP's role in assessing climate impacts with explicit representation of uncertainties in climate scenarios and simulations using crop and economic models. An intercomparison of wheat model simulations near Obregón, Mexico reveals inter-model differences in yield sensitivity to [CO2] with model uncertainty holding approximately steady as concentrations rise, while uncertainty related to choice of crop model increases with

  11. Improving glycopeptide synthesis: a convenient protocol for the preparation of beta-glycosylamines and the synthesis of glycopeptides.

    PubMed

    Hackenberger, Christian P R; O'Reilly, Mary K; Imperiali, Barbara

    2005-04-29

    [reaction: see text] Herein we apply a recently introduced protocol using ammonium carbamate in methanol to the amination of crude chitobiose leading to 1,beta-aminochitobiose. This simple, one-step procedure allows a facile preparation of unstable glycosylamines in contrast to the commonly implemented ammonium bicarbonate based amination of water-soluble carbohydrates. The new amination protocol leads to an improved synthesis of the key chitobiosyl-asparagine building block for the SPPS of glycopeptides. The utility of the method is demonstrated with the synthesis of a 39-amino acid glycoprotein.

  12. Innovative Approaches Using Lichen Enriched Media to Improve Isolation and Culturability of Lichen Associated Bacteria.

    PubMed

    Biosca, Elena G; Flores, Raquel; Santander, Ricardo D; Díez-Gil, José Luis; Barreno, Eva

    2016-01-01

    Lichens, self-supporting mutualistic associations between a fungal partner and one or more photosynthetic partners, also harbor non-photosynthetic bacteria. The diversity and contribution of these bacteria to the functioning of lichen symbiosis have recently begun to be studied, often by culture-independent techniques due to difficulties in their isolation and culture. However, culturing as yet unculturable lichenic bacteria is critical to unravel their potential functional roles in lichen symbiogenesis, to explore and exploit their biotechnological potential and for the description of new taxa. Our objective was to improve the recovery of lichen associated bacteria by developing novel isolation and culture approaches, initially using the lichen Pseudevernia furfuracea. We evaluated the effect of newly developed media enriched with novel lichen extracts, as well as the influence of thalli washing time and different disinfection and processing protocols of thalli. The developed methodology included: i) the use of lichen enriched media to mimic lichen nutrients, supplemented with the fungicide natamycin; ii) an extended washing of thalli to increase the recovery of ectolichenic bacteria, thus allowing the disinfection of thalli to be discarded, hence enhancing endolichenic bacteria recovery; and iii) the use of an antioxidant buffer to prevent or reduce oxidative stress during thalli disruption. The optimized methodology allowed significant increases in the number and diversity of culturable bacteria associated with P. furfuracea, and it was also successfully applied to the lichens Ramalina farinacea and Parmotrema pseudotinctorum. Furthermore, we provide, for the first time, data on the abundance of culturable ecto- and endolichenic bacteria that naturally colonize P. furfuracea, R. farinacea and P. pseudotinctorum, some of which were only able to grow on lichen enriched media. This innovative methodology is also applicable to other microorganisms inhabiting these

  13. Innovative Approaches Using Lichen Enriched Media to Improve Isolation and Culturability of Lichen Associated Bacteria

    PubMed Central

    Biosca, Elena G.; Flores, Raquel; Santander, Ricardo D.; Díez-Gil, José Luis; Barreno, Eva

    2016-01-01

    Lichens, self-supporting mutualistic associations between a fungal partner and one or more photosynthetic partners, also harbor non-photosynthetic bacteria. The diversity and contribution of these bacteria to the functioning of lichen symbiosis have recently begun to be studied, often by culture-independent techniques due to difficulties in their isolation and culture. However, culturing as yet unculturable lichenic bacteria is critical to unravel their potential functional roles in lichen symbiogenesis, to explore and exploit their biotechnological potential and for the description of new taxa. Our objective was to improve the recovery of lichen associated bacteria by developing novel isolation and culture approaches, initially using the lichen Pseudevernia furfuracea. We evaluated the effect of newly developed media enriched with novel lichen extracts, as well as the influence of thalli washing time and different disinfection and processing protocols of thalli. The developed methodology included: i) the use of lichen enriched media to mimic lichen nutrients, supplemented with the fungicide natamycin; ii) an extended washing of thalli to increase the recovery of ectolichenic bacteria, thus allowing the disinfection of thalli to be discarded, hence enhancing endolichenic bacteria recovery; and iii) the use of an antioxidant buffer to prevent or reduce oxidative stress during thalli disruption. The optimized methodology allowed significant increases in the number and diversity of culturable bacteria associated with P. furfuracea, and it was also successfully applied to the lichens Ramalina farinacea and Parmotrema pseudotinctorum. Furthermore, we provide, for the first time, data on the abundance of culturable ecto- and endolichenic bacteria that naturally colonize P. furfuracea, R. farinacea and P. pseudotinctorum, some of which were only able to grow on lichen enriched media. This innovative methodology is also applicable to other microorganisms inhabiting these

  14. Butterfly survival on an isolated island by improved grip.

    PubMed

    Duplouy, Anne; Hanski, Ilkka

    2013-04-23

    On small isolated islands, natural selection is expected to reduce the dispersal capacity of organisms, as short distances do not require a high rate of dispersal, which might lead to accidental emigration from the population. In addition, individuals foregoing the high cost of maintaining flight capacity may instead allocate resources to other functions. However, in butterflies and many other insects, flight is necessary not only for dispersal but also for most other activities. A weakly flying individual would probably do worse and have an elevated rather than reduced probability of accidental emigration. Here, we report results consistent with the hypothesis that a butterfly population on an isolated island, instead of having lost its flight capacity, has evolved better grip to resist the force of wind and to avoid being blown off the island. Our study suggests that local adaptation has occurred in this population in spite of its very small size (Ne ∼ 100), complete isolation, low genetic variation and high genetic load.

  15. Communications protocol

    NASA Technical Reports Server (NTRS)

    Zhou, Xiaoming (Inventor); Baras, John S. (Inventor)

    2010-01-01

    The present invention relates to an improved communications protocol which increases the efficiency of transmission in return channels on a multi-channel slotted Alohas system by incorporating advanced error correction algorithms, selective retransmission protocols and the use of reserved channels to satisfy the retransmission requests.

  16. Improving adherence to web-based cessation programs: a randomized controlled trial study protocol

    PubMed Central

    2013-01-01

    Background Reducing smoking prevalence is a public health priority that can save more lives and money than almost any other known preventive intervention. Internet interventions have the potential for enormous public health impact given their broad reach and effectiveness. However, most users engage only minimally with even the best designed websites, diminishing their impact due to an insufficient ‘dose’. Two approaches to improve adherence to Internet cessation programs are integrating smokers into an online social network and providing free nicotine replacement therapy (NRT). Active participation in online communities is associated with higher rates of cessation. Integrating smokers into an online social network can increase support and may also increase utilization of cessation tools and NRT. Removing barriers to NRT may increase uptake and adherence, and may also increase use of online cessation tools as smokers look for information and support while quitting. The combination of both strategies may exert the most powerful effects on adherence compared to either strategy alone. Methods/Design This study compares the efficacy of a smoking cessation website (WEB) alone and in conjunction with free NRT and a social network (SN) protocol designed to integrate participants into the online community. Using a 2 (SN, no SN) x 2 (NRT, no NRT) randomized, controlled factorial design with repeated measures at baseline, 3 months, and 9 months, this study will recruit N = 4,000 new members of an internet cessation program and randomize them to: 1) WEB, 2) WEB + SN, 3) WEB + NRT, or 4) WEB + SN + NRT. Hypotheses are that all interventions will outperform WEB and that WEB + SN + NRT will outperform WEB + NRT and WEB + SN on 30-day point prevalence abstinence at 9 months. Exploratory analyses will examine theory-driven hypotheses about the mediators and moderators of outcome. Discussion Addressing adherence in internet cessation programs is critical and timely to leverage

  17. Isolation of 1E4 IgM Anti-Fasciola hepatica Rediae Monoclonal Antibody from Ascites: Comparison of Two Purification Protocols.

    PubMed

    Alba, Annia; Marcet, Ricardo; Otero, Oscar; Hernández, Hilda M; Figueredo, Mabel; Sarracent, Jorge

    2016-02-01

    Purification of immunoglobulin M (IgM) antibodies could be challenging, and is often characterized by the optimization of the purification protocol to best suit the particular features of the molecule. Here, two different schemes were compared to purify, from ascites, the 1E4 IgM monoclonal antibody (mAb) previously raised against the stage of redia of the trematode Fasciola hepatica. This immunoglobulin is used as capture antibody in an immunoenzymatic assay to detect parasite ongoing infection in its intermediate hosts. The first purification protocol of the 1E4 mAb involved two chromatographic steps: an affinity chromatography on a Concanavalin A matrix followed by size exclusion chromatography. An immunoaffinity chromatography was selected as the second protocol for one-step purification of the antibody using the crude extract of adult parasites coupled to a commercial matrix. Immunoreactivity of the fractions during purification schemes was assessed by indirect immunoenzymatic assays against the crude extract of F. hepatica rediae, while purity was estimated by protein electrophoresis. Losses on the recovery of the antibody isolated by the first purification protocol occurred due to protein precipitation during the concentration of the sample and to low resolution of the size exclusion molecular chromatography step regarding this particular immunoglobulin. The immunoaffinity chromatography using F. hepatica antigens as ligands proved to be the most suitable protocol yielding a pure and immunoreactive antibody. The purification protocols used are discussed regarding efficiency and difficulties.

  18. Improving the performance of community health workers in humanitarian emergencies: a realist evaluation protocol for the PIECES programme

    PubMed Central

    Gilmore, Brynne; Adams, Ben Jack; Bartoloni, Alex; Alhaydar, Bana; McAuliffe, Eilish; Raven, Joanna; Taegtmeyer, Miriam; Vallières, Frédérique

    2016-01-01

    Introduction Understanding what enhances the motivation and performance of community health workers (CHWs) in humanitarian emergencies represents a key research gap within the field of human resources for health. This paper presents the research protocol for the Performance ImprovEment of CHWs in Emergency Settings (PIECES) research programme. Enhancing Learning and Research in Humanitarian Action (ELRHA) funded the development of this protocol as part of their Health in Humanitarian Crises (R2HC) call (No.19839). PIECES aims to understand what factors improve the performance of CHWs in level III humanitarian emergencies. Methods and analysis The suggested protocol uses a realist evaluation with multiple cases across the 3 country sites: Turkey, Iraq and Lebanon. Working with International Medical Corps (IMC), an initial programme theory was elicited through literature and document reviews, semistructured interviews and focus groups with IMC programme managers and CHWs. Based on this initial theory, this protocol proposes a combination of semistructured interviews, life histories and critical incident narratives, surveys and latent variable modelling of key constructs to explain how contextual factors work to trigger mechanisms for specific outcomes relating to IMC's 300+ CHWs' performance. Participants will also include programme staff, CHWs and programme beneficiaries. Realist approaches will be used to better understand ‘what works, for whom and under what conditions’ for improving CHW performance within humanitarian contexts. Ethics and dissemination Trinity College Dublin's Health Policy and Management/Centre for Global Health Research Ethics Committee gave ethical approval for the protocol development phase. For the full research project, additional ethical approval will be sought from: Université St. Joseph (Lebanon), the Ethics Committee of the Ministry of Health in Baghdad (Iraq) and the Middle East Technical University (Turkey). Dissemination

  19. Improving the performance of community health workers in humanitarian emergencies: a realist evaluation protocol for the PIECES programme.

    PubMed

    Gilmore, Brynne; Adams, Ben Jack; Bartoloni, Alex; Alhaydar, Bana; McAuliffe, Eilish; Raven, Joanna; Taegtmeyer, Miriam; Vallières, Frédérique

    2016-08-16

    Understanding what enhances the motivation and performance of community health workers (CHWs) in humanitarian emergencies represents a key research gap within the field of human resources for health. This paper presents the research protocol for the Performance ImprovEment of CHWs in Emergency Settings (PIECES) research programme. Enhancing Learning and Research in Humanitarian Action (ELRHA) funded the development of this protocol as part of their Health in Humanitarian Crises (R2HC) call (No.19839). PIECES aims to understand what factors improve the performance of CHWs in level III humanitarian emergencies. The suggested protocol uses a realist evaluation with multiple cases across the 3 country sites: Turkey, Iraq and Lebanon. Working with International Medical Corps (IMC), an initial programme theory was elicited through literature and document reviews, semistructured interviews and focus groups with IMC programme managers and CHWs. Based on this initial theory, this protocol proposes a combination of semistructured interviews, life histories and critical incident narratives, surveys and latent variable modelling of key constructs to explain how contextual factors work to trigger mechanisms for specific outcomes relating to IMC's 300+ CHWs' performance. Participants will also include programme staff, CHWs and programme beneficiaries. Realist approaches will be used to better understand 'what works, for whom and under what conditions' for improving CHW performance within humanitarian contexts. Trinity College Dublin's Health Policy and Management/Centre for Global Health Research Ethics Committee gave ethical approval for the protocol development phase. For the full research project, additional ethical approval will be sought from: Université St. Joseph (Lebanon), the Ethics Committee of the Ministry of Health in Baghdad (Iraq) and the Middle East Technical University (Turkey). Dissemination activities will involve a mixture of research feedback, policy briefs

  20. Cryptanalysis and improvement of authentication and key agreement protocols for telecare medicine information systems.

    PubMed

    Islam, S K Hafizul; Khan, Muhammad Khurram

    2014-10-01

    Recently, many authentication protocols have been presented using smartcard for the telecare medicine information system (TMIS). In 2014, Xu et al. put forward a two-factor mutual authentication with key agreement protocol using elliptic curve cryptography (ECC). However, the authors have proved that the protocol is not appropriate for practical use as it has many problems (1) it fails to achieve strong authentication in login and authentication phases; (2) it fails to update the password correctly in the password change phase; (3) it fails to provide the revocation of lost/stolen smartcard; and (4) it fails to protect the strong replay attack. We then devised an anonymous and provably secure two-factor authentication protocol based on ECC. Our protocol is analyzed with the random oracle model and demonstrated to be formally secured against the hardness assumption of computational Diffie-Hellman problem. The performance evaluation demonstrated that our protocol outperforms from the perspective of security, functionality and computation costs over other existing designs.

  1. Study of accent-based music speech protocol development for improving voice problems in stroke patients with mixed dysarthria.

    PubMed

    Kim, Soo Ji; Jo, Uiri

    2013-01-01

    Based on the anatomical and functional commonality between singing and speech, various types of musical elements have been employed in music therapy research for speech rehabilitation. This study was to develop an accent-based music speech protocol to address voice problems of stroke patients with mixed dysarthria. Subjects were 6 stroke patients with mixed dysarthria and they received individual music therapy sessions. Each session was conducted for 30 minutes and 12 sessions including pre- and post-test were administered for each patient. For examining the protocol efficacy, the measures of maximum phonation time (MPT), fundamental frequency (F0), average intensity (dB), jitter, shimmer, noise to harmonics ratio (NHR), and diadochokinesis (DDK) were compared between pre and post-test and analyzed with a paired sample t-test. The results showed that the measures of MPT, F0, dB, and sequential motion rates (SMR) were significantly increased after administering the protocol. Also, there were statistically significant differences in the measures of shimmer, and alternating motion rates (AMR) of the syllable /K$\\inve$/ between pre- and post-test. The results indicated that the accent-based music speech protocol may improve speech motor coordination including respiration, phonation, articulation, resonance, and prosody of patients with dysarthria. This suggests the possibility of utilizing the music speech protocol to maximize immediate treatment effects in the course of a long-term treatment for patients with dysarthria.

  2. Improving preterm infant outcomes: implementing an evidence-based oral feeding advancement protocol in the neonatal intensive care unit.

    PubMed

    Kish, Mary Z

    2014-10-01

    The ability of a preterm infant to exclusively oral feed is a necessary standard for discharge readiness from the neonatal intensive care unit (NICU). Many of the interventions related to oral feeding advancement currently employed for preterm infants in the NICU are based on individual nursing observations and judgment. Studies involving standardized feeding protocols for oral feeding advancement have been shown to decrease variability in feeding practices, facilitate shortened transition times from gavage to oral feedings, improve bottle feeding performance, and significantly decrease the length of stay (LOS) in the NICU. This project critically evaluated the implementation of an oral feeding advancement protocol in a 74-bed level III NICU in an attempt to standardize the process of advancing oral feedings in medically stable preterm infants. A comprehensive review of the literature identified key features for successful oral feeding in preterm infants. Strong levels of evidence suggested an association between both nonnutritive sucking (NNS) opportunities and standardized feeding advancement protocols with successful oral feeding in preterm infants. These findings prompted a pilot practice change using a feeding advancement protocol and consisted of NNS and standardized oral feeding advancement opportunities. Time to exclusive oral feedings and LOS were compared pre- and postprotocol implementation during more than a 2-month evaluation period. Infants using NNS and the standardized oral feeding advancement protocol had an observed reduction in time to exclusive oral feedings and LOS, although statistical significance was not achieved.

  3. Improved protocol for rapid identification of certain spa types using high resolution melting curve analysis.

    PubMed

    Mayerhofer, Benjamin; Stöger, Anna; Pietzka, Ariane T; Fernandez, Haizpea Lasa; Prewein, Bernhard; Sorschag, Sieglinde; Kunert, Renate; Allerberger, Franz; Ruppitsch, Werner

    2015-01-01

    Methicillin-resistant Staphylococcus aureus is one of the most significant pathogens associated with health care. For efficient surveillance, control and outbreak investigation, S. aureus typing is essential. A high resolution melting curve analysis was developed and evaluated for rapid identification of the most frequent spa types found in an Austrian hospital consortium covering 2,435 beds. Among 557 methicillin-resistant Staphylococcus aureus isolates 38 different spa types were identified by sequence analysis of the hypervariable region X of the protein A gene (spa). Identification of spa types through their characteristic high resolution melting curve profiles was considerably improved by double spiking with genomic DNA from spa type t030 and spa type t003 and allowed unambiguous and fast identification of the ten most frequent spa types t001 (58%), t003 (12%), t190 (9%), t041 (5%), t022 (2%), t032 (2%), t008 (2%), t002 (1%), t5712 (1%) and t2203 (1%), representing 93% of all isolates within this hospital consortium. The performance of the assay was evaluated by testing samples with unknown spa types from the daily routine and by testing three different high resolution melting curve analysis real-time PCR instruments. The ten most frequent spa types were identified from all samples and on all instruments with 100% specificity and 100% sensitivity. Compared to classical spa typing by sequence analysis, this gene scanning assay is faster, cheaper and can be performed in a single closed tube assay format. Therefore it is an optimal screening tool to detect the most frequent endemic spa types and to exclude non-endemic spa types within a hospital.

  4. Improved Protocol for Rapid Identification of Certain Spa Types Using High Resolution Melting Curve Analysis

    PubMed Central

    Mayerhofer, Benjamin; Stöger, Anna; Pietzka, Ariane T.; Fernandez, Haizpea Lasa; Prewein, Bernhard; Sorschag, Sieglinde; Kunert, Renate; Allerberger, Franz; Ruppitsch, Werner

    2015-01-01

    Methicillin-resistant Staphylococcus aureus is one of the most significant pathogens associated with health care. For efficient surveillance, control and outbreak investigation, S. aureus typing is essential. A high resolution melting curve analysis was developed and evaluated for rapid identification of the most frequent spa types found in an Austrian hospital consortium covering 2,435 beds. Among 557 methicillin-resistant Staphylococcus aureus isolates 38 different spa types were identified by sequence analysis of the hypervariable region X of the protein A gene (spa). Identification of spa types through their characteristic high resolution melting curve profiles was considerably improved by double spiking with genomic DNA from spa type t030 and spa type t003 and allowed unambiguous and fast identification of the ten most frequent spa types t001 (58%), t003 (12%), t190 (9%), t041 (5%), t022 (2%), t032 (2%), t008 (2%), t002 (1%), t5712 (1%) and t2203 (1%), representing 93% of all isolates within this hospital consortium. The performance of the assay was evaluated by testing samples with unknown spa types from the daily routine and by testing three different high resolution melting curve analysis real-time PCR instruments. The ten most frequent spa types were identified from all samples and on all instruments with 100% specificity and 100% sensitivity. Compared to classical spa typing by sequence analysis, this gene scanning assay is faster, cheaper and can be performed in a single closed tube assay format. Therefore it is an optimal screening tool to detect the most frequent endemic spa types and to exclude non-endemic spa types within a hospital. PMID:25768007

  5. An improved qPCR protocol for rapid detection and quantification of Clostridium difficile in cattle feces.

    PubMed

    Bandelj, Petra; Logar, Katarina; Usenik, Alenka M; Vengust, Modest; Ocepek, Matjaz

    2013-04-01

    Clostridium difficile (CD) can cause a significant and transmissible disease in animals and humans, with poorly understood epidemiology. Animals have been suggested as a possible source of infection and environment contamination. It is necessary that a precise and rapid diagnostic tool is available for the detection of CD from clinical and/or environmental samples. A quantitative real-time PCR (qPCR) protocol for CD detection defined by Penders et al. (FEMS Microbiol Lett, 243, 2005, 141-147) was modified. The modified protocol, supported by a novel extraction method, was tested on CD-spiked cattle feces and clinical fecal samples from calves. Quantification was performed targeting CD 16S rRNA gene. Three different commonly used TaqMan universal PCR master mixes were also compared. Results indicate that the modified protocol is very sensitive with an LOD of 7.72 CD cells per g CD-spiked feces. The protocol is capable of precise quantification with an LOQ of 77.2 CD cells per g CD-spiked feces, R(2) between 0.9957 and 0.9968, isolation efficiency from 87.89% to 90.96%, and an interassay CV ranging from 3.71% to 9.57%. The qPCR protocol for the detection and quantification of CD from animal feces investigated and described in this article using MIQE guidelines has the lowest detection and quantification limits published to date. Therefore, it can be implemented for precise epidemiological investigations of CD infections in animals and humans. © 2013 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  6. A consensus protocol: Image-improved therapeutic guidelines for limited adult Hodgkin's disease.

    PubMed

    Busetto, Mario; Sotti, Guido; Zorat, Pierluigi; Salvagno, Luigi; Dal Fior, Sandro; Gaion, Fernando; Soraru, Mariella

    2004-01-01

    to improve the homogeneity of treatments. This protocol includes a list of adverse treatment effects (chemo- and/or radiotherapy) together with follow-up guidelines for the early detection of secondary cancers in previously irradiated patients.

  7. Use of a patient hand hygiene protocol to reduce hospital-acquired infections and improve nurses' hand washing.

    PubMed

    Fox, Cherie; Wavra, Teresa; Drake, Diane Ash; Mulligan, Debbie; Bennett, Yvonne Pacheco; Nelson, Carla; Kirkwood, Peggy; Jones, Louise; Bader, Mary Kay

    2015-05-01

    Critically ill patients are at marked risk of hospital-acquired infections, which increase patients' morbidity and mortality. Registered nurses are the main health care providers of physical care, including hygiene to reduce and prevent hospital-acquired infections, for hospitalized critically ill patients. To investigate a new patient hand hygiene protocol designed to reduce hospital-acquired infection rates and improve nurses' hand-washing compliance in an intensive care unit. A preexperimental study design was used to compare 12-month rates of 2 common hospital-acquired infections, central catheter-associated bloodstream infection and catheter-associated urinary tract infection, and nurses' hand-washing compliance measured before and during use of the protocol. Reductions in 12-month infection rates were reported for both types of infections, but neither reduction was statistically significant. Mean 12-month nurse hand-washing compliance also improved, but not significantly. A hand hygiene protocol for patients in the intensive care unit was associated with reductions in hospital-acquired infections and improvements in nurses' hand-washing compliance. Prevention of such infections requires continuous quality improvement efforts to monitor lasting effectiveness as well as investigation of strategies to eliminate these infections. ©2015 American Association of Critical-Care Nurses.

  8. A novel multi-locus sequence typing (MLST) protocol for Leuconostoc lactis isolates from traditional dairy products in China and Mongolia.

    PubMed

    Dan, Tong; Liu, Wenjun; Sun, Zhihong; Lv, Qiang; Xu, Haiyan; Song, Yuqin; Zhang, Heping

    2014-06-09

    Economically, Leuconostoc lactis is one of the most important species in the genus Leuconostoc. It plays an important role in the food industry including the production of dextrans and bacteriocins. Currently, traditional molecular typing approaches for characterisation of this species at the isolate level are either unavailable or are not sufficiently reliable for practical use. Multilocus sequence typing (MLST) is a robust and reliable method for characterising bacterial and fungal species at the molecular level. In this study, a novel MLST protocol was developed for 50 L. lactis isolates from Mongolia and China. Sequences from eight targeted genes (groEL, carB, recA, pheS, murC, pyrG, rpoB and uvrC) were obtained. Sequence analysis indicated 20 different sequence types (STs), with 13 of them being represented by a single isolate. Phylogenetic analysis based on the sequences of eight MLST loci indicated that the isolates belonged to two major groups, A (34 isolates) and B (16 isolates). Linkage disequilibrium analyses indicated that recombination occurred at a low frequency in L. lactis, indicating a clonal population structure. Split-decomposition analysis indicated that intraspecies recombination played a role in generating genotypic diversity amongst isolates. Our results indicated that MLST is a valuable tool for typing L. lactis isolates that can be used for further monitoring of evolutionary changes and population genetics.

  9. A novel multi-locus sequence typing (MLST) protocol for Leuconostoc lactis isolates from traditional dairy products in China and Mongolia

    PubMed Central

    2014-01-01

    Background Economically, Leuconostoc lactis is one of the most important species in the genus Leuconostoc. It plays an important role in the food industry including the production of dextrans and bacteriocins. Currently, traditional molecular typing approaches for characterisation of this species at the isolate level are either unavailable or are not sufficiently reliable for practical use. Multilocus sequence typing (MLST) is a robust and reliable method for characterising bacterial and fungal species at the molecular level. In this study, a novel MLST protocol was developed for 50 L. lactis isolates from Mongolia and China. Results Sequences from eight targeted genes (groEL, carB, recA, pheS, murC, pyrG, rpoB and uvrC) were obtained. Sequence analysis indicated 20 different sequence types (STs), with 13 of them being represented by a single isolate. Phylogenetic analysis based on the sequences of eight MLST loci indicated that the isolates belonged to two major groups, A (34 isolates) and B (16 isolates). Linkage disequilibrium analyses indicated that recombination occurred at a low frequency in L. lactis, indicating a clonal population structure. Split-decomposition analysis indicated that intraspecies recombination played a role in generating genotypic diversity amongst isolates. Conclusions Our results indicated that MLST is a valuable tool for typing L. lactis isolates that can be used for further monitoring of evolutionary changes and population genetics. PMID:24912963

  10. Improving accessibility of trust guidelines and protocols at the Great Western Hospital, Swindon

    PubMed Central

    Robertson, Ian; Smith, Abigail; Tucker, Jennifer; Cilia, Erica; Chen, Kangni; Marion, Rose; Nesbitt, Julian; Ramcharitar, Steve; Cathiavadi Greamspet, Mala

    2014-01-01

    Trust guidelines and policies outline recommendations for the management of common clinical and non-clinical situations, serving to standardise best practice. Prior to this project, there was no consolidated location for these documents. Lack of organisational structure and inadequate search functionality within the trust intranet led to time wasted locating information, acting outside of recognised best practice, and ultimately potentially compromising patient safety. We surveyed 55 junior doctors, 95% of respondents were dependent on guidelines on a daily basis. 20% spending greater than 5 minutes to locate protocols and 38% unable to locate some relevant documents at all. We analysed the time taken for junior doctors to locate six randomly selected protocols. Pre-intervention mean time was 133 seconds (on six occasions doctors were unable to locate the guideline). All trust guidelines and protocols currently available on the intranet were collated, consolidated, and renamed according to content. These were then re-alphabetised and new search terms linked to each document. Existing links were then uploaded and a single web page made available via the trust intranet homepage. The new page was publicised by email, posters and interdepartmental presentations. In our post intervention survey, 97% of respondents were aware of the project and had made use of the page. All protocols were located during re-testing with 90% of those resurveyed stating it was easier to locate protocols. Overall, a reduction in the time and number of clicks required to locate protocols was demonstrated: mean time 16 seconds vs 133 seconds pre-intervention (n=60). 53% of guidelines located in <30s and 86% <2 minutes. PMID:26732607

  11. Improving accessibility of trust guidelines and protocols at the Great Western Hospital, Swindon.

    PubMed

    Robertson, Ian; Smith, Abigail; Tucker, Jennifer; Cilia, Erica; Chen, Kangni; Marion, Rose; Nesbitt, Julian; Ramcharitar, Steve; Cathiavadi Greamspet, Mala

    2014-01-01

    Trust guidelines and policies outline recommendations for the management of common clinical and non-clinical situations, serving to standardise best practice. Prior to this project, there was no consolidated location for these documents. Lack of organisational structure and inadequate search functionality within the trust intranet led to time wasted locating information, acting outside of recognised best practice, and ultimately potentially compromising patient safety. We surveyed 55 junior doctors, 95% of respondents were dependent on guidelines on a daily basis. 20% spending greater than 5 minutes to locate protocols and 38% unable to locate some relevant documents at all. We analysed the time taken for junior doctors to locate six randomly selected protocols. Pre-intervention mean time was 133 seconds (on six occasions doctors were unable to locate the guideline). All trust guidelines and protocols currently available on the intranet were collated, consolidated, and renamed according to content. These were then re-alphabetised and new search terms linked to each document. Existing links were then uploaded and a single web page made available via the trust intranet homepage. The new page was publicised by email, posters and interdepartmental presentations. In our post intervention survey, 97% of respondents were aware of the project and had made use of the page. All protocols were located during re-testing with 90% of those resurveyed stating it was easier to locate protocols. Overall, a reduction in the time and number of clicks required to locate protocols was demonstrated: mean time 16 seconds vs 133 seconds pre-intervention (n=60). 53% of guidelines located in <30s and 86% <2 minutes.

  12. How blockchain-timestamped protocols could improve the trustworthiness of medical science

    PubMed Central

    Irving, Greg; Holden, John

    2016-01-01

    Trust in scientific research is diminished by evidence that data are being manipulated. Outcome switching, data dredging and selective publication are some of the problems that undermine the integrity of published research. Methods for using blockchain to provide proof of pre-specified endpoints in clinical trial protocols were first reported by Carlisle. We wished to empirically test such an approach using a clinical trial protocol where outcome switching has previously been reported. Here we confirm the use of blockchain as a low cost, independently verifiable method to audit and confirm the reliability of scientific studies. PMID:27239273

  13. How blockchain-timestamped protocols could improve the trustworthiness of medical science.

    PubMed

    Irving, Greg; Holden, John

    2016-01-01

    Trust in scientific research is diminished by evidence that data are being manipulated. Outcome switching, data dredging and selective publication are some of the problems that undermine the integrity of published research. Methods for using blockchain to provide proof of pre-specified endpoints in clinical trial protocols were first reported by Carlisle. We wished to empirically test such an approach using a clinical trial protocol where outcome switching has previously been reported. Here we confirm the use of blockchain as a low cost, independently verifiable method to audit and confirm the reliability of scientific studies.

  14. Improving Station Performance by Building Isolation Walls in the Tunnel

    NASA Astrophysics Data System (ADS)

    Jia, Yan; Horn, Nikolaus; Leohardt, Roman

    2014-05-01

    Conrad Observatory is situated far away from roads and industrial areas on the Trafelberg in Lower Austria. At the end of the seismic tunnel, the main seismic instrument of the Observatory with a station code CONA is located. This station is one of the most important seismic stations in the Austrian Seismic Network (network code OE). The seismic observatory consists of a 145m long gallery and an underground laboratory building with several working areas. About 25 meters away from the station CONA, six temporary seismic stations were implemented for research purposes. Two of them were installed with the same equipment as CONA, while the remaining four stations were set up with digitizers having lower noise and higher resolution (Q330HR) and sensors with the same type (STS-2). In order to prevent possible disturbances by air pressure and temperature fluctuation, three walls were built inside of the tunnel. The first wall is located ca 63 meters from the tunnel entrance, while a set of double walls with a distance of 1.5 meters is placed about 53 meters from the first isolation wall but between the station CONA and the six temporary stations. To assess impact of the isolation walls on noise reduction and detection performance, investigations are conducted in two steps. The first study is carried out by comparing the noise level and detection performance between the station CONA behind the double walls and the stations in front of the double walls for verifying the noise isolation by the double walls. To evaluate the effect of the single wall, station noise level and detection performance were studied by comparing the results before and after the installation of the wall. Results and discussions will be presented. Additional experiment is conducted by filling insulation material inside of the aluminium boxes of the sensors (above and around the sensors). This should help us to determine an optimal insulation of the sensors with respect to pressure and temperature

  15. Does surgical stabilization improve outcomes in patients with isolated multiple distracted and painful non-flail rib fractures?

    PubMed

    Girsowicz, Elie; Falcoz, Pierre-Emmanuel; Santelmo, Nicola; Massard, Gilbert

    2012-03-01

    A best evidence topic was constructed according to a structured protocol. The question addressed was whether surgical stabilization is effective in improving the outcomes of patients with isolated multiple distracted and painful non-flail rib fractures. Of the 356 papers found using a report search, nine presented the best evidence to answer the clinical question. The authors, journal, date and country of publication, study type, group studied, relevant outcomes and results of these papers are given. We conclude that, on the whole, the nine retrieved studies clearly support the use of surgical stabilization in the management of isolated multiple non-flail and painful rib fractures for improving patient outcomes. The interest and benefit was shown not only in terms of pain (McGill pain questionnaire) and respiratory function (forced vital capacity, forced expiratory volume in 1 s and carbon monoxide diffusing capacity), but also in improved quality of life (RAND 36-Item Health Survey) and reduced socio-professional disability. Indeed, most of the authors justified surgical management based on the fact that the results of surgical stabilization showed improvement in short- and long-term patient outcomes, with fast reduction in pain and disability, as well as lower average wait before recommencing normal activities. Hence, the current evidence shows surgical stabilization to be safe and effective in alleviating post-operative pain and in improving patient recovery, thus enhancing the outcome after isolated multiple rib fractures. However, given the little published evidence, prospective trials are necessary to confirm these encouraging results.

  16. An improved method for isolating viruses from asymptomatic carrier fish

    USGS Publications Warehouse

    Amend, Donald F.; Pietsch, John P.

    1972-01-01

    This paper describes a method using elevated levels of penicillin, streptomycin, and nystatin instead of filters to control bacteria and mold contaminants in specimens processed for virus isolation. Filters were shown to significantly reduce the virus concentration. Virus and tissue cultures were not affected by this procedure. In field tests nearly three times more specimens were positive for virus with this method than with the widely used filter technique. Moreover, the cost of materials was less. This method is recommended for inspection and certification purposes.

  17. Disseminating quality improvement: study protocol for a large cluster-randomized trial

    PubMed Central

    2011-01-01

    Background Dissemination is a critical facet of implementing quality improvement in organizations. As a field, addiction treatment has produced effective interventions but disseminated them slowly and reached only a fraction of people needing treatment. This study investigates four methods of disseminating quality improvement (QI) to addiction treatment programs in the U.S. It is, to our knowledge, the largest study of organizational change ever conducted in healthcare. The trial seeks to determine the most cost-effective method of disseminating quality improvement in addiction treatment. Methods The study is evaluating the costs and effectiveness of different QI approaches by randomizing 201 addiction-treatment programs to four interventions. Each intervention used a web-based learning kit plus monthly phone calls, coaching, face-to-face meetings, or the combination of all three. Effectiveness is defined as reducing waiting time (days between first contact and treatment), increasing program admissions, and increasing continuation in treatment. Opportunity costs will be estimated for the resources associated with providing the services. Outcomes The study has three primary outcomes: waiting time, annual program admissions, and continuation in treatment. Secondary outcomes include: voluntary employee turnover, treatment completion, and operating margin. We are also seeking to understand the role of mediators, moderators, and other factors related to an organization's success in making changes. Analysis We are fitting a mixed-effect regression model to each program's average monthly waiting time and continuation rates (based on aggregated client records), including terms to isolate state and intervention effects. Admissions to treatment are aggregated to a yearly level to compensate for seasonality. We will order the interventions by cost to compare them pair-wise to the lowest cost intervention (monthly phone calls). All randomized sites with outcome data will be

  18. Towards an Improved LAI Collection Protocol via Simulated and Field-Based PAR Sensing.

    PubMed

    Yao, Wei; Kelbe, David; Leeuwen, Martin van; Romanczyk, Paul; Aardt, Jan van

    2016-07-14

    In support of NASA's next-generation spectrometer-the Hyperspectral Infrared Imager (HyspIRI)-we are working towards assessing sub-pixel vegetation structure from imaging spectroscopy data. Of particular interest is Leaf Area Index (LAI), which is an informative, yet notoriously challenging parameter to efficiently measure in situ. While photosynthetically-active radiation (PAR) sensors have been validated for measuring crop LAI, there is limited literature on the efficacy of PAR-based LAI measurement in the forest environment. This study (i) validates PAR-based LAI measurement in forest environments, and (ii) proposes a suitable collection protocol, which balances efficiency with measurement variation, e.g., due to sun flecks and various-sized canopy gaps. A synthetic PAR sensor model was developed in the Digital Imaging and Remote Sensing Image Generation (DIRSIG) model and used to validate LAI measurement based on first-principles and explicitly-known leaf geometry. Simulated collection parameters were adjusted to empirically identify optimal collection protocols. These collection protocols were then validated in the field by correlating PAR-based LAI measurement to the normalized difference vegetation index (NDVI) extracted from the "classic" Airborne Visible Infrared Imaging Spectrometer (AVIRIS-C) data ( R 2 was 0.61). The results indicate that our proposed collecting protocol is suitable for measuring the LAI of sparse forest (LAI < 3-5 ( m 2 / m 2 )).

  19. Modified CTAB and TRIzol protocols improve RNA extraction from chemically complex Embryophyta1

    PubMed Central

    Jordon-Thaden, Ingrid E.; Chanderbali, Andre S.; Gitzendanner, Matthew A.; Soltis, Douglas E.

    2015-01-01

    Premise of the study: Here we present a series of protocols for RNA extraction across a diverse array of plants; we focus on woody, aromatic, aquatic, and other chemically complex taxa. Methods and Results: Ninety-one taxa were subjected to RNA extraction with three methods presented here: (1) TRIzol/TURBO DNA-free kits using the manufacturer’s protocol with the addition of sarkosyl; (2) a combination method using cetyltrimethylammonium bromide (CTAB) and TRIzol/sarkosyl/TURBO DNA-free; and (3) a combination of CTAB and QIAGEN RNeasy Plant Mini Kit. Bench-ready protocols are given. Conclusions: After an iterative process of working with chemically complex taxa, we conclude that the use of TRIzol supplemented with sarkosyl and the TURBO DNA-free kit is an effective, efficient, and robust method for obtaining RNA from 100 mg of leaf tissue of land plant species (Embryophyta) examined. Our protocols can be used to provide RNA of suitable stability, quantity, and quality for transcriptome sequencing. PMID:25995975

  20. Towards an Improved LAI Collection Protocol via Simulated and Field-Based PAR Sensing

    PubMed Central

    Yao, Wei; Kelbe, David; van Leeuwen, Martin; Romanczyk, Paul; van Aardt, Jan

    2016-01-01

    In support of NASA’s next-generation spectrometer—the Hyperspectral Infrared Imager (HyspIRI)—we are working towards assessing sub-pixel vegetation structure from imaging spectroscopy data. Of particular interest is Leaf Area Index (LAI), which is an informative, yet notoriously challenging parameter to efficiently measure in situ. While photosynthetically-active radiation (PAR) sensors have been validated for measuring crop LAI, there is limited literature on the efficacy of PAR-based LAI measurement in the forest environment. This study (i) validates PAR-based LAI measurement in forest environments, and (ii) proposes a suitable collection protocol, which balances efficiency with measurement variation, e.g., due to sun flecks and various-sized canopy gaps. A synthetic PAR sensor model was developed in the Digital Imaging and Remote Sensing Image Generation (DIRSIG) model and used to validate LAI measurement based on first-principles and explicitly-known leaf geometry. Simulated collection parameters were adjusted to empirically identify optimal collection protocols. These collection protocols were then validated in the field by correlating PAR-based LAI measurement to the normalized difference vegetation index (NDVI) extracted from the “classic” Airborne Visible Infrared Imaging Spectrometer (AVIRIS-C) data (R2 was 0.61). The results indicate that our proposed collecting protocol is suitable for measuring the LAI of sparse forest (LAI < 3–5 (m2/m2)). PMID:27428969

  1. Towards an improved LAI collection protocol via simulated field-based PAR sensing

    SciTech Connect

    Yao, Wei; Van Leeuwen, Martin; Romanczyk, Paul; van Aardt, Jan; Kelbe, David

    2016-07-14

    In support of NASA’s next-generation spectrometer—the Hyperspectral Infrared Imager (HyspIRI)—we are working towards assessing sub-pixel vegetation structure from imaging spectroscopy data. Of particular interest is Leaf Area Index (LAI), which is an informative, yet notoriously challenging parameter to efficiently measure in situ. While photosynthetically-active radiation (PAR) sensors have been validated for measuring crop LAI, there is limited literature on the efficacy of PAR-based LAI measurement in the forest environment. This study (i) validates PAR-based LAI measurement in forest environments, and (ii) proposes a suitable collection protocol, which balances efficiency with measurement variation, e.g., due to sun flecks and various-sized canopy gaps. A synthetic PAR sensor model was developed in the Digital Imaging and Remote Sensing Image Generation (DIRSIG) model and used to validate LAI measurement based on first-principles and explicitly-known leaf geometry. Simulated collection parameters were adjusted to empirically identify optimal collection protocols. Furthermore, these collection protocols were then validated in the field by correlating PAR-based LAI measurement to the normalized difference vegetation index (NDVI) extracted from the “classic” Airborne Visible Infrared Imaging Spectrometer (AVIRIS-C) data (R2 was 0.61). The results indicate that our proposed collecting protocol is suitable for measuring the LAI of sparse forest (LAI < 3–5 ( m2/m2)).

  2. Towards an improved LAI collection protocol via simulated field-based PAR sensing

    DOE PAGES

    Yao, Wei; Van Leeuwen, Martin; Romanczyk, Paul; ...

    2016-07-14

    In support of NASA’s next-generation spectrometer—the Hyperspectral Infrared Imager (HyspIRI)—we are working towards assessing sub-pixel vegetation structure from imaging spectroscopy data. Of particular interest is Leaf Area Index (LAI), which is an informative, yet notoriously challenging parameter to efficiently measure in situ. While photosynthetically-active radiation (PAR) sensors have been validated for measuring crop LAI, there is limited literature on the efficacy of PAR-based LAI measurement in the forest environment. This study (i) validates PAR-based LAI measurement in forest environments, and (ii) proposes a suitable collection protocol, which balances efficiency with measurement variation, e.g., due to sun flecks and various-sized canopymore » gaps. A synthetic PAR sensor model was developed in the Digital Imaging and Remote Sensing Image Generation (DIRSIG) model and used to validate LAI measurement based on first-principles and explicitly-known leaf geometry. Simulated collection parameters were adjusted to empirically identify optimal collection protocols. Furthermore, these collection protocols were then validated in the field by correlating PAR-based LAI measurement to the normalized difference vegetation index (NDVI) extracted from the “classic” Airborne Visible Infrared Imaging Spectrometer (AVIRIS-C) data (R2 was 0.61). The results indicate that our proposed collecting protocol is suitable for measuring the LAI of sparse forest (LAI < 3–5 ( m2/m2)).« less

  3. Bedside electronic capture of clinical observations and automated clinical alerts to improve compliance with an Early Warning Score protocol.

    PubMed

    Jones, Steve; Mullally, Miki; Ingleby, Sarah; Buist, Michael; Bailey, Michael; Eddleston, Jane M

    2011-06-01

    Failure to comply with clinical protocols and failure of communication to ensure delivery of the most appropriate timely clinical responses to patients whose conditions are acutely deteriorating have been shown to be significant causative factors associated with inhospital adverse events. To determine whether automated clinical alerts increase compliance with an Early Warning Score (EWS) protocol and improve patient outcomes. We performed a historically controlled study of bedside electronic capture of observations and automated clinical alerts. The primary outcome measure was hospital length of stay (LOS); secondary outcome measures were compliance with the EWS protocol, cardiac arrest incidence, critical care utilisation and hospital mortality. Between baseline and intervention, 1481 consecutive patients were recruited generating 13 668 observation sets. There was a reduction in hospital LOS between the baseline and alert phase (9.7 days v 6.9 days, P < 0.001). EWS accuracy improved from 81% to 100% with electronic calculation. Clinical attendance to patients with EWS 3, 4 or 5 increased from 29% at baseline to 78% with automated alerts (P < 0.001). For patients with an EWS > 5, clinical attendance increased from 67% at baseline to 96% with automatic alerts (P < 0.001). Electronic recording of patient observations linked to a computer system that calculates patient risk and then issues automatic graded alerts can improve clinical attendance to unstable general medical ward patients.

  4. Cryptanalysis and improvement of quantum broadcast communication and authentication protocol with a quantum one-time pad

    NASA Astrophysics Data System (ADS)

    Liu, Zhi-Hao; Chen, Han-Wu

    2016-08-01

    The security of quantum broadcast communication (QBC) and authentication protocol based on Greenberger-Horne-Zeilinger (GHZ) state and quantum one-time pad is analyzed. It is shown that there are some security issues in this protocol. Firstly, an external eavesdropper can take the intercept-measure-resend attack strategy to eavesdrop on 0.369 bit of every bit of the identity string of each receiver without being detected. Meanwhile, 0.524 bit of every bit of the secret message can be eavesdropped on without being detected. Secondly, an inner receiver can take the intercept-measure-resend attack strategy to eavesdrop on half of the identity string of the other’s definitely without being checked. In addition, an alternative attack called the CNOT-operation attack is discussed. As for the multi-party QBC protocol, the attack efficiency increases with the increase of the number of users. Finally, the QBC protocol is improved to a secure one. Project supported by the National Natural Science Foundation of China (Grant Nos. 61502101 and 61170321), the Natural Science Foundation of Jiangsu Province, China (Grant No. BK20140651), the Research Fund for the Doctoral Program of Higher Education, China (Grant No. 20110092110024), and the Project Funded by PAPD and CICAEET.

  5. Improving post-stroke dysphagia outcomes through a standardized and multidisciplinary protocol: an exploratory cohort study.

    PubMed

    Gandolfi, Marialuisa; Smania, Nicola; Bisoffi, Giulia; Squaquara, Teresa; Zuccher, Paola; Mazzucco, Sara

    2014-12-01

    Stroke is a major cause of dysphagia. Few studies to date have reported on standardized multidisciplinary protocolized approaches to the management of post-stroke dysphagia. The aim of this retrospective cohort study was to evaluate the impact of a standardized multidisciplinary protocol on clinical outcomes in patients with post-stroke dysphagia. We performed retrospective chart reviews of patients with post-stroke dysphagia admitted to the neurological ward of Verona University Hospital from 2004 to 2008. Outcomes after usual treatment for dysphagia (T- group) were compared versus outcomes after treatment under a standardized diagnostic and rehabilitative multidisciplinary protocol (T+ group). Outcome measures were death, pneumonia on X-ray, need for respiratory support, and proportion of patients on tube feeding at discharge. Of the 378 patients admitted with stroke, 84 had dysphagia and were enrolled in the study. A significantly lower risk of in-hospital death (odds ratio [OR] 0.20 [0.53-0.78]), pneumonia (OR 0.33 [0.10-1.03]), need for respiratory support (OR 0.48 [0.14-1.66]), and tube feeding at discharge (OR 0.30 [0.09-0.91]) was recorded for the T+ group (N = 39) as compared to the T- group (N = 45). The adjusted OR showed no difference between the two groups for in-hospital death and tube feeding at discharge. Use of a standardized multidisciplinary protocolized approach to the management of post-stroke dysphagia may significantly reduce rates of aspiration pneumonia, in-hospital mortality, and tube feeding in dysphagic stroke survivors. Consistent with the study's exploratory purposes, our findings suggest that the multidisciplinary protocol applied in this study offers an effective model of management of post-stroke dysphagia.

  6. A simple and effective protocol for fast isolation of human Tenon's fibroblasts from a single trabeculectomy biopsy - a comparison of cell behaviour in different culture media.

    PubMed

    Przekora, Agata; Zarnowski, Tomasz; Ginalska, Grazyna

    2017-01-01

    Human Tenon's fibroblasts (HTFs) play a crucial role in wound healing. They cause postoperative scarring of the filtering bleb and are thus responsible for trabeculectomy failure. This study aimed to find an effective and fast protocol for HTF isolation from trabeculectomy biopsies. The protocol was compared with the commonly recommended HTF isolation procedure, which uses Dulbecco's modified Eagle's medium (DMEM). We used Eagle's minimum essential medium (EMEM) enriched with fibroblast growth factor (FGF), which selectively promoted the proliferation of HTF cells. A secondary goal was to compare HTF morphology, metabolism and growth during parallel cultivation of the isolated cells in FGF-enriched EMEM and DMEM. Standard procedures for HTF isolation from tissue biopsies require a 20- to 30-day culture of the explants to obtain the first monolayer. Our protocol yielded the first monolayer after approx. 15 days. More importantly, the majority of the cells were fibroblasts with only individual epithelium-derived cells present. Using FGF-enriched EMEM allowed 1.3 × 10(6) vimentin-positive fibroblasts to be obtained from a single biopsy within approx. 25 days. Using DMEM resulted in isolation failure and required exchange to FGF-enriched medium to recover the fibroblast culture. HTFs maintained in FGF-enriched EMEM also showed faster proliferation and a different type I collagen production ability compared to HTFs cultured in DMEM. Thus, FGF-enriched EMEM is recommended for fast propagation of HTFs unless the aim of the study is to assess the effect of a tested agent on proliferation ability or type I collagen production. Our fast protocol for HTF isolation allows easy setup of cell banks by researchers under laboratory conditions and could be very useful during testing of novel ophthalmologic anti-fibrotic agents in vitro. Molecular analysis of HTFs isolated from patients with known treatment histories may provide valuable information on the effects of some

  7. A simplified and cost-effective enrichment protocol for the isolation of Campylobacter spp. from retail broiler meat without microaerobic incubation

    PubMed Central

    2011-01-01

    Background To simplify the methodology for the isolation of Campylobacter spp. from retail broiler meat, we evaluated 108 samples (breasts and thighs) using an unpaired sample design. The enrichment broths were incubated under aerobic conditions (subsamples A) and for comparison under microaerobic conditions (subsamples M) as recommended by current reference protocols. Sensors were used to measure the dissolved oxygen (DO) in the broth and the percentage of oxygen (O2) in the head space of the bags used for enrichment. Campylobacter isolates were identified with multiplex PCR assays and typed using pulsed-field gel electrophoresis (PFGE). Ribosomal intergenic spacer analyses (RISA) and denaturing gradient gel electrophoresis (DGGE) were used to study the bacterial communities of subsamples M and A after 48 h enrichment. Results The number of Campylobacter positive subsamples were similar for A and M when all samples were combined (P = 0.81) and when samples were analyzed by product (breast: P = 0.75; thigh: P = 1.00). Oxygen sensors showed that DO values in the broth were around 6 ppm and O2 values in the head space were 14-16% throughout incubation. PFGE demonstrated high genomic similarity of isolates in the majority of the samples in which isolates were obtained from subsamples A and M. RISA and DGGE results showed a large variability in the bacterial populations that could be attributed to sample-to-sample variations and not enrichment conditions (aerobic or microaerobic). These data also suggested that current sampling protocols are not optimized to determine the true number of Campylobacter positive samples in retail boiler meat. Conclusions Decreased DO in enrichment broths is naturally achieved. This simplified, cost-effective enrichment protocol with aerobic incubation could be incorporated into reference methods for the isolation of Campylobacter spp. from retail broiler meat. PMID:21812946

  8. Improving the characterization of endothelial progenitor cell subsets by an optimized FACS protocol.

    PubMed

    Huizer, Karin; Mustafa, Dana A M; Spelt, J Clarissa; Kros, Johan M; Sacchetti, Andrea

    2017-01-01

    The characterization of circulating endothelial progenitor cells (EPCs) is fundamental to any study related to angiogenesis. Unfortunately, current literature lacks consistency in the definition of EPC subsets due to variations in isolation strategies and inconsistencies in the use of lineage markers. Here we address critical points in the identification of hematopoietic progenitor cells (HPCs), circulating endothelial cells (CECs), and culture-generated outgrowth endothelial cells (OECs) from blood samples of healthy adults (AB) and umbilical cord (UCB). Peripheral blood mononuclear cells (PBMCs) were enriched using a Ficoll-based gradient followed by an optimized staining and gating strategy to enrich for the target cells. Sorted EPC populations were subjected to RT-PCR for tracing the expression of markers beyond the limits of cell surface-based immunophenotyping. Using CD34, CD133 and c-kit staining, combined with FSC and SSC, we succeeded in the accurate and reproducible identification of four HPC subgroups and found significant differences in the respective populations in AB vs. UCB. Co-expression analysis of endothelial markers on HPCs revealed a complex pattern characterized by various subpopulations. CECs were identified by using CD34, KDR, CD45, and additional endothelial markers, and were subdivided according to their apoptotic state and expression of c-kit. Comparison of UCB-CECs vs. AB-CECs revealed significant differences in CD34 and KDR levels. OECs were grown from PBMC-fractions We found that viable c-kit+ CECs are a candidate circulating precursor for CECs. RT-PCR to angiogenic factors and receptors revealed that all EPC subsets expressed angiogenesis-related molecules. Taken together, the improvements in immunophenotyping and gating strategies resulted in accurate identification and comparison of better defined cell populations in a single procedure.

  9. Improve isolation materials and recommendation of residential buildings in Kazakhstan

    NASA Astrophysics Data System (ADS)

    Kim, Yuliya; Sun, Cheng; Nuguzhinov, Zhmagul

    2017-04-01

    The article shows the problem of high volumes of energy use in Kazakhstan and introduction of energy-efficient technologies, production and causes of the problem. To improve the thermal comfort, needed primarily to improve the thermal insulation properties of materials. This article examines the characteristics of thermal insulation materials and their application in the construction process in Kazakhstan. The material properties are calculated and shown as formulas, which can use in the future in the design of residential buildings. The construction studied of passive technology in the residential area. By the design process and the combination of design variables selected option.

  10. Immunodepletion and Hypoxia Preconditioning of Mouse Compact Bone Cells as a Novel Protocol to Isolate Highly Immunosuppressive Mesenchymal Stem Cells.

    PubMed

    Sivanathan, Kisha Nandini; Gronthos, Stan; Grey, Shane T; Rojas-Canales, Darling; Coates, Patrick T

    2017-04-01

    Compact bones (CB) are major reservoirs of mouse mesenchymal stem cells (mMSC). Here, we established a protocol to isolate MSC from CB and tested their immunosuppressive potential. Collagenase type II digestion of BM-flushed CB from C57B/6 mice was performed to liberate mMSC precursors from bone surfaces to establish nondepleted mMSC. CB cells were also immunodepleted based on the expression of CD45 (leukocytes) and TER119 (erythroid cells) to eliminate hematopoietic cells. CD45(-)TER119(-) CB cells were subsequently used to generate depleted mMSC. CB nondepleted and depleted mMSC progenitors were cultured under hypoxic conditions to establish primary mMSC cultures. CB depleted mMSC compared to nondepleted mMSC showed greater cell numbers at subculturing and had increased functional ability to differentiate into adipocytes and osteoblasts. CB depleted mMSC had high purity and expressed key mMSC markers (>85% Sca-1, CD29, CD90) with no mature hematopoietic contaminating cells (<5% CD45, CD11b) when subcultured to passage 5 (P5). Nondepleted mMSC cultures, however, were less pure and heterogenous with <72% Sca-1(+), CD29(+), and CD90(+) cells at early passages (P1 or P2), along with high percentages of contaminating CD11b(+) (35.6%) and CD45(+) (39.2%) cells that persisted in culture long term. Depleted and nondepleted mMSC nevertheless exhibited similar potency to suppress total (CD3(+)), CD4(+) and CD8(+) T cell proliferation, in a dendritic cell allostimulatory one-way mixed lymphocyte reaction. CB depleted mMSC, pretreated with proinflammatory cytokines IFN-γ, TNF-α, and IL-17A, showed superior suppression of CD8(+) T cell, but not CD4(+) T cell proliferation, relative to untreated-mMSC. In conclusion, CB depleted mMSC established under hypoxic conditions and treated with selective cytokines represent a novel source of potent immunosuppressive MSC. As these cells have enhanced immune modulatory function, they may represent a superior product for use in clinical

  11. Improvement of the SEP protocol based on community structure of node degree

    NASA Astrophysics Data System (ADS)

    Li, Donglin; Wei, Suyuan

    2017-05-01

    Analyzing the Stable election protocol (SEP) in wireless sensor networks and aiming at the problem of inhomogeneous cluster-heads distribution and unreasonable cluster-heads selectivity and single hop transmission in the SEP, a SEP Protocol based on community structure of node degree (SEP-CSND) is proposed. In this algorithm, network node deployed by using grid deployment model, and the connection between nodes established by setting up the communication threshold. The community structure constructed by node degree, then cluster head is elected in the community structure. On the basis of SEP, the node's residual energy and node degree is added in cluster-heads election. The information is transmitted with mode of multiple hops between network nodes. The simulation experiments showed that compared to the classical LEACH and SEP, this algorithm balances the energy consumption of the entire network and significantly prolongs network lifetime.

  12. The Engagement Interview Protocol (EIP): improving the acceptance of mental health treatment among Chinese immigrants

    PubMed Central

    Yeung, Albert; Trinh, Nhi-Ha T.; Chang, Trina E.; Fava, Maurizio

    2014-01-01

    Many depressed Chinese immigrants are unfamiliar with Western psychiatric terminology and have high levels of stigma toward psychiatric illnesses, making it difficult to engage them into psychiatric treatment. We have designed the Engagement Interview Protocol (EIP), a semi-standardized protocol that incorporates cultural components to a standard psychiatric evaluation. The EIP elicits patients’ narratives and uses anthropological questions to explore patients’ illness beliefs, which are integrated with patients’ information on medical and psychiatric history, psychosocial background and mental status examination so that treatment options can be negotiated in a culturally sensitive manner. In our field testing on depressed Chinese immigrants, the EIP model was found to be a practical tool that can be completed within the allotted one-hour time frame and was highly effective in facilitating the enrollment of patients in treatment for depression. The EIP is a concise, time-effective, user-friendly protocol that can be used both in research and real-world clinical settings with diverse patient populations. PMID:25076983

  13. An improved technique for isolating codominant compound microsatellite markers.

    PubMed

    Lian, Chunlan L; Abdul Wadud, Md; Geng, Qifang; Shimatani, Kenichiro; Hogetsu, Taizo

    2006-07-01

    An approach for developing codominant polymorphic markers (compound microsatellite (SSR) markers), with substantial time and cost savings, is introduced in this paper. In this technique, fragments flanked by a compound SSR sequence at one end were amplified from the constructed DNA library using compound SSR primer (AC)6(AG)5 or (TC)6(AC)5 and an adaptor primer for the suppression-PCR. A locus-specific primer was designed from the sequence flanking the compound SSR. The primer pairs of the locus-specific and compound SSR primers were used as a compound SSR marker. Because only one locus-specific primer was needed for design of each marker and only a common compound SSR primer was needed as the fluorescence-labeled primer for analyzing all the compound SSR markers, this approach substantially reduced the cost of developing codominant markers and analyzing their polymorphism. We have demonstrated this technique for Dendropanax trifidus and easily developed 11 codominant markers with high polymorphism for D. trifidus. Use of the technique for successful isolation of codominant compound SSR markers for several other plant species is currently in progress.

  14. An optimised protocol to isolate high-quality genomic DNA from seed tissues streamlines the workflow to obtain direct estimates of seed dispersal distances in gymnosperms.

    PubMed

    García, C; Escribano-Ávila, G

    2016-05-01

    Genotyping of maternally derived seed tissues from georefered seeds that moved away from their source tree yield direct estimates of seed dispersal distances when the location and the genotype of the fruiting tree are available. These estimates are instrumental in forecasting the response of plant communities to drivers of global change, such as fragmentation or the expansion of invasive species. Obtaining robust assessments of seed dispersal distances requires comparing reliable multilocus genotypes of maternally derived seed tissues and fruiting trees, as previously shown for angiosperm species. However, robust estimates of seed dispersal distances based on direct methods are rare in non-model gymnosperms due to the difficulty in isolating high quality DNA from inconspicuous maternally derived seed tissues. These tissues tend to yield low DNA quantities that increase the frequency of genotyping errors. Here, we deliver a step-by-step visual protocol used to identify and isolate different seed tissues of interest for dispersal studies: embryos (2n, bi-parentally derived), seed coats (2n, maternally derived), and megagametophytes (n, maternally derived). We also provide an optimised lab protocol used to obtain multilocus genotypes from the target seed tissue. These broadly applicable protocols proved successful both in avoiding contamination among different seed tissues and providing reliable multilocus genotypes.

  15. In vitro skin irritation testing: Improving the sensitivity of the EpiDerm skin irritation test protocol.

    PubMed

    Kandárová, Helena; Hayden, Patrick; Klausner, Mitch; Kubilus, Joseph; Kearney, Paul; Sheasgreen, John

    2009-12-01

    irritants, the sensitivity of the Modified EpiDerm SIT increased to above 90%. The measurement of interleukin (IL)-1alpha release did not further contribute to improvement of the method. The results demonstrate that the modified EpiDerm SIT protocol has the required sensitivity and specificity to be accepted as a stand alone method for complete replacement of the in vivo rabbit test. 2009 FRAME.

  16. A major haemorrhage protocol improves the delivery of blood component therapy and reduces waste in trauma massive transfusion.

    PubMed

    Khan, Sirat; Allard, Shubha; Weaver, Anne; Barber, Colin; Davenport, Ross; Brohi, Karim

    2013-05-01

    Major haemorrhage protocols (MHP) are required as part of damage control resuscitation regimens in modern trauma care. The primary objectives of this study were to ascertain whether a MHP improved blood product administration and reduced waste compared to traditional massive transfusion protocols (MTP). Datasets on adult trauma admissions 1 year prior and 1 year post implementation of a MHP at a Level 1 trauma centre were obtained from the trauma registry. Demographic and clinical data were collected prospectively including mechanism of injury, physiological observations, ICU admission and length of stay. The volume of blood components (packed red blood cells, platelets, cryoprecipitate and fresh frozen plasma) issued, transfused, returned to stock and wasted within the first 24h was gathered retrospectively. Over the 2-year study period 2986 patient records were available for analysis. 40 patients required a 10+ Units of packed red blood ells transfusion in the MTP group vs. 56 patients post MHP implementation. The administration of blood component therapy improved significantly post MHP implementation. FFP:PRBC transfusion improved from 1:3 to 1:2 (p<0.01) and CRYO:PRBC improved from 1:10 to 1:7 (p<0.05). We reported a significant reduction in the waste of platelets from 14% to 2% (p<0.01). Outcomes had improved: Median hospital length of stay was reduced from 54 days to 26 days (p<0.05). Implementation of a MHP results in improved delivery of blood components and a reduction in the waste of blood products compared to the older model of MTP. In combination with educational programmes MHP can significantly improve blood product administration and patient outcomes in trauma haemorrhage. Level III diagnostic test study. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

  17. Variable Reference Alignment: an improved peak alignment protocol for NMR spectral data with large inter-sample variation

    PubMed Central

    MacKinnon, Neil; Ge, Wencheng; Khan, Amjad P.; Somashekar, Bagganahalli S.; Tripathi, Pratima; Siddiqui, Javed; Wei, John T.; Chinnaiyan, Arul M.; Rajendiran, Thekkelnaycke M.; Ramamoorthy, Ayyalusamy

    2012-01-01

    In an effort to address the variable correspondence problem across large sample cohorts common in metabolomic/metabonomic studies, we have developed a pre-alignment protocol that aims to generate spectral segments sharing a common target spectrum. Under the assumption that a single reference spectrum will not correctly represent all spectra of a data set, the goal of this approach is to perform local alignment corrections on spectral regions which share a common ‘most similar’ spectrum. A natural beneficial outcome of this procedure is the automatic definition of spectral segments, a feature that is not common to all alignment methods. This protocol is shown to specifically improve the quality of alignment in 1H NMR data sets exhibiting large inter-sample compositional variation (e.g. pH, ionic strength). As a proof-of-principle demonstration, we have utilized two recently developed alignment algorithms specific to NMR data, recursive segment-wise peak alignment and interval correlated shifting and applied them to two data sets comprised of 15 aqueous cell line extract and 20 human urine 1H NMR profiles. Application of this protocol represents a fundamental shift from current alignment methodologies that seek to correct misalignments utilizing a single representative spectrum, with the added benefit that it can be appended to any alignment algorithm. PMID:22616856

  18. Improved cryoEM-Guided Iterative Molecular Dynamics–Rosetta Protein Structure Refinement Protocol for High Precision Protein Structure Prediction

    PubMed Central

    2016-01-01

    Many excellent methods exist that incorporate cryo-electron microscopy (cryoEM) data to constrain computational protein structure prediction and refinement. Previously, it was shown that iteration of two such orthogonal sampling and scoring methods – Rosetta and molecular dynamics (MD) simulations – facilitated exploration of conformational space in principle. Here, we go beyond a proof-of-concept study and address significant remaining limitations of the iterative MD–Rosetta protein structure refinement protocol. Specifically, all parts of the iterative refinement protocol are now guided by medium-resolution cryoEM density maps, and previous knowledge about the native structure of the protein is no longer necessary. Models are identified solely based on score or simulation time. All four benchmark proteins showed substantial improvement through three rounds of the iterative refinement protocol. The best-scoring final models of two proteins had sub-Ångstrom RMSD to the native structure over residues in secondary structure elements. Molecular dynamics was most efficient in refining secondary structure elements and was thus highly complementary to the Rosetta refinement which is most powerful in refining side chains and loop regions. PMID:25883538

  19. Improved cryoEM-Guided Iterative Molecular Dynamics--Rosetta Protein Structure Refinement Protocol for High Precision Protein Structure Prediction.

    PubMed

    Lindert, Steffen; McCammon, J Andrew

    2015-03-10

    Many excellent methods exist that incorporate cryo-electron microscopy (cryoEM) data to constrain computational protein structure prediction and refinement. Previously, it was shown that iteration of two such orthogonal sampling and scoring methods – Rosetta and molecular dynamics (MD) simulations – facilitated exploration of conformational space in principle. Here, we go beyond a proof-of-concept study and address significant remaining limitations of the iterative MD–Rosetta protein structure refinement protocol. Specifically, all parts of the iterative refinement protocol are now guided by medium-resolution cryoEM density maps, and previous knowledge about the native structure of the protein is no longer necessary. Models are identified solely based on score or simulation time. All four benchmark proteins showed substantial improvement through three rounds of the iterative refinement protocol. The best-scoring final models of two proteins had sub-Ångstrom RMSD to the native structure over residues in secondary structure elements. Molecular dynamics was most efficient in refining secondary structure elements and was thus highly complementary to the Rosetta refinement which is most powerful in refining side chains and loop regions.

  20. IMPROVED COMPUTATIONAL NEUTRONICS METHODS AND VALIDATION PROTOCOLS FOR THE ADVANCED TEST REACTOR

    SciTech Connect

    David W. Nigg; Joseph W. Nielsen; Benjamin M. Chase; Ronnie K. Murray; Kevin A. Steuhm

    2012-04-01

    The Idaho National Laboratory (INL) is in the process of modernizing the various reactor physics modeling and simulation tools used to support operation and safety assurance of the Advanced Test Reactor (ATR). Key accomplishments so far have encompassed both computational as well as experimental work. A new suite of stochastic and deterministic transport theory based reactor physics codes and their supporting nuclear data libraries (HELIOS, KENO6/SCALE, NEWT/SCALE, ATTILA, and an extended implementation of MCNP5) has been installed at the INL. Corresponding models of the ATR and ATRC are now operational with all five codes, demonstrating the basic feasibility of the new code packages for their intended purpose. Of particular importance, a set of as-run core depletion HELIOS calculations for all ATR cycles since August 2009 was successfully completed during 2011. This demonstration supported a decision late in the year to proceed with the phased incorporation of the HELIOS methodology into the ATR fuel cycle management process beginning in 2012. On the experimental side of the project, new hardware was fabricated, measurement protocols were finalized, and the first four of six planned physics code validation experiments based on neutron activation spectrometry were conducted at the ATRC facility. Data analysis for the first three experiments, focused on characterization of the neutron spectrum in one of the ATR flux traps, has been completed. The six experiments will ultimately form the basis for a flexible, easily-repeatable ATR physics code validation protocol that is consistent with applicable ASTM standards.

  1. Improved protocol and data analysis for accelerated shelf-life estimation of solid dosage forms.

    PubMed

    Waterman, Kenneth C; Carella, Anthony J; Gumkowski, Michael J; Lukulay, Patrick; MacDonald, Bruce C; Roy, Michael C; Shamblin, Sheri L

    2007-04-01

    To propose and test a new accelerated aging protocol for solid-state, small molecule pharmaceuticals which provides faster predictions for drug substance and drug product shelf-life. The concept of an isoconversion paradigm, where times in different temperature and humidity-controlled stability chambers are set to provide a critical degradant level, is introduced for solid-state pharmaceuticals. Reliable estimates for temperature and relative humidity effects are handled using a humidity-corrected Arrhenius equation, where temperature and relative humidity are assumed to be orthogonal. Imprecision is incorporated into a Monte-Carlo simulation to propagate the variations inherent in the experiment. In early development phases, greater imprecision in predictions is tolerated to allow faster screening with reduced sampling. Early development data are then used to design appropriate test conditions for more reliable later stability estimations. Examples are reported showing that predicted shelf-life values for lower temperatures and different relative humidities are consistent with the measured shelf-life values at those conditions. The new protocols and analyses provide accurate and precise shelf-life estimations in a reduced time from current state of the art.

  2. Lingual orthodontics for children and adolescents: improvement of the indirect bonding protocol.

    PubMed

    Beyling, Frauke; Schwestka-Polly, Rainer; Wiechmann, Dirk

    2013-09-11

    Demineralization of the dental enamel is a finding associated with fixed orthodontic treatment. When an indirect bonding procedure is used in children and adolescents the area beneath the bracket base may be affected. To evaluate if the addition of an extra layer of a hydrophilic resin, to a conventional indirect bonding protocol, can reduce the incidence of demineralization beneath the bracket base. 40 patients under 18 years of age were treated with completely customized lingual appliances. Two different bonding protocols were used either with or without the application of an additional layer of hydrophilic resin. Demineralization beneath the bracket base, after de-bonding, was evaluated by standardized intra-oral photographs. The addition of an extra layer of a hydrophilic resin helps to reduce the number of demineralized areas beneath the bracket bases significantly (three times less). The severity of the few remaining defects were minor and without any clinical consequence. When bonding a completely customized lingual appliance in children and adolescents, an extra layer of a hydrophilic resin should be added to the teeth.

  3. A New Cecal Slurry Preparation Protocol with Improved Long-Term Reproducibility for Animal Models of Sepsis

    PubMed Central

    Starr, Marlene E.; Steele, Allison M.; Saito, Mizuki; Hacker, Bill J.; Evers, B. Mark; Saito, Hiroshi

    2014-01-01

    Sepsis, a life-threatening systemic inflammatory response syndrome induced by infection, is widely studied using laboratory animal models. While cecal-ligation and puncture (CLP) is considered the gold standard model for sepsis research, it may not be preferable for experiments comparing animals of different size or under different dietary regimens. By comparing cecum size, shape, and cecal content characteristics in mice under different experimental conditions (aging, diabetes, pancreatitis), we show that cecum variability could be problematic for some CLP experiments. The cecal slurry (CS) injection model, in which the cecal contents of a laboratory animal are injected intraperitoneally to other animals, is an alternative method for inducing polymicrobial sepsis; however, the CS must be freshly prepared under conventional protocols, which is a major disadvantage with respect to reproducibility and convenience. The objective of this study was to develop an improved CS preparation protocol that allows for long-term storage of CS with reproducible results. Using our new CS preparation protocol we found that bacterial viability is maintained for at least 6 months when the CS is prepared in 15% glycerol-PBS and stored at -80°C. To test sepsis-inducing efficacy of stored CS stocks, various amounts of CS were injected to young (4–6 months old), middle-aged (12–14 months old), and aged (24–26 months old) male C57BL/6 mice. Dose- and age-dependent mortality was observed with high reproducibility. Circulating bacteria levels strongly correlated with mortality suggesting an infection-mediated death. Further, injection with heat-inactivated CS resulted in acute hypothermia without mortality, indicating that CS-mediated death is not due to endotoxic shock. This new CS preparation protocol results in CS stocks which are durable for freezing preservation without loss of bacterial viability, allowing experiments to be performed more conveniently and with higher

  4. Improving the efficiency of isolated microspore culture in six-row spring barley: I-optimization of key physical factors.

    PubMed

    Esteves, Patricio; Belzile, François

    2014-06-01

    An improved isolated microspore culture protocol alleviating the recalcitrance typically observed in six-row spring barley was developed by optimizing four key physical factors to increase embryogenesis and reduce albinism. Doubled haploid (DH) plants are completely homozygous individuals that can be generated in just a few months via androgenesis in vitro. DHs are useful tools in genetic research and in plant breeding. Isolated microspore culture (IMC) is the most efficient way to produce DHs, but a strong genotype dependency imposes limitations to its wide application. Six-row, spring barley genotypes are considered as particularly recalcitrant due to a low frequency of embryogenesis and a high rate of albinism. Seeking to develop an efficient IMC protocol for this type of barley, we explored four important factors: (1) the harvest stage of immature spikes, (2) the type of pretreatment applied, (3) the osmotic potential in the induction medium, and (4) the plating density of microspores. This work was first performed using four barley genotypes: two typical six-row spring cultivars (ACCA and Léger), a two-row spring (Gobernadora) and a two-row winter (Igri) cultivar. First, by optimizing the harvest stage for each genotype we obtained a twofold to fourfold increase in the yield of embryogenic microspores. Second, two pretreatments (0.3 M mannitol for 2 days, or a combination of cold and heat over 15 days) both performed significantly better than the commonly used cold pretreatment (28 days at 4 °C). Third, an induction medium-containing mannitol (32 g/l) doubled green plant regeneration. Fourth, a plating density of 10(6) microspores/ml yielded the highest number of green regenerated plants. Our most important findings were then confirmed using sets of F1s from a six-row, spring-type breeding program.

  5. Improved pose and affinity predictions using different protocols tailored on the basis of data availability

    NASA Astrophysics Data System (ADS)

    Prathipati, Philip; Nagao, Chioko; Ahmad, Shandar; Mizuguchi, Kenji

    2016-09-01

    The D3R 2015 grand drug design challenge provided a set of blinded challenges for evaluating the applicability of our protocols for pose and affinity prediction. In the present study, we report the application of two different strategies for the two D3R protein targets HSP90 and MAP4K4. HSP90 is a well-studied target system with numerous co-crystal structures and SAR data. Furthermore the D3R HSP90 test compounds showed high structural similarity to existing HSP90 inhibitors in BindingDB. Thus, we adopted an integrated docking and scoring approach involving a combination of both pharmacophoric and heavy atom similarity alignments, local minimization and quantitative structure activity relationships modeling, resulting in the reasonable prediction of pose [with the root mean square deviation (RMSD) values of 1.75 Å for mean pose 1, 1.417 Å for the mean best pose and 1.85 Å for the mean all poses] and affinity (ROC AUC = 0.702 at 7.5 pIC50 cut-off and R = 0.45 for 180 compounds). The second protein, MAP4K4, represents a novel system with limited SAR and co-crystal structure data and little structural similarity of the D3R MAP4K4 test compounds to known MAP4K4 ligands. For this system, we implemented an exhaustive pose and affinity prediction protocol involving docking and scoring using the PLANTS software which considers side chain flexibility together with protein-ligand fingerprints analysis assisting in pose prioritization. This protocol through fares poorly in pose prediction (with the RMSD values of 4.346 Å for mean pose 1, 4.69 Å for mean best pose and 4.75 Å for mean all poses) and produced reasonable affinity prediction (AUC = 0.728 at 7.5 pIC50 cut-off and R = 0.67 for 18 compounds, ranked 1st among 80 submissions).

  6. Recovery of human DNA profiles from poached deer remains part 2: improved recovery protocol without the need for LCN analysis.

    PubMed

    Tobe, Shanan S; Bailey, Stuart; Govan, James; Welch, Lindsey A

    2013-03-01

    Although poaching is a common wildlife crime, the high and prohibitive cost of specialised animal testing means that many cases are left un-investigated. We previously described a novel approach to wildlife crime investigation that looked at the identification of human DNA on poached animal remains (Tobe, Govan and Welch, 2011). Human DNA was successfully isolated and amplified from simulated poaching incidents, however a low template protocol was required which made this method unsuitable for use in many laboratories. We now report on an optimised recovery and amplification protocol which removes the need for low template analysis. Samples from 10 deer (40 samples total - one from each leg) analysed in the original study were re-analysed in the current study with an additional 11 deer samples. Four samples analysed using Chelex did not show any results and a new method was devised whereby the available DNA was concentrated. By combining the DNA extracts from all tapings of the same deer remains followed by concentration, the recovered quantity of human DNA was found to be 29.5pg±43.2pg, 31× greater than the previous study. The use of the Investigator Decaplex SE (QIAGEN) STR kit provided better results in the form of more complete profiles than did the AmpFℓSTR® SGM Plus® kit at 30cycles (Applied Biosystems). Re-analysis of the samples from the initial study using the new, optimised protocol resulted in an average increase of 18% of recovered alleles. Over 17 samples, 71% of the samples analysed using the optimised protocol showed sufficient amplification for comparison to a reference profile and gave match probabilities ranging from 7.7690×10(-05) to 2.2706×10(-14). The removal of low template analysis means this optimised method provides evidence of high probative value and is suitable for immediate use in forensic laboratories. All methods and techniques used are standard and are compatible with current SOPs. As no high cost non-human DNA analysis is

  7. An Improved Mobility-Based Control Protocol for Tolerating Clone Failures in Wireless Sensor Networks

    PubMed Central

    Zhou, Yuping; Xiong, Naixue; Tan, Mingxin; Huang, Rufeng; Kleonbet, Jon

    2016-01-01

    Nowadays, with the ubiquitous presence of the Internet of Things industry, the application of emerging sensor networks has become a focus of public attention. Unattended sensor nodes can be comprised and cloned to destroy the network topology. This paper proposes a novel distributed protocol and management technique for the detection of mobile replicas to tolerate node failures. In our scheme, sensors’ location claims are forwarded to obtain samples only when the corresponding witnesses meet. Meanwhile, sequential tests of statistical hypotheses are applied to further detect the cloned node by witnesses. The combination of randomized detection based on encountering and sequential tests drastically reduces the routing overhead and false positive/negative rate for detection. Theoretical analysis and simulation results show the detection efficiency and reasonable overhead of the proposed method. PMID:27886054

  8. An Improved Mobility-Based Control Protocol for Tolerating Clone Failures in Wireless Sensor Networks.

    PubMed

    Zhou, Yuping; Xiong, Naixue; Tan, Mingxin; Huang, Rufeng; Kleonbet, Jon

    2016-11-23

    Nowadays, with the ubiquitous presence of the Internet of Things industry, the application of emerging sensor networks has become a focus of public attention. Unattended sensor nodes can be comprised and cloned to destroy the network topology. This paper proposes a novel distributed protocol and management technique for the detection of mobile replicas to tolerate node failures. In our scheme, sensors' location claims are forwarded to obtain samples only when the corresponding witnesses meet. Meanwhile, sequential tests of statistical hypotheses are applied to further detect the cloned node by witnesses. The combination of randomized detection based on encountering and sequential tests drastically reduces the routing overhead and false positive/negative rate for detection. Theoretical analysis and simulation results show the detection efficiency and reasonable overhead of the proposed method.

  9. Study on an improved variable stiffness tuned mass damper based on conical magnetorheological elastomer isolators

    NASA Astrophysics Data System (ADS)

    Wang, Qi; Dong, Xufeng; Li, Luyu; Ou, Jinping

    2017-10-01

    Use of a variable stiffness tuned mass damper (VSTMD) is an effective approach to reduce the dynamic responses of a structure with shifting natural frequencies. Magnetorheological elastomer (MRE) isolators can be used to build VSTMD due to their tunable stiffness by applying a magnetic field. However, conventional MRE isolators show deformation limits, huge energy consumption and uneconomic production. Focused on developing a MRE VSTMD system to improve this situation, a conical MRE isolator has been proposed and tested. Compared to conventional MRE isolators, the conical isolator shows much higher efficiency, better overall stability, greater deformability and a larger tunable range. The experimental results indicate that the prototype can provide a 46.29% increase in frequency and a 75 N control force range with a 25 W power source. The quick responding MRE VSTMD system has the potential to accurately provide the desired stiffness in two directions to achieve a better structure control.

  10. A Protocol for Improved Measurement of Arterial Flow Rate in Preclinical Ultrasound.

    PubMed

    Kenwright, D A; Thomson, A J W; Hadoke, P W F; Anderson, T; Moran, C M; Gray, G A; Hoskins, P R

    2015-11-01

    To describe a protocol for the measurement of blood flow rate in small animals and to compare flow rate measurements against measurements made using a transit time flowmeter. Measurements were made in rat and mice using a Visualsonics Vevo 770 scanner. The flow rate in carotid and femoral arteries was calculated from the time-average maximum velocity and vessel diameter. A correction factor was applied to correct for the overestimation of velocity arising from geometric spectral broadening. Invasive flow rate measurements were made using a Transonics system. Measurements were achieved in rat carotid and femoral arteries and in mouse carotid arteries. Image quality in the mouse femoral artery was too poor to obtain diameter measurements. The applied correction factor in practice was 0.71-0.77. The diameter varied by 6-18% during the cardiac cycle. There was no overall difference in the flow rate measured using ultrasound and using transit-time flowmeters. The flow rates were comparable with those previously reported in the literature. There was wide variation in flow rates in the same artery in individual animals. Transit-time measurements were associated with changes of a factor of 10 during the typical 40 min measurement period, associated with probe movement, vessel spasm, vessel kinking and other effects. A protocol for the measurement of flow rate in arteries in small animals has been described and successfully used in rat carotid and femoral arteries and in mouse carotid arteries. The availability of a noninvasive procedure for flow rate measurement avoids the problems with changes in flow associated with an invasive procedure.

  11. Improving Packet Delivery Performance of Publish/Subscribe Protocols in Wireless Sensor Networks

    PubMed Central

    Davis, Ernesto García; Calveras, Anna; Demirkol, Ilker

    2013-01-01

    MQTT-S and CoAP are two protocols able to use the publish/subscribe model in Wireless Sensor Networks (WSNs). The high scalability provided by the publish/subscribe model may incur a high packet loss and therefore requires an efficient reliability mechanism to cope with this situation. The reliability mechanism of MQTT-S and CoAP employs a method which defines a fixed value for the retransmission timeout (RTO). This article argues that this method is not efficient for deploying publish/subscribe in WSN, because it may be unable to recover a packet, therefore resulting in a lower packet delivery ratio (PDR) at the subscriber nodes. This article proposes and evaluates an adaptive RTO method, which consists in using a Smooth Round-trip Time and multiplying it by a constant parameter (K). Thanks to this method, the reliability mechanism of MQTT-S and CoAP would be able to react properly to packet loss and would also be lightweight in terms of energy, memory and computing for sensor nodes where these resources are critical. We present a detailed evaluation of the effects of the K value on the calculation of the adaptive RTO method. We also establish the setting for obtaining the highest PDR on the subscriber nodes for single-hop and multi-hop scenarios. The results for single-hop scenario show that use of the appropriate K value for the adaptive RTO method increases the PDR up to 76% for MQTT-S and up to 38% for CoAP when compared with the use of fixed RTO method for both protocols, respectively. Meanwhile the same comparison for multi-hop scenario, the adaptive RTO method increases the PDR up to 36% for MQTT-S and up to 14% for CoAP. PMID:23291579

  12. Improving packet delivery performance of publish/subscribe protocols in wireless sensor networks.

    PubMed

    Davis, Ernesto García; Calveras, Anna; Demirkol, Ilker

    2013-01-04

    MQTT-S and CoAP are two protocols able to use the publish/subscribe model in Wireless Sensor Networks (WSNs). The high scalability provided by the publish/subscribe model may incur a high packet loss and therefore requires an efficient reliability mechanism to cope with this situation. The reliability mechanism of MQTT-S and CoAP employs a method which defines a fixed value for the retransmission timeout (RTO). This article argues that this method is not efficient for deploying publish/subscribe in WSN, because it may be unable to recover a packet, therefore resulting in a lower packet delivery ratio (PDR) at the subscriber nodes. This article proposes and evaluates an adaptive RTO method, which consists in using a Smooth Round-trip Time and multiplying it by a constant parameter (K). Thanks to this method, the reliability mechanism of MQTT-S and CoAP would be able to react properly to packet loss and would also be lightweight in terms of energy, memory and computing for sensor nodes where these resources are critical. We present a detailed evaluation of the effects of the K value on the calculation of the adaptive RTO method. We also establish the setting for obtaining the highest PDR on the subscriber nodes for single-hop and multi-hop scenarios. The results for single-hop scenario show that use of the appropriate K value for the adaptive RTO method increases the PDR up to 76% for MQTT-S and up to 38% for CoAP when compared with the use of fixed RTO method for both protocols, respectively. Meanwhile the same comparison for multi-hop scenario, the adaptive RTO method increases the PDR up to 36% for MQTT-S and up to 14% for CoAP.

  13. A Protocol for Improved Measurement of Arterial Flow Rate in Preclinical Ultrasound

    PubMed Central

    Kenwright, D. A.; Thomson, A. J. W.; Hadoke, P. W. F.; Anderson, T.; Moran, C. M.; Gray, G. A.; Hoskins, P. R.

    2015-01-01

    Purpose: To describe a protocol for the measurement of blood flow rate in small animals and to compare flow rate measurements against measurements made using a transit time flowmeter. Materials and Methods: Measurements were made in rat and mice using a Visualsonics Vevo 770 scanner. The flow rate in carotid and femoral arteries was calculated from the time-average maximum velocity and vessel diameter. A correction factor was applied to correct for the overestimation of velocity arising from geometric spectral broadening. Invasive flow rate measurements were made using a Transonics system. Results: Measurements were achieved in rat carotid and femoral arteries and in mouse carotid arteries. Image quality in the mouse femoral artery was too poor to obtain diameter measurements. The applied correction factor in practice was 0.71–0.77. The diameter varied by 6–18% during the cardiac cycle. There was no overall difference in the flow rate measured using ultrasound and using transit-time flowmeters. The flow rates were comparable with those previously reported in the literature. There was wide variation in flow rates in the same artery in individual animals. Transit-time measurements were associated with changes of a factor of 10 during the typical 40 min measurement period, associated with probe movement, vessel spasm, vessel kinking and other effects. Conclusion: A protocol for the measurement of flow rate in arteries in small animals has been described and successfully used in rat carotid and femoral arteries and in mouse carotid arteries. The availability of a noninvasive procedure for flow rate measurement avoids the problems with changes in flow associated with an invasive procedure. PMID:27689153

  14. A learning protocol improves the validity of the Borg 6-20 RPE scale during indoor cycling.

    PubMed

    Soriano-Maldonado, A; Romero, L; Femia, P; Roero, C; Ruiz, J R; Gutierrez, A

    2014-05-01

    The present study assessed the effectiveness of an RPE learning protocol for improving the validity of the Borg 6-20 RPE scale for self-regulating exercise intensity during Indoor Cycling (IC). 16 healthy adults performed 5 IC sessions. Heart rate (HR) was continuously monitored. In sessions 1 and 5, participants perceptually regulated exercise intensity to produce pre-determined RPE levels equivalent to estimated HR levels. Sessions 2, 3 and 4 focused on the RPE learning protocol. The RPE-measured HR correlation was 0.59 in session 1 and 0.67 in session 5 (both P<0.001). The intraclass correlation coefficient between measured HR and estimated HR was 0.85 in session 1 and 0.90 in session 5. There was bias in session 1 (3.69 bpm [SD=12.44]; P<0.001) but not in session 5 (0.99 bpm [SD=9.7]; P=0.178; mean HR difference session 5 vs. 1: -2.70 bpm [95% CI: -4.49 to -0.91]; P=0.003; Cohen's d=0.24). There was association between the HR difference and the magnitude of the measurement in session 1 (β=0.208; P<0.01) but not in session 5 (β=-0.072; P=0.34). These preliminary results suggested that an RPE learning protocol might slightly improve the validity of the Borg 6-20 RPE scale for exercise intensity self-regulation during IC sessions in healthy adults. © Georg Thieme Verlag KG Stuttgart · New York.

  15. Improvement and scale-down of a Trichoderma reesei shake flask protocol to microtiter plates enables high-throughput screening.

    PubMed

    Giese, Heiner; Kruithof, Paulien; Meier, Kristina; Sieben, Michaela; Antonov, Elena; Hommes, Ronald W J; Büchs, Jochen

    2014-12-01

    Nowadays, high-throughput screening is essential for determining the best microbial strains and fermentation conditions. Although microtiter plates allow higher throughput in screening than shake flasks, they do not guarantee sufficient oxygen supply if operated at unsuitable conditions. This is especially the case in viscous fermentations, potentially leading to poor liquid movement and surface growth. Therefore, in this study, two aims were pursued. First, an industrial Trichoderma reesei shake flask protocol is improved with respect to oxygen supply and production. Second, this improved shake flask protocol is scaled down into microtiter plate under consideration of similar oxygen supply. For this purpose, the respiration activity monitoring system (RAMOS) was applied. An approach based on a sulfite system was introduced to ensure equal maximum oxygen transfer capacities (OTRmax) in microtiter plates and shake flasks. OTRmax-values of 250 mL shake flasks and 24-well microtiter plates were determined in a wide range of operating conditions. These sulfite datasets were used to identify operating conditions leading to the same oxygen supply for T. reesei in shake flasks and 24-well microtiter plates. For 24-well microtiter plates, the shake flask OTRmax of 20 mmol/L/h of an industrial protocol was obtained under the following optimal operating conditions: 1 mL filling volume per well, 200 rpm shaking frequency and 50 mm shaking diameter. With these conditions almost identical oxygen transfer rates and product concentrations were measured in both scales. The proposed approach is a fast and accurate means to scale-down established screening procedures into microtiter plates to achieve high-throughput.

  16. AN IMPROVED CELL ISOLATION METHOD FOR FLOW CYTOMETRIC AND FUNCTIONAL ANALYSES OF CUTANEOUS WOUND LEUKOCYTES

    PubMed Central

    Brubaker, Aleah L.; Schneider, David F.; Palmer, Jessica L.; Faunce, Douglas E.; Kovacs, Elizabeth J.

    2011-01-01

    Isolation of leukocytes from full-thickness excisional wounds has proven to be a difficult process that results in poor cell yield and holds significant limitations for functional assays. Given the increased interest in the isolation, characterization and functional measurements of wound-derived cell populations, herein we describe a method for preparing wound cell suspensions with an improved yield that enables both phenotypic and functional assessments. PMID:21889511

  17. Improving the Sandia Test Protocols with Advanced Inverter Functionality Testing of INV3, VV11, FW21, and L/HVRT

    SciTech Connect

    Johnson, Jay Dean

    2013-11-01

    Sandia National Laboratories has created a test protocol for IEC TR 61850-90-7 advanced distributed energy resource (DER) functions, titled "Test Protocols for Advanced Inverter Interoperability Functions," often referred to as the Sandia Test Protocols. This document is currently in draft form, but has been shared with stakeholders around the world with the ultimate goal of collaborating to create a consensus set of test protocols which can be then incorporated into an International Electrotechnical Commission (IEC) and/or Underwriters Laboratories (UL) certification standard. The protocols are designed to ensure functional interoperability of DER (primarily photovoltaic (PV) inverters and energy storage systems) as specified by the IEC technical report through communication and electrical tests. In this report, Sandia exercises the electrical characterization portion of the test protocols for four functions: constant power factor (INV3), volt-var (VV11), frequency-watt (FW21), and Low and High Voltage Ride Through (L/HVRT). The goal of the tests reported here was not to characterize the performance of the equipment under test (EUT), but rather to (a) exercise the draft Sandia Test Protocols in order to identify any revisions needed in test procedures, conditions, or equipment and (b) gain experience with state-of-the-art DER equipment to determine if the tests put unrealistic or overly aggressive requirements on EUT operation. In performing the work according to the current versions of the protocols, Sandia was able to identify weaknesses in the current versions and suggest improvements to the test protocols.

  18. A decade of improvements in Mimiviridae and Marseilleviridae isolation from amoeba.

    PubMed

    Pagnier, Isabelle; Reteno, Dorine-Gaelle Ikanga; Saadi, Hanene; Boughalmi, Mondher; Gaia, Morgan; Slimani, Meriem; Ngounga, Tatsiana; Bekliz, Meriem; Colson, Philippe; Raoult, Didier; La Scola, Bernard

    2013-01-01

    Since the isolation of the first giant virus, the Mimivirus, by T.J. Rowbotham in a cooling tower in Bradford, UK, and after its characterisation by our group in 2003, we have continued to develop novel strategies to isolate additional strains. By first focusing on cooling towers using our original time-consuming procedure, we were able to isolate a new lineage of giant virus called Marseillevirus and a new Mimivirus strain called Mamavirus. In the following years, we have accumulated the world's largest unique collection of giant viruses by improving the use of antibiotic combinations to avoid bacterial contamination of amoeba, developing strategies of preliminary screening of samples by molecular methods, and using a high-throughput isolation method developed by our group. Based on the inoculation of nearly 7,000 samples, our collection currently contains 43 strains of Mimiviridae (14 in lineage A, 6 in lineage B, and 23 in lineage C) and 17 strains of Marseilleviridae isolated from various environments, including 3 of human origin. This study details the procedures used to build this collection and paves the way for the high-throughput isolation of new isolates to improve the record of giant virus distribution in the environment and the determination of their pangenome.

  19. Improvement of Treatment Outcomes after Implementation of a Massive Transfusion Protocol: A Level I Trauma Center Experience.

    PubMed

    Nunn, Andrew; Fischer, Peter; Sing, Ronald; Templin, Megan; Avery, Michael; Christmas, A Britton

    2017-04-01

    We assessed the effectiveness of the implementation of an institutional massive transfusion protocol (MTP) for resuscitation with a 1:1:1 transfusion ratio of packed red blood cell (PRBC), fresh frozen plasma, and platelet units. In a Level I trauma center database, all trauma admissions (2004-2012) that received massive transfusions (≥10 units PRBCs in the first 24 hours) were reviewed retrospectively. Demographic data, transfusion ratios, and outcomes were compared before (PRE) and after (POST) MTP implementation in May 2008. Age, sex, and mechanism of injury were similar between 239 PRE and 208 POST trauma patients requiring massive transfusion. Transfusion ratios of fresh frozen plasma:PRBC and platelet:PRBC increased after MTP implementation. Among survivors, MTP implementation shortened hospital length of stay from 31 to 26 days (P = 0.04) and intensive care unit length of stay from 31 to 26 days (P = 0.02). Linear regression identified treatment after (versus before) implementation of MTP as an independent predictor of decreased ventilator days after adjusting for age, Glasgow Coma Scale, and chest Abbreviated Injury Score (P < 0.0001). Modest improvement in ratios likely does not account for all significant improvements in outcomes. Implementing a standardized protocol likely impacts automation, efficiency, and/or timeliness of product delivery.

  20. An optimised protocol for isolation of RNA from small sections of laser-capture microdissected FFPE tissue amenable for next-generation sequencing.

    PubMed

    Amini, Parisa; Ettlin, Julia; Opitz, Lennart; Clementi, Elena; Malbon, Alexandra; Markkanen, Enni

    2017-08-23

    Formalin-fixed paraffin embedded (FFPE) tissue constitutes a vast treasury of samples for biomedical research. Thus far however, extraction of RNA from FFPE tissue has proved challenging due to chemical RNA-protein crosslinking and RNA fragmentation, both of which heavily impact on RNA quantity and quality for downstream analysis. With very small sample sizes, e.g. when performing Laser-capture microdissection (LCM) to isolate specific subpopulations of cells, recovery of sufficient RNA for analysis with reverse-transcription quantitative PCR (RT-qPCR) or next-generation sequencing (NGS) becomes very cumbersome and difficult. We excised matched cancer-associated stroma (CAS) and normal stroma from clinical specimen of FFPE canine mammary tumours using LCM, and compared the commonly used protease-based RNA isolation procedure with an adapted novel technique that additionally incorporates a focused ultrasonication step. We successfully adapted a protocol that uses focused ultrasonication to isolate RNA from small amounts of deparaffinised, stained, clinical LCM samples. Using this approach, we found that total RNA yields could be increased by 8- to 12-fold compared to a commonly used protease-based extraction technique. Surprisingly, RNA extracted using this new approach was qualitatively at least equal if not superior compared to the old approach, as Cq values in RT-qPCR were on average 2.3-fold lower using the new method. Finally, we demonstrate that RNA extracted using the new method performs comparably in NGS as well. We present a successful isolation protocol for extraction of RNA from difficult and limiting FFPE tissue samples that enables successful analysis of small sections of clinically relevant specimen. The possibility to study gene expression signatures in specific small sections of archival FFPE tissue, which often entail large amounts of highly relevant clinical follow-up data, unlocks a new dimension of hitherto difficult-to-analyse samples which now

  1. Monitoring universal protocol compliance through real-time clandestine observation by medical students results in performance improvement.

    PubMed

    Logan, Catherine A; Cressey, Brienne D; Wu, Roger Y; Janicki, Adam J; Chen, Cyril X; Bolourchi, Meena L; Hodnett, Jessica L; Stratigis, John D; Mackey, William C; Fairchild, David G

    2012-01-01

    To measure universal protocol compliance through real-time, clandestine observation by medical students compared with chart audit reviews, and to enable medical students the opportunity to become conscious of the importance of medical errors and safety initiatives. With endorsement from Tufts Medical Center's (TMC's) Chief Medical Officer and Surgeon-in-Chief, 8 medical students performed clandestine observation audits of 98 cases from April to August 2009. A compliance checklist was based on TMC's presurgical checklist. Our initial results led to interventions to improve our universal protocol procedures, including modifications to the operating room white board and presurgical checklist, and specific feedback to surgical departments. One year later, 6 medical students performed observations of 100 cases from June to August 2010. Tufts Medical Center, Boston, Massachusetts, which is an academic medical center and the principal teaching hospital for Tufts University School of Medicine. An operating room coordinator placed the medical students into 1 of our 25 operating rooms with students entering under the premise of observing the anesthesiologist for clinical education. The observations were performed Monday to Friday between 7 am and 4 pm. Although observations were not randomized, no single service or type of surgery was targeted for observation. A broad range of departments was observed. In 8.2% of cases, the surgical site was unmarked. A Time Out occurred in 89.7% of cases. The entire surgical team was attentive during the time out in 82% of cases. The presurgical checklist was incomplete before incision in 13 cases. Images were displayed in 82% of cases. The operating room "white board" was filled out completely in 49% of cases. Team introductions occurred in 13 cases. One year later, compliance increased in all Universal Protocol dimensions. Direct, real-time observation by medical students provides an accurate and granular assessment of compliance with

  2. Protocol Improvements for Low Concentration DNA-Based Bioaerosol Sampling and Analysis

    PubMed Central

    Ng, Chun Kiat; Miller, Dana; Cao, Bin

    2015-01-01

    Introduction As bioaerosol research attracts increasing attention, there is a need for additional efforts that focus on method development to deal with different environmental samples. Bioaerosol environmental samples typically have very low biomass concentrations in the air, which often leaves researchers with limited options in choosing the downstream analysis steps, especially when culture-independent methods are intended. Objectives This study investigates the impacts of three important factors that can influence the performance of culture-independent DNA-based analysis in dealing with bioaerosol environmental samples engaged in this study. The factors are: 1) enhanced high temperature sonication during DNA extraction; 2) effect of sampling duration on DNA recoverability; and 3) an alternative method for concentrating composite samples. In this study, DNA extracted from samples was analysed using the Qubit fluorometer (for direct total DNA measurement) and quantitative polymerase chain reaction (qPCR). Results and Findings The findings suggest that additional lysis from high temperature sonication is crucial: DNA yields from both high and low biomass samples increased up to 600% when the protocol included 30-min sonication at 65°C. Long air sampling duration on a filter media was shown to have a negative impact on DNA recoverability with up to 98% of DNA lost over a 20-h sampling period. Pooling DNA from separate samples during extraction was proven to be feasible with margins of error below 30%. PMID:26619279

  3. Protocol: Optimised methodology for isolation of nuclei from leaves of species in the Solanaceae and Rosaceae families

    PubMed Central

    2013-01-01

    In this study, a protocol is described for rapid preparation of an enriched, reasonably pure fraction of nuclear proteins from the leaves of tobacco (Nicotiana tabacum), potato (Solanum tuberosum) and apple (Malus domestica). The protocol gives reproducible results and can be carried out quickly in 2 hours. Tissue extracts clarified with filtration were treated with non-ionic detergent (Triton X-100) to lyse membranes of contaminating organelles. Nuclei were collected from a 60% Percoll layer of density gradient following low-speed centrifugation. Western blot analysis using antibodies to marker proteins of organelles indicated that the nuclear protein fractions were highly enriched and free or nearly free of proteins from the endoplasmic reticulum and chloroplasts. PMID:23886449

  4. Network protocol changes can improve DisCom WAN performance : evaluating TCP modifications and SCTP in the ASC tri-lab environment.

    SciTech Connect

    Tolendino, Lawrence F.; Hu, Tan Chang

    2005-06-01

    The Advanced Simulation and Computing (ASC) Distance Computing (DisCom) Wide Area Network (WAN) is a high performance, long distance network environment that is based on the ubiquitous TCP/IP protocol set. However, the Transmission Control Protocol (TCP) and the algorithms that govern its operation were defined almost two decades ago for a network environment vastly different from the DisCom WAN. In this paper we explore and evaluate possible modifications to TCP that purport to improve TCP performance in environments like the DisCom WAN. We also examine a much newer protocol, SCTP (Stream Control Transmission Protocol) that claims to provide reliable network transport while also implementing multi-streaming, multi-homing capabilities that are appealing in the DisCom high performance network environment. We provide performance comparisons and recommendations for continued development that will lead to network communications protocol implementations capable of supporting the coming ASC Petaflop computing environments.

  5. Improvement in quality of life and sexual functioning in a comorbid sample after the unified protocol transdiagnostic group treatment.

    PubMed

    de Ornelas Maia, Ana Claudia Corrêa; Sanford, Jenny; Boettcher, Hannah; Nardi, Antonio E; Barlow, David

    2017-10-01

    Patients with multiple mental disorders often experience sexual dysfunction and reduced quality of life. The unified protocol (UP) is a transdiagnostic treatment for emotional disorders that has the potential to improve quality of life and sexual functioning via improved emotion management. The present study evaluates changes in quality of life and sexual functioning in a highly comorbid sample treated with the UP in a group format. Forty-eight patients were randomly assigned to either a UP active-treatment group or a medication-only control group. Treatment was delivered in 14 sessions over the course of 4 months. Symptoms of anxiety and depression were assessed using the Beck Anxiety Inventory and Beck Depression Inventory. Sexual functioning was assessed by the Arizona Sexual Experience Scale (ASEX), and quality of life was assessed by the World Health Organization Quality of Life-BREF scale (WHOQOL-BREF). Quality of life, anxiety and depression all significantly improved among participants treated with the UP. Some improvement in sexual functioning was also noted. The results support the efficacy of the UP in improving quality of life and sexual functioning in comorbid patients. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Equine chorionic gonadotropin improves the efficacy of a timed artificial insemination protocol in buffalo during the nonbreeding season.

    PubMed

    Carvalho, N A T; Soares, J G; Porto Filho, R M; Gimenes, L U; Souza, D C; Nichi, M; Sales, J S; Baruselli, P S

    2013-02-01

    Two experiments were conducted to evaluate the effects of equine chorionic gonadotropin (eCG) treatment on ovarian follicular response, luteal function, and pregnancy in buffaloes subjected to a timed artificial insemination (TAI) protocol during the nonbreeding season. In experiment 1, 59 buffalo cows were randomly assigned to two groups (with and without eCG). On the first day of the synchronization protocol (Day 0), cows received an intravaginal progesterone (P4) device plus 2.0 mg estradiol benzoate im. On Day 9, the P4 device was removed, all cows were given 0.150 mg PGF(2α) im, and half were given 400 IU eCG im. On Day 11, all cows were given 10 μg of buserelin acetate im (GnRH). Transrectal ultrasonography of the ovaries was performed on Days 0 and 9 to determine the presence and diameter of the largest follicle; between Days 11 and 14 (12 hours apart), to evaluate the dominant follicle diameter and the interval from device removal to ovulation; and on Days 16, 20, and 24 to measure CL diameter. Blood samples were collected on Days 16, 20, and 24 to measure serum P4. In experiment 2, 256 buffaloes were assigned to the same treatments described in experiment 1, and TAI was performed 16 hours after GnRH treatment. Pregnancy diagnosis was performed by ultrasonography 30 days after TAI. Treatment with eCG increased the maximum diameter of dominant follicles (P = 0.09), ovulation rate (P = 0.05), CL diameter (P = 0.03), and P4 concentrations (P = 0.01) 4 days after TAI, and pregnancy per AI (52.7%, 68/129 vs. 39.4%, 50/127; P = 0.03). Therefore, eCG improved ovarian follicular response, luteal function during the subsequent diestrus, and fertility for buffalo subjected to a TAI synchronization protocol during the nonbreeding season.

  7. Use of a five-day progesterone-based timed AI protocol to determine if flunixin meglumine improves pregnancy per timed AI in dairy heifers.

    PubMed

    Rabaglino, M B; Risco, C A; Thatcher, M-J; Lima, F; Santos, J E P; Thatcher, W W

    2010-06-01

    Two experiments were conducted to test the hypothesis that the 5 d Co-Synch + CIDR (Controlled Internal Drug Release insert containing progesterone) protocol could be applied as an efficient timed AI (TAI) protocol in dairy heifers, and that treatment with flunixin meglumine (FM) during the period of CL maintenance would increase pregnancy per TAI (P/TAI) and late survival of embryos. Objectives were: 1) in Experiment 1, to compare P/TAI with the 5 d Co-Synch+CIDR protocol to a PGF(2alpha)/GnRH protocol; and 2) in Experiment 2, to determine if FM administered 15.5 and 16 d after first TAI would increase P/TAI, using the 5 d Co-Synch+CIDR protocol with a new or previously used (5 d) CIDR insert. In Experiment 1, 248 heifers were assigned randomly to either the PGF(2alpha)/GnRH protocol (n=120) or the 5 d Co-Synch+CIDR protocol (n=128). Pregnancy per TAI did not differ between the 5 d Co-Synch+CIDR protocol (53.1%) and the PGF(2alpha)/GnRH protocol (45.8%; P=0.22). In Experiment 2, 325 heifers synchronized with the 5 d Co-Synch+CIDR protocol were assigned randomly to receive two injections of FM (FM group; n=158) at 15.5 and 16 d after TAI, or to remain as untreated controls (n=165). Pregnancy per TAI in Experiment 2 was 59.4 and 59.5% at 45 d for control and FM groups, respectively, with no differences between groups (P=0.83). The 5 d Co-Synch+CIDR protocol resulted in an acceptable P/TAI in dairy heifers. However, FM did not improve P/TAI in dairy heifers. 2010 Elsevier Inc. All rights reserved.

  8. Strategies to improve the quality of life of persons post-stroke: protocol of a systematic review.

    PubMed

    Munce, Sarah E P; Perrier, Laure; Shin, Saeha; Adhihetty, Chamila; Pitzul, Kristen; Nelson, Michelle L A; Bayley, Mark T

    2017-09-07

    While many outcomes post-stroke (e.g., depression) have been previously investigated, there is no complete data on the impact of a variety of quality improvement strategies on the quality of life and physical and psychological well-being of individuals post-stroke. The current paper outlines a systematic review protocol on the impact of quality improvement strategies on quality of life as well as physical and psychological well-being of individuals with stroke. MEDLINE, CINAHL, EMBASE, and PsycINFO databases will be searched. Two independent reviewers will conduct all levels of screening, data abstraction, and quality appraisal. Only randomized controlled trials that report on the impact of quality improvement strategies on quality of life outcomes in people with stroke will be included. The secondary outcomes will be physical and psychological well-being. Quality improvement strategies include audit and feedback, case management, team changes, electronic patient registries, clinician education, clinical reminders, facilitated relay of clinical information to clinicians, patient education, (promotion of) self-management, patient reminder systems, and continuous quality improvement. Studies published since 2000 will be included to increase the relevancy of findings. Results will be grouped according to the target group of the varying quality improvement strategies (i.e., health system, health care professionals, or patients) and/or by any other noteworthy grouping variables, such as etiology of stroke or by sex. This systematic review will identify those quality improvement strategies aimed at the health system, health care professionals, and patients that impact the quality of life of individuals with stroke. Improving awareness and utilization of such strategies may enhance uptake of stroke best practices and reduce inappropriate health care utilization costs. PROSPERO, CRD42017064141.

  9. An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater

    PubMed Central

    Kim, Jinyong; Oh, Euna; Banting, Graham S.; Braithwaite, Shannon; Chui, Linda; Ashbolt, Nicholas J.; Neumann, Norman F.; Jeon, Byeonghwa

    2016-01-01

    Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater. PMID:27617011

  10. Implementing performance improvement in New Zealand emergency departments: the six hour time target policy national research project protocol

    PubMed Central

    2012-01-01

    Background In May 2009, the New Zealand government announced a new policy aimed at improving the quality of Emergency Department care and whole hospital performance. Governments have increasingly looked to time targets as a mechanism for improving hospital performance and from a whole system perspective, using the Emergency Department waiting time as a performance measure has the potential to see improvements in the wider health system. However, the imposition of targets may have significant adverse consequences. There is little empirical work examining how the performance of the wider hospital system is affected by such a target. This project aims to answer the following questions: How has the introduction of the target affected broader hospital performance over time, and what accounts for these changes? Which initiatives and strategies have been successful in moving hospitals towards the target without compromising the quality of other care processes and patient outcomes? Is there a difference in outcomes between different ethnic and age groups? Which initiatives and strategies have the greatest potential to be transferred across organisational contexts? Methods/design The study design is mixed methods; combining qualitative research into the behaviour and practices of specific case study hospitals with quantitative data on clinical outcomes and process measures of performance over the period 2006-2012. All research activity is guided by a Kaupapa Māori Research methodological approach. A dynamic systems model of acute patient flows was created to frame the study. Consequences of the target (positive and negative) will be explored by integrating analyses and insights gained from the quantitative and qualitative streams of the study. Discussion At the time of submission of this protocol, the project has been underway for 12 months. This time was necessary to finalise both the case study sites and the secondary outcomes through key stakeholder consultation. We

  11. A cluster randomized trial on improving nurses' detection and management of elder abuse and neglect (I-NEED): study protocol.

    PubMed

    Loh, Debbie Ann; Choo, Wan Yuen; Hairi, Noran Naqiah; Othman, Sajaratulnisah; Mohd Hairi, Farizah; Mohd Mydin, Fadzilah Hanum; Jaafar, Siti Nur Illiani; Tan, Maw Pin; Mohd Ali, Zainudin; Abdul Aziz, Suriyati; Ramli, Rohaya; Mohamad, Rosmala; Lal Mohammad, Zaiton; Hassan, Norlela; Brownell, Patricia; Bulgiba, Awang

    2015-11-01

    The aim of this study was to describe a trial protocol of an educational intervention for nurses to improve their awareness and practice in detecting and managing elder abuse and neglect. Knowledgeable and skilful nurses are crucial amidst the growing numbers of maltreated older patients. This trial is a multi-site, three-armed, community-based cluster randomized controlled trial with 6-months follow-up. This study will involve 390 community and registered nurses from government health clinics in Negeri Sembilan, Malaysia (protocol approved in October 2013). This three-phased study, premised on the Precede-Proceed Model, comprises baseline focus group discussion and survey (Phase 1), development of training module (Phase 2) and implementation and evaluation of the training (Phase 3). Eligible participants will be randomized to the control group (continuous nursing education), intervention group A (face-to-face intensive training programme) or group B (face-to-face intensive training programme and an educational video). Outcome measures include improvement in knowledge and awareness on elder abuse and neglect and the number of cases identified and managed during follow-up. Data will be collected at baseline, immediate postintervention, 3- and 6-month follow-up. Findings from this study will provide empirical support for the development of a training module for nurses on the detection and management of elder abuse and neglect, towards improving healthcare delivery and the well-being of vulnerable older adults. This study is funded by the University of Malaya Research Grant (RP001C-13HTM), (FL002-13SBS) and University of Malaya Grand Challenge (PEACE) Grant (GC001C-14HTM) awarded in May 2013, July 2013 and September 2014. © 2015 John Wiley & Sons Ltd.

  12. Improving the methods for isolation of monocyte and establishing macrophage cell culture in caprine model.

    PubMed

    Shaqinah, Nurrul Nasruddin; Mazlina, Mazlan; Zamri-Saad, M; Hazilawati, Hamzah; Jasni, Sabri

    2016-08-01

    Monocytes are widely used for immunological research, especially in the study of innate immune system. Although methods for isolation of human monocytes have been established, the procedure for non-human monocyte has not been well developed. This paper describes an improved method for isolation of monocyte and the subsequent macrophage cultivation from caprine blood. Monocytes were isolated from 16 ml of heparinized caprine blood using double density methods; the Ficoll and Percoll. The number of monocytes obtained was 5.12 ± 0.89 × 10(7) cells/ml at 70 % purity. The isolated monocytes were maintained in 10 % fetal bovine serum-enriched Dulbecco's Modified Eagle Medium for maturation to form macrophage cell culture. At the end of the experiment, the harvested macrophage was 2.48 ± 0.33 × 10(6) cells/ml.

  13. Development of a training protocol to improve reading performance in peripheral vision

    PubMed Central

    Yu, Deyue; Legge, Gordon E.; Park, Heejung; Gage, Emily; Chung, Susana T. L.

    2009-01-01

    People with central-field loss must use peripheral vision for reading. Previous studies have shown that reading performance in peripheral vision can improve with extensive practice on a trigram letter-recognition task. The present study compared training on this task with training on two other character-based tasks (lexical decision and RSVP (Rapid Serial Visual Presentation) reading) which might plausibly produce more improvement in peripheral reading speed. Twenty-eight normally sighted young adults were trained at 10° in the lower visual field in a pre/post design. All three training methods produced significant improvements in reading speed, with average gains of 39% for lexical-decision, 54% for trigram letter-recognition, and 72% for RSVP training. Although the RSVP training was most effective, the lexical-decision task has the advantage of easy self administration making it more practical for home-based training. PMID:19819251

  14. Trial protocol OPPTIMUM– Does progesterone prophylaxis for the prevention of preterm labour improve outcome?

    PubMed Central

    2012-01-01

    Background Preterm birth is a global problem, with a prevalence of 8 to 12% depending on location. Several large trials and systematic reviews have shown progestogens to be effective in preventing or delaying preterm birth in selected high risk women with a singleton pregnancy (including those with a short cervix or previous preterm birth). Although an improvement in short term neonatal outcomes has been shown in some trials these have not consistently been confirmed in meta-analyses. Additionally data on longer term outcomes is limited to a single trial where no difference in outcomes was demonstrated at four years of age of the child, despite those in the “progesterone” group having a lower incidence of preterm birth. Methods/Design The OPPTIMUM study is a double blind randomized placebo controlled trial to determine whether progesterone prophylaxis to prevent preterm birth has long term neonatal or infant benefit. Specifically it will study whether, in women with singleton pregnancy and at high risk of preterm labour, prophylactic vaginal natural progesterone, 200 mg daily from 22 – 34 weeks gestation, compared to placebo, improves obstetric outcome by lengthening pregnancy thus reducing the incidence of preterm delivery (before 34 weeks), improves neonatal outcome by reducing a composite of death and major morbidity, and leads to improved childhood cognitive and neurosensory outcomes at two years of age. Recruitment began in 2009 and is scheduled to close in Spring 2013. As of May 2012, over 800 women had been randomized in 60 sites. Discussion OPPTIMUM will provide further evidence on the effectiveness of vaginal progesterone for prevention of preterm birth and improvement of neonatal outcomes in selected groups of women with singleton pregnancy at high risk of preterm birth. Additionally it will determine whether any reduction in the incidence of preterm birth is accompanied by improved childhood outcome. Trial registration ISRCTN14568373 PMID

  15. One-shock versus three-shock defibrillation protocol significantly improves outcome in a porcine model of prolonged ventricular fibrillation cardiac arrest.

    PubMed

    Tang, Wanchun; Snyder, David; Wang, Jinglan; Huang, Lei; Chang, Yun-Te; Sun, Shijie; Weil, Max Harry

    2006-06-13

    The success of resuscitation with a 1-shock versus the conventional 3-shock defibrillation protocol was investigated subject to the range of treatment variation imposed by automated external defibrillators (AEDs). Ventricular fibrillation was induced in 44 domestic pigs. After 7 minutes of untreated VF, animals were randomized among 4 groups representing all combinations of the 1- versus 3-shock protocol and 2 different AED regimens (AED1, AED2). Because few AEDs support a 1-shock protocol, manual defibrillators were used to replicate the AED treatment regimen: electrical waveform, dose sequence, and cardiopulmonary resuscitation (CPR) interruption intervals. Initial shock(s) were delivered, followed by 60 seconds of CPR, and the treatment was repeated until resuscitation was successful or for 15 minutes. The 1-shock protocol was associated with improved outcome, reducing CPR interruptions from 45% to 34% of total resuscitation time (P=0.019) and increasing survival from 64% to 100% (P=0.004). Survival was 91% for AED1 versus 36% for AED2 (P=0.024) with a 3-shock protocol but was increased to 100% for both by adoption of a 1-shock protocol. Improvements in postresuscitation left ventricular ejection fraction and stroke volume were observed with AED1 compared with AED2 (difference of means, 15% and 28% of baseline respectively, P<0.001) regardless of defibrillation protocol. Adoption of a 1-shock versus a 3-shock resuscitation protocol improved survival and minimized outcome differences imposed by variations in AED design and implementation. When a conventional 3-shock defibrillation protocol was used, however, the choice of AED had a significant impact on resuscitation outcome.

  16. Attempting to improve function and quality of life using the FTM Protocol: case report.

    PubMed

    Butler, Andrew; Blanton, Sarah; Rowe, Veronica; Wolf, Steven

    2006-09-01

    The Functional Tone Management (FTM) arm training program uses repetitive task practice, stretching, and electrical stimulation as patients with moderate upper extremity hemiparesis wear a dynamic hand orthosis to retrain grasp and release of objects. The case study quantitatively evaluates the extent to which FTM training improved function and quality of life in a patient who met criteria for which the device was intended. The participant, a 44-year-old male in the chronic stage of stroke recovery with moderate right upper extremity motor impairment, was trained for 10 days over a 2-week period by a licensed clinician with extensive experience in FTM training methods. Measurements were taken over 3 baseline visits, then immediately preintervention, postintervention, and at a 3-month follow-up. The outcome measures, following the International Classification of Functioning, Disability, and Health (ICF) Model, included the: Fugl-Meyer assessment (FMA) (upper extremity portion), UE range of motion (ROM), Modified Ashworth scale, Wolf Motor Function Test (WMFT), and the Motor Activity Log (MAL). Health related quality of life was measured using the Stroke Impact Scale (SIS). The participant showed limited gains in upper extremity function immediately postintervention. Increases in AROM occurred in forearm supination and wrist flexion and extension, but no improvements were noted in finger ROM. A slight decrease in tone was found in forearm pronators (1 to 0) and wrist flexors (1+ to 1). WMFT values for the more affected upper extremity did not change immediately after training, but a 17% reduction in time occurred at follow-up, with improvements, most notably in the tasks of lifting a pencil and lifting a paper clip. No changes occurred in UE-FMA scores immediately following the intervention, however, a 17% improvement was measured at followup. A slight improvement in MAL scores was noted but not to a level of independent functional use of the impaired upper extremity

  17. Efficacy and cost effectiveness of telemedicine for improving access to care in the Paris region: study protocols for eight trials.

    PubMed

    Charrier, Nathanael; Zarca, Kevin; Durand-Zaleski, Isabelle; Calinaud, Christine

    2016-02-08

    With the development of information and communication technologies, telemedicine has been proposed as a way to improve patient management by facilitating access to appropriate diagnosis and treatment. The Paris Ile de France Regional Health Agency is currently funding a comprehensive program of telemedicine experiments. This article describes the protocols for the evaluation of the implementation of telemedicine in the Paris region. Over 2,500 patients have been included in eight studies addressing the use of telemedicine in the context of specific diseases or settings. Two projects are randomized controlled trials, while the six other projects are based on before-after designs (differences in differences studies). Based on the MAST model and the French national framework, we identified endpoints to assess the impact of telemedicine on five dimensions: clinical effectiveness, cost-effectiveness, security of the application, patient satisfaction and quality of life and perception of professionals. Telemedicine encompasses a wide range of services and stakeholders, and thus study protocols must be tailored to the specific constraints and interests of the users. NCT02110433 (03/07/2014), NCT02157740 (05/27/2014), NCT02374697 (02/05/2015), NCT02157727 (05/27/2014), NCT02229279 (08/28/2014), NCT02368769 (02/05/2015), NCT02164747 (NCT02164747), NCT02309905 (11/27/2014).

  18. Improvement of Electrical Stimulation Protocol for Simultaneous Measurement of Extracellular Potential with On-Chip Multi-Electrode Array System

    NASA Astrophysics Data System (ADS)

    Kaneko, Tomoyuki; Nomura, Fumimasa; Hattori, Akihiro; Yasuda, Kenji

    2012-06-01

    Cardiotoxicity testing with a multi-electrode array (MEA) system requires the stable beating of cardiomyocytes for the measurement of the field potential duration (FPD), because different spontaneous beating rates cause different responses of FPD prolongation induced by drugs, and the beating rate change effected by drugs complicates the FPD prolongation assessment. We have developed an on-chip MEA system with electrical stimulation for the measurement of the FPD during the stable beating of human embryonic stem (ES) cell-derived cardiomyocyte clusters. Using a conventional bipolar stimulation protocol, we observed such large artifacts in electrical stimulation that we could not estimate the FPD quantitatively. Therefore, we improved the stimulation protocol by using sequential rectangular pulses in which the positive and negative stimulation voltages and number of pulses could be changed flexibly. The balanced voltages and number of pulses for sequential rectangular pulses enabled the recording of small negative artifacts only, which hardly affected the FPD measurement of human-ES-cell-derived cardiomyocyte clusters. These conditions of electrical stimulation are expected to find applications for the control of constant beating for cardiotoxicity testing.

  19. A modified ultra-long pituitary downregulation protocol improved endometrial receptivity and clinical outcome for infertile patients with polycystic ovarian syndrome.

    PubMed

    Gong, Fei; Li, Xihong; Zhang, Shunji; Ma, Hainan; Cai, Sufen; Li, Juan; Lin, G E; Lu, Guangxiu

    2015-11-01

    There are currently various protocols for in vitro fertilization (IVF). For patients with polycystic ovarian syndrome (PCOS), an optimized protocol for the downregulation of pituitary follicle stimulating hormone and luteinizing hormone via gonadotropin-releasing hormone agonist (GnRHa) remains a challenge. In the present study, the primary endpoint of an ultra-long and a conventional long GnRHa protocol for intracytoplasmic sperm injection/IVF treatments of patients with PCOS was retrospectively compared. In the modified ultra-long protocol group, endometrial thickness, morphology, and blood flow were significantly improved, as compared with in the conventional long protocol group. Furthermore, the serum progestogen (P) concentrations and P/estrogen (E2) [(Px1,000/E2)] ratio on the day of human chorionic gonadotrophin administration were significantly decreased in the modified ultra-long downregulation group, whereas the pregnancy and implantation rates were significantly higher. There were no significant differences in the average number of obtained oocytes, good quality embryo rates, cancel rates, fertilization rates, abortion rates, serious ovarian hyperstimulation syndrome incidences, ectopic pregnancy rates or gonadotropin (Gn) dosages between the two groups. These results suggest that the modified ultra-long protocol plus human menopausal Gn medication may be superior to the conventional long protocol, and may lead to improved implantation and pregnancy outcomes for infertile patients with PCOS.

  20. A modified ultra-long pituitary downregulation protocol improved endometrial receptivity and clinical outcome for infertile patients with polycystic ovarian syndrome

    PubMed Central

    GONG, FEI; LI, XIHONG; ZHANG, SHUNJI; MA, HAINAN; CAI, SUFEN; LI, JUAN; LIN, GE; LU, GUANGXIU

    2015-01-01

    There are currently various protocols for in vitro fertilization (IVF). For patients with polycystic ovarian syndrome (PCOS), an optimized protocol for the downregulation of pituitary follicle stimulating hormone and luteinizing hormone via gonadotropin-releasing hormone agonist (GnRHa) remains a challenge. In the present study, the primary endpoint of an ultra-long and a conventional long GnRHa protocol for intracytoplasmic sperm injection/IVF treatments of patients with PCOS was retrospectively compared. In the modified ultra-long protocol group, endometrial thickness, morphology, and blood flow were significantly improved, as compared with in the conventional long protocol group. Furthermore, the serum progestogen (P) concentrations and P/estrogen (E2) [(Px1,000/E2)] ratio on the day of human chorionic gonadotrophin administration were significantly decreased in the modified ultra-long downregulation group, whereas the pregnancy and implantation rates were significantly higher. There were no significant differences in the average number of obtained oocytes, good quality embryo rates, cancel rates, fertilization rates, abortion rates, serious ovarian hyperstimulation syndrome incidences, ectopic pregnancy rates or gonadotropin (Gn) dosages between the two groups. These results suggest that the modified ultra-long protocol plus human menopausal Gn medication may be superior to the conventional long protocol, and may lead to improved implantation and pregnancy outcomes for infertile patients with PCOS. PMID:26640563

  1. Development of an improved vaccine evaluation protocol to compare the efficacy of Newcastle disease vaccines

    USDA-ARS?s Scientific Manuscript database

    The failure to control and to eradicate Newcastle Disease (ND) with vaccination alone in countries where the etiological agent of the disease, virulent Newcastle Disease Virus (vNDV) is endemic underscores the need to improve the efficacy of currently available NDV vaccines and vaccination approache...

  2. The agricultural model intercomparison and improvement project (AgMIP): Protocols and pilot studies

    USDA-ARS?s Scientific Manuscript database

    The Agricultural Model Intercomparison and Improvement Project (AgMIP) is a distributed climate-scenario simulation research activity for historical period model intercomparison and future climate change conditions with participation of multiple crop and agricultural economic model groups around the...

  3. MObile Technology for Improved Family Planning Services (MOTIF): study protocol for a randomised controlled trial.

    PubMed

    Smith, Chris; Vannak, Uk; Sokhey, Ly; Ngo, Thoai D; Gold, Judy; Khut, Khemrin; Edwards, Phil; Rathavy, Tung; Free, Caroline

    2013-12-12

    Providing women with contraceptive methods following abortion is important to reduce repeat abortion rates, yet evidence for effective post-abortion family planning interventions are limited. This protocol outlines the evaluation of a mobile phone-based intervention using voice messages to support post-abortion family planning in Cambodia. A single blind randomised controlled trial of 500 participants. Clients aged 18 or over, attending for abortion at four Marie Stopes International clinics in Cambodia, owning a mobile phone and not wishing to have a child at the current time are randomised to the mobile phone-based intervention or control (standard care) with a 1:1 allocation ratio.The intervention comprises a series of six automated voice messages to remind clients about available family planning methods and provide a conduit for additional support. Clients can respond to message prompts to request a phone call from a counsellor, or alternatively to state they have no problems. Clients requesting to talk to a counsellor, or who do not respond to the message prompts, receive a call from a Marie Stopes International Cambodia counsellor who provides individualised advice and support regarding family planning. The duration of the intervention is 3 months. The control group receive existing standard of care without the additional mobile phone-based support.We hypothesise that the intervention will remind clients about contraceptive methods available, identify problems with side effects early and provide support, and therefore increase use of post-abortion family planning, while reducing discontinuation and unsafe method switching.Participants are assessed at baseline and at 4 months. The primary outcome measure is use of an effective modern contraceptive method at 4 months post abortion. Secondary outcome measures include contraception use, pregnancy and repeat abortion over the 4-month post-abortion period.Risk ratios will be used as the measure of effect of the

  4. MObile Technology for Improved Family Planning Services (MOTIF): study protocol for a randomised controlled trial

    PubMed Central

    2013-01-01

    Background Providing women with contraceptive methods following abortion is important to reduce repeat abortion rates, yet evidence for effective post-abortion family planning interventions are limited. This protocol outlines the evaluation of a mobile phone-based intervention using voice messages to support post-abortion family planning in Cambodia. Methods/Design A single blind randomised controlled trial of 500 participants. Clients aged 18 or over, attending for abortion at four Marie Stopes International clinics in Cambodia, owning a mobile phone and not wishing to have a child at the current time are randomised to the mobile phone-based intervention or control (standard care) with a 1:1 allocation ratio. The intervention comprises a series of six automated voice messages to remind clients about available family planning methods and provide a conduit for additional support. Clients can respond to message prompts to request a phone call from a counsellor, or alternatively to state they have no problems. Clients requesting to talk to a counsellor, or who do not respond to the message prompts, receive a call from a Marie Stopes International Cambodia counsellor who provides individualised advice and support regarding family planning. The duration of the intervention is 3 months. The control group receive existing standard of care without the additional mobile phone-based support. We hypothesise that the intervention will remind clients about contraceptive methods available, identify problems with side effects early and provide support, and therefore increase use of post-abortion family planning, while reducing discontinuation and unsafe method switching. Participants are assessed at baseline and at 4 months. The primary outcome measure is use of an effective modern contraceptive method at 4 months post abortion. Secondary outcome measures include contraception use, pregnancy and repeat abortion over the 4-month post-abortion period. Risk ratios will be used as

  5. An improved microinjection protocol for the transfer of Wolbachia pipientis between infected and uninfected strains of the flour beetle Tribolium confusum.

    PubMed

    Chang, N W; Wade, M J

    1996-07-01

    The lethality of halocarbon and other oils to hatching larvae of the flour beetle Tribolium confusum limits existing microinjection protocols, because postinjection survivorship is only 5.6% of the eggs injected. We report the development of an oil-free protocol that improves survivorship fivefold. We used this protocol to transfect the cytoplasmic endosymbiont Wolbachia pipientis from infected donor eggs to uninfected host eggs and observed reproductive incompatibility in 40% of the surviving, injected eggs. Compared with mock injected controls (35.9% survival) or microinjection of uninfected cytoplasm (32.3% survival), injection of Wolbachia-infected cytoplasm into host eggs significantly reduced egg-to-adult survival (25.1%).

  6. Study protocol: national research partnership to improve primary health care performance and outcomes for Indigenous peoples

    PubMed Central

    2010-01-01

    Background Strengthening primary health care is critical to reducing health inequity between Indigenous and non-Indigenous Australians. The Audit and Best practice for Chronic Disease Extension (ABCDE) project has facilitated the implementation of modern Continuous Quality Improvement (CQI) approaches in Indigenous community health care centres across Australia. The project demonstrated improvements in health centre systems, delivery of primary care services and in patient intermediate outcomes. It has also highlighted substantial variation in quality of care. Through a partnership between academic researchers, service providers and policy makers, we are now implementing a study which aims to 1) explore the factors associated with variation in clinical performance; 2) examine specific strategies that have been effective in improving primary care clinical performance; and 3) work with health service staff, management and policy makers to enhance the effective implementation of successful strategies. Methods/Design The study will be conducted in Indigenous community health centres from at least six States/Territories (Northern Territory, Western Australia, New South Wales, South Australia, Queensland and Victoria) over a five year period. A research hub will be established in each region to support collection and reporting of quantitative and qualitative clinical and health centre system performance data, to investigate factors affecting variation in quality of care and to facilitate effective translation of research evidence into policy and practice. The project is supported by a web-based information system, providing automated analysis and reporting of clinical care performance to health centre staff and management. Discussion By linking researchers directly to users of research (service providers, managers and policy makers), the partnership is well placed to generate new knowledge on effective strategies for improving the quality of primary health care and

  7. One-day pulsed-field gel electrophoresis protocol for rapid determination of emetic Bacillus cereus isolates.

    PubMed

    Kaminska, Paulina S; Fiedoruk, Krzysztof; Jankowska, Dominika; Mahillon, Jacques; Nowosad, Karol; Drewicka, Ewa; Zambrzycka, Monika; Swiecicka, Izabela

    2015-04-01

    Bacillus cereus, the Gram-positive and spore-forming ubiquitous bacterium, may cause emesis as the result of food intoxication with cereulide, a heat-stable emetic toxin. Rapid determination of cereulide-positive B. cereus isolates is of highest importance due to consequences of this intoxication for human health and life. Here we present a 1-day pulsed-field gel electrophoresis for emetic B. cereus isolates, which allows rapid and efficient determination of their genomic relatedness and helps determining the source of intoxication in case of outbreaks caused by these bacilli.

  8. Improvement of Longevity and Viability of Sperm Cells Isolated from Pollen of Zea mays L. 1

    PubMed Central

    Zhang, Guichang; Williams, Connie M.; Campenot, Mary K.; McGann, Locksley E.; Cass, David D.

    1992-01-01

    Our previous studies showed that the common maize (Zea mays L.) sperm isolation medium (Brewbaker and Kwack salts in 0.44 m sucrose without buffering) caused cell lysis in vitro. In an attempt to remedy this situation, 6 sugars, 10 buffers, 5 pH values, and 3 membrane protective agents were screened to improve longevity and viability of isolated Zea mays sperm cells as estimated by hemacytometry and flow cytometry. Use of 0.55 m galactose in the isolation solution increased sperm yield by 2.5-fold compared with sucrose, and suspension of isolated sperm cells in the galactose solution gave the best longevity among the six sugars. Buffering the galactose solution with 2 mm 2-(N-morpholino)ethanesulfonic acid significantly improved longevity, whereas other buffers had no effect or decreased the longevity and/or viability. Among the five pH values tested (5.0, 6.0, 6.7, 7.0, and 8.0), pH 6.7 appeared to be optimal for maintenance of both longevity and viability. Screening of membrane protectants showed that cysteine caused a rapid decrease in cell viability and increased lysis, whereas dithiothreitol increased the cell numbers but lowered their viability. Addition of 0.1% bovine serum albumin increased cell numbers and viability, and about 70% of the cells remained viable after 72 h of suspension. Cell longevity and viability were also improved in 0.44 m sucrose when the solution was conditioned with 2-(N-morpholino)ethanesulfonic acid and bovine serum albumin. Use of 2-(N-morpholino)ethanesulfonic acid and bovine serum albumin inthe isolation and suspension medium significantly improved the viability and longevity of sperm cells isolated from Zea mays pollen. PMID:16652985

  9. An improved procedure for the isolation of intact chloroplasts of high photosynthetic capacity.

    PubMed Central

    Cerović, Z G; Plesnicar, M

    1984-01-01

    An improved procedure for the mechanical isolation of chloroplasts of high degree of intactness (90-95%) and photosynthetic capacity (25-50 mmol of O2/s per mol of chlorophyll) is described. The combination of pea plants (Pisum sativum L.) as starting material and the high reproducibility of the procedure readily and cheaply yields reliable intact chloroplasts for photosynthetic studies. PMID:6497861

  10. Systematic review protocol of interventions to improve the psychological well-being of general practitioners.

    PubMed

    Murray, Marylou; Murray, Lois; Donnelly, Michael

    2015-09-22

    The challenges and complexities faced by general practitioners are increasing, and there are concerns about their well-being. Consequently, attention has been directed towards developing and evaluating interventions and strategies to improve general practitioner well-being and their capacity to cope with workplace challenges. This systematic review aims to evaluate research evidence regarding the effectiveness of interventions designed to improve general practitioner well-being. Eligible studies will include programmes developed to improve psychological well-being that have assessed outcomes using validated tools pertaining to well-being and related outcomes. Only programmes that have been evaluated using controlled study designs will be reviewed. An appropriately developed search strategy will be applied to six electronic databases: the Cochrane Database of Systematic Reviews, MEDLINE, Embase, CINAHL, PsycINFO and Web of Science. Studies will be screened in two stages by two independent reviewers. A third reviewer will arbitrate when required. Pre-specified inclusion and exclusion criteria will be assessed during a pilot phase early on in the review process. The Cochrane data extraction form will be adapted and applied to each eligible study by two independent reviewers, and each study will be appraised critically using standardised checklists from the Cochrane Handbook. Methodological quality will be taken into account in the analysis of the data and the synthesis of results. A narrative synthesis will be undertaken if data is unsuited to a meta-analysis. The systematic review will be reported according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidance. This will be the first systematic review on this topic, and the evidence synthesis will aid decision-making by general practitioners, policy makers and planners regarding ways in which to improve GP well-being. Findings will be disseminated at general practitioner meetings

  11. A Comprehensive Protocol for Improving the Description of Saprolegniales (Oomycota): Two Practical Examples (Saprolegnia aenigmatica sp. nov. and Saprolegnia racemosa sp. nov.)

    PubMed Central

    Sandoval-Sierra, Jose Vladimir; Diéguez-Uribeondo, Javier

    2015-01-01

    The description, identification and classification of organisms are the pillar in biodiversity and evolutionary studies. The fungal-like organism Saprolegnia contains important animal pathogens. However, its taxonomy is weak, making it difficult to perform further studies. This problem mainly arises from the unavailability of suitable holotypes. We propose a standardized protocol for describing Saprolegnia spp. that includes good cultural practices and proper holotype preservation. In order to illustrate this new proposal, we describe two species, Saprolegnia aenigmatica sp. nov. and Saprolegnia racemosa sp. nov., based on the recently described molecular operational taxonomic units (MOTUs), phylogenetic relationships, and the analyses of morphological features. We show that they belong to two different MOTUs that are grouped into two sister clades. Morphologically, we find that S. racemosa exhibits a species-specific character, i.e., aggrupation of oogonia in racemes, while S. aenigmatica does not have any specific characters. Analyses of a combined set of characters, i.e., length and breadth of sporangia, length/breadth ratio (l/b) of oogonia, cyst and oospore diameter, and the number of oospores per oogomium, allow distinguishing these two species. To improve Saprolegnia taxonomy, we propose to incorporate into the protologue: (i) several isolates of the new species; (ii) the rDNA sequences to compare them to data-bases of Saprolegnia sequences of reference; (iii) a phylogenetic analysis to check relationships with other species; (iv) to preserve holotypes in absolute ethanol and to include lyophilized material from holotype; and (v) the ex-type as a pure culture from single-spore isolates stored in at least two different collections. PMID:26186364

  12. An improved protocol for sequencing of repetitive genomic regions and structural variations using mutagenesis and next generation sequencing.

    PubMed

    Sipos, Botond; Massingham, Tim; Stütz, Adrian M; Goldman, Nick

    2012-01-01

    The rise of Next Generation Sequencing (NGS) technologies has transformed de novo genome sequencing into an accessible research tool, but obtaining high quality eukaryotic genome assemblies remains a challenge, mostly due to the abundance of repetitive elements. These also make it difficult to study nucleotide polymorphism in repetitive regions, including certain types of structural variations. One solution proposed for resolving such regions is Sequence Assembly aided by Mutagenesis (SAM), which relies on the fact that introducing enough random mutations breaks the repetitive structure, making assembly possible. Sequencing many different mutated copies permits the sequence of the repetitive region to be inferred by consensus methods. However, this approach relies on molecular cloning in order to isolate and amplify individual mutant copies, making it hard to scale-up the approach for use in conjunction with high-throughput sequencing technologies. To address this problem, we propose NG-SAM, a modified version of the SAM protocol that relies on PCR and dilution steps only, coupled to a NGS workflow. NG-SAM therefore has the potential to be scaled-up, e.g. using emerging microfluidics technologies. We built a realistic simulation pipeline to study the feasibility of NG-SAM, and our results suggest that under appropriate experimental conditions the approach might be successfully put into practice. Moreover, our simulations suggest that NG-SAM is capable of reconstructing robustly a wide range of potential target sequences of varying lengths and repetitive structures.

  13. Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols.

    PubMed

    Figueroa, Gloria; Parira, Tiyash; Laverde, Alejandra; Casteleiro, Gianna; El-Mabhouh, Amal; Nair, Madhavan; Agudelo, Marisela

    2016-10-18

    Dendritic cells (DCs) are antigen presenting cells of the immune system that play a crucial role in lymphocyte responses, host defense mechanisms, and pathogenesis of inflammation. Isolation and study of DCs have been important in biological research because of their distinctive features. Although they are essential key mediators of the immune system, DCs are very rare in blood, accounting for approximately 0.1 - 1% of total blood mononuclear cells. Therefore, alternatives for isolation methods rely on the differentiation of DCs from monocytes isolated from peripheral blood mononuclear cells (PBMCs). The utilization of proper isolation techniques that combine simplicity, affordability, high purity, and high yield of cells is imperative to consider. In the current study, two distinct methods for the generation of DCs will be compared. Monocytes were selected by adherence or negatively enriched using magnetic separation procedure followed by differentiation into DCs with IL-4 and GM-CSF. Monocyte and MDDC viability, proliferation, and phenotype were assessed using viability dyes, MTT assay, and CD11c/ CD14 surface marker analysis by imaging flow cytometry. Although the magnetic separation method yielded a significant higher percentage of monocytes with higher proliferative capacity when compared to the adhesion method, the findings have demonstrated the ability of both techniques to simultaneously generate monocytes that are capable of proliferating and differentiating into viable CD11c+ MDDCs after seven days in culture. Both methods yielded > 70% CD11c+ MDDCs. Therefore, our results provide insights that contribute to the development of reliable methods for isolation and characterization of human DCs.

  14. Efficiency of four secondary enrichment protocols in differentiation and isolation of Listeria spp. and Listeria monocytogenes from smoked fish processing chains.

    PubMed

    Duarte, G; Vaz-Velho, M; Capell, C; Gibbs, P

    1999-11-15

    Four secondary enrichment protocols (conventional methods: UVM II, Fraser 24 h and Fraser 48 h: Impedimetric method: Listeria electrical detection medium) were studied for their ability to isolate Listeria spp. and Listeria monocytogenes from fish and environmental samples collected along the processing chain of cold-smoked fish. From all methods, Listeria spp. and L. monocytogenes were respectively present in 56 and 34 of 315 samples analysed. Fraser broth incubated for 48 h gave the fewest false negative Listeria spp. results [4/56; (7.1%)], but concurrently only 15/34 (44.1%) samples were correctly identified as containing L. monocytogenes, Listeria electrical detection (LED) medium detected only 36/56 (64.3%) Listeria spp. positive samples. Despite this lower isolation rate, LED identified 20/34 (58.8%) L. monocytogenes positive samples correctly and gave fewer false positive results. The overall conclusion was that more than one isolation method is needed to accurately estimate L. monocytogenes contamination rates.

  15. Implementation of observational pain management protocol to improve pain management for long-term institutionalized older care residents with dementia: study protocol for a cluster-randomized controlled trial.

    PubMed

    Liu, Justina Yat Wa; Lai, Claudia

    2014-03-13

    Systematic use of observational pain tools has been advocated as a means to improve pain management for care home residents with dementia. Pain experts suggest that any observational tool should be used as part of a comprehensive pain management protocol, which should include score interpretation and verification with appropriately suggested treatments. The Observational Pain Management Protocol (Protocol) was therefore developed. This study aims to investigate the extent to which the implementation of this Protocol can improve pain management in care home residents with dementia. In this two-group, single-blinded, cluster-randomized controlled trial, 122 care home residents with dementia and pain-related diagnoses will be recruited from eight care homes (that is 15 to 16 residents from each care home). Invitations will be sent to all local care homes who meet the home selection criteria. The eight care homes will be randomly selected from all care homes that agree to join this trial. They will then be randomized to either the control or experimental conditions. Participants from each care home will be placed into their home's corresponding group to avoid 'contamination' effects across participants. Each intervention cycle will take 16 weeks (that is, baseline assessment and care home staff training for 4 weeks and Protocol implementation for 12 weeks). The Protocol will guide the pain management of the participants in the experimental care homes. Meanwhile, the control care homes will continue their usual pain management strategies. Intervention effects will be measured weekly during the protocol implementation period and compared with the baseline measurements, as well as between the experimental and control conditions. Although similar pain protocols have been suggested previously, the recommendations were based on experts' opinions rather than evaluation of research studies. The feasibility and effectiveness of this kind of pain management protocol, tailored to

  16. Text Messaging to Improve Hypertension Medication Adherence in African Americans: BPMED Intervention Development and Study Protocol

    PubMed Central

    Artinian, Nancy T; Schwiebert, Loren; Yarandi, Hossein; Levy, Phillip D

    2015-01-01

    Background Hypertension (HTN) is a major public health concern in the United States, with almost 78 million Americans age 20 years and over suffering from the condition. Moreover, HTN is a key risk factor for health disease and stroke. African Americans disproportionately shoulder the burdens of HTN, with greater prevalence, disease severity, earlier onset, and more HTN-related complications than age-matched whites. Medication adherence for the treatment of HTN is poor, with estimates indicating that only about half of hypertensive patients are adherent to prescribed medication regimens. Although no single intervention for improving medication adherence has emerged as superior to others, text message medication reminders have the potential to help improve medication adherence in African Americans with uncontrolled HTN as mobile phone adoption is very high in this population. Objective The purpose of this two-phased study was to develop (Phase I) and test in a randomized controlled trial (RCT) (Phase II) a text message system, BPMED, to improve the quality of medication management through increasing medication adherence in African Americans with uncontrolled HTN. Methods In Phase I, we recruited 16 target end-users from a primary care clinic, to assist in the development of BPMED through participating in one of three focus groups. Focus groups sought to gain patient perspectives on HTN, medication adherence, mobile phone use, and the use of text messaging to support medication adherence. Potential intervention designs were presented to participants, and feedback on the designs was solicited. In Phase II, we conducted two pilot RCTs to determine the feasibility, acceptability, and preliminary efficacy of BPMED in primary care and emergency department settings. Both pilot studies recruited approximately 60 participants, who were randomized equally between usual care and the BPMED intervention. Results Although data collection is now complete, data analysis from the

  17. Improved protocol for high-quality co-extraction of DNA and RNA from rumen digesta.

    PubMed

    Popova, M; Martin, C; Morgavi, D P

    2010-07-01

    We report an improved method for total nucleic acids extraction from rumen content samples. The method employs bead beating, and phenol-chloroform extraction followed by saline-alcohol precipitation. Total nucleic acids and RNA yield and purity were assessed by spectrophotometric measurements; RNA integrity was estimated using Agilent RNA 6000 Nano Kit on an Agilent 2100 Bioanalyzer. The method provided total nucleic acids and RNA extracts of good quantity and quality. The extraction is not time consuming and it is valuable for ecological studies of rumen microbial community structure and gene expression.

  18. Rational approaches to improving the isolation of endophytic actinobacteria from Australian native trees.

    PubMed

    Kaewkla, Onuma; Franco, Christopher M M

    2013-02-01

    In recent years, new actinobacterial species have been isolated as endophytes of plants and shrubs and are sought after both for their role as potential producers of new drug candidates for the pharmaceutical industry and as biocontrol inoculants for sustainable agriculture. Molecular-based approaches to the study of microbial ecology generally reveal a broader microbial diversity than can be obtained by cultivation methods. This study aimed to improve the success of isolating individual members of the actinobacterial population as pure cultures as well as improving the ability to characterise the large numbers obtained in pure culture. To achieve this objective, our study successfully employed rational and holistic approaches including the use of isolation media with low concentrations of nutrients normally available to the microorganism in the plant, plating larger quantities of plant sample, incubating isolation plates for up to 16 weeks, excising colonies when they are visible and choosing Australian endemic trees as the source of the actinobacteria. A hierarchy of polyphasic methods based on culture morphology, amplified 16S rRNA gene restriction analysis and limited sequencing was used to classify all 576 actinobacterial isolates from leaf, stem and root samples of two eucalypts: a Grey Box and Red Gum, a native apricot tree and a native pine tree. The classification revealed that, in addition to 413 Streptomyces spp., isolates belonged to 16 other actinobacterial genera: Actinomadura (two strains), Actinomycetospora (six), Actinopolymorpha (two), Amycolatopsis (six), Gordonia (one), Kribbella (25), Micromonospora (six), Nocardia (ten), Nocardioides (11), Nocardiopsis (one), Nonomuraea (one), Polymorphospora (two), Promicromonospora (51), Pseudonocardia (36), Williamsia (two) and a novel genus Flindersiella (one). In order to prove novelty, 12 strains were characterised fully to the species level based on polyphasic taxonomy. One strain represented a novel

  19. Improving the performance of heterogeneous multi-core processors by modifying the cache coherence protocol

    NASA Astrophysics Data System (ADS)

    Fang, Juan; Hao, Xiaoting; Fan, Qingwen; Chang, Zeqing; Song, Shuying

    2017-05-01

    In the Heterogeneous multi-core architecture, CPU and GPU processor are integrated on the same chip, which poses a new challenge to the last-level cache management. In this architecture, the CPU application and the GPU application execute concurrently, accessing the last-level cache. CPU and GPU have different memory access characteristics, so that they have differences in the sensitivity of last-level cache (LLC) capacity. For many CPU applications, a reduced share of the LLC could lead to significant performance degradation. On the contrary, GPU applications can tolerate increase in memory access latency when there is sufficient thread-level parallelism. Taking into account the GPU program memory latency tolerance characteristics, this paper presents a method that let GPU applications can access to memory directly, leaving lots of LLC space for CPU applications, in improving the performance of CPU applications and does not affect the performance of GPU applications. When the CPU application is cache sensitive, and the GPU application is insensitive to the cache, the overall performance of the system is improved significantly.

  20. A Mediterranean Diet to Improve Cardiovascular and Cognitive Health: Protocol for a Randomised Controlled Intervention Study.

    PubMed

    Wade, Alexandra T; Davis, Courtney R; Dyer, Kathryn A; Hodgson, Jonathan M; Woodman, Richard J; Keage, Hannah A D; Murphy, Karen J

    2017-02-16

    The Mediterranean diet has demonstrated efficacy for improving cardiovascular and cognitive health. However, a traditional Mediterranean diet delivers fewer serves of dairy and less dietary calcium than is currently recommended in Australia, which may limit long-term sustainability. The present study aims to evaluate whether a Mediterranean diet with adequate dairy and calcium can improve cardiovascular and cognitive function in an at-risk population, and thereby reduce risk of cardiovascular disease (CVD) and cognitive decline. A randomised, controlled, parallel, crossover design trial will compare a Mediterranean diet supplemented with dairy foods against a low-fat control diet. Forty participants with systolic blood pressure above 120 mmHg and at least two other risk factors of CVD will undertake each dietary intervention for eight weeks, with an eight-week washout period between interventions. Systolic blood pressure will be the primary measure of interest. Secondary outcomes will include measures of cardiometabolic health, dietary compliance, cognitive function, assessed using the Cambridge Neuropsychological Test Automated Battery (CANTAB), psychological well-being and dementia risk. This research will provide empirical evidence as to whether the Mediterranean diet can be modified to provide recommended dairy and calcium intakes while continuing to deliver positive effects for cardiovascular and cognitive health. The findings will hold relevance for the field of preventative healthcare and may contribute to revisions of national dietary guidelines.

  1. A Mediterranean Diet to Improve Cardiovascular and Cognitive Health: Protocol for a Randomised Controlled Intervention Study

    PubMed Central

    Wade, Alexandra T.; Davis, Courtney R.; Dyer, Kathryn A.; Hodgson, Jonathan M.; Woodman, Richard J.; Keage, Hannah A. D.; Murphy, Karen J.

    2017-01-01

    The Mediterranean diet has demonstrated efficacy for improving cardiovascular and cognitive health. However, a traditional Mediterranean diet delivers fewer serves of dairy and less dietary calcium than is currently recommended in Australia, which may limit long-term sustainability. The present study aims to evaluate whether a Mediterranean diet with adequate dairy and calcium can improve cardiovascular and cognitive function in an at-risk population, and thereby reduce risk of cardiovascular disease (CVD) and cognitive decline. A randomised, controlled, parallel, crossover design trial will compare a Mediterranean diet supplemented with dairy foods against a low-fat control diet. Forty participants with systolic blood pressure above 120 mmHg and at least two other risk factors of CVD will undertake each dietary intervention for eight weeks, with an eight-week washout period between interventions. Systolic blood pressure will be the primary measure of interest. Secondary outcomes will include measures of cardiometabolic health, dietary compliance, cognitive function, assessed using the Cambridge Neuropsychological Test Automated Battery (CANTAB), psychological well-being and dementia risk. This research will provide empirical evidence as to whether the Mediterranean diet can be modified to provide recommended dairy and calcium intakes while continuing to deliver positive effects for cardiovascular and cognitive health. The findings will hold relevance for the field of preventative healthcare and may contribute to revisions of national dietary guidelines. PMID:28212320

  2. Improving the prevention of cardiovascular disease in primary health care: the model for prevention study protocol.

    PubMed

    Volker, Nerida; Davey, Rachel C; Cochrane, Thomas; Williams, Lauren T; Clancy, Tanya

    2014-07-09

    Cardiovascular disease (CVD) is the leading cause of death globally, and accounted for nearly 31% of all deaths in Australia in 2011. The primary health care sector is at the frontline for addressing CVD, however, an evidence-to-practice gap exists in CVD risk assessment and management. General practice plays a key role in CVD risk assessment and management, but this sector cannot provide ongoing lifestyle change support in isolation. Community-based lifestyle modification services and programs provided outside the general practice setting have a key role in supporting and sustaining health behavior change. Fostering linkages between the health sector and community-based lifestyle services, and creating sustainable systems that support these sectors is important. The objective of the study Model for Prevention (MoFoP) is to take a case study approach to examine a CVD risk reduction intervention in primary health care, with the aim of identifying the key elements required for an effective and sustainable approach to coordinate CVD risk reduction across the health and community sectors. These elements will be used to consider a new systems-based model for the prevention of CVD that informs future practice. The MoFoP study will use a mixed methods approach, comprising two complementary research elements: (1) a case study, and (2) a pre/post quasi-experimental design. The case study will consider the organizations and systems involved in a CVD risk reduction intervention as a single case. The pre/post experimental design will be used for HeartLink, the intervention being tested, where a single cohort of patients between 45 and 74 years of age (or between 35 and 74 years of age if Aboriginal or Torres Strait Islander) considered to be at high risk for a CVD event will be recruited through general practice, provided with enhanced usual care and additional health behavior change support. A range of quantitative and qualitative data will be collected. This will include

  3. Triton Hodge Test: Improved Protocol for Modified Hodge Test for Enhanced Detection of NDM and Other Carbapenemase Producers

    PubMed Central

    Pasteran, Fernando; Gonzalez, Lisandro J.; Albornoz, Ezequiel; Bahr, Guillermo; Vila, Alejandro J.

    2015-01-01

    Accurate detection of carbapenemase-producing Gram-negative bacilli is of utmost importance for the control of nosocomial spread and the initiation of appropriate antimicrobial therapy. The modified Hodge test (MHT), a carbapenem inactivation assay, has shown poor sensitivity in detecting the worldwide spread of New Delhi metallo-β-lactamase (NDM). Recent studies demonstrated that NDM is a lipoprotein anchored to the outer membrane in Gram-negative bacteria, unlike all other known carbapenemases. Here we report that membrane anchoring of β-lactamases precludes detection of carbapenemase activity by the MHT. We also show that this limitation can be overcome by the addition of Triton X-100 during the test, which allows detection of NDM. We propose an improved version of the assay, called the Triton Hodge test (THT), which allows detection of membrane-bound carbapenemases with the addition of this nonionic surfactant. This test was challenged with a panel of 185 clinical isolates (145 carrying known carbapenemase-encoding genes and 40 carbapenemase nonproducers). The THT displayed test sensitivity of >90% against NDM-producing clinical isolates, while improving performance against other carbapenemases. Ertapenem provided the highest sensitivity (97 to 100%, depending on the type of carbapenemase), followed by meropenem (92.5 to 100%). Test specificity was not affected by the addition of Triton (87.5% and 92.5% with ertapenem and meropenem, respectively). This simple inexpensive test confers a large improvement to the sensitivity of the MHT for the detection of NDM and other carbapenemases. PMID:26719442

  4. Triton Hodge Test: Improved Protocol for Modified Hodge Test for Enhanced Detection of NDM and Other Carbapenemase Producers.

    PubMed

    Pasteran, Fernando; Gonzalez, Lisandro J; Albornoz, Ezequiel; Bahr, Guillermo; Vila, Alejandro J; Corso, Alejandra

    2016-03-01

    Accurate detection of carbapenemase-producing Gram-negative bacilli is of utmost importance for the control of nosocomial spread and the initiation of appropriate antimicrobial therapy. The modified Hodge test (MHT), a carbapenem inactivation assay, has shown poor sensitivity in detecting the worldwide spread of New Delhi metallo-β-lactamase (NDM). Recent studies demonstrated that NDM is a lipoprotein anchored to the outer membrane in Gram-negative bacteria, unlike all other known carbapenemases. Here we report that membrane anchoring of β-lactamases precludes detection of carbapenemase activity by the MHT. We also show that this limitation can be overcome by the addition of Triton X-100 during the test, which allows detection of NDM. We propose an improved version of the assay, called the Triton Hodge test (THT), which allows detection of membrane-bound carbapenemases with the addition of this nonionic surfactant. This test was challenged with a panel of 185 clinical isolates (145 carrying known carbapenemase-encoding genes and 40 carbapenemase nonproducers). The THT displayed test sensitivity of >90% against NDM-producing clinical isolates, while improving performance against other carbapenemases. Ertapenem provided the highest sensitivity (97 to 100%, depending on the type of carbapenemase), followed by meropenem (92.5 to 100%). Test specificity was not affected by the addition of Triton (87.5% and 92.5% with ertapenem and meropenem, respectively). This simple inexpensive test confers a large improvement to the sensitivity of the MHT for the detection of NDM and other carbapenemases.

  5. Improving community ambulation after hip fracture: protocol for a randomised, controlled trial.

    PubMed

    Orwig, D; Mangione, K K; Baumgarten, M; Terrin, M; Fortinsky, R; Kenny, A M; Gruber-Baldini, A L; Beamer, B; Tosteson, Ana; Shardell, M; Magder, L; Binder, E; Koval, K; Resnick, B; Craik, R L; Magaziner, J

    2017-01-01

    interventions. Participants are randomly assigned to one of two multi-component treatment groups: PUSH or PULSE. PUSH is based on aerobic conditioning, specificity of training, and muscle overload, while PULSE includes transcutaneous electrical nerve stimulation, flexibility activities, and active range of motion exercises. Participants in both groups receive 32 visits in their place of residence from a study physiotherapist (two visits per week on non-consecutive days for 16 weeks). The physiotherapists' adherence to the treatment protocol, and the participants' receipt of the prescribed activities are assessed. Participants also receive counselling from a registered dietician and vitamin D, calcium and multivitamin supplements during the 16-week intervention period. The primary outcome (community ambulation) is the ability to walk 300 m or more in 6minutes, as assessed by the 6-minute walk test, at 16 weeks after randomisation. Other measures at 16 and 40 weeks include cost-effectiveness, endurance, dynamic balance, walking speed, quadriceps strength, lower extremity function, activities of daily living, balance confidence, quality of life, physical activity, depressive symptoms, increase of ≥ 50 m in distance walked in 6minutes, cognitive status, and nutritional status. Analyses for all aims will be performed according to the intention-to-treat paradigm. Except for testing of the primary hypothesis, all statistical tests will be two-sided and not adjusted for multiple comparisons. The test of the primary hypothesis (comparing groups on the proportion who are community ambulators at 16 weeks after randomisation) will be based on a one-sided 0.025-level hypothesis test using a procedure consisting of four interim analyses and one final analysis with critical values chosen by a Hwang-Shih-Decani alpha-spending function. Analyses will be performed to test group differences on other outcome measures and to examine the differential impact of PUSH relative to PULSE in subgroups

  6. Improving community ambulation after hip fracture: protocol for a randomised, controlled trial

    PubMed Central

    Orwig, D; Mangione, KK; Baumgarten, M; Terrin, M; Fortinsky, R; Kenny, AM; Gruber-Baldini, AL; Beamer, B; Tosteson, ANA; Shardell, M; Magder, L; Binder, E; Koval, K; Resnick, B; Craik, RL; Magaziner, J

    2017-01-01

    , or if the participant has low potential to benefit from the interventions. Interventions Participants are randomly assigned to one of two multi-component treatment groups: PUSH or PULSE. PUSH is based on aerobic conditioning, specificity of training, and muscle overload, while PULSE includes transcutaneous electrical nerve stimulation, flexibility activities, and active range of motion exercises. Participants in both groups receive 32 visits in their place of residence from a study physiotherapist (two visits per week on non-consecutive days for 16 weeks). The physiotherapists’ adherence to the treatment protocol, and the participants’ receipt of the prescribed activities are assessed. Participants also receive counselling from a registered dietician and vitamin D, calcium and multivitamin supplements during the 16-week intervention period. Measurements The primary outcome (community ambulation) is the ability to walk 300 m or more in 6 minutes, as assessed by the 6-minute walk test, at 16 weeks after randomisation. Other measures at 16 and 40 weeks include cost-effectiveness, endurance, dynamic balance, walking speed, quadriceps strength, lower extremity function, activities of daily living, balance confidence, quality of life, physical activity, depressive symptoms, increase of ≥ 50 m in distance walked in 6 minutes, cognitive status, and nutritional status. Analysis Analyses for all aims will be performed according to the intention-to-treat paradigm. Except for testing of the primary hypothesis, all statistical tests will be two-sided and not adjusted for multiple comparisons. The test of the primary hypothesis (comparing groups on the proportion who are community ambulators at 16 weeks after randomisation) will be based on a one-sided 0.025-level hypothesis test using a procedure consisting of four interim analyses and one final analysis with critical values chosen by a Hwang-Shih-Decani alpha-spending function. Analyses will be performed to test group

  7. Working to improve survival and health for babies born very preterm: the WISH project protocol.

    PubMed

    Crowther, Caroline A; Middleton, Philippa F; Bain, Emily; Ashwood, Pat; Bubner, Tanya; Flenady, Vicki; Morris, Jonathan; McIntyre, Sarah

    2013-12-19

    Babies born very preterm (before 30 weeks gestation) are at high risk of dying in their first weeks of life, and those who survive are at risk of developing cerebral palsy in childhood. Recent high-quality evidence has shown that giving women magnesium sulphate immediately prior to very early birth can significantly increase the chances of their babies surviving free of cerebral palsy. In 2010 Australian and New Zealand clinical practice guidelines recommended this therapy. The WISH (Working to Improve Survival and Health for babies born very preterm) Project aims to bi-nationally improve and monitor the use of this therapy to reduce the risk of very preterm babies dying or having cerebral palsy. The WISH Project is a prospective cohort study. The 25 Australian and New Zealand tertiary level maternity hospitals will be provided with a package of active implementation strategies to guide the introduction and local adaptation of guideline recommendations. Surveys will be conducted at individual hospitals to evaluate outcomes related to local implementation progress and the use and value of the WISH implementation strategies. For the hospitals participating in the 'WISH audit of uptake and health outcomes data collection', the primary health outcomes (assessed through case note review, and 24 month corrected age questionnaires) will be: the proportion of eligible women receiving antenatal magnesium sulphate; and rates of death prior to primary hospital discharge and cerebral palsy at two years corrected age in infants born to eligible mothers. For hospitals wishing to assess factors influencing translation locally, barriers and facilitators will be measured through interviews with health care professionals, to further guide implementation strategies. Study outcomes for the early phase of the project (Year 1) will be compared with the later intervention phase (Years 2 and 3). The WISH Project will offer insight into the effectiveness of a multifaceted implementation

  8. Working to improve survival and health for babies born very preterm: the WISH project protocol

    PubMed Central

    2013-01-01

    Background Babies born very preterm (before 30 weeks gestation) are at high risk of dying in their first weeks of life, and those who survive are at risk of developing cerebral palsy in childhood. Recent high-quality evidence has shown that giving women magnesium sulphate immediately prior to very early birth can significantly increase the chances of their babies surviving free of cerebral palsy. In 2010 Australian and New Zealand clinical practice guidelines recommended this therapy. The WISH (Working to Improve Survival and Health for babies born very preterm) Project aims to bi-nationally improve and monitor the use of this therapy to reduce the risk of very preterm babies dying or having cerebral palsy. Methods/Design The WISH Project is a prospective cohort study. The 25 Australian and New Zealand tertiary level maternity hospitals will be provided with a package of active implementation strategies to guide the introduction and local adaptation of guideline recommendations. Surveys will be conducted at individual hospitals to evaluate outcomes related to local implementation progress and the use and value of the WISH implementation strategies. For the hospitals participating in the ‘WISH audit of uptake and health outcomes data collection’, the primary health outcomes (assessed through case note review, and 24 month corrected age questionnaires) will be: the proportion of eligible women receiving antenatal magnesium sulphate; and rates of death prior to primary hospital discharge and cerebral palsy at two years corrected age in infants born to eligible mothers. For hospitals wishing to assess factors influencing translation locally, barriers and facilitators will be measured through interviews with health care professionals, to further guide implementation strategies. Study outcomes for the early phase of the project (Year 1) will be compared with the later intervention phase (Years 2 and 3). Discussion The WISH Project will offer insight into the

  9. Intervention to improve social and family support for caregivers of dependent patients: ICIAS study protocol

    PubMed Central

    2014-01-01

    Background Despite the existence of formal professional support services, informal support (mainly family members) continues to be the main source of eldercare, especially for those who are dependent or disabled. Professionals on the primary health care are the ideal choice to educate, provide psychological support, and help to mobilize social resources available to the informal caregiver. Controversy remains concerning the efficiency of multiple interventions, taking a holistic approach to both the patient and caregiver, and optimum utilization of the available community resources. .For this reason our goal is to assess whether an intervention designed to improve the social support for caregivers effectively decreases caregivers burden and improves their quality of life. Methods/design Design: Controlled, multicentre, community intervention trial, with patients and their caregivers randomized to the intervention or control group according to their assigned Primary Health Care Team (PHCT). Study area: Primary Health Care network (9 PHCTs). Study participants: Primary informal caregivers of patients receiving home health care from participating PHCTs. Sample: Required sample size is 282 caregivers (141 from PHCTs randomized to the intervention group and 141 from PHCTs randomized to the control group. Intervention: a) PHCT professionals: standardized training to implement caregivers intervention. b) Caregivers: 1 individualized counselling session, 1 family session, and 4 educational group sessions conducted by participating PHCT professionals; in addition to usual home health care visits, periodic telephone follow-up contact and unlimited telephone support. Control: Caregivers and dependent patients: usual home health care, consisting of bimonthly scheduled visits, follow-up as needed, and additional attention upon request. Data analysis Dependent variables: Caregiver burden (short-form Zarit test), caregivers’ social support (Medical Outcomes Study), and

  10. The Microgravity Isolation Mount (MGIM): A Columbus facility for improving the microgravity quality of payloads

    NASA Technical Reports Server (NTRS)

    Owen, R. G.; Jones, D. I.; Owens, A. R.; Roberts, G.; Hadfield, P.

    1992-01-01

    The Microgravity Isolation Mount (MGIM) is a facility for providing active vibration isolation for sensitive experiments on the Columbus Attached Laboratory and the Columbus Free-Flying Laboratory. The facility is designed to be accommodated in a standard Columbus rack, and it iterfaces with existing rack utility services. The design is based on a non-contact strategy, whereby the payload 'floats' inside the rack, and its position is controlled by a number of magnetic actuators. The main advantage of using this non-contact strategy is the improved microgravity quality available. The overall design of the facility and a description of its elements are given.

  11. The Microgravity Isolation Mount (MGIM): A Columbus facility for improving the microgravity quality of payloads

    NASA Technical Reports Server (NTRS)

    Owen, R. G.; Jones, D. I.; Owens, A. R.; Roberts, G.; Hadfield, P.

    1992-01-01

    The Microgravity Isolation Mount (MGIM) is a facility for providing active vibration isolation for sensitive experiments on the Columbus Attached Laboratory and the Columbus Free-Flying Laboratory. The facility is designed to be accommodated in a standard Columbus rack, and it iterfaces with existing rack utility services. The design is based on a non-contact strategy, whereby the payload 'floats' inside the rack, and its position is controlled by a number of magnetic actuators. The main advantage of using this non-contact strategy is the improved microgravity quality available. The overall design of the facility and a description of its elements are given.

  12. Protocol for using mixed methods and process improvement methodologies to explore primary care receptionist work.

    PubMed

    Litchfield, Ian; Gale, Nicola; Burrows, Michael; Greenfield, Sheila

    2016-11-16

    The need to cope with an increasingly ageing and multimorbid population has seen a shift towards preventive health and effective management of chronic disease. This places general practice at the forefront of health service provision with an increased demand that impacts on all members of the practice team. As these pressures grow, systems become more complex and tasks delegated across a broader range of staff groups. These include receptionists who play an essential role in the successful functioning of the surgery and are a major influence on patient satisfaction. However, they do so without formal recognition of the clinical implications of their work or with any requirements for training and qualifications. Our work consists of three phases. The first will survey receptionists using the validated Work Design Questionnaire to help us understand more precisely the parameters of their role; the second involves the use of iterative focus groups to help define the systems and processes within which they work. The third and final phase will produce recommendations to increase the efficiency and safety of the key practice processes involving receptionists and identify the areas and where receptionists require targeted support. In doing so, we aim to increase job satisfaction of receptionists, improve practice efficiency and produce better outcomes for patients. Our work will be disseminated using conferences, workshops, trade journals, electronic media and through a series of publications in the peer reviewed literature. At the very least, our work will serve to prompt discussion on the clinical role of receptionists and assess the advantages of using value streams in conjunction with related tools for process improvement. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  13. Improving physician hand hygiene compliance using behavioural theories: a study protocol

    PubMed Central

    2013-01-01

    Background Healthcare-associated infections affect 10% of patients in Canadian acute-care hospitals and are significant and preventable causes of morbidity and mortality among hospitalized patients. Hand hygiene is among the simplest and most effective preventive measures to reduce these infections. However, compliance with hand hygiene among healthcare workers, specifically among physicians, is consistently suboptimal. We aim to first identify the barriers and enablers to physician hand hygiene compliance, and then to develop and pilot a theory-based knowledge translation intervention to increase physicians’ compliance with best hand hygiene practice. Design The study consists of three phases. In Phase 1, we will identify barriers and enablers to hand hygiene compliance by physicians. This will include: key informant interviews with physicians and residents using a structured interview guide, informed by the Theoretical Domains Framework; nonparticipant observation of physician/resident hand hygiene audit sessions; and focus groups with hand hygiene experts. In Phase 2, we will conduct intervention mapping to develop a theory-based knowledge translation intervention to improve physician hand hygiene compliance. Finally, in Phase 3, we will pilot the knowledge translation intervention in four patient care units. Discussion In this study, we will use a behavioural theory approach to obtain a better understanding of the barriers and enablers to physician hand hygiene compliance. This will provide a comprehensive framework on which to develop knowledge translation interventions that may be more successful in improving hand hygiene practice. Upon completion of this study, we will refine the piloted knowledge translation intervention so it can be tested in a multi-site cluster randomized controlled trial. PMID:23379466

  14. Protocol for using mixed methods and process improvement methodologies to explore primary care receptionist work

    PubMed Central

    Litchfield, Ian; Gale, Nicola; Burrows, Michael; Greenfield, Sheila

    2016-01-01

    Introduction The need to cope with an increasingly ageing and multimorbid population has seen a shift towards preventive health and effective management of chronic disease. This places general practice at the forefront of health service provision with an increased demand that impacts on all members of the practice team. As these pressures grow, systems become more complex and tasks delegated across a broader range of staff groups. These include receptionists who play an essential role in the successful functioning of the surgery and are a major influence on patient satisfaction. However, they do so without formal recognition of the clinical implications of their work or with any requirements for training and qualifications. Methods and analysis Our work consists of three phases. The first will survey receptionists using the validated Work Design Questionnaire to help us understand more precisely the parameters of their role; the second involves the use of iterative focus groups to help define the systems and processes within which they work. The third and final phase will produce recommendations to increase the efficiency and safety of the key practice processes involving receptionists and identify the areas and where receptionists require targeted support. In doing so, we aim to increase job satisfaction of receptionists, improve practice efficiency and produce better outcomes for patients. Ethics and dissemination Our work will be disseminated using conferences, workshops, trade journals, electronic media and through a series of publications in the peer reviewed literature. At the very least, our work will serve to prompt discussion on the clinical role of receptionists and assess the advantages of using value streams in conjunction with related tools for process improvement. PMID:27852720

  15. Manual-Protocol Inspired Technique for Improving Automated MR Image Segmentation during Label Fusion

    PubMed Central

    Bhagwat, Nikhil; Pipitone, Jon; Winterburn, Julie L.; Guo, Ting; Duerden, Emma G.; Voineskos, Aristotle N.; Lepage, Martin; Miller, Steven P.; Pruessner, Jens C.; Chakravarty, M. Mallar

    2016-01-01

    Recent advances in multi-atlas based algorithms address many of the previous limitations in model-based and probabilistic segmentation methods. However, at the label fusion stage, a majority of algorithms focus primarily on optimizing weight-maps associated with the atlas library based on a theoretical objective function that approximates the segmentation error. In contrast, we propose a novel method—Autocorrecting Walks over Localized Markov Random Fields (AWoL-MRF)—that aims at mimicking the sequential process of manual segmentation, which is the gold-standard for virtually all the segmentation methods. AWoL-MRF begins with a set of candidate labels generated by a multi-atlas segmentation pipeline as an initial label distribution and refines low confidence regions based on a localized Markov random field (L-MRF) model using a novel sequential inference process (walks). We show that AWoL-MRF produces state-of-the-art results with superior accuracy and robustness with a small atlas library compared to existing methods. We validate the proposed approach by performing hippocampal segmentations on three independent datasets: (1) Alzheimer's Disease Neuroimaging Database (ADNI); (2) First Episode Psychosis patient cohort; and (3) A cohort of preterm neonates scanned early in life and at term-equivalent age. We assess the improvement in the performance qualitatively as well as quantitatively by comparing AWoL-MRF with majority vote, STAPLE, and Joint Label Fusion methods. AWoL-MRF reaches a maximum accuracy of 0.881 (dataset 1), 0.897 (dataset 2), and 0.807 (dataset 3) based on Dice similarity coefficient metric, offering significant performance improvements with a smaller atlas library (< 10) over compared methods. We also evaluate the diagnostic utility of AWoL-MRF by analyzing the volume differences per disease category in the ADNI1: Complete Screening dataset. We have made the source code for AWoL-MRF public at: https://github.com/CobraLab/AWoL-MRF. PMID

  16. Improved protocol for somatic embryogenesis and calcium alginate encapsulation in Anethum graveolens L.: a medicinal herb.

    PubMed

    Dhir, Richa; Shekhawat, G S; Alam, Afroz

    2014-08-01

    An improved procedure has been developed for efficient somatic embryogenesis in Anethum graveolens. Green friable embryogenic callus was obtained from hypocotyl segments on medium augmented with 2,4-dichlorophenoxyacetic acid (2,4-D). The highest embryogenic callus induction frequency of 87 % was obtained on Murashige and Skoog (MS) medium containing 1.13 μM 2,4-D. At lower concentration of 2,4-D (0.34 μM) callus turned dark in color and slow growing. Embryogenic cultures (76 %) responded with a mean number of 43 globular and 18 heart stage embryos. Somatic embryo maturation and subsequent conversion into plantlets took place on MS lacking growth regulators. Maximum number of somatic embryos developed on MS medium was 128.3 (per flask) and a plantlet conversion of 82 % was observed. Calcium alginate beads were produced by encapsulating somatic embryos. Highest percent germination (83 %) was observed on 0.8 % agar solidified MS medium with the plantlets acquiring an average length of 2.1 cm. Encapsulated somatic embryos could be stored at 4 °C up to 60 days with a conversion frequency of 49.3 %. Highest protein and proline content has been observed in embryogenic callus with small globular embryos. During morphological differentiation of the somatic embryos, changes in the antioxidant enzymatic system were observed. Superoxide dismutase (SOD) activity increased during initial stages and decreased catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) activities were detected.

  17. Learning from experience: improving the process of internationally educated nurses' application for registration - a study protocol.

    PubMed

    Giblin, Cathy; Lemermeyer, Gillian; Cummings, Greta; Wang, Mengzhe; Kwan, Jennifer Anne

    2016-03-01

    This study aims to improve the efficiency of the application for registration process for internationally educated nurses seeking licensure to practice. The licensure and employment of internationally educated nurses has been one strategy to address the global nursing shortage. However, little is known about which application characteristics relate to success in obtaining licensure. This project uses evidence from a retrospective statistical analysis of four years of internationally educated nurse application data to inform the development and implementation of changes to policies and practices at the College and Association of Registered Nurses of Alberta. Analysis of application data will also be conducted to evaluate the impact of the changes on outcomes and timelines. The project encompasses four phases, with funding from Health Canada's Internationally Educated Health Professionals Initiative approved in March 2011. Phase One focuses on a statistical analysis of application data to identify characteristics associated with success in the application process and quantify timelines for the phases of the process. The resulting knowledge will inform Phase Two, the development and implementation of policy and practice changes. Further analysis will be completed in Phase Three, comparing outcomes and timelines between pre- and postimplementation data using statistical analyses and exemplar-based statistics. Phase Four will focus on dissemination and knowledge transfer, potentially leading to further changes. Findings, policy and practice adaptations and implementation evaluation will provide evidence about the internationally educated nurse application for registration process to Registered Nurse regulators, educational institutions, internationally educated nurses, employers and governments. © 2015 John Wiley & Sons Ltd.

  18. New 'monolithic' templates and improved protocols for soft lithography and microchip fabrication

    NASA Astrophysics Data System (ADS)

    Pallandre, Antoine; Pal, Debjani; de Lambert, Bertrand; Viovy, Jean-Louis; Fütterer, Claus

    2006-05-01

    We report a new method for fast prototyping and fabrication of polydimethylsiloxane (PDMS) and plastic microfluidic chips. These methods share in common the preparation of monolithic masters which includes the fabrication of the planar support, the 'negative pattern' of the microchannels and the fluidic connectors. The monolithic templates are extremely robust compared to conventional ones made of silicon and SU-8, and easier to produce and cheaper than all-silicon or electroplated templates. In contrast to the above-mentioned methods, our process allows one to cast both micrometre- (e.g. the microchannel) and millimetre-sized structures (e.g. the fluidic connection to the outer world) in a single fabrication step. The 'monolithic template' strategy can be used to fabricate both elastomeric (e.g. poly(dimethyl siloxane (PDMS)) polyester thermoset masters and glassy polymeric (e.g. cyclic olefin copolymer (COC)) devices. In this study we also report on one step fabrication of elastomer chips and on surface modifications of the above mentioned monolithically fabricated masters in order to improve separation of the chip from the template.

  19. Roles of Smartphone App Use in Improving Social Capital and Reducing Social Isolation.

    PubMed

    Cho, Jaehee

    2015-06-01

    This study investigated the relationships among smartphone app use, social capital, and social isolation. It focused on two different smartphone apps--communication and social networking site (SNS) apps--and their effects on bonding and bridging social capital. Generational differences in smartphone use were also considered. Results from hierarchical regression analyses indicated that individuals' use of communication apps was helpful for increasing social capital and that this effect of using communication apps was stronger among those of the millennial generation than among older users. Moreover, bonding and bridging social capital was found to reduce individuals' social isolation significantly. These results imply the notable role of smartphone apps in reducing social isolation and improving the personal lives of individuals.

  20. A cluster randomized trial of an organizational process improvement intervention for improving the assessment and case planning of offenders: a Study Protocol.

    PubMed

    Shafer, Michael S; Prendergast, Michael; Melnick, Gerald; Stein, Lynda A; Welsh, Wayne N

    2014-01-01

    The Organizational Process Improvement Intervention (OPII), conducted by the NIDA-funded Criminal Justice Drug Abuse Treatment Studies consortium of nine research centers, examined an organizational intervention to improve the processes used in correctional settings to assess substance abusing offenders, develop case plans, transfer this information to community-based treatment agencies, and monitor the services provided by these community based treatment agencies. A multi-site cluster randomized design was used to evaluate an inter-agency organizational process improvement intervention among dyads of correctional agencies and community based treatment agencies. Linked correctional and community based agencies were clustered among nine (9) research centers and randomly assigned to an early or delayed intervention condition. Participants included administrators, managers, and line staff from the participating agencies; some participants served on interagency change teams while other participants performed agency tasks related to offender services. A manualized organizational intervention that includes the use of external organizational coaches was applied to create and support interagency change teams that proceeded through a four-step process over a planned intervention period of 12 months. The primary outcome of the process improvement intervention was to improve processes associated with the assessment, case planning, service referral and service provision processes within the linked organizations. Providing substance abuse offenders with coordinated treatment and access to community-based services is critical to reducing offender recidivism. Results from this study protocol will provide new and critical information on strategies and processes that improve the assessment and case planning for such offenders as they transition between correctional and community based systems and settings. Further, this study extends current knowledge of and methods for, the study

  1. The effectiveness of orthopedic patient education in improving patient outcomes: a systematic review protocol.

    PubMed

    Majid, Norhaini; Lee, Susan; Plummer, Virginia

    2015-01-01

    and empowerment were all improved as a result of patient education. While Johansson et al. included studies up to 2003, the proposed systematic review will include studies from 2003 to 2013.Kruzik also reported benefits of decreased length of stay, reduced pain medication requested post-operatively together with increased patient and family member satisfaction. Bastable reported benefits of patient education, including increased patient satisfaction, improved quality of life, enhanced continuity of care, decreased anxiety, fewer complications, promotion of adherence to the plan of care, maximized independence, and empowerment.Types of education involved in this systematic review are patient education, pre-operative education, and discharge education. Only one study discussed discharge education and the outcome from this review. The outcomes that have been discussed in this systematic review include length of stay, satisfaction, pain level,, 21 cost of care, functional abilities, knowledge, anxiety, and quality of life.Major outcomes discussed in these studies are length of stay, pain,, 21 functional abilities, and anxiety. Most of these studies found significant results of reduced length of stay on those patients who received patient education compared to patient who did not received any patient education or information. Studies support the positive outcomes around length of stay.Common orthopedic conditions that have been discussed are osteoarthritis with total arthroplasty either hip or knee or joint replacement. A study on spinal surgery patients and the effects of pre-operative education, which stated that although there are many studies on the effectiveness on patient education, there are missing data on spinal surgery. This study found that the implementation of patient education has positive impacts upon patient satisfaction especially in managing pain.This review will look specifically at the effectiveness of orthopedic patient education for length of stay

  2. MHealth to Improve Measles Immunization in Guinea-Bissau: Study Protocol for a Randomized Controlled Trial

    PubMed Central

    Ravn, Henrik; Batista, Celso Soares Pereira; Rodrigues, Amabelia

    2016-01-01

    Background Recent studies have revealed a low measles vaccination (MV) rate in the Republic of Guinea-Bissau (West Africa) that has not increased in accordance with the increasing coverage of other vaccinations. Measles is the deadliest of all childhood rash/fever illnesses and spreads easily, implying that if the vaccination coverage is declining there is a significant risk of new measles outbreaks [27]. Meanwhile, mobile health (mHealth; the use of mobile phones for health interventions) has generated much enthusiasm, and shown potential in improving health service delivery in other contexts. Objective The aim of this study is to evaluate the efficiency of mHealth as a tool for improving MV coverage while contributing to the mHealth evidence base. Methods This study will take place at three health centers in different regions of Guinea-Bissau. Participants, defined as mothers of the children receiving the MV, will be enrolled when they arrive with their children at the health center to receive the Bacillus Calmette-Guérin vaccination, usually within one month of the child’s birth. Enrolment will continue until a study population of 990 children has been reached. The participants will be randomly assigned to a control arm or one of two intervention arms. Each of the three groups will have 330 participants, distributed equally between health centers. Participants in the first intervention arm will receive a scheduled short message service (SMS) text message reminding them of the MV. Participants in the second intervention arm will receive a voice call in addition to the SMS message, while the control arm will receive no interventions. The MV is scheduled to be administered at 9 months of age. Although the vaccine would still be effective after 12 months, local policy in Guinea-Bissau prevents children aged >12 months from receiving the vaccination, and thus the study will follow-up with participants after the children reach 12 months of age. Children who have

  3. Efficacy of interventions to improve physical activity levels in individuals with stroke: a systematic review protocol

    PubMed Central

    Aguiar, Larissa Tavares; Martins, Júlia Caetano; Nadeau, Sylvie; Britto, Raquel Rodrigues; Teixeira-Salmela, Luci F; Faria, Christina D C M

    2017-01-01

    Introduction Stroke is a leading health problem worldwide and an important cause of disability. Stroke survivors show low levels of physical activity, and increases in physical activity levels may improve function and health status. Therefore, the aims are to identify which interventions that have been employed to increase physical activity levels with stroke survivors, to verify their efficacy and to identify the gaps in the literature. Methods and analysis A systematic review of randomised controlled trials that investigated the efficacy of interventions aiming at increasing physical activity levels of stroke survivors will be conducted. Electronic searches will be performed in the MEDLINE, Physiotherapy Evidence Database (PEDro), Excerpta Medica (EMBASE), Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS) and Scientific Electronic Library Online (SCIELO) databases. Hand searches of the reference lists of the included studies or relevant reviews will also be employed. Two independent reviewers will screen all the retrieved titles, abstracts and full texts. A third reviewer will be referred to solve any disagreements. The quality of the included studies will be assessed by the PEDro Rating Scale. This systematic review will also include a qualitative synthesis. Meta-analyses will be performed, if the studies are sufficiently homogeneous. This review will follow the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) statement. The quality of the evidence regarding physical activity will be assessed, according to the Grading of Recommendations Assessment, Development and Evaluation (GRADE). Discussion This systematic review will provide information on which interventions are effective for increasing physical activity levels of stroke survivors. This evidence may be important for clinical decision-making and will allow the identification of gaps in the literature that may be useful for the definition of future research

  4. Efficacy of interventions to improve physical activity levels in individuals with stroke: a systematic review protocol.

    PubMed

    Aguiar, Larissa Tavares; Martins, Júlia Caetano; Nadeau, Sylvie; Britto, Raquel Rodrigues; Teixeira-Salmela, Luci F; Faria, Christina D C M

    2017-01-05

    Stroke is a leading health problem worldwide and an important cause of disability. Stroke survivors show low levels of physical activity, and increases in physical activity levels may improve function and health status. Therefore, the aims are to identify which interventions that have been employed to increase physical activity levels with stroke survivors, to verify their efficacy and to identify the gaps in the literature. A systematic review of randomised controlled trials that investigated the efficacy of interventions aiming at increasing physical activity levels of stroke survivors will be conducted. Electronic searches will be performed in the MEDLINE, Physiotherapy Evidence Database (PEDro), Excerpta Medica (EMBASE), Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS) and Scientific Electronic Library Online (SCIELO) databases. Hand searches of the reference lists of the included studies or relevant reviews will also be employed. Two independent reviewers will screen all the retrieved titles, abstracts and full texts. A third reviewer will be referred to solve any disagreements. The quality of the included studies will be assessed by the PEDro Rating Scale. This systematic review will also include a qualitative synthesis. Meta-analyses will be performed, if the studies are sufficiently homogeneous. This review will follow the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) statement. The quality of the evidence regarding physical activity will be assessed, according to the Grading of Recommendations Assessment, Development and Evaluation (GRADE). This systematic review will provide information on which interventions are effective for increasing physical activity levels of stroke survivors. This evidence may be important for clinical decision-making and will allow the identification of gaps in the literature that may be useful for the definition of future research goals and the planning of new trials. CRD

  5. Facile mechanical shaking method is an improved isolation approach for islet preparation and transplantation

    PubMed Central

    Yin, Nina; Chen, Tao; Yu, Yuling; Han, Yongming; Yan, Fei; Zheng, Zhou; Chen, Zebin

    2016-01-01

    Successful islet isolation is crucial for islet transplantation and cell treatment for type 1 diabetes. Current isolation methods are able to obtain 500–1,000 islets per rat, which results in a waste of ≥50% of total islets. In the present study, a facile mechanical shaking method for improving islet yield (up to 1,500 per rat) was developed and summarized, which was demonstrated to be more effective than the existing well-established stationary method. The present results showed that isolated islets have a maximum yield of 1,326±152 when shaking for 15 min for the fully-cannulated pancreas. For both fully-cannulated and half-cannulated pancreas in the presence of rat DNAse inhibitor, the optimal shaking time was amended to 20 min with a further increased yield of 1,344±134 and 1,286±124 islets, respectively. Furthermore, the majority of the isolated islets were morphologically intact with a well-defined surface and almost no central necrotic zone, which suggested that the condition of islets obtained via the mechanical shaking method was consistent with the stationary method. Islet size distribution was also calculated and it was demonstrated that islets from the stationary method exhibited the same size distribution as the non-cannulated group, which had more larger islets than the fully-cannulated and half-cannulated groups isolated via the shaking method. In addition, the results of glucose challenge showed that the refraction index of each group was >2.5, which indicated the well-preserved function of isolated islets. Furthermore, the transplanted islets exhibited a therapeutic effect after 1 day of transplantation; however, they failed to control blood glucose levels after ~7 days of transplantation. In conclusion, these results demonstrated that the facile mechanical shaking method may markedly improve the yield of rat islet isolation, and in vitro and in vivo investigation demonstrated the well-preserved function of isolated islets in the control of

  6. Randomized controlled trial protocol: balance training with rhythmical cues to improve and maintain balance control in Parkinson's disease.

    PubMed

    Capato, Tamine Teixeira da Costa; Tornai, Juliana; Ávila, Patrícia; Barbosa, Egberto Reis; Piemonte, Maria Elisa Pimentel

    2015-09-07

    Postural instability is a particularly incapacitating disorder, whose loss of motor independence by Parkinson´s Disease (PD) patients marks a significant stage of disease onset. Evidence suggests that deficits in automatic motor control, sensory integration and attention are associated with the lack of balance in PD. Physiotherapy together with medication play an important role in the treatment of this state, although no consensus has been reached on the best treatment modality. The aim of this randomized controlled trial protocol is to evaluate the effects of balance training with rhythmical (BRT), which is a motor program to improve balance associated with rhythmical auditory cues (RACs). This study is ongoing in the stage 1. A total of 150 PD patients at H&Y stages II-III and asymptomatic for depression and dementia are enrolled in a single-blind randomized study. Randomization is achieved via a computer-generated random-sequence table. All patients should also present a fall history. They will be assigned into one of three groups, and their balance and gait will be assessed before and after 10 training sessions, and after 4 and 30 weeks subsequent to the end of the training. The BRT group will receive a motor program to improve balance associated with RACs, the MT group will perform motor training with the same aims as those in the BRT group but without RACs, and the control group (CG) will be trained only in orientations. The exercise program specific to balance is of 5 weeks' duration with two sessions per week, 45 min each, and consists of general physiotherapy exercises. Each session will be divided into five warm-up minutes-30 min for the main part and 10 min for the cool down. The training progresses and intensifies each week depending on the individual's performance. The subjects should be able to execute 10 repetitions of the exercise sequences correctly to progress to the next movement. This randomized study protocol will evaluate the effects of

  7. Quality improvement competency gaps in primary care in Albanian, polish and slovenian contexts: a study protocol.

    PubMed

    Czabanowska, Katarzyna; Burazeri, Genc; Klemenc-Ketis, Zalika; Kijowska, Violetta; Tomasik, Tomasz; Brand, Helmut

    2012-12-01

    Nowadays, general practitioners (GPs) and family doctors (FDs) face increasing demands, as a consequence of complex patients' expectations, developments in science and technology, and limitations within healthcare systems which can result in competency gaps. Therefore, there is a need to identify which competencies in quality improvement (QI) are most important for GPs and FDs to possess in order to meet the demands of contemporary health care practice. To date, however, little information is available on the self-assessment of competencies related to QI among GPs and FDs. To deal with these issues, a project on QI in continuous medical education was launched in 2011. The project aims to broaden the GPs'/ FDs' continuous education offer, its quality and attractiveness, as well as provide them with opportunities for vocational advancement and enable the development of common, European frame of reference for GPs'/FDs' occupational competencies. The third work package of the project consists of the validation research of the questionnaire developed on the basis of the competency framework in QI for GPs/FDs in Europe. A cross-sectional study will be carried out using the self-assessment QI questionnaire which was originally developed in English and subsequently it was cross-culturally adapted in Slovenian, Albanian and Polish settings by use of a pilot study on a conveniently selected group of FDs/GPs (N=10) in each participating country. The final version of the questionnaire will be administered to large samples in each country involved in the survey. Two weeks after the first administration of the questionnaire, a second round, with the same procedure and including the same group of respondents, will follow. Psychometric tests will be conducted including internal consistency (after the initial and subsequent application of the instrument) and stability over time (two-week test-retest reliability). This self-assessment study will demonstrate the complex environment

  8. Telerehabilitation to improve outcomes for people with stroke: study protocol for a randomised controlled trial

    PubMed Central

    2012-01-01

    contribute to a functional decline following discharge from formal rehabilitation. Best practice guidelines recommend a prolonged period of rehabilitation, however this is expensive and therefore not undertaken in most publicly funded centres. An effective, cost-effective, and preference-sensitive therapy using basic technology to assist programme delivery may improve patient autonomy as they leave formal rehabilitation and return home. Trial registration ACTRN12612000464864 PMID:23216861

  9. Improving maternity care using a personal health record: study protocol for a stepped-wedge, randomised, controlled trial.

    PubMed

    Groenen, Carola J M; Faber, Marjan J; Kremer, Jan A M; Vandenbussche, Frank P H A; van Duijnhoven, Noortje T L

    2016-04-16

    A personal health record (PHR) is an online application through which individuals can access, manage, and share their health information in a private, secure, and confidential environment. Personal health records empower patients, facilitate collaboration among healthcare professionals, and improve health outcomes. Given these anticipated positive effects, we want to implement a PHR, named MyPregn@ncy, in a Dutch maternity care setting and to evaluate its effects in routine care. This paper presents the study protocol. The effects of implementing a PHR in maternity care on patients and professionals will be identified in a stepped-wedge, cluster-randomised, controlled trial. The study will be performed in the region of Nijmegen, a Dutch area with an average of 4,500 births a year and more than 230 healthcare professionals involved in maternity care. Data analyses will describe the effects of MyPregn@ncy on health outcomes in maternity care, quality of care from the patients' perspectives, and collaboration among healthcare professionals. Additionally, a process evaluation of the implementation of MyPregn@ncy will be performed. Data will be collected using data from the Dutch perinatal registry, questionnaires, interviews, and log data. The study is expected to yield new information about the effects, strengths, possibilities, and challenges to the implementation and usage of a PHR in routine maternal care settings. Results may lead to new insights and improvements in the quality of maternal and perinatal care. Netherlands Trial Register: NTR4063.

  10. Improved Detection by Next-Generation Sequencing of Pyrazinamide Resistance in Mycobacterium tuberculosis Isolates.

    PubMed

    Maningi, Nontuthuko E; Daum, Luke T; Rodriguez, John D; Mphahlele, Matsie; Peters, Remco P H; Fischer, Gerald W; Chambers, James P; Fourie, P Bernard

    2015-12-01

    The technical limitations of common tests used for detecting pyrazinamide (PZA) resistance in Mycobacterium tuberculosis isolates pose challenges for comprehensive and accurate descriptions of drug resistance in patients with multidrug-resistant tuberculosis (MDR-TB). In this study, a 606-bp fragment (comprising the pncA coding region plus the promoter) was sequenced using Ion Torrent next-generation sequencing (NGS) to detect associated PZA resistance mutations in 88 recultured MDR-TB isolates from an archived series collected in 2001. These 88 isolates were previously Sanger sequenced, with 55 (61%) designated as carrying the wild-type pncA gene and 33 (37%) showing mutations. PZA susceptibility of the isolates was also determined using the Bactec 460 TB system and the Wayne test. In this study, isolates were recultured and susceptibility testing was performed in Bactec 960 MGIT. Concordance between NGS and MGIT results was 93% (n = 88), and concordance values between the Bactec 460, the Wayne test, or pncA gene Sanger sequencing and NGS results were 82% (n = 88), 83% (n = 88), and 89% (n = 88), respectively. NGS confirmed the majority of pncA mutations detected by Sanger sequencing but revealed several new and mixed-strain mutations that resolved discordancy in other phenotypic results. Importantly, in 53% (18/34) of these isolates, pncA mutations were located in the 151 to 360 region and warrant further exploration. In these isolates, with their known resistance to rifampin, NGS of pncA improved PZA resistance detection sensitivity to 97% and specificity to 94% using NGS as the gold standard and helped to resolve discordant results from conventional methodologies.

  11. Vibration control of platform structures with magnetorheological elastomer isolators based on an improved SAVS law

    NASA Astrophysics Data System (ADS)

    Xu, Zhao-Dong; Suo, Si; Lu, Yong

    2016-06-01

    This paper presents a study on the vibration control of platform structures with magnetorheological elastomer (MRE) isolators. Firstly, a novel MRE isolator design is put forward based on the mechanical properties of MREs, and subsequently a single-degree-of-freedom (SDOF) dynamic model and a multiple-degree-of-freedom (MDOF) dynamic model for platform systems incorporating such isolators are developed. In order to overcome the shortcomings of the conventional on-off control law, an improved semi-active variable stiffness (SAVS) control law is proposed. The proposed SAVS scheme makes full use of the continuously variable stiffness of MREs, and it takes into account the influence of the sampling interval such that the field-dependent restoring force is made to do negative work during the whole sampling interval as far as possible. The results of numerical simulations demonstrate that the improved SAVS control law can achieve better vibration-control effectiveness than the on-off control law. The comparative results are discussed through examining the mechanisms of these two control laws in light of the power spectral density and the energy input. For an MDOF platform a simplified approach is proposed to combine the local response signals with an equivalent SDOF representation to generate the control parameters for individual isolators, and the effectiveness of such a scheme is also verified through numerical simulation.

  12. Study protocol: a randomised controlled trial of a theory-based online intervention to improve sun safety among Australian adults

    PubMed Central

    2014-01-01

    Background The effects of exposure to ultraviolet radiation are a significant concern in Australia which has one of the highest incidences of skin cancer in the world. Despite most skin cancers being preventable by encouraging consistent adoption of sun-protective behaviours, incidence rates are not decreasing. There is a dearth of research examining the factors involved in engaging in sun-protective behaviours. Further, online multi-behavioural theory-based interventions have yet to be explored fully as a medium for improving sun-protective behaviour in adults. This paper presents the study protocol of a randomised controlled trial of an online intervention based on the Theory of Planned Behaviour (TPB) that aims to improve sun safety among Australian adults. Methods/Design Approximately 420 adults aged 18 and over and predominantly from Queensland, Australia, will be recruited and randomised to the intervention (n = 200), information only (n = 200) or the control group (n = 20). The intervention focuses on encouraging supportive attitudes and beliefs toward sun-protective behaviour, fostering perceptions of normative support for sun protection, and increasing perceptions of control/self-efficacy over sun protection. The intervention will be delivered online over a single session. Data will be collected immediately prior to the intervention (Time 1), immediately following the intervention (Time 1b), and one week (Time 2) and one month (Time 3) post-intervention. Primary outcomes are intentions to sun protect and sun-protective behaviour. Secondary outcomes are the participants’ attitudes toward sun protection, perceptions of normative support for sun protection (i.e. subjective norms, group norms, personal norms and image norms) and perceptions of control/self-efficacy toward sun protection. Discussion The study will contribute to an understanding of the effectiveness of a TPB-based online intervention to improve Australian adults’ sun

  13. Evaluating the implementation of a national clinical programme for diabetes to standardise and improve services: a realist evaluation protocol.

    PubMed

    McHugh, S; Tracey, M L; Riordan, F; O'Neill, K; Mays, N; Kearney, P M

    2016-07-28

    Over the last three decades in response to the growing burden of diabetes, countries worldwide have developed national and regional multifaceted programmes to improve the monitoring and management of diabetes and to enhance the coordination of care within and across settings. In Ireland in 2010, against a backdrop of limited dedicated strategic planning and engrained variation in the type and level of diabetes care, a national programme was established to standardise and improve care for people with diabetes in Ireland, known as the National Diabetes Programme (NDP). The NDP comprises a range of organisational and service delivery changes to support evidence-based practices and policies. This realist evaluation protocol sets out the approach that will be used to identify and explain which aspects of the programme are working, for whom and in what circumstances to produce the outcomes intended. This mixed method realist evaluation will develop theories about the relationship between the context, mechanisms and outcomes of the diabetes programme. In stage 1, to identify the official programme theories, documentary analysis and qualitative interviews were conducted with national stakeholders involved in the design, development and management of the programme. In stage 2, as part of a multiple case study design with one case per administrative region in the health system, qualitative interviews are being conducted with frontline staff and service users to explore their responses to, and reasoning about, the programme's resources (mechanisms). Finally, administrative data will be used to examine intermediate implementation outcomes such as service uptake, acceptability, and fidelity to models of care. This evaluation is using the principles of realist evaluation to examine the implementation of a national programme to standardise and improve services for people with diabetes in Ireland. The concurrence of implementation and evaluation has enabled us to produce formative

  14. Study protocol: a randomised controlled trial of a theory-based online intervention to improve sun safety among Australian adults.

    PubMed

    Cleary, Cathy M; White, Katherine M; Young, Ross McD; Hawkes, Anna L; Leske, Stuart; Starfelt, Louise C; Wihardjo, Kylie

    2014-03-07

    The effects of exposure to ultraviolet radiation are a significant concern in Australia which has one of the highest incidences of skin cancer in the world. Despite most skin cancers being preventable by encouraging consistent adoption of sun-protective behaviours, incidence rates are not decreasing. There is a dearth of research examining the factors involved in engaging in sun-protective behaviours. Further, online multi-behavioural theory-based interventions have yet to be explored fully as a medium for improving sun-protective behaviour in adults. This paper presents the study protocol of a randomised controlled trial of an online intervention based on the Theory of Planned Behaviour (TPB) that aims to improve sun safety among Australian adults. Approximately 420 adults aged 18 and over and predominantly from Queensland, Australia, will be recruited and randomised to the intervention (n = 200), information only (n = 200) or the control group (n = 20). The intervention focuses on encouraging supportive attitudes and beliefs toward sun-protective behaviour, fostering perceptions of normative support for sun protection, and increasing perceptions of control/self-efficacy over sun protection. The intervention will be delivered online over a single session. Data will be collected immediately prior to the intervention (Time 1), immediately following the intervention (Time 1b), and one week (Time 2) and one month (Time 3) post-intervention. Primary outcomes are intentions to sun protect and sun-protective behaviour. Secondary outcomes are the participants' attitudes toward sun protection, perceptions of normative support for sun protection (i.e. subjective norms, group norms, personal norms and image norms) and perceptions of control/self-efficacy toward sun protection. The study will contribute to an understanding of the effectiveness of a TPB-based online intervention to improve Australian adults' sun-protective behaviour. Australian and New Zealand Trials Registry

  15. The effect and process evaluations of the national quality improvement programme for palliative care: the study protocol

    PubMed Central

    2014-01-01

    Background The nationwide integration of palliative care best practices into general care settings is challenging but important in improving the quality of palliative care. This is why the Dutch National Quality Improvement Programme for Palliative Care has recently been launched. This four-year programme consists of about 70 implementation trajectories of best practices. A large evaluation study has been set up to evaluate this national programme and separate implementation trajectories. Methods/Design This paper presents the protocol of the evaluation study consisting of a quantitative effect evaluation and a qualitative process evaluation. The effect evaluation has a pre-test post-test design, with measurements before implementation (month 0) and after implementation (month 9) of a best practice. Patients are eligible if they have a life expectancy of less than six months and/or if they are undergoing palliative treatment and provided they are physically and mentally capable of responding to questionnaires. Bereaved relatives are eligible if they have been involved in the care of a deceased patient who died after a sickbed between six weeks and six months ago. Three types of measurement instruments are used: (1) numerical rating scales for six symptoms (pain, fatigue, breathlessness, obstipation, sadness and anxiety), (2) the Consumer Quality Index Palliative Care - patient version and (3) the version for bereaved relatives. The process evaluation consists of analysing implementation plans and reports of the implementation, and individual and group interviews with healthcare professionals. This will be done nine to eleven months after the start of the implementation of a best practice. Discussion This mixed-method evaluation study gives more insight into the effects of the total programme and the separate implementation trajectories. However, evaluation of large quality improvement programmes is complicated due to changing, non-controlled environments. Therefore

  16. A filtration-based protocol to isolate human plasma membrane-derived vesicles and exosomes from blood plasma.

    PubMed

    Grant, Ryan; Ansa-Addo, Ephraim; Stratton, Dan; Antwi-Baffour, Samuel; Jorfi, Samireh; Kholia, Sharad; Krige, Lizelle; Lange, Sigrun; Inal, Jameel

    2011-08-31

    The methods of Plasma Membrane-derived Vesicle (PMV) isolation and quantification vary considerably in the literature and a new standard needs to be defined. This study describes a novel filtration method to isolate PMVs in plasma, which avoids high speed centrifugation, and to quantify them using a Becton Dickinson (BD) FACS Calibur™ flow cytometer, as annexin V-positive vesicles, larger than 0.2 μm in diameter. Essentially microvesicles (which comprise a mixture of PMVs and exosomes) from citrate plasma were sonicated to break up clumped exosomes, and filtered using Millipore 0.1 μm pore size Hydrophilic Durapore membranes in Swinnex 13 mm filter holders. Phosphatidylserine-positive PMVs detected with annexin V-PE were quantified using combined labelling and gating strategies in conjunction with Polysciences Polybead Microspheres (0.2 μm) and BDTrucount tubes. The PMV absolute count was calculated on the analysis template using the Trucount tube lot number information and expressed in PMV count/ml. Having estimated a normal reference range (0.51×10(5)-2.82×10(5) PMVs/ml) from a small sample of human donors, using the developed method, the effect of certain variables was investigated. Variations such as freezing of samples and gender status did not significantly alter the PMV absolute count, and with age plasma PMV levels were only marginally reduced. Smokers appeared to have reduced PMV levels. Nicotine, as for calpeptin was shown to dose-dependently (from 10 up to 50 μM) reduce levels of early apoptosis in THP-1 monocytes and to decrease the level of PMV release. Fasting individuals had 2-3 fold higher PMV absolute counts compared to non-fasting subjects.

  17. An improvement in isolation and preservation of clinical strains of Helicobacter pylori.

    PubMed

    Farshad, Shohreh; Japoni, Aziz; Shahidi, Maneli Amin; Hosseini, Marziyeh; Alborzi, Abdolvahab

    2011-01-01

    Isolation of H. pylori from gastric mucosal biopsy specimens is a prerequisite for further studies addressing drug susceptibility testing, analysis and characterization of virulence factors, molecular epidemiology studying or other comparative studies. In this study, we used a modified enriched culture medium with short incubation time to improve the isolation rate of H. pylori from the clinical specimens. Between October 2008 and October 2009, 266 dyspeptic patients attending the endoscopy ward of Motahhary Clinic of Shiraz University of Medical Sciences, were investigated. The biopsy samples were cultured on two selective media called M1, which we used in our previous studies, and a modified medium called M2. The cultures were kept in a microaerophilic atmosphere at 37 degrees C. The plates were inspected first on day 1, and then on daily basis for a total of 10 days. The isolates were confirmed as H. pylori by colony morphology and positive oxidase, catalase and rapid urease tests. We used the same media and culture conditions to subculture the isolates for several times. Specimens were considered to be H. pylori positive if either the culture or two of the three diagnostic methods yielded positive results. The isolation rate of H. pylori strains from the samples was significantly higher on M2 in comparison with M1 medium (p<0.05). The bacterial growth on M2 was observed after a significantly shorter time (p<0.05), i.e., after incubation for about 24 hrs. Following these procedures, the preservation time could be extended beyond 6 months without a significant loss of viability. The modified culture technique enabled a shorter incubation time and a higher isolation rate for H.pylori obtained from clinical samples.

  18. Safety and Feasibility of a Protocolized Approach to In-Bed Cycling Exercise in the Intensive Care Unit: Quality Improvement Project.

    PubMed

    Kimawi, Ibtehal; Lamberjack, Bryanna; Nelliot, Archana; Toonstra, Amy Lee; Zanni, Jennifer; Huang, Minxuan; Mantheiy, Earl; Kho, Michelle E; Needham, Dale M

    2017-03-31

    : In-bed, supine cycle ergometry as a part of early rehabilitation in the intensive care unit (ICU) appears to be safe, feasible, and beneficial, but no standardized protocol exists. A standardized protocol may help guide use of cycle ergometry in the ICU. We investigated whether a standardized protocol for in-bed cycling is safe and feasible, and results in cycling for a longer duration and achieves a higher resistance. A quality improvement (QI) project was conducted. : A 35-minute in-bed cycling protocol was implemented in a single medical intensive care unit (MICU) over a 7-month quality improvement (QI) period compared to pre-existing, prospectively collected data from an 18-month pre-QI period. : 106 MICU patients received 260 cycling sessions in the QI period vs. 178 MICU patients receiving 498 sessions in the pre-QI period. The protocol was used in 249 (96%) of cycling sessions. The QI group cycled for longer median (IQR) duration (35 [25 - 35] vs. 25 [18 - 30] minutes, P<0.001) and more frequently achieved a resistance level greater than gear 0 (47% vs. 17% of sessions, P<0.001). There were 4 (1.5%) transient physiologic abnormalities during the QI period, and 1 (0.2%) during the pre-QI period (P = 0.031). Patient outcomes were not evaluated to understand if the protocol has clinical benefits. : Use of a protocolized approach for in-bed cycling appears safe and feasible, and results in cycling for longer duration and achieved higher resistance.

  19. Optimizing isolation protocol of organic carbon and elemental carbon for 14C analysis using fine particulate samples

    NASA Astrophysics Data System (ADS)

    Liu, Junwen; Li, Jun; Ding, Ping; Zhang, Yanlin; Liu, Di; Shen, Chengde; Zhang, Gan

    2017-04-01

    Radiocarbon (14C) analysis is a unique tool that can be used to directly apportion organic carbon (OC) and elemental carbon (EC) into fossil and non-fossil fractions. In this study, a coupled carbon analyzer and high-vacuum setup was established to collect atmospheric OC and EC. We thoroughly investigated the correlations between 14C levels and mass recoveries of OC and EC using urban PM2.5 samples collected from a city in central China and found that: (1) the 14C signal of the OC fraction collected in the helium phase of the EUSSAR_2 protocol (200 °C for 120 s, 300 °C for 150 s, 450 °C for 180 s, and 650 °C for 180 s) was representative of the entire OC fraction, with a relative error of approximately 6%, and (2) after thermal treatments of 120 s at 200 °C, 150 s at 300 °C, and 180 s at 475 °C in an oxidative atmosphere (10% oxygen, 90% helium) and 180 s at 650 °C in helium, the remaining EC fraction sufficiently represented the 14C level of the entire EC, with a relative error of <10%. The average recovery of the OC and EC fractions for 14C analysis was 64± 7% (n = 5) and 87 ± 5% (n = 5), respectively. The fraction of modern carbon in the OC and EC of reference material (RM) 8785 was 0.564 ± 0.013 and 0.238 ± 0.006, respectively. Analysis of 14C levels in four selected PM2.5 samples in Xinxiang, China revealed that the relative contribution of fossil sources in OC and EC in the PM2.5 samples were 50.5± 5.8% and 81.4± 2.6%, respectively, which are comparable to findings in previous studies conducted in other Chinese cities. We confirmed that most urban EC derives from fossil fuel combustion processes, whereas both fossil and non-fossil sources have comparable and important impacts on OC. Our results suggested that water-soluble organic carbon (WSOC) and its pyrolytic carbon can be completely removed before EC collection via the method employed in this study.

  20. Evaluation of a suicide prevention training curriculum for substance abuse treatment providers based on Treatment Improvement Protocol Number 50.

    PubMed

    Conner, Kenneth R; Wood, Jane; Pisani, Anthony R; Kemp, Janet

    2013-01-01

    Substance use disorders (SUDs) confer risk for suicide yet there are no empirically supported suicide prevention training curricula tailored to SUD treatment providers. We assessed the efficacy of a 2-hour training that featured a suicide prevention training video produced by the Department of Veterans Affairs. The video was based on Treatment Improvement Protocol Number 50 (TIP 50) a practical manual to manage suicide risk produced by the Substance Abuse and Mental Health Services Administration. The training was provided in small groups to 273 SUD treatment providers in 18 states. Results were evaluated using self-report assessments obtained at pre-test, post-test, and 2-month follow-up. Statistically significant changes (p < .001) within subjects were obtained on self-efficacy, knowledge, and frequency of suicide prevention practice behaviors. The positive results together with the brevity of the training and its ease of implementation indicate high potential for widespread adoption and the importance of further study. Published by Elsevier Inc.

  1. A Qualitative Study of the Treatment Improvement Protocols (TIPs): An Assessment of the Use of TIPs by Individuals Affiliated with the Addiction Technology Transfer Centers (ATTCs).

    ERIC Educational Resources Information Center

    Hayashi, Susan W.; Suzuki, Marcia; Hubbard, Susan M.; Huang, Judy Y.; Cobb, Anita M.

    2003-01-01

    Evaluated the Addiction Technology Transfer Centers (ATTCs) of the Center for Substance Abuse Treatment (CSAT) as a means of diffusion of innovations, focusing on use of the Treatment Improvement Protocols (TIPs). Qualitative studies at 6 ATTCs that included 57 interviews show that the CSAT is at the forefront of providing resources to the…

  2. Improving active seismic isolation in aLIGO using a ground rotation sensor

    NASA Astrophysics Data System (ADS)

    Venkateswara, Krishna; Hagedorn, Charles; Ross, Michael; Gundlach, Jens

    2016-03-01

    The active seismic isolation in Advanced LIGO achieves a factor of 10 -104 isolation from ground displacement in the frequency range from 0.1-10 Hz enabling stable low noise interferometer operation. It uses seismometers on the ground and the optics platform in feedback loops to reduce the transmission of ground motion to the platform. However, due to the inability of a seismometer to distinguish between horizontal acceleration and rotation (coupling through gravity), wind-induced tilt limits the performance of the active isolation in the 10-500 mHz frequency range, thereby reducing the duty-cycle of the detectors. We describe a ground rotation sensor, consisting of a low frequency beam-balance and an autocollimator readout with better than 0.4 nrad/rt(Hz) sensitivity above 10 mHz, which can be used to subtract tilt-noise from a horizontal seismometer, thus improving the active seismic isolation system. This work was supported by NSF Grant: 1306613.

  3. Data for improvement and clinical excellence: protocol for an audit with feedback intervention in long-term care

    PubMed Central

    2010-01-01

    Background There is considerable evidence about the effectiveness of audit coupled with feedback, although few audit with feedback interventions have been conducted in long-term care (LTC) settings to date. In general, the effects have been found to be modest at best, although in settings where there has been little history of audit and feedback, the effects may be greater, at least initially. The primary purpose of the Data for Improvement and Clinical Excellence (DICE) Long-Term Care project is to assess the effects of an audit with feedback intervention delivered monthly over 13 months in four LTC facilities. The research questions we addressed are: 1. What effects do feedback reports have on processes and outcomes over time? 2. How do different provider groups in LTC and home care respond to feedback reports based on data targeted at improving quality of care? Methods/design The research team conducting this study comprises researchers and decision makers in continuing care in the province of Alberta, Canada. The intervention consists of monthly feedback reports in nine LTC units in four facilities in Edmonton, Alberta. Data for the feedback reports comes from the Resident Assessment Instrument Minimum Data Set (RAI) version 2.0, a standardized instrument mandated for use in LTC facilities throughout Alberta. Feedback reports consist of one page, front and back, presenting both graphic and textual information. Reports are delivered to all staff working in the four LTC facilities. The primary evaluation uses a controlled interrupted time series design both adjusted and unadjusted for covariates. The concurrent process evaluation uses observation and self-report to assess uptake of the feedback reports. Following the project phase described in this protocol, a similar intervention will be conducted in home care settings in Alberta. Depending on project findings, if they are judged useful by decision makers participating in this research team, we plan

  4. Improving quality and outcomes of stroke care in hospitals: Protocol and statistical analysis plan for the Stroke123 implementation study.

    PubMed

    Cadilhac, Dominique A; Andrew, Nadine E; Kilkenny, Monique F; Hill, Kelvin; Grabsch, Brenda; Lannin, Natasha A; Thrift, Amanda G; Anderson, Craig S; Donnan, Geoffrey A; Middleton, Sandy; Grimley, Rohan

    2017-01-01

    Rationale The effectiveness of clinician-focused interventions to improve stroke care is uncertain. Aims To determine whether an organizational intervention can improve the quality of stroke care over usual care. Sample size estimates To detect an absolute 10% difference in overall performance (composite outcome), a minimum of 21 hospitals and 843 patients per group was determined. Methods and design Before and after controlled design in hospitals in Queensland, Australia. Intervention Externally facilitated program (StrokeLink) using outreach workshops incorporating clinical performance feedback, patient outcomes (survival, quality of life at 90-180 days), local barrier assessments to best practice care, action planning, and ongoing support. Descriptive and multivariable analyses adjusted for patient correlations by hospital (intention-to-treat method). Context Concurrent implementation of financial incentives to increase stroke unit access and use of the Australian Stroke Clinical Registry for performance monitoring. Study outcome(s) Primary outcome: net change in composite score (i.e. total number of process indicators achieved divided by the sum of eligible indicators for each cohort). change in individual indicators, change in composite score comparing hospitals that did or did not develop action plans (per-protocol analysis), impact on 90-180-day health outcomes. Sensitivity analyses: hospital self-rated status, alternate cross-sectional audit data (Stroke Foundation). To account for temporal effects, comparison of Queensland hospital performance relative to other Australian hospitals will also be undertaken. Discussion Twenty-one hospitals were recruited; however, one was unable to participate within the study time frame. Workshops were held between 11 March 2014 and 7 November 2014. Data are ready for analysis.

  5. Understanding how and why audits work: protocol for a realist review of audit programmes to improve hospital care.

    PubMed

    Hut-Mossel, Lisanne; Welker, Gera; Ahaus, Kees; Gans, Rijk

    2017-06-14

    Many types of audits are commonly used in hospital care to promote quality improvements. However, the evidence on the effectiveness of audits is mixed. The objectives of this proposed realist review are (1) to understand how and why audits might, or might not, work in terms of delivering the intended outcome of improved quality of hospital care and (2) to examine under what circumstances audits could potentially be effective. This protocol will provide the rationale for using a realist review approach and outline the method. This review will be conducted using an iterative four-stage approach. The first and second steps have already been executed. The first step was to develop an initial programme theory based on the literature that explains how audits are supposed to work. Second, a systematic literature search was conducted using relevant databases. Third, data will be extracted and coded for concepts relating to context, outcomes and their interrelatedness. Finally, the data will be synthesised in a five-step process: (1) organising the extracted data into evidence tables, (2) theming, (3) formulating chains of inference from the identified themes, (4) linking the chains of inference and formulating CMO configurations and (5) refining the initial programme theory. The reporting of the review will follow the 'Realist and Meta-Review Evidence Synthesis: Evolving Standards' (RAMESES) publication standards. This review does not require formal ethical approval. A better understanding of how and why these audits work, and how context impacts their effectiveness, will inform stakeholders in deciding how to tailor and implement audits within their local context. We will use a range of dissemination strategies to ensure that findings from this realist review are broadly disseminated to academic and non-academic audiences. CRD42016039882. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial

  6. Understanding how and why audits work: protocol for a realist review of audit programmes to improve hospital care

    PubMed Central

    Hut-Mossel, Lisanne; Welker, Gera; Ahaus, Kees; Gans, Rijk

    2017-01-01

    Introduction Many types of audits are commonly used in hospital care to promote quality improvements. However, the evidence on the effectiveness of audits is mixed. The objectives of this proposed realist review are (1) to understand how and why audits might, or might not, work in terms of delivering the intended outcome of improved quality of hospital care and (2) to examine under what circumstances audits could potentially be effective. This protocol will provide the rationale for using a realist review approach and outline the method. Methods and analysis This review will be conducted using an iterative four-stage approach. The first and second steps have already been executed. The first step was to develop an initial programme theory based on the literature that explains how audits are supposed to work. Second, a systematic literature search was conducted using relevant databases. Third, data will be extracted and coded for concepts relating to context, outcomes and their interrelatedness. Finally, the data will be synthesised in a five-step process: (1) organising the extracted data into evidence tables, (2) theming, (3) formulating chains of inference from the identified themes, (4) linking the chains of inference and formulating CMO configurations and (5) refining the initial programme theory. The reporting of the review will follow the ‘Realist and Meta-Review Evidence Synthesis: Evolving Standards’ (RAMESES) publication standards. Ethics and dissemination This review does not require formal ethical approval. A better understanding of how and why these audits work, and how context impacts their effectiveness, will inform stakeholders in deciding how to tailor and implement audits within their local context. We will use a range of dissemination strategies to ensure that findings from this realist review are broadly disseminated to academic and non-academic audiences. PROSPERO registration number CRD42016039882. PMID:28615270

  7. Improvement in the quality of the cardiac vein images by optimizing the scan protocol of multidetector-row computed tomography.

    PubMed

    Hara, Tetsuya; Yamashiro, Kohei; Okajima, Katsunori; Hayashi, Takatoshi; Kajiya, Teishi

    2009-11-01

    The present study aimed at optimizing the scan protocol for multidetector-row computed tomography (MDCT) to adequately visualize coronary veins. Circulation time (Cir.T) was defined as the time period from the injection of contrast media into the coronary artery to the pervasion of the contrast media into the coronary sinus as observed by coronary angiography. We investigated the relation between the Cir.T and echocardiographic parameters in 64 patients. The left ventricular end-diastolic diameter (LVDd) and left ventricular end-systolic diameter (LVDs) were correlated with the Cir.T (r = 0.58, P < 0.0001, and r = 0.60, P < 0.0001 respectively). In addition, the left ventricular ejection fraction (LVEF) was negatively correlated with the Cir.T (r = 0.48, P < 0.0001). The average Cir. T was longer in patients with LVEF < 35% (8.0 s vs 6.7 s; P < 0.05) or LVDd > 55 mm (7.9 s vs 6.2 s; P < 0.05) than in the other patients. The quality of the MDCT images of the coronary veins obtained at different scan timings (coronary artery phase and 10 s or 15 s after the coronary artery phase) were graded and classified into four categories (0 = worst, 3 = best) in 25 patients with LVEF < 35%. The delays of 10 and 15 s after the coronary artery phase significantly improved the mean image quality (P < 0.05). The Cir.T was prolonged in patients with low LVEF and LV dilation. An appropriate delay improved the quality of the MDCT images of the coronary veins in patients with LV dysfunction.

  8. Effectiveness of group reminiscence for improving wellbeing of institutionalized elderly adults: study protocol for a randomized controlled trial.

    PubMed

    Gaggioli, Andrea; Scaratti, Chiara; Morganti, Luca; Stramba-Badiale, Marco; Agostoni, Monica; Spatola, Chiara A M; Molinari, Enrico; Cipresso, Pietro; Riva, Giuseppe

    2014-10-25

    Group reminiscence therapy is a brief and structured intervention in which participants share personal past events with peers. This approach has been shown to be promising for improving wellbeing and reducing depressive symptoms among institutionalized older adults. However, despite the considerable interest in reminiscence group therapy, controlled studies to determine its specific benefits as compared to generic social interactions with peers (group conversations about everyday subjects) are still lacking. We have designed a randomized controlled trial aimed at comparing the effects of group reminiscence therapy with those of group recreational activity on the psychological wellbeing of an institutionalized sample of older adults. The study includes two groups of 20 hospitalized elderly participants: the experimental group and the control group. Participants included in the experimental group will receive six sessions of group reminiscence therapy, while the control group will participate in a recreational group discussion. A repeated-measures design will be used post-intervention and three months post-intervention to evaluate changes in self-reported outcome measures of depressive symptoms, self-esteem, life satisfaction, and quality of life from baseline. The protocol of a study aimed at examining the specific effects of group reminiscence therapy on psychological wellbeing, depression, and quality of life among institutionalized elderly people is described. It is expected that the outcomes of this trial will contribute to our knowledge about the process of group reminiscence, evaluate its effectiveness in improving psychological wellbeing of institutionalized individuals, and identify the best conditions for optimizing this approach. This trial was registered with ClinicalTrials.gov (registration number: NCT02077153) on 31 January 2014.

  9. Rivastigmine improves isolation rearing-induced prepulse inhibition deficits via muscarinic acetylcholine receptors in mice.

    PubMed

    Higashino, Kosuke; Ago, Yukio; Umeki, Takahiro; Hasebe, Shigeru; Onaka, Yusuke; Hashimoto, Hitoshi; Takuma, Kazuhiro; Matsuda, Toshio

    2016-02-01

    The acetylcholinesterase inhibitors donepezil, galantamine, and rivastigmine are used for the treatment of Alzheimer's disease. We previously demonstrated that donepezil and galantamine differentially affect isolation rearing-induced prepulse inhibition (PPI) deficits and that this might be due to differential effects on brain muscarinic acetylcholine (mACh) receptor function in mice. We examined the effects of rivastigmine on isolation rearing-induced PPI deficits, brain ACh levels, and mACh receptor function in mice. Acoustic startle responses were measured in a startle chamber. Microdialysis was performed, and the levels of dopamine and ACh in the prefrontal cortex were measured. Rivastigmine (0.3 mg/kg) improved PPI deficits, and this improvement was antagonized by the mACh receptor antagonist telenzepine but not by the nicotinic ACh receptor antagonist mecamylamine. Rivastigmine increased extracellular ACh levels by approximately 2-3-fold, less than the increase produced by galantamine. Rivastigmine enhanced the effect of the mACh receptor agonist N-desmethylclozapine on prefrontal dopamine release, a marker of mACh receptor function, and this increase was blocked by telenzepine. In contrast, galantamine did not affect N-desmethylclozapine-induced dopamine release. Furthermore, rivastigmine did not affect cortical dopamine release induced by the serotonin1A receptor agonist osemozotan, suggesting that the effect of rivastigmine has specificity for mACh receptors. Taken together with our previous finding that marked increases in ACh levels are required for the PPI deficit improvement induced by galantamine, our present results suggest that rivastigmine improves isolation rearing-induced PPI deficits by increasing ACh levels and by concomitantly enhancing mACh receptor function.

  10. Improved isolation of Chlamydia trachomatis from a low-prevalence population by using polyethylene glycol.

    PubMed

    Gibson, J P; Egerer, R M; Wiedbrauk, D L

    1993-02-01

    The effect of polyethylene glycol (PEG) on the isolation of Chlamydia trachomatis was evaluated in our laboratory. Initial range-finding experiments demonstrated that the number of chlamydial inclusion bodies increased with increasing PEG concentrations. However, PEG concentrations above 10.5% became progressively more toxic to the McCoy cell monolayers. When 50 frozen clinical Chlamydia isolates were inoculated onto McCoy cell cultures with and without 7% PEG, the PEG-treated cultures produced three- to fivefold more chlamydial inclusions than cultures without PEG. This enhancement was also observed when 1,144 fresh clinical specimens from a low-prevalence population were tested. With fresh clinical specimens, PEG-treated cultures produced two- to sixfold more inclusions than standard cultures. The addition of 7% PEG to the chlamydial overlay medium significantly increased the number of inclusions in each culture, improved the sensitivity of the culture, and decreased the probability of missing a weakly positive specimen.

  11. Isolation of rat cardiac sarcoplasmic reticulum with improved Ca2+ uptake and ryanodine binding.

    PubMed

    Feher, J J; Davis, M D

    1991-03-01

    The instability of the oxalate-supported Ca2+ uptake activity of rat cardiac sarcoplasmic reticulum (CSR) in ventricular homogenates most likely accounts for the low specific activity of the rate of oxalate-supported Ca2+ uptake in previously reported fractions of isolated rat CSR. We have found that CSR vesicles with improved Ca2+ transport capabilities can be isolated if 1 M KCl is used to stabilize the CSR activity and to allow the extraction of the CSR from the cellular debris. The average rate of Ca2+ uptake by the isolated rat CSR in the presence of 10 mM oxalate at 37 degrees C was 0.45 mumols/min-mg in the absence of CSR Ca2+ channel blockers and 0.87 mumols/min-mg in the presence of 10 microM ruthenium red. The Ca(2+)-dependent ATPase activity under the conditions of oxlate-supported uptake was 1.25 mumols/min-mg and 0.84 mumols/min-mg in the absence and presence of 10 microM ruthenium red, respectively. The rat CSR vesicles bound 3H-ryanodine with a Kd of 1.45 nM and a Bmax of 3.7 pmol mg. The level of phosphorylated intermediate was 0.30 nmol/mg. The values Bmax, EP and Ca(2+)-ATPase activity are from one-third to one-half of those previously reported for isolated canine CSR vesicles. These results suggest that the isolated rat CSR may be quite similar to dog CSR.

  12. Incubating Isolated Mouse EDL Muscles with Creatine Improves Force Production and Twitch Kinetics in Fatigue Due to Reduction in Ionic Strength

    PubMed Central

    Head, Stewart I.; Greenaway, Bronwen; Chan, Stephen

    2011-01-01

    Background Creatine supplementation can improve performance during high intensity exercise in humans and improve muscle strength in certain myopathies. In this present study, we investigated the direct effects of acute creatine incubation on isolated mouse fast-twitch EDL muscles, and examined how these effects change with fatigue. Methods and Results The extensor digitorum longus muscle from mice aged 12–14 weeks was isolated and stimulated with field electrodes to measure force characteristics in 3 different states: (i) before fatigue; (ii) immediately after a fatigue protocol; and (iii) after recovery. These served as the control measurements for the muscle. The muscle was then incubated in a creatine solution and washed. The measurement of force characteristics in the 3 different states was then repeated. In un-fatigued muscle, creatine incubation increased the maximal tetanic force. In fatigued muscle, creatine treatment increased the force produced at all frequencies of stimulation. Incubation also increased the rate of twitch relaxation and twitch contraction in fatigued muscle. During repetitive fatiguing stimulation, creatine-treated muscles took 55.1±9.5% longer than control muscles to lose half of their original force. Measurement of weight changes showed that creatine incubation increased EDL muscle mass by 7%. Conclusion Acute creatine application improves force production in isolated fast-twitch EDL muscle, and these improvements are particularly apparent when the muscle is fatigued. One likely mechanism for this improvement is an increase in Ca2+ sensitivity of contractile proteins as a result of ionic strength decreases following creatine incubation. PMID:21850234

  13. Using Green Star Metrics to Optimize the Greenness of Literature Protocols for Syntheses

    ERIC Educational Resources Information Center

    Duarte, Rita C. C.; Ribeiro, M. Gabriela T. C.; Machado, Adélio A. S. C.

    2015-01-01

    A procedure to improve the greenness of a synthesis, without performing laboratory work, using alternative protocols available in the literature is presented. The greenness evaluation involves the separate assessment of the different steps described in the available protocols--reaction, isolation, and purification--as well as the global process,…

  14. Using Green Star Metrics to Optimize the Greenness of Literature Protocols for Syntheses

    ERIC Educational Resources Information Center

    Duarte, Rita C. C.; Ribeiro, M. Gabriela T. C.; Machado, Adélio A. S. C.

    2015-01-01

    A procedure to improve the greenness of a synthesis, without performing laboratory work, using alternative protocols available in the literature is presented. The greenness evaluation involves the separate assessment of the different steps described in the available protocols--reaction, isolation, and purification--as well as the global process,…

  15. Improving patient-centeredness of fertility care using a multifaceted approach: study protocol for a randomized controlled trial.

    PubMed

    Huppelschoten, Aleida G; van Duijnhoven, Noortje T L; Hermens, Rosella P M G; Verhaak, Chris; Kremer, Jan A M; Nelen, Willianne L D M

    2012-09-24

    Beside traditional outcomes of safety and (cost-)effectiveness, the Institute of Medicine states patient-centeredness as an independent outcome indicator to evaluate the quality of healthcare. Providing patient-centered care is important because patients want to be heard for their ideas and concerns. Healthcare areas associated with high emotions and intensive treatment periods could especially benefit from patient-centered care. How care can become optimally improved in patient-centeredness is unknown. Therefore, we will conduct a study in the context of Dutch fertility care to determine the effects of a multifaceted approach on patient-centeredness, patients' quality of life (QoL) and levels of distress. Our aims are to investigate the effectiveness of a multifaceted approach and to identify determinants of a change in the level of patient-centeredness, patients' QoL and distress levels. This paper presents the study protocol. In a cluster-randomized trial in 32 Dutch fertility clinics the effects of a multifaceted approach will be determined on the level of patient-centeredness (Patient-centredness Questionnaire - Infertility), patients' QoL (FertiQoL) and levels of distress (SCREENIVF). The multifaceted approach includes audit and feedback, educational outreach visits and patient-mediated interventions. Potential determinants of a change in patient-centeredness, patients' QoL and levels of distress will be collected by an addendum to the patients' questionnaire and a professionals' questionnaire. The latter includes the Organizational Culture Assessment Instrument about the clinic's culture as a possible determinant of an increase in patient-centered care. The study is expected to yield important new evidence about the effects of a multifaceted approach on levels of patient-centeredness, patients' QoL and distress in fertility care. Furthermore, determinants associated with a change in these outcome measures will be studied. With knowledge of these results

  16. Steps toward improving diet and exercise for cancer survivors (STRIDE): a quasi-randomised controlled trial protocol.

    PubMed

    Frensham, Lauren J; Zarnowiecki, Dorota M; Parfitt, Gaynor; Stanley, Rebecca M; Dollman, James

    2014-06-13

    Cancer survivorship rates have increased in developed countries largely due to population ageing and improvements in cancer care. Survivorship is a neglected phase of cancer treatment and is often associated with adverse physical and psychological effects. There is a need for broadly accessible, non-pharmacological measures that may prolong disease-free survival, reduce or alleviate co-morbidities and enhance quality of life. The aim of the Steps TowaRd Improving Diet and Exercise (STRIDE) study is to evaluate the effectiveness of an online-delivered physical activity intervention for increasing walking in cancer survivors living in metropolitan and rural areas of South Australia. This is a quasi-randomised controlled trial. The intervention period is 12-weeks with 3-month follow-up. The trial will be conducted at a university setting and community health services in South Australia. Participants will be insufficiently active and aged 18 years or older. Participants will be randomly assigned to either the intervention or control group. All participants will receive a pedometer but only the intervention group will have access to the STRIDE website where they will report steps, affect and ratings of perceived exertion (RPE) during exercise daily. Researchers will use these variables to individualise weekly step goals to increase walking.The primary outcome measure is steps per day. The secondary outcomes are a) health measures (anthropometric and physiological), b) dietary habits (consumption of core foods and non-core foods) and c) quality of life (QOL) including physical, psychological and social wellbeing. Measures will be collected at baseline, post-intervention and 3-month follow-up. This protocol describes the implementation of a trial using an online resource to assist cancer survivors to become more physically active. It is an innovative tool that uses ratings of perceived exertion and daily affect to create individualised step goals for cancer survivors. The

  17. A multifaceted quality improvement intervention for CVD risk management in Australian primary healthcare: a protocol for a process evaluation.

    PubMed

    Patel, Bindu; Patel, Anushka; Jan, Stephen; Usherwood, Tim; Harris, Mark; Panaretto, Katie; Zwar, Nicholas; Redfern, Julie; Jansen, Jesse; Doust, Jenny; Peiris, David

    2014-12-17

    Cardiovascular disease (CVD) is the leading cause of death and disability worldwide. Despite the widespread availability of evidence-based clinical guidelines and validated risk predication equations for prevention and management of CVD, their translation into routine practice is limited. We developed a multifaceted quality improvement intervention for CVD risk management which incorporates electronic decision support, patient risk communication tools, computerised audit and feedback tools, and monthly, peer-ranked performance feedback via a web portal. The intervention was implemented in a cluster randomised controlled trial in 60 primary healthcare services in Australia. Overall, there were improvements in risk factor recording and in prescribing of recommended treatments among under-treated individuals, but it is unclear how this intervention was used in practice and what factors promoted or hindered its use. This information is necessary to optimise intervention impact and maximally implement it in a post-trial context. In this study protocol, we outline our methods to conduct a theory-based, process evaluation of the intervention. Our aims are to understand how, why, and for whom the intervention produced the observed outcomes and to develop effective strategies for translation and dissemination. We will conduct four discrete but inter-related studies taking a mixed methods approach. Our quantitative studies will examine (1) the longer term effectiveness of the intervention post-trial, (2) patient and health service level correlates with trial outcomes, and (3) the health economic impact of implementing the intervention at scale. The qualitative studies will (1) identify healthcare provider perspectives on implementation barriers and enablers and (2) use video ethnography and patient semi-structured interviews to understand how cardiovascular risk is communicated in the doctor/patient interaction both with and without the use of intervention. We will also

  18. Improvement of phytoplankton culture isolation using single cell sorting by flow cytometry.

    PubMed

    Marie, Dominique; Le Gall, Florence; Edern, Roseline; Gourvil, Priscillia; Vaulot, Daniel

    2017-04-01

    Flow cytometry provides a tool to physically sort single algal cells in order to obtain clonal cultures. During sorting, cells are submitted to physical stress factors such as high fluidic pressure, exposure to the laser beam, electrostatic charges, deflection through high voltage fields, and collisions with container surfaces. All of these can damage the cells of interest and success rates for initiation of cultures from flow-sorted cells are generally very low. We found that the addition of bovine serum albumin in the culture medium into which cells were sorted drastically improved the success of initiation of pico- and nano-eukaryotic phytoplankton strains. Adding a mixture of antibiotics (Penicillin, Neomycin, Streptomycin) to the medium in order to slow down bacterial growth further improved culture development. This approach was successfully used to isolate taxonomically diverse strains, including novel taxa, from a fresh sample obtained in the English Channel and from enrichment cultures established during an Atlantic meridional transect cruise. We anticipate that these improvements will be useful to clone or purify existing cultures and to isolate novel cultures from oceanic samples. © 2016 Phycological Society of America.

  19. Downhill Running-Based Overtraining Protocol Improves Hepatic Insulin Signaling Pathway without Concomitant Decrease of Inflammatory Proteins

    PubMed Central

    Pauli, José R.; Cintra, Dennys E.; de Souza, Claudio T.; Ropelle, Eduardo R.; R. da Silva, Adelino S.

    2015-01-01

    The purpose of this study was to verify the effects of overtraining (OT) on insulin, inflammatory and gluconeogenesis signaling pathways in the livers of mice. Rodents were divided into control (CT), overtrained by downhill running (OTR/down), overtrained by uphill running (OTR/up) and overtrained by running without inclination (OTR). Rotarod, incremental load, exhaustive and grip force tests were used to evaluate performance. Thirty-six hours after a grip force test, the livers were extracted for subsequent protein analyses. The phosphorylation of insulin receptor beta (pIRbeta), glycogen synthase kinase 3 beta (pGSK3beta) and forkhead box O1 (pFoxo1) increased in OTR/down versus CT. pGSK3beta was higher in OTR/up versus CT, and pFoxo1 was higher in OTR/up and OTR versus CT. Phosphorylation of protein kinase B (pAkt) and insulin receptor substrate 1 (pIRS–1) were higher in OTR/up versus CT and OTR/down. The phosphorylation of IκB kinase alpha and beta (pIKKalpha/beta) was higher in all OT protocols versus CT, and the phosphorylation of stress-activated protein kinases/Jun amino-terminal kinases (pSAPK-JNK) was higher in OTR/down versus CT. Protein levels of peroxisome proliferator-activated receptor-gamma coactivator 1alpha (PGC-1alpha) and hepatocyte nuclear factor 4alpha (HNF-4alpha) were higher in OTR versus CT. In summary, OTR/down improved the major proteins of insulin signaling pathway but up-regulated TRB3, an Akt inhibitor, and its association with Akt. PMID:26445495

  20. Downhill Running-Based Overtraining Protocol Improves Hepatic Insulin Signaling Pathway without Concomitant Decrease of Inflammatory Proteins.

    PubMed

    da Rocha, Alisson L; Pereira, Bruno C; Pauli, José R; Cintra, Dennys E; de Souza, Claudio T; Ropelle, Eduardo R; da Silva, Adelino S R

    2015-01-01

    The purpose of this study was to verify the effects of overtraining (OT) on insulin, inflammatory and gluconeogenesis signaling pathways in the livers of mice. Rodents were divided into control (CT), overtrained by downhill running (OTR/down), overtrained by uphill running (OTR/up) and overtrained by running without inclination (OTR). Rotarod, incremental load, exhaustive and grip force tests were used to evaluate performance. Thirty-six hours after a grip force test, the livers were extracted for subsequent protein analyses. The phosphorylation of insulin receptor beta (pIRbeta), glycogen synthase kinase 3 beta (pGSK3beta) and forkhead box O1 (pFoxo1) increased in OTR/down versus CT. pGSK3beta was higher in OTR/up versus CT, and pFoxo1 was higher in OTR/up and OTR versus CT. Phosphorylation of protein kinase B (pAkt) and insulin receptor substrate 1 (pIRS-1) were higher in OTR/up versus CT and OTR/down. The phosphorylation of IκB kinase alpha and beta (pIKKalpha/beta) was higher in all OT protocols versus CT, and the phosphorylation of stress-activated protein kinases/Jun amino-terminal kinases (pSAPK-JNK) was higher in OTR/down versus CT. Protein levels of peroxisome proliferator-activated receptor-gamma coactivator 1alpha (PGC-1alpha) and hepatocyte nuclear factor 4alpha (HNF-4alpha) were higher in OTR versus CT. In summary, OTR/down improved the major proteins of insulin signaling pathway but up-regulated TRB3, an Akt inhibitor, and its association with Akt.

  1. A Structured Forensic Interview Protocol Improves the Quality and Informativeness of Investigative Interviews with Children: A Review of Research Using the NICHD Investigative Interview Protocol

    ERIC Educational Resources Information Center

    Lamb, Michael; Orbach, Yael; Hershkowitz, Irit; Esplin, Phillip W.; Horowitz, Dvora

    2007-01-01

    Objective: To show how the results of research on children's memory, communicative skills, social knowledge, and social tendencies can be translated into guidelines that improve the quality of forensic interviews of children. Method: We review studies designed to evaluate children's capacities as witnesses, explain the development of the…

  2. Improved isolation of archeomagnetic signals by combined low temperature and alternating field demagnetization

    NASA Astrophysics Data System (ADS)

    Borradaile, Graham J.; Lagroix, France; Trimble, Dale

    2001-09-01

    Conventional alternating field (AF) demagnetization of the magnetite-bearing claystone foundations of a Saxon or late medieval lime kiln in Lincolnshire, England fail to isolate stable characteristic remanences, or remanences compatible with possible contemporary geomagnetic field orientations. Consolidation of the material prevented thermal demagnetization. When low temperature demagnetization (LTD) precedes AF demagnetization, however, the vector plots show a stable characteristic (primary) component. Magnetic anisotropy measurements show that the LTD did not significantly disturb the mineral fabric of the claystone, that the mineral fabric did not deflect the palaeofield, and that AF demagnetization did not induce a field-impressed anisotropy during the experiments. Anisotropy of low-field magnetic susceptibility (AMS) is affected by all minerals, and therefore the anisotropy of the magnetite was isolated by measuring anisotropy of anhysteretic remanence (AARM); this is of more relevance in evaluating the potential for palaeofield deflection. Thus, we conclude that LTD preceding AF demagnetization is responsible for improving the isolation of a characteristic remanence, which then favours a late medieval age for the kiln foundation.

  3. Association between Pseudonocardia symbionts and Atta leaf-cutting ants suggested by improved isolation methods.

    PubMed

    Marsh, Sarah E; Poulsen, Michael; Gorosito, Norma B; Pinto-Tomás, Adrián; Masiulionis, Virginia E; Currie, Cameron R

    2013-03-01

    Fungus-growing ants associate with multiple symbiotic microbes, including Actinobacteria for production of antibiotics. The best studied of these bacteria are within the genus Pseudonocardia, which in most fungus-growing ants are conspicuously visible on the external cuticle of workers. However, given that fungus-growing ants in the genus Atta do not carry visible Actinobacteria on their cuticle, it is unclear if this genus engages in the symbiosis with Pseudonocardia. Here we explore whether improving culturing techniques can allow for successful isolation of Pseudonocardia from Atta cephalotes leaf-cutting ants. We obtained Pseudonocardia from 9 of 11 isolation method/colony component combinations from all 5 colonies intensively sampled. The most efficient technique was bead-beating workers in phosphate buffer solution, then plating the suspension on carboxymethylcellulose medium. Placing these strains in a fungus-growing ant-associated Pseudonocardia phylogeny revealed that while some strains grouped with clades of Pseudonocardia associated with other genera of fungus-growing ants, a large portion of the isolates fell into two novel phylogenetic clades previously not identified from this ant-microbe symbiosis. Our findings suggest that Pseudonocardia may be associated with Atta fungus-growing ants, potentially internalized, and that localizing the symbiont and exploring its role is necessary to shed further light on the association.

  4. Improving safety of salami by application of bacteriocins produced by an autochthonous Lactobacillus curvatus isolate.

    PubMed

    de Souza Barbosa, Matheus; Todorov, Svetoslav Dimitrov; Ivanova, Iskra; Chobert, Jean-Marc; Haertlé, Thomas; de Melo Franco, Bernadette Dora Gombossy

    2015-04-01

    The aims of this study were to isolate LAB with anti-Listeria activity from salami samples, characterize the bacteriocin/s produced by selected isolates, semi-purify them and evaluate their effectiveness for the control of Listeria monocytogenes during manufacturing of salami in a pilot scale. Two isolates (differentiated by RAPD-PCR) presented activity against 22 out of 23 L. monocytogenes strains for bacteriocin MBSa2, while the bacteriocin MBSa3 inhibited all 23 strains in addition to several other Gram-positive bacteria for both antimicrobials and were identified as Lactobacillus curvatus based on 16S rRNA sequencing. A three-step purification procedure indicated that both strains produced the same two active peptides (4457.9 Da and 4360.1 Da), homlogous to sakacins P and X, respectively. Addition of the semi-purified bacteriocins produced by Lb. curvatus MBSa2 to the batter for production of salami, experimentally contaminated with L. monocytogenes (10(4)-10(5) CFU/g), caused 2 log and 1.5 log reductions in the counts of the pathogen in the product after 10 and 20 days respectively, highlighting the interest for application of these bacteriocins to improve safety of salami during its manufacture.

  5. False star detection and isolation during star tracking based on improved chi-square tests

    NASA Astrophysics Data System (ADS)

    Zhang, Hao; Niu, Yanxiong; Lu, Jiazhen; Yang, Yanqiang; Su, Guohua

    2017-08-01

    The star sensor is a precise attitude measurement device for a spacecraft. Star tracking is the main and key working mode for a star sensor. However, during star tracking, false stars become an inevitable interference for star sensor applications, which may result in declined measurement accuracy. A false star detection and isolation algorithm in star tracking based on improved chi-square tests is proposed in this paper. Two estimations are established based on a Kalman filter and a priori information, respectively. The false star detection is operated through adopting the global state chi-square test in a Kalman filter. The false star isolation is achieved using a local state chi-square test. Semi-physical experiments under different trajectories with various false stars are designed for verification. Experiment results show that various false stars can be detected and isolated from navigation stars during star tracking, and the attitude measurement accuracy is hardly influenced by false stars. The proposed algorithm is proved to have an excellent performance in terms of speed, stability, and robustness.

  6. False star detection and isolation during star tracking based on improved chi-square tests.

    PubMed

    Zhang, Hao; Niu, Yanxiong; Lu, Jiazhen; Yang, Yanqiang; Su, Guohua

    2017-08-01

    The star sensor is a precise attitude measurement device for a spacecraft. Star tracking is the main and key working mode for a star sensor. However, during star tracking, false stars become an inevitable interference for star sensor applications, which may result in declined measurement accuracy. A false star detection and isolation algorithm in star tracking based on improved chi-square tests is proposed in this paper. Two estimations are established based on a Kalman filter and a priori information, respectively. The false star detection is operated through adopting the global state chi-square test in a Kalman filter. The false star isolation is achieved using a local state chi-square test. Semi-physical experiments under different trajectories with various false stars are designed for verification. Experiment results show that various false stars can be detected and isolated from navigation stars during star tracking, and the attitude measurement accuracy is hardly influenced by false stars. The proposed algorithm is proved to have an excellent performance in terms of speed, stability, and robustness.

  7. Salvinorins D-F, new neoclerodane diterpenoids from Salvia divinorum, and an improved method for the isolation of salvinorin A.

    PubMed

    Munro, Thomas A; Rizzacasa, Mark A

    2003-05-01

    Three new neoclerodane diterpenoids, salvinorins D-F (4-6), have been isolated from the leaves of Salvia divinorum. The structures were elucidated by chemical and spectroscopic methods, particularly 1D and 2D NMR. A simplified isolation method using chromatography on activated carbon also gave improved yields of the controlled substance salvinorin A (1) and of salvinorin C (3).

  8. PREDOMOS study, impact of a social intervention program for socially isolated elderly cancer patients: study protocol for a randomized controlled trial.

    PubMed

    Crétel-Durand, Elodie; Nouguerède, Emilie; Le Caer, Hervé; Rousseau, Frédérique; Retornaz, Frédérique; Guillem, Olivier; Couderc, Anne-Laure; Greillier, Laurent; Norguet, Emmanuelle; Cécile, Maud; Boulahssass, Rabia; Le Caer, Francoise; Tournier, Sandrine; Butaud, Chantal; Guillet, Pierre; Nahon, Sophie; Poudens, Laure; Kirscher, Sylvie; Loubière, Sandrine; Diaz, Nadine; Dhorne, Jean; Auquier, Pascal; Baumstarck, Karine

    2017-04-12

    Cancer incidence and social isolation increase along with advanced age, and social isolation potentiates the relative risk of death by cancer. Once spotted, social isolation can be averted with the intervention of a multidisciplinary team. Techniques of automation and remote assistance have already demonstrated their positive impact on falls prevention and quality of life (QoL), though little is known about their impact on socially isolated elderly patients supported for cancer. The primary objective of the PREDOMOS study is to evaluate the impact of establishing a Program of Social intervention associated with techniques of Domotic and Remote assistance (PS-DR) on the improvement of QoL of elderly isolated patients, treated for locally advanced or metastatic cancer. The secondary objectives include treatment failure, tolerance, survival, and autonomy. This trial is a multicenter, prospective, randomized, placebo-controlled, open-label, two-parallel group study. The setting is 10 French oncogeriatric centers. Inclusion criteria are patients aged at least 70 years with a social isolation risk and a histological diagnosis of cancer, locally advanced or metastatic disease. The groups are (1) the control group, receiving usual care; (2) the experimental group, receiving usual care associating with monthly social assistance, domotic, and remote assistance. Participants are randomized in a 1:1 allocation ratio. Evaluation times involve inclusion (randomization) and follow-up (12 months). The primary endpoint is QoL at 3 months (via European Organization for Research and Treatment of Cancer (EORTC) QLQ C30); secondary endpoints are social isolation, time to treatment failure, toxicity, dose response-intensity, survival, autonomy, and QoL at 6 months. For the sample size, 320 individuals are required to obtain 90% power to detect a 10-point difference (standard deviation 25) in QoL score between the two groups (20% loss to follow-up patients expected). The randomized

  9. Data for improvement and clinical excellence: protocol for an audit with feedback intervention in home care and supportive living

    PubMed Central

    2012-01-01

    Background Although considerable evidence exists about the effectiveness of audit coupled with feedback, very few audit-with-feedback interventions have been done in either home care or supportive living settings to date. With little history of audit and feedback in home care or supportive living there is potential for greater effects, at least initially. This study extends the work of an earlier study designed to assess the effects of an audit-with-feedback intervention. It will be delivered quarterly over a one-year period in seven home care offices and 11 supportive living sites. The research questions are the same as in the first study but in a different environment. They are as follows: 1. What effects do feedback reports have on processes and outcomes over time? 2. How do different provider groups in home care and supportive living sites respond to feedback reports based on quality indicator data? Methods The research team conducting this study includes researchers and decision makers in continuing care in the province of Alberta, Canada. The intervention consists of quarterly feedback reports in 19 home care offices and supportive living sites across Alberta. Data for the feedback reports are based on the Resident Assessment Instrument Home Care tool, a standardized instrument mandated for use in home care and supportive living environments throughout Alberta. The feedback reports consist of one page, printed front and back, presenting both graphic and textual information. Reports are delivered to all employees working in each site. The primary evaluation uses a controlled interrupted time-series design, both adjusted and unadjusted for covariates. The concurrent process evaluation includes observation, focus groups, and self-reports to assess uptake of the feedback reports. The project described in this protocol follows a similar intervention conducted in our previous study, Data for Improvement and Clinical Excellence--Long-Term Care. We will offer

  10. Acupuncture to improve live birth rates for women undergoing in vitro fertilization: a protocol for a randomized controlled trial.

    PubMed

    Smith, Caroline A; de Lacey, Sheryl; Chapman, Michael; Ratcliffe, Julie; Norman, Robert J; Johnson, Neil; Sacks, Gavin; Lyttleton, Jane; Boothroyd, Clare

    2012-05-18

    IVF is a costly treatment option for women, their partners, and the public. Therefore new therapies that improve reproductive and health outcomes are highly desirable. There is a growing body of research evaluating the effect of acupuncture administered during IVF, and specifically on the day of embryo transfer (ET). Many trials are heterogeneous and results inconsistent. There remains insufficient evidence to determine if acupuncture can enhance live birth rates when used as an adjunct to IVF treatment.The study will determine the clinical effectiveness of acupuncture with improving the proportion of women undergoing IVF having live births. Other objectives include: determination of the cost effectiveness of IVF with acupuncture; and examination of the personal and social context of acupuncture in IVF patients, and examining the reasons why the acupuncture may or may not have worked. We will conduct a randomized controlled trial of acupuncture compared to placebo acupuncture.Inclusion criteria include: women aged less than 43 years; undergoing a fresh IVF or ICSI cycle; and restricted to women with the potential for a lower live birth rate defined as two or more previous unsuccessful ETs; and unsuccessful clinical pregnancies of quality embryos deemed by the embryologist to have been suitable for freezing by standard criteria. Women will be randomized to acupuncture or placebo acupuncture. Treatment is administered on days 6 to 8 of the stimulated cycle and two treatments on the day of ET. A non-randomized cohort of women not using acupuncture will be recruited to the study. The primary study outcome is the proportion of women reporting a live birth. Secondary outcomes include the proportion of women reporting a clinical pregnancy miscarriage prior to 12 weeks, quality of life, and self-efficacy. The sample size of the study is 1,168 women, with the aim of detecting a 7% difference in live births between groups (P = 0.05, 80% power). There remains a need for

  11. Protocol for a prospective observational study to improve prehospital notification of injured patients presenting to trauma centres in India

    PubMed Central

    Mitra, Biswadev; Mathew, Joseph; Gupta, Amit; Cameron, Peter; O'Reilly, Gerard; Soni, Kapil Dev; Kaushik, Gaurav; Howard, Teresa; Fahey, Madonna; Stephenson, Michael; Kumar, Vineet; Vyas, Sharad; Dharap, Satish; Patel, Pankaj; Thakor, Advait; Sharma, Naveen; Walker, Tony; Misra, Mahesh Chandra; Gruen, Russell; Fitzgerald, Mark

    2017-01-01

    Introduction Prehospital notification of injured patients enables prompt and timely care in hospital through adequate preparation of trauma teams, space, equipment and consumables necessary for resuscitation, and may improve outcomes. In India, anecdotal reports suggest that prehospital notification, in those few places where it occurs, is unstructured and not linked to a well-defined hospital response. The aim of this manuscript is to describe, in detail, a study protocol for the evaluation of a formalised approach to prehospital notification. Methods and analysis This is a longitudinal prospective cohort study of injured patients being transported by ambulance to major trauma centres in India. In the preintervention phase, prospective data on patients will be collected on prehospital assessment, notification, inhospital assessment, management and outcomes and recorded in a new tailored multihospital trauma registry. All injured patients arriving by ambulance and allocated to a red or yellow priority category will be eligible for inclusion. The intervention will be a prehospital notification application to be used by ambulance clinicians to notify emergency departments of the impending arrival of a patient. The proportion of eligible patients arriving to hospital after notification will be the primary outcome measure. Secondary outcomes evaluated will be availability of a trauma cubicle, presence of a trauma team on patient arrival, time to first chest X-ray and inhospital mortality. Progress Ethical approval has been obtained from the All India Institute of Medical Sciences, New Delhi and site-specific approval granted by relevant trauma services. The trial has also been registered with the Monash University Human Research and Ethics Committee; Project number: CF16/1814 – 2016000929. Results will be fed back to prehospital and hospital clinicians via a series of reports and presentations. These will be used to facilitate discussions about service redesign and

  12. Protocol for a prospective observational study to improve prehospital notification of injured patients presenting to trauma centres in India.

    PubMed

    Mitra, Biswadev; Mathew, Joseph; Gupta, Amit; Cameron, Peter; O'Reilly, Gerard; Soni, Kapil Dev; Kaushik, Gaurav; Howard, Teresa; Fahey, Madonna; Stephenson, Michael; Kumar, Vineet; Vyas, Sharad; Dharap, Satish; Patel, Pankaj; Thakor, Advait; Sharma, Naveen; Walker, Tony; Misra, Mahesh Chandra; Gruen, Russell; Fitzgerald, Mark

    2017-07-17

    Prehospital notification of injured patients enables prompt and timely care in hospital through adequate preparation of trauma teams, space, equipment and consumables necessary for resuscitation, and may improve outcomes. In India, anecdotal reports suggest that prehospital notification, in those few places where it occurs, is unstructured and not linked to a well-defined hospital response. The aim of this manuscript is to describe, in detail, a study protocol for the evaluation of a formalised approach to prehospital notification. This is a longitudinal prospective cohort study of injured patients being transported by ambulance to major trauma centres in India. In the preintervention phase, prospective data on patients will be collected on prehospital assessment, notification, inhospital assessment, management and outcomes and recorded in a new tailored multihospital trauma registry. All injured patients arriving by ambulance and allocated to a red or yellow priority category will be eligible for inclusion. The intervention will be a prehospital notification application to be used by ambulance clinicians to notify emergency departments of the impending arrival of a patient. The proportion of eligible patients arriving to hospital after notification will be the primary outcome measure. Secondary outcomes evaluated will be availability of a trauma cubicle, presence of a trauma team on patient arrival, time to first chest X-ray and inhospital mortality. Ethical approval has been obtained from the All India Institute of Medical Sciences, New Delhi and site-specific approval granted by relevant trauma services. The trial has also been registered with the Monash University Human Research and Ethics Committee; Project number: CF16/1814 - 2016000929. Results will be fed back to prehospital and hospital clinicians via a series of reports and presentations. These will be used to facilitate discussions about service redesign and implementation. It is expected that evidence

  13. Improved oil recovery using bacteria isolated from North Sea petroleum reservoirs

    SciTech Connect

    Davey, R.A.; Lappin-Scott, H.

    1995-12-31

    During secondary oil recovery, water is injected into the formation to sweep out the residual oil. The injected water, however, follows the path of least resistance through the high-permeability zones, leaving oil in the low-permeability zones. Selective plugging of these their zones would divert the waterflood to the residual oil and thus increase the life of the well. Bacteria have been suggested as an alternative plugging agent to the current method of polymer injection. Starved bacteria can penetrate deeply into rock formations where they attach to the rock surfaces, and given the right nutrients can grow and produce exo-polymer, reducing the permeability of these zones. The application of microbial enhanced oil recovery has only been applied to shallow, cool, onshore fields to date. This study has focused on the ability of bacteria to enhance oil recovery offshore in the North Sea, where the environment can be considered extreme. A screen of produced water from oil reservoirs (and other extreme subterranean environments) was undertaken, and two bacteria were chosen for further work. These two isolates were able to grow and survive in the presence of saline formation waters at a range of temperatures above 50{degrees}C as facultative anaerobes. When a solution of isolates was passed through sandpacks and nutrients were added, significant reductions in permeabilities were achieved. This was confirmed in Clashach sandstone at 255 bar, when a reduction of 88% in permeability was obtained. Both isolates can survive nutrient starvation, which may improve penetration through the reservoir. Thus, the isolates show potential for field trials in the North Sea as plugging agents.

  14. Improved hypothermic short-term storage of isolated mouse islets by adding serum to preservation solutions.

    PubMed

    Kimura, Yasuko; Okitsu, Teru; Xibao, Liu; Teramae, Hiroki; Okonogi, Atsuhito; Toyoda, Kentaro; Uemoto, Shinji; Fukushima, Masanori

    2013-01-01

    Preserving isolated islets at low temperature appears attractive because it can keep islet quantity comparable to freshly isolated islets. In this study, we evaluated the effect of serum as an additive to preservation solutions on islet quality after short-term hypothermic storage. Isolated mouse islets were preserved at 4°C in University of Wisconsin solution (UW) alone, UW with serum, M-Kyoto solution (MK) alone or MK with serum. We then assessed islet quantity, morphology, viability and function in vitro as well as in vivo. Islet quantity after storage in all four solutions was well maintained for up to 120 h. However, islets functioned for different duration; glucose-stimulated insulin release assay revealed that the duration was 72 h when islets were stored in UW with serum and MK with serum, but only 24 h in UW alone, and the islet function disappeared immediately in MK alone. Viability assay confirmed that more than 70% islet cells survived for up to 48 h when islets are preserved in UW with serum and MK with serum, but the viability decreased rapidly in UW alone and MK alone. In in vivo bioassays using 48-h preserved isogeneic islets, all recipient mice restored normal blood glucose concentrations by transplants preserved in UW with serum or MK with serum, whereas 33.3% recipients and no recipient restored diabetes by transplants preserved in UW alone and in MK alone respectively. Adding serum to both UW and MK improves their capability to store isolated islets by maintaining islet functional viability.

  15. Improved hypothermic short-term storage of isolated mouse islets by adding serum to preservation solutions

    PubMed Central

    Kimura, Yasuko; Okitsu, Teru; Xibao, Liu; Teramae, Hiroki; Okonogi, Atsuhito; Toyoda, Kentaro; Uemoto, Shinji; Fukushima, Masanori

    2013-01-01

    Preserving isolated islets at low temperature appears attractive because it can keep islet quantity comparable to freshly isolated islets. In this study, we evaluated the effect of serum as an additive to preservation solutions on islet quality after short-term hypothermic storage. Isolated mouse islets were preserved at 4°C in University of Wisconsin solution (UW) alone, UW with serum, M-Kyoto solution (MK) alone or MK with serum. We then assessed islet quantity, morphology, viability and function in vitro as well as in vivo. Islet quantity after storage in all four solutions was well maintained for up to 120 h. However, islets functioned for different duration; glucose-stimulated insulin release assay revealed that the duration was 72 h when islets were stored in UW with serum and MK with serum, but only 24 h in UW alone, and the islet function disappeared immediately in MK alone. Viability assay confirmed that more than 70% islet cells survived for up to 48 h when islets are preserved in UW with serum and MK with serum, but the viability decreased rapidly in UW alone and MK alone. In in vivo bioassays using 48-h preserved isogeneic islets, all recipient mice restored normal blood glucose concentrations by transplants preserved in UW with serum or MK with serum, whereas 33.3% recipients and no recipient restored diabetes by transplants preserved in UW alone and in MK alone respectively. Adding serum to both UW and MK improves their capability to store isolated islets by maintaining islet functional viability. PMID:23552019

  16. Isolation of Ashbya gossypii mutant for an improved riboflavin production targeting for biorefinery technology.

    PubMed

    Park, E Y; Zhang, J H; Tajima, S; Dwiarti, L

    2007-08-01

    To isolate a strain overproducing riboflavin and to improve riboflavin production for practical use in a biorefinery technology. Ashbya gossypii spores were mutagenized by exposure to UV light and mutant ZP4 strain, producing riboflavin threefold the riboflavin that of the wild-type strain, was isolated by the first and second screenings. Proteomic analysis of ZP4 strain showed the expression patterns of eight types of genes related to riboflavin biosynthesis different from those of the wild-type strain and those enzyme activities were investigated. When activated bleaching earth (ABE) containing 75 g l(-1) rapeseed oil was added in the culture of the ZP4 strain with oxygen-enriched air supplied, riboflavin concentration increased to 8.7 g l(-1) at 5 days of culture. Riboflavin production yield was 0.17 g g(-1) of consumed oil, which was eightfold higher than that of the wild-type strain. The results show that the mutant ZP4 strain shows potential for improving riboflavin production for practical utilization using vegetable oil as the sole carbon source. Our results indicate that the mutant ZP4 strain shows potential for producing riboflavin from vegetable oil, and therefore will be contributed to biorefinery technology.

  17. Enhanced high-energy protocol using a fractional bipolar radiofrequency device combined with bipolar radiofrequency and infrared light for improving facial skin appearance and wrinkles.

    PubMed

    Gold, Michael H; Biesman, Brian S; Taylor, Mark

    2017-02-08

    Fractional bipolar radiofrequency treatment and treatment with bipolar radiofrequency combined with infrared light have been shown in previous trials to safely and effectively improve the appearance of facial wrinkles. To evaluate a high-energy protocol with combined bipolar radiofrequency and infrared light energies for improvement in photoaged facial skin. Seventy-two patients presenting with mild to moderate facial wrinkles underwent a single full-face treatment (n=54) or two treatments (n=18) at 6-week intervals. Independent blinded assessment and investigator assessment were performed, using the Fitzpatrick Wrinkle and Elastosis Scale (0-9) and the Global Aesthetic Improvement scale. Patients also completed a self-assessment questionnaire concerning satisfaction with the treatment. All patients achieved some degree of improvement in their wrinkles and skin appearance, following a single treatment or two treatments with the enhanced-energy protocol. Blinded evaluation demonstrated 71% and 70% of the patients showing improvement of one unit or greater on the Fitzpatrick Scale, at the 12-week and 24-week follow-ups post-treatment, respectively. Similar results were reported by investigators. Under the Global Aesthetic Improvement Scale, investigators observed 87%, 91% and 81% of patients showing improvement at the 6-, 12-, and 24-week post-treatment end, respectively. Patients tolerated the treatments well and were satisfied with the clinical results. The enhanced-energy treatment protocol, with fractional bipolar radiofrequency treatment and treatment with bipolar radiofrequency combined with infrared light applications, yields significant improvement of skin texture, wrinkling, and overall appearance following a single treatment. The results appear gradually over time and are maintained for at least 6 months' post-treatment. © 2017 Wiley Periodicals, Inc.

  18. Do treatment protocols improve end results? A study of survival of patients with multiple myeloma in Finland.

    PubMed

    Karjalainen, S; Palva, I

    1989-10-28

    diagnosis. The patients in the trial area benefited from the clinical trials, which suggests that the use of a treatment protocol improves the end results of treatment. In other words, the results favour a systematic treatment schedule in preference to a schedule determined by the free choice of a clinician.

  19. Evaluation of a minimal sedation protocol using ICU sedative consumption as a monitoring tool: a quality improvement multicenter project.

    PubMed

    Ranzani, Otavio T; Simpson, Evelyn Senna; Augusto, Talita Barbosa; Cappi, Sylas Bezerra; Noritomi, Danilo Teixeira

    2014-10-24

    Oversedation frequently occurs in ICUs. We aimed to evaluate a minimal sedation policy, using sedative consumption as a monitoring tool, in a network of ICUs targeting decrement of oversedation and mechanical ventilation (MV) duration. A prospective quality improvement project was conducted in ten ICUs within a network of nonteaching hospitals in Brazil during a 2-year period (2010 to 2012). In the first 12 months (the preintervention period), we conducted an audit to identify sedation practice and barriers to current guideline-based practice regarding sedation. In the postintervention period, we implemented a multifaceted program, including multidisciplinary daily rounds, and monthly audits focusing on sedative consumption, feedback and benchmarking purposes. To analyze the effect of the campaign, we fit an interrupted time series (ITS). To account for variability among the network ICUs, we fit a hierarchical model. During the study period, 21% of patients received MV (4,851/22,963). In the postintervention period, the length of MV was lower (3.91 ± 6.2 days versus 3.15 ± 4.6 days; mean difference, -0.76 (95% CI, -1.10; -0.43), P <0.001) and 28 ventilator-free days were higher (16.07 ± 12.2 days versus 18.33 ± 11.6 days; mean difference, 2.30 (95% CI, 1.57; 3.00), P <0.001) than in the preintervention period. Midazolam consumption (in milligrams per day of MV) decreased from 329 ± 70 mg/day to 163 ± 115 mg/day (mean difference, -167 (95% CI, -246; -87), P <0.001). In contrast, consumption of propofol (P = 0.007), dexmedetomidine (P = 0.017) and haloperidol (P = 0.002) increased in the postintervention period, without changes in the consumption of fentanyl. Through ITS, age (P = 0.574) and Simplified Acute Physiology Score III (P = 0.176) remained stable. The length of MV showed a secular effect (secular trend β(1) = -0.055, P = 0.012) and a strong decrease immediately after the intervention (intervention β(2) = -0.976, P <0.001). The impact was

  20. The Ontological Perspectives of the Semantic Web and the Metadata Harvesting Protocol: Applications of Metadata for Improving Web Search.

    ERIC Educational Resources Information Center

    Fast, Karl V.; Campbell, D. Grant

    2001-01-01

    Compares the implied ontological frameworks of the Open Archives Initiative Protocol for Metadata Harvesting and the World Wide Web Consortium's Semantic Web. Discusses current search engine technology, semantic markup, indexing principles of special libraries and online databases, and componentization and the distinction between data and…

  1. The Ontological Perspectives of the Semantic Web and the Metadata Harvesting Protocol: Applications of Metadata for Improving Web Search.

    ERIC Educational Resources Information Center

    Fast, Karl V.; Campbell, D. Grant

    2001-01-01

    Compares the implied ontological frameworks of the Open Archives Initiative Protocol for Metadata Harvesting and the World Wide Web Consortium's Semantic Web. Discusses current search engine technology, semantic markup, indexing principles of special libraries and online databases, and componentization and the distinction between data and…

  2. Assessment of a decision making protocol to improve the efficacy of VELscope™ in general dental practice: a prospective evaluation.

    PubMed

    Bhatia, Nirav; Matias, Marie Anne T; Farah, Camile S

    2014-10-01

    VELscope™ is a device designed to help detect potentially malignant disorders of the oral mucosa at an early stage using direct tissue autofluorescence. Previous research indicates a high rate of false positives using this device. This study assesses a decision making protocol for the detection of oral mucosal lesions using conventional oral examination and VELscope™ in a general dental practice setting. 305 patients presenting for general dental treatment were screened by a general dental practitioner (GDP) for oral mucosal lesions using incandescent light (conventional oral examination - COE), followed by VELscope™ and then by correlating the findings from these two examinations. A decision making protocol was followed. Patients were either reviewed or referred to an Oral Medicine specialist (OMS) for consultation, and biopsy was undertaken as required for definitive diagnosis. 146 patients presented with at least one oral mucosal lesion, and a total of 222 lesions were detected. COE detected 161 oral mucosal lesions and an additional 61 lesions were detected with VELscope™. COE alone showed a sensitivity of 44.0% and specificity of 99.0% while VELscope™ alone showed a sensitivity of 64.0% and specificity of 54.3%. Using the decision making protocol, the sensitivity and specificity were 73.9% and 97.9% respectively. Using the decision making protocol proposed in this study allows for the detection of additional oral mucosal lesions requiring specialist referral by incorporating VELscope™ into routine general dental practice, without compromising patient care. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Short-term supervised inpatient physiotherapy exercise protocol improves cardiac autonomic function after coronary artery bypass graft surgery--a randomised controlled trial.

    PubMed

    Mendes, Renata Gonçalves; Simões, Rodrigo Polaquini; De Souza Melo Costa, Fernando; Pantoni, Camila Bianca Falasco; Di Thommazo, Luciana; Luzzi, Sérgio; Catai, Aparecida Maria; Arena, Ross; Borghi-Silva, Audrey

    2010-01-01

    Coronary artery bypass grafting (CABG) is accompanied by severe impairment of cardiac autonomous regulation (CAR). This study aimed to determine whether a short-term physiotherapy exercise protocol post-CABG, during inpatient cardiac rehabilitation (CR), might improve CAR. Seventy-four patients eligible for CABG were recruited and randomised into physiotherapy exercise group (EG) or physiotherapy usual care group (UCG). EG patients underwent a short-term supervised inpatient physiotherapy exercise protocol consisting of an early mobilisation with progressive exercises plus usual care (respiratory exercises). UCG only received respiratory exercises. Forty-seven patients (24 EG and 23 UGC) completed the study. Outcome measures of CAR included linear and non-linear measures of heart rate variability (HRV) assessed before discharge. By hospital discharge, EG presented significantly higher parasympathetic HRV values [rMSSD, high frequency (HF), SD1)], global power (STD RR, SD2), non-linear HRV indexes [detrended fluctuation analysis (DFA)alpha1, DFAalpha2, approximate entropy (ApEn)] and mean RR compared to UCG (p<0.05). Conversely, higher values of mean HR, low frequency (LF) (sympathetic activity) and the LF/HF (global sympatho-vagal balance) were found in the UCG. A short-term supervised physiotherapy exercise protocol during inpatient CR improves CAR at the time of discharge. Thus, exercise-based inpatient CR might be an effective non-pharmacological tool to improve autonomic cardiac tone in patient's post-CABG.

  4. Improved methods in Agrobacterium-mediated transformation of almond using positive (mannose/pmi) or negative (kanamycin resistance) selection-based protocols.

    PubMed

    Ramesh, Sunita A; Kaiser, Brent N; Franks, Tricia; Collins, Graham; Sedgley, Margaret

    2006-08-01

    A protocol for Agrobacterium-mediated transformation with either kanamycin or mannose selection was developed for leaf explants of the cultivar Prunus dulcis cv. Ne Plus Ultra. Regenerating shoots were selected on medium containing 15 muM kanamycin (negative selection), while in the positive selection strategy, shoots were selected on 2.5 g/l mannose supplemented with 15 g/l sucrose. Transformation efficiencies based on PCR analysis of individual putative transformed shoots from independent lines relative to the initial numbers of leaf explants tested were 5.6% for kanamycin/nptII and 6.8% for mannose/pmi selection, respectively. Southern blot analysis on six randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene in each, and five randomly chosen lines identified to contain the pmi transgene by PCR showed positive hybridisation to a pmi DNA probe. The positive (mannose/pmi) and the negative (kanamycin) selection protocols used in this study have greatly improved transformation efficiency in almond, which were confirmed with PCR and Southern blot. This study also demonstrates that in almond the mannose/pmi selection protocol is appropriate and can result in higher transformation efficiencies over that of kanamycin/nptII selection protocols.

  5. Superparamagnetic iron oxide-enhanced liver MRI with SHU 555 A (RESOVIST): New protocol infusion to improve arterial phase evaluation--a prospective study.

    PubMed

    Grazioli, Luigi; Bondioni, Maria Pia; Romanini, Laura; Frittoli, Barbara; Gambarini, Sebastiana; Donato, Francesco; Santoro, Lucia; Colagrande, Stefano

    2009-03-01

    To compare the arterial enhancement of hypervascular hepatic lesions by T1-weighted 3D-GRE (gradient-recalled echo) fat-sat sequence after slow (0.5 mL/sec) and fast (2 mL/sec) RESOVIST infusion. We prospectively enrolled 71 patients with hypervascular hepatic lesions to undergo dynamic magnetic resonance imaging (MRI) examination with RESOVIST. A total of 92 benign and malignant lesions, 44 of which histologically confirmed, were examined. Three blinded and independent readers visually assessed the arterial enhancement using a score from 0 (none) to 3 (maximum), the latter score comparable to that achievable by MultiHance administration. Out of the 92 hypervascular lesions, 41, 31, and 20 nodules were examined using the slow, fast, and both protocols, respectively. Relevant enhancement (scores 2-3) was found in 42% vs. 14.5% of cases for slow and fast protocols, respectively. Intraindividual comparison evaluation confirmed the better results obtained by slow than fast protocol (25% vs. 10%), with statistically relevant difference in distribution of scores (P=0.0004). The slow protocol showed values between 0 and 3 with an arithmetic mean of 1.1; the fast one, on the other hand, showed values between 0 and 2 with an arithmetic mean of 0.66. Slow infusion improves arterial enhancement after RESOVIST administration. Copyright (c) 2009 Wiley-Liss, Inc.

  6. Isolation and identification of bacteria to improve the strength of concrete.

    PubMed

    Krishnapriya, S; Venkatesh Babu, D L; G, Prince Arulraj

    2015-05-01

    The objective of this research work is to isolate and identify calcite precipitating bacteria and to check the suitability of these bacteria for use in concrete to improve its strength. Bacteria to be incorporated in concrete should be alkali resistant to endure the high pH of concrete and endospore forming to withstand the mechanical stresses induced in concrete during mixing. They must exhibit high urease activity to precipitate calcium carbonate in the form of calcite. Bacterial strains were isolated from alkaline soil samples of a cement factory and were tested for urease activity, potential to form endospores and precipitation of calcium carbonate. Based on these results, three isolates were selected and identified by 16S rRNA gene sequencing. They were identified as Bacillus megaterium BSKAU, Bacillus licheniformis BSKNAU and Bacillus flexus BSKNAU. The results were compared with B. megaterium MTCC 1684 obtained from Microbial Type Culture Collection and Gene Bank, Chandigarh, India. Experimental work was carried out to assess the influence of bacteria on the compressive strength and tests revealed that bacterial concrete specimens showed enhancement in compressive strength. The efficiency of bacteria toward crack healing was also tested. Substantial increase in strength and complete healing of cracks was observed in concrete specimens cast with B. megaterium BSKAU, B. licheniformis BSKNAU and B. megaterium MTCC 1684. This indicates the suitability of these bacterial strains for use in concrete. The enhancement of strength and healing of cracks can be attributed to the filling of cracks in concrete by calcite which was visualized by scanning electron microscope.

  7. Development of an Improved Selective and Differential Medium for Isolation of Salmonella spp.

    PubMed Central

    Park, Sang-Hyun; Ryu, Sangryeol

    2012-01-01

    We describe an improved selective, differential, and cost-effective medium, XA medium, which contains d-arabinose, to facilitate the selective isolation of Salmonella spp. The sensitivity and the specificity of XA medium were compared to those of xylose lysine desoxycholate agar (XLD) using stock cultures and naturally contaminated food samples. XA medium and XLD were evaluated with a total of 82 Salmonella and 69 non-Salmonella stock cultures. Of 82 strains of Salmonella spp. tested, 76 produced a characteristic black colony on XA medium and XLD. The remaining 6 strains belonged to Salmonella enterica serovars Berta (n = 1), Paratyphi A (n = 1), Gallinarum (n = 2), and Pullorum (n = 2). The sensitivities of XA medium and XLD were identical (92.7%). Citrobacter freundii (n = 21) and Proteus mirabilis (n = 21) stock cultures produced black colonies on XLD, whereas only 4 strains of P. mirabilis appeared as black colonies on XA medium. In the second phase of the study, a total of 180 food samples were cultured onto XA medium and XLD after selective enrichment. The sensitivities of XA medium and XLD were equal (100%), and a total of 6 Salmonella strains were isolated from the 180 food samples. The specificity of XA medium (92.0%) was superior to that of XLD (73.0%), with a total of 14 and 47 false-positive results found on XA medium and XLD, respectively. On the basis of its good specificity, XA medium is useful for the isolation of Salmonella spp. from food samples. PMID:22814469

  8. Enriched environment increases neurogenesis and improves social memory persistence in socially isolated adult mice.

    PubMed

    Monteiro, Brisa M M; Moreira, Fabrício A; Massensini, André R; Moraes, Márcio F D; Pereira, Grace S

    2014-02-01

    Social memory consists of the information necessary to identify and recognize cospecifics and is essential to many forms of social interaction. Social memory persistence is strongly modulated by the animal's experiences. We have shown in previous studies that social isolation (SI) in adulthood impairs social memory persistence and that an enriched environment (EE) prevents this impairment. However, the mechanisms involved in the effects of SI and EE on social memory persistence remain unknown. We hypothesized that the mechanism by which SI and EE affect social memory persistence is through their modulation of neurogenesis. To investigate this hypothesis, adult mice were submitted to 7 days of one of the following conditions: group-housing in a standard (GH) or enriched environment (GH+EE); social isolation in standard (SI) or enriched environment (SI+EE). We observed an increase in the number of newborn neurons in the dentate gyrus of the hippocampus (DG) and glomerular layer of the olfactory bulb (OB) in both GH+EE and SI+EE mice. However, this increase of newborn neurons in the granule cell layer of the OB was restricted to the GH+EE group. Furthermore, both SI and SI+EE groups showed less neurogenesis in the mitral layer of the OB. Interestingly, the performance of the SI mice in the buried food-finding task was inferior to that of the GH mice. To further analyze whether increased neurogenesis is in fact the mechanism by which the EE improves social memory persistence in SI mice, we administered the mitotic inhibitor AraC or saline directly into the lateral ventricles of the SI+EE mice. We found that the AraC treatment decreased cell proliferation in both the DG and OB, and impaired social memory persistence in the SI+EE mice. Taken together, our results strongly suggest that neurogenesis is what supports social memory persistence in socially isolated mice.

  9. Improved tandem affinity purification tag and methods for isolation of protein heterocomplexes from plants.

    PubMed

    Rohila, Jai S; Chen, Mei; Cerny, Ronald; Fromm, Michael E

    2004-04-01

    A synthetic gene encoding the tandem affinity purification (TAP) tag has been constructed, and the TAP tag assayed for its effects on expression levels and subcellular localization by fusion to green fluorescent protein (GFP) as well as for its effects on steroid-dependent translocation to the nucleus and transcription when fused to a hybrid glucocorticoid receptor. A nuclear localization signal (NLS) was detected in the calmodulin-binding protein (CBP) domain and removed by mutation to improve the usefulness of the TAP tag. Additionally, purification improvements were made, including inhibition of a co-purifying protease, and adding a protein cross-linking step to increase the recovery of interacting proteins. The improved synthetic TAP tag gene and methods were used to isolate proteins interacting with the hybrid glucocorticoid receptor and to identify them by mass spectrometry. The two proteins identified, HSP70 and HSP90, are known to interact with the glucocorticoid receptor in vivo in mammalian cells and in vitro in plants.

  10. The Use of Variable Q1 Isolation Windows Improves Selectivity in LC-SWATH-MS Acquisition.

    PubMed

    Zhang, Ying; Bilbao, Aivett; Bruderer, Tobias; Luban, Jeremy; Strambio-De-Castillia, Caterina; Lisacek, Frédérique; Hopfgartner, Gérard; Varesio, Emmanuel

    2015-10-02

    As tryptic peptides and metabolites are not equally distributed along the mass range, the probability of cross fragment ion interference is higher in certain windows when fixed Q1 SWATH windows are applied. We evaluated the benefits of utilizing variable Q1 SWATH windows with regards to selectivity improvement. Variable windows based on equalizing the distribution of either the precursor ion population (PIP) or the total ion current (TIC) within each window were generated by an in-house software, swathTUNER. These two variable Q1 SWATH window strategies outperformed, with respect to quantification and identification, the basic approach using a fixed window width (FIX) for proteomic profiling of human monocyte-derived dendritic cells (MDDCs). Thus, 13.8 and 8.4% additional peptide precursors, which resulted in 13.1 and 10.0% more proteins, were confidently identified by SWATH using the strategy PIP and TIC, respectively, in the MDDC proteomic sample. On the basis of the spectral library purity score, some improvement warranted by variable Q1 windows was also observed, albeit to a lesser extent, in the metabolomic profiling of human urine. We show that the novel concept of "scheduled SWATH" proposed here, which incorporates (i) variable isolation windows and (ii) precursor retention time segmentation further improves both peptide and metabolite identifications.

  11. An improved protocol and a new grinding device for extraction of genomic DNA from microorganisms by a two-step extraction procedure.

    PubMed

    Zhang, S S; Chen, D; Lu, Q

    2012-05-21

    Current protocols to extract genomic DNA from microorganisms are still laborious, tedious and costly, especially for the species with thick cell walls. In order to improve the effectiveness of extracting DNA from microbial samples, a novel protocol, defined as two-step extraction method, along with an improved tissue-grinding device, was developed. The protocol included two steps, disruption of microbial cells or spores by grinding the sample together with silica sand in a new device and extraction of DNA with an effective buffer containing cell lysis chemicals. The device was prepared by using a commercial electric mini-grinder, adapted with a grinding stone, and a sample cup processed by lathing from a polytetrafluoroethylene rod. We tested the method with vegetative cells of four microbial species and two microbial spores that have thick cell walls and are therefore hard to process; these included Escherichia coli JM109, Bacillus subtilis WB600, Sacchromyces cerevisiae INVSc1, Trichoderma viride AS3.3711, and the spores of S. cerevisiae and T. viride, respectively, representing Gram-positive bacteria, Gram-negative bacteria, yeast, filamentous fungi. We found that this new method and device extracted usable quantities of genomic DNA from the samples. The DNA fragments that were extracted exceeded 23 kb. The target sequences up to about 5 kb were successfully and exclusively amplified by PCR using extracted DNA as the template. In addition, the DNA extraction was finalized within 1.5 h. Thus, we conclude that this two-step extraction method is an effective and improved protocol for extraction of genomic DNA from microbial samples.

  12. Isolation and Characterization of Brewer's Yeast Variants with Improved Fermentation Performance under High-Gravity Conditions▿

    PubMed Central

    Blieck, Lies; Toye, Geert; Dumortier, Françoise; Verstrepen, Kevin J.; Delvaux, Freddy R.; Thevelein, Johan M.; Van Dijck, Patrick

    2007-01-01

    To save energy, space, and time, today's breweries make use of high-gravity brewing in which concentrated medium (wort) is fermented, resulting in a product with higher ethanol content. After fermentation, the product is diluted to obtain beer with the desired alcohol content. While economically desirable, the use of wort with an even higher sugar concentration is limited by the inability of brewer's yeast (Saccharomyces pastorianus) to efficiently ferment such concentrated medium. Here, we describe a successful strategy to obtain yeast variants with significantly improved fermentation capacity under high-gravity conditions. We isolated better-performing variants of the industrial lager strain CMBS33 by subjecting a pool of UV-induced variants to consecutive rounds of fermentation in very-high-gravity wort (>22° Plato). Two variants (GT336 and GT344) showing faster fermentation rates and/or more-complete attenuation as well as improved viability under high ethanol conditions were identified. The variants displayed the same advantages in a pilot-scale stirred fermenter under high-gravity conditions at 11°C. Microarray analysis identified several genes whose altered expression may be responsible for the superior performance of the variants. The role of some of these candidate genes was confirmed by genetic transformation. Our study shows that proper selection conditions allow the isolation of variants of commercial brewer's yeast with superior fermentation characteristics. Moreover, it is the first study to identify genes that affect fermentation performance under high-gravity conditions. The results are of interest to the beer and bioethanol industries, where the use of more-concentrated medium is economically advantageous. PMID:17158628

  13. Isolation and characterization of brewer's yeast variants with improved fermentation performance under high-gravity conditions.

    PubMed

    Blieck, Lies; Toye, Geert; Dumortier, Françoise; Verstrepen, Kevin J; Delvaux, Freddy R; Thevelein, Johan M; Van Dijck, Patrick

    2007-02-01

    To save energy, space, and time, today's breweries make use of high-gravity brewing in which concentrated medium (wort) is fermented, resulting in a product with higher ethanol content. After fermentation, the product is diluted to obtain beer with the desired alcohol content. While economically desirable, the use of wort with an even higher sugar concentration is limited by the inability of brewer's yeast (Saccharomyces pastorianus) to efficiently ferment such concentrated medium. Here, we describe a successful strategy to obtain yeast variants with significantly improved fermentation capacity under high-gravity conditions. We isolated better-performing variants of the industrial lager strain CMBS33 by subjecting a pool of UV-induced variants to consecutive rounds of fermentation in very-high-gravity wort (>22 degrees Plato). Two variants (GT336 and GT344) showing faster fermentation rates and/or more-complete attenuation as well as improved viability under high ethanol conditions were identified. The variants displayed the same advantages in a pilot-scale stirred fermenter under high-gravity conditions at 11 degrees C. Microarray analysis identified several genes whose altered expression may be responsible for the superior performance of the variants. The role of some of these candidate genes was confirmed by genetic transformation. Our study shows that proper selection conditions allow the isolation of variants of commercial brewer's yeast with superior fermentation characteristics. Moreover, it is the first study to identify genes that affect fermentation performance under high-gravity conditions. The results are of interest to the beer and bioethanol industries, where the use of more-concentrated medium is economically advantageous.

  14. A service-level action research intervention to improve identification and treatment of cannabis and related mental health issues in young Indigenous Australians: a study protocol.

    PubMed

    Bohanna, India; Bird, Katrina; Copeland, Jan; Roberts, Nicholas; Clough, Alan

    2014-07-31

    Regular cannabis use is associated with negative mental health impacts including psychosis, depression and anxiety. Rates of cannabis use have increased in Aboriginal and Torres Strait Islander communities in northern Australia within the last two decades, presenting a significant increased risk to young people's mental health in these regions. Improved screening, early detection and treatment for cannabis-related mental health issues are urgently required. This paper describes a service-level action research intervention and evaluation protocol for use in the few services where it is possible to engage young Aboriginal and Torres Strait Islander Australians. The protocol is being developed in two services where youth mental health is core business: a primary healthcare centre and a youth service in the Cairns and hinterland region, far north Queensland. The protocol calls first for baseline data to be collected using staff and client surveys; network mapping; and analysis of screening, treatment and referral rates. The protocol's intervention phase is driven by service needs identified from baseline data. Intervention strategies focus on implementing/enhancing cannabis screening instruments and processes in line with current best practice; enhancing networks with external drug and mental health services; developing culturally acceptable training and resources; developing activities aiming to reduce cannabis use in young Aboriginal and Torres Strait Islander clients using the services. The protocol requires implementation of the multilevel intervention within each service for 1 year, with follow-up data then collected and compared to baseline. Process evaluation identifies the more effective intervention strategies and documents the challenges to be overcome for full implementation. Ethics approval was provided by The James Cook University, Human Research Ethics Committee. Ethics Approval Number H5322. Peer-reviewed publications will also be used to disseminate

  15. A service-level action research intervention to improve identification and treatment of cannabis and related mental health issues in young Indigenous Australians: a study protocol

    PubMed Central

    Bohanna, India; Bird, Katrina; Copeland, Jan; Roberts, Nicholas; Clough, Alan

    2014-01-01

    Introduction Regular cannabis use is associated with negative mental health impacts including psychosis, depression and anxiety. Rates of cannabis use have increased in Aboriginal and Torres Strait Islander communities in northern Australia within the last two decades, presenting a significant increased risk to young people's mental health in these regions. Improved screening, early detection and treatment for cannabis-related mental health issues are urgently required. This paper describes a service-level action research intervention and evaluation protocol for use in the few services where it is possible to engage young Aboriginal and Torres Strait Islander Australians. Methods/Design The protocol is being developed in two services where youth mental health is core business: a primary healthcare centre and a youth service in the Cairns and hinterland region, far north Queensland. The protocol calls first for baseline data to be collected using staff and client surveys; network mapping; and analysis of screening, treatment and referral rates. The protocol's intervention phase is driven by service needs identified from baseline data. Intervention strategies focus on implementing/enhancing cannabis screening instruments and processes in line with current best practice; enhancing networks with external drug and mental health services; developing culturally acceptable training and resources; developing activities aiming to reduce cannabis use in young Aboriginal and Torres Strait Islander clients using the services. The protocol requires implementation of the multilevel intervention within each service for 1 year, with follow-up data then collected and compared to baseline. Process evaluation identifies the more effective intervention strategies and documents the challenges to be overcome for full implementation. Ethics and dissemination Ethics approval was provided by The James Cook University, Human Research Ethics Committee. Ethics Approval Number H5322. Peer

  16. Improved functional properties of glycosylated soy protein isolate using D-glucose and xanthan gum.

    PubMed

    Li, Ruiqi; Hettiarachchy, Navam; Rayaprolu, Srinivas; Davis, Mike; Eswaranandam, Satchithanandam; Jha, Alok; Chen, Pengyin

    2015-09-01

    Functional properties of the soy protein need to improve to have better applications in food industry. Alkali extracted and acid precipitated soy protein isolate (SPI) was glycosylated using D-glucose (G) and Xanthan gum (X) via Maillard reaction to improve solubility. The effects of SPI to G and SPI to X ratios (SPI:G = 2:1, 1:1, and 1:2; SPI:X = 100:1 and 10:1) and incubation time (0, 6, 12, and 24 h) on the solubility and functional properties of glycosylated SPI were evaluated. The SPI:G ratio of 1:2 yielded a maximum degree of glycosylation of 71.1 %. The solubility of SPI after glycosylation significantly increased (P < 0.05) at pH 4.0-8.0 compared to SPI alone. Although the emulsion stability of glycosylated SPIs has not significantly increased (P > 0.05), the emulsifying activity improved significantly (P < 0.05). Glycosylation with SPI-X at a ratio of 10: 1 showed maximum emulsifying activity of 191.6 m(2)/g (SPI alone: 66.3 m(2)/g). Moreover, the SPI:X (ratio of 100:1) showed the maximum foaming activity (205 mL) compared to SPI alone (155 mL). The foaming stability of SPI (2.6 %) increased to 5.5 and 8.2 % when using xanthan gum at the ratio of 100:1 and 10:1, respectively. Glycosylated SPI with enhanced emulsifying and foaming properties has potential to improve the functional quality of the food products.

  17. Optimized Protocols for In Vitro Maturation of Rat Oocytes Dramatically Improve Their Developmental Competence to a Level Similar to That of Ovulated Oocytes.

    PubMed

    Jiao, Guang-Zhong; Cui, Wei; Yang, Rui; Lin, Juan; Gong, Shuai; Lian, Hua-Yu; Sun, Ming-Ju; Tan, Jing-He

    2016-02-01

    The developmental capacity of in vitro-matured (IVM) oocytes is markedly lower than that of their in vivo-matured (IVO) counterparts, suggesting the need for optimization of IVM protocols in different species. There are few studies on IVM of rat oocytes, and there are even fewer attempts to improve ooplasmic maturation compared to those reported in other species. Furthermore, rat oocytes are well known to undergo spontaneous activation (SA) after leaving the oviduct; however, whether IVM rat oocytes have lower SA rates than IVO oocytes and can potentially be used for nuclear transfer is unknown. In this study, we investigated the effects of maturation protocols on cytoplasmic maturation of IVM rat oocytes and observed the possibility to reduce SA by using IVM rat oocytes. Ooplasmic maturation was assessed using multiple markers, including pre- and postimplantation development, meiotic progression, CG redistribution, redox state, and the expression of developmental potential- and apoptosis-related genes. The results showed that the best protocol consisting of modified Tissue Culture Medium-199 (TCM-199) supplemented with cysteamine/cystine and the cumulus cell monolayer dramatically improved the developmental competence of rat oocytes and supported both pre- and postimplantation development and other ooplasmic maturation makers to levels similar to that observed in ovulated oocytes. Rates of SA were significantly lower in IVM oocytes than in IVO oocytes when observed at the same intervals after nuclear maturation. In conclusion, we have optimized protocols for IVM of rat oocytes that sustain ooplasmic maturation to a level similar to ovulated oocytes. The results suggest that IVM rat oocytes might be used to reduce SA for rat cloning.

  18. Shock wave therapy associated with eccentric strengthening versus isolated eccentric strengthening for Achilles insertional tendinopathy treatment: a double-blinded randomised clinical trial protocol.

    PubMed

    Mansur, Nacime Salomão Barbachan; Faloppa, Flávio; Belloti, João Carlos; Ingham, Sheila J McNeill; Matsunaga, Fabio Teruo; Santos, Paulo Roberto Dias Dos; Santos, Bruno Schiefer Dos; Carrazzone, Oreste Lemos; Peixoto, Gabriel; Aoyama, Bruno Takeshi; Tamaoki, Marcel Jun Sugawara

    2017-01-27

    There is no consensus regarding the treatment of Achilles insertional tendinopathies. Eccentric training remains the main choice in the conservative treatment of this illness; however, the good results in the management of non-insertional Achilles tendinopathy were not replicated in the insertional condition. Low energy shock wave therapy has been described as an alternative to these patients, but has yet to be empirically tested. Shock wave therapy, adjunctive to the eccentric strengthening protocol, will improve measures of pain and function. Double blind, placebo-controlled, parallel groups, randomised clinical trial. 93 patients with a diagnosis of chronic insertional tendinopathy, referred from primary or secondary healthcare services, will be assessed and enrolled in this study. They will be divided into two groups (randomised by sequentially numbered identical envelopes, which will be administered serially to participants), one containing the combination of low energy shock wave and eccentric exercises, as treatment and the other comprehending the exercises and the placebo treatment (an apparatus placed in the therapeutic head). The assessments will occur in 2, 4, 6, 12 and 24 weeks. Patients will be evaluated primarily by the Victorian Institute of Sport Assessment-Achilles questionnaire and secondarily by the visual analogue scale, Algometry, the American Orthopedic Foot and Ankle Society scale, the Foot and Ankle Outcome Score and the 12-item Short Form Health Survey. We will use comparison of two proportions via relative frequency analysis, the Pearson Correlation the χ(2) test and the analysis of variance for statistical analyses. This study intends to demonstrate if the association of the eccentric exercise programme with the shock wave therapy can produce good results regarding the treatment of the Achilles insertional tendinopathy. In an attempt to prevent the high costs and complications associated with the surgical intervention, we will try to

  19. Shock wave therapy associated with eccentric strengthening versus isolated eccentric strengthening for Achilles insertional tendinopathy treatment: a double-blinded randomised clinical trial protocol

    PubMed Central

    Mansur, Nacime Salomão Barbachan; Faloppa, Flávio; Belloti, João Carlos; Ingham, Sheila J McNeill; Matsunaga, Fabio Teruo; dos Santos, Paulo Roberto Dias; dos Santos, Bruno Schiefer; Carrazzone, Oreste Lemos; Peixoto, Gabriel; Aoyama, Bruno Takeshi; Tamaoki, Marcel Jun Sugawara

    2017-01-01

    Background There is no consensus regarding the treatment of Achilles insertional tendinopathies. Eccentric training remains the main choice in the conservative treatment of this illness; however, the good results in the management of non-insertional Achilles tendinopathy were not replicated in the insertional condition. Low energy shock wave therapy has been described as an alternative to these patients, but has yet to be empirically tested. Hypothesis Shock wave therapy, adjunctive to the eccentric strengthening protocol, will improve measures of pain and function. Design Double blind, placebo-controlled, parallel groups, randomised clinical trial. Materials and methods 93 patients with a diagnosis of chronic insertional tendinopathy, referred from primary or secondary healthcare services, will be assessed and enrolled in this study. They will be divided into two groups (randomised by sequentially numbered identical envelopes, which will be administered serially to participants), one containing the combination of low energy shock wave and eccentric exercises, as treatment and the other comprehending the exercises and the placebo treatment (an apparatus placed in the therapeutic head). The assessments will occur in 2, 4, 6, 12 and 24 weeks. Patients will be evaluated primarily by the Victorian Institute of Sport Assessment-Achilles questionnaire and secondarily by the visual analogue scale, Algometry, the American Orthopedic Foot and Ankle Society scale, the Foot and Ankle Outcome Score and the 12-item Short Form Health Survey. We will use comparison of two proportions via relative frequency analysis, the Pearson Correlation the χ2 test and the analysis of variance for statistical analyses. Discussion This study intends to demonstrate if the association of the eccentric exercise programme with the shock wave therapy can produce good results regarding the treatment of the Achilles insertional tendinopathy. In an attempt to prevent the high costs and complications

  20. Effect of hydrogen peroxide on improving the heat stability of whey protein isolate solutions.

    PubMed

    Sutariya, Suresh; Patel, Hasmukh

    2017-05-15

    Whey protein isolate (WPI) solutions (12.8%w/w protein) were treated with varying concentrations of H2O2 in the range of 0-0.144 H2O2 to protein ratios (HTPR) by the addition of the required quantity of H2O2 and deionized water. The samples were analyzed for heat stability, rheological properties, denaturation level of β-lactoglobulin (β-LG) and α-lactalbumin (α-LA). The samples treated with H2O2 concentration >0.072 (HTPR) showed significant improvement in the heat stability, and decreased whey protein denaturation and aggregation. The WPI solution treated with H2O2 (>0.072 HTPR) remained in the liquid state after heat treatment at 120°C, whereas the control samples formed gel upon heat treatment. Detailed analysis of these samples suggested that the improvement in the heat stability of H2O2 treated WPI solution was attributed to the significant reduction in the sulfhydryl-disulfide interchange reaction during denaturation of β-LG and α-LA. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Improvement of emulsifying properties of oat protein isolate-dextran conjugates by glycation.

    PubMed

    Zhang, Bei; Guo, Xiaona; Zhu, Kexue; Peng, Wei; Zhou, Huiming

    2015-01-01

    In order to improve the emulsifying properties of oat protein, oat protein isolate (OPI)-dextran (Dex) conjugates were prepared by glycation reaction. Emulsifying properties of emulsions stabilized by native OPI (OPIN), OPI-Dex conjugates (ODC) and heated OPI (OPIH) were characterized by zeta-potential, mean droplet size and microstructure. The results showed that the covalent attachment of OPI and dextran was confirmed by determining degree of graft and SDS-PAGE. OPI-Dex conjugates were capable of forming a finer emulsion, which exhibited smaller average particle size and better storage stability under different homogenization pressures (30, 60, 90 MPa) compared with OPIN and OPIH. When assessed in different pH and ionic strength, emulsions stabilized by OPI-Dex conjugates resulted in improved emulsion stability to environmental stresses. Confocal laser scanning microscopy depicted more uniform and smaller oil droplets that had a reduced tendency to coalesce for emulsions prepared with ODC. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Improvements in tongue strength and pressure-generation precision following a tongue-pressure training protocol in older individuals with dysphagia: Three case reports

    PubMed Central

    Yeates, Erin M; Molfenter, Sonja M; Steele, Catriona M

    2008-01-01

    Dysphagia, or difficulty swallowing, often occurs secondary to conditions such as stroke, head injury or progressive disease, many of which increase in frequency with advancing age. Sarcopenia, the gradual loss of muscle bulk and strength, can place older individuals at greater risk for dysphagia. Data are reported for three older participants in a pilot trial of a tongue-pressure training therapy. During the experimental therapy protocol, participants performed isometric strength exercises for the tongue as well as tongue pressure accuracy tasks. Biofeedback was provided using the Iowa Oral Performance Instrument (IOPI), an instrument that measures tongue pressure. Treatment outcome measures show increased isometric tongue strength, improved tongue pressure generation accuracy, improved bolus control on videofluoroscopy, and improved functional dietary intake by mouth. These preliminary results indicate that, for these three adults with dysphagia, tongue-pressure training was beneficial for improving both instrumental and functional aspects of swallowing. The experimental treatment protocol holds promise as a rehabilitative tool for various dysphagia populations. PMID:19281066

  3. Effectiveness of a Vestibular Rehabilitation Protocol to Improve the Health-Related Quality of Life and Postural Balance in Patients with Vertigo.

    PubMed

    Tsukamoto, Heloísa Freiria; Costa, Viviane de Souza Pinho; Silva, Rubens Alexandre da; Pelosi, Gislaine Garcia; Marchiori, Luciana Lozza de Moraes; Vaz, Cláudia Regina Sanches; Fernandes, Karen Barros Parron

    2015-07-01

    Introduction Dizziness can be characterized as a balance disorder that causes discomfort, leading to several functional limitations. Currently, vestibular rehabilitation has been highlighted as a possible treatment. Objective Analyze the effects of completing a vestibular rehabilitation treatment protocol on quality of life and postural balance in patients with vestibular complaints, as well as to compare these effects between the patients taking or not taking antivertigo drugs. Methods A nonrandomized controlled trial was performed with 20 patients previously diagnosed with vestibular diseases. Information regarding vertigo symptoms, quality of life as assessed through the Dizziness Handicap Inventory, visual analog scale of dizziness, and stabilometry using force platform was collected. Patients were treated for 12 weeks by a custom protocol. The sample was divided into two groups according to the use (medicated group, n = 9) or not (control group, n = 11) of antivertigo drugs. Results There was improvement in quality of life (p < 0.001) and intensity of dizziness (p = 0.003) with the intervention. An improvement of postural balance was observed through functional tests. However, no statistically significant difference was noted in stabilometry. When both groups were compared, no statistically significant differences between the variations of the variables analyzed were found in the re-evaluation session. Conclusion Quality of life and postural balance are improved with intervention. However, this improvement is not associated with pharmacologic treatment.

  4. Effectiveness of a Vestibular Rehabilitation Protocol to Improve the Health-Related Quality of Life and Postural Balance in Patients with Vertigo

    PubMed Central

    Tsukamoto, Heloísa Freiria; Costa, Viviane de Souza Pinho; Silva, Rubens Alexandre da; Pelosi, Gislaine Garcia; Marchiori, Luciana Lozza de Moraes; Vaz, Cláudia Regina Sanches; Fernandes, Karen Barros Parron

    2015-01-01

    Introduction Dizziness can be characterized as a balance disorder that causes discomfort, leading to several functional limitations. Currently, vestibular rehabilitation has been highlighted as a possible treatment. Objective Analyze the effects of completing a vestibular rehabilitation treatment protocol on quality of life and postural balance in patients with vestibular complaints, as well as to compare these effects between the patients taking or not taking antivertigo drugs. Methods A nonrandomized controlled trial was performed with 20 patients previously diagnosed with vestibular diseases. Information regarding vertigo symptoms, quality of life as assessed through the Dizziness Handicap Inventory, visual analog scale of dizziness, and stabilometry using force platform was collected. Patients were treated for 12 weeks by a custom protocol. The sample was divided into two groups according to the use (medicated group, n = 9) or not (control group, n = 11) of antivertigo drugs. Results There was improvement in quality of life (p < 0.001) and intensity of dizziness (p = 0.003) with the intervention. An improvement of postural balance was observed through functional tests. However, no statistically significant difference was noted in stabilometry. When both groups were compared, no statistically significant differences between the variations of the variables analyzed were found in the re-evaluation session. Conclusion Quality of life and postural balance are improved with intervention. However, this improvement is not associated with pharmacologic treatment. PMID:26157499

  5. Sclareol isolated from Salvia officinalis improves facial wrinkles via an antiphotoaging mechanism.

    PubMed

    Park, Ji-Eun; Lee, Kyung-Eun; Jung, Eunsun; Kang, Seunghyun; Kim, Youn Joon

    2016-12-01

    Ultraviolet (UV) irradiation triggers skin photoaging processes, which disrupt the normal three-dimensional integrity of skin. UV-induced oxidative stress, both directly and indirectly, stimulates complex signaling pathways. UV radiation activates skin cell surface receptors on a molecular level and triggers severe changes in extracellular matrix (ECM) proteins, resulting in skin photoaging. Sclareol isolated from Salvia officinalis is widely used as a fragrance material. Sclareol is known to exert various biological activities, but its antiphotoaging effect has not been elucidated to date. Therefore, we evaluated wrinkle improvement efficacy of sclareol. Human dermal fibroblast cell line (Hs68) and a reconstructed human epidermis (RHE) model were used to evaluate the antiphotoaging effect of sclareol in vitro. A clinical study treated with 0.02% sclareol-containing cream was conducted to identify the ability of sclareol to improve wrinkles. First, sclareol enhanced cellular proliferation and blocked UVB-induced cell death. Sclareol inhibited the UVB-induced mRNA expression of matrix metalloproteinases (MMPs) by regulating the protein expression of AP-1 constituents. In RHE model, sclareol recovered the UVB-induced decrease in epidermal thickness and the expression of proliferating cell nuclear antigen (PCNA). In clinical trial, visually assessed changes and several wrinkle parameters were considered to be statistically different between the test and control groups at 12 weeks. In this study, sclareol inhibited various photoaging phenomena in human fibroblasts and RHE model. In addition, sclareol-containing cream improved wrinkles in a clinical trial. Taken together, sclareol alleviates facial wrinkle formation via an antiphotoaging mechanism and may be an effective candidate ingredient. © 2016 Wiley Periodicals, Inc.

  6. Continuous Improvement and the Safety Case for the Waste Isolation Pilot Plant Geologic Repository - 13467

    SciTech Connect

    Van Luik, Abraham; Patterson, Russell; Nelson, Roger; Leigh, Christi

    2013-07-01

    The Waste Isolation Pilot Plant (WIPP) is a geologic repository 2150 feet (650 m) below the surface of the Chihuahuan desert near Carlsbad, New Mexico. WIPP permanently disposes of transuranic waste from national defense programs. Every five years, the U.S. Department of Energy (DOE) submits an application to the U.S. Environmental Protection Agency (EPA) to request regulatory-compliance re-certification of the facility for another five years. Every ten years, DOE submits an application to the New Mexico Environment Department (NMED) for the renewal of its hazardous waste disposal permit. The content of the applications made by DOE to the EPA for re-certification, and to the NMED for permit-renewal, reflect any optimization changes made to the facility, with regulatory concurrence if warranted by the nature of the change. DOE points to such changes as evidence for its having taken seriously its 'continuous improvement' operations and management philosophy. Another opportunity for continuous improvement is to look at any delta that may exist between the re-certification and re-permitting cases for system safety and the consensus advice on the nature and content of a safety case as being developed and published by the Nuclear Energy Agency's Integration Group for the Safety Case (IGSC) expert group. DOE at WIPP, with the aid of its Science Advisor and teammate, Sandia National Laboratories, is in the process of discerning what can be done, in a reasonably paced and cost-conscious manner, to continually improve the case for repository safety that is being made to the two primary regulators on a recurring basis. This paper will discuss some aspects of that delta and potential paths forward to addressing them. (authors)

  7. Use of Human Hybridoma Technology To Isolate Human Monoclonal Antibodies.

    PubMed

    Smith, Scott A; Crowe, James E

    2015-02-01

    The human hybridoma technique offers an important approach for isolation of human monoclonal antibodies. A diversity of approaches can be used with varying success. Recent technical advances in expanding the starting number of human antigen-specific B cells, improving fusion efficiency, and isolating new myeloma partners and new cell cloning methods have enabled the development of protocols that make the isolation of human monoclonal antibodies from blood samples feasible. Undoubtedly, additional innovations that could improve efficiency are possible.

  8. Performance improvements of MOEMS-based diffractive arrays: address isolation and optical switching

    NASA Astrophysics Data System (ADS)

    Panaman, Ganesh; Madison, Seth; Sano, Michael; Castracane, James

    2005-01-01

    Micro-Opto-Electro-Mechanical Systems (MOEMS) have found a variety of applications in fields such as telecommunications, spectroscopy and display technology. MOEMS-based optical switching is currently under investigation for the increased flexibility that such devices provide for reconfiguration of the I/O network for inter-chip communication applications. This potential not only adds an additional degree of freedom for adjustment of transmitter/receiver links but also allows for fine alignment of individual channels in the network link. Further, this use of diffractive arrays for specific applications combines beam steering/adjustment capabilities with the inherent wavelength dependence of the diffractive approach for channel separation and de-multiplexing. Research and development has been concentrated on the progression from single MOEMS components to parallel arrays integrated with optical source arrays for a successful feasibility demonstration. Successful development of such an approach will have a major impact of the next generation communication protocols. This paper will focus on the current status of the MOEMS research program for Free Space Optical inter-chip communication at the College of NanoScale Science and Engineering, University at Albany-SUNY (CNSE). New versions of diffractive arrays stemming from the basic MEMS Compound Grating (MCG; patent #5,999,319) have been produced through various fabrication methods including the MUMPs process1. Most MEMS components relying on electrostatic actuation tend to require high actuation voltages (>20V) compared to the typical 5V levels prevalent in conventional integrated circuits. The specific goal is to yield improved performance while minimizing the power consumption of the components. Structural modifications through the variation in the ruling/electrode spacing distance and array wiring layout through individually addressable gratings have been studied to understand effects on the actuation voltage and

  9. Improving care for victims: a study protocol of the evaluation of a centre for sexual and family violence

    PubMed Central

    Zijlstra, E; LoFoWong, S; Hutschemaekers, G; Lagro-Janssen, A

    2016-01-01

    Introduction Worldwide, sexual and family violence are highly prevalent problems. Victims of sexual and family violence often do not seek formal help in the acute phase. When they do seek help, they encounter a system of scattered care. For this reason, a centre for sexual and family violence was launched in Nijmegen, the Netherlands. The centre provides multidisciplinary care for victims of acute sexual and/or family violence. With the study described in this study protocol, we want to evaluate the implementation process and the reach of the Center for Sexual and Family Violence Nijmegen (CSFVN). Methods and analysis We will conduct a mixed-methods study including quantitative and qualitative methods of data collection and analysis. Data about the implementation process will be obtained via semistructured interviews and focus group discussions. Content analysis will be done in software program Atlas.ti. Analysis of file data will be undertaken to assess the reach of the CSFVN (patient characteristics and characteristics of the care they received). The data will be analysed in SPSS. Ethics and dissemination The Medical Ethics Committee of the Radboud University Nijmegen Medical Center approved the study protocol under file number 2012–1218. Dissemination will be done by submitting scientific articles to academic peer-reviewed journals. We will present the results at relevant international, national and local conferences and meetings. We will send press releases to relevant media. We will share the results with the network of assault centres in the Netherlands. PMID:27619828

  10. Patient education interventions to improve physical activity in patients with intermittent claudication: a protocol for a systematic mixed-studies review

    PubMed Central

    Abaraogu, Ukachukwu Okoroafor; Dall, Philippa Margaret; Seenan, Christopher Andrew

    2016-01-01

    Introduction Peripheral arterial disease (PAD) and intermittent claudication (IC) decrease an individual's capacity to engage in physical activity (PA) with potentially negative effects on PA behaviour. Strategies to improve PA among this population may provide a range of positive health benefits. We present a protocol to assess the components of patient education interventions that improve PA capacity and PA behaviour in patients with PAD and IC. Methods and analysis Published peer-reviewed studies will be searched in the following databases: CINAHL, the Cochrane Library, OVID, ProQuest, AMED, MEDLINE, PsycINFO, Web of Science Core Collection and PEDro, to identify literature investigating the effect of patient education on PA of patients with PAD and IC, or studies that investigated patients' perceptions or experience with these interventions. Two authors will independently perform screening for study eligibility, result synthesis and then appraise study quality. For interventions without follow-up, primary outcome measures will include change in PA capacity, or change in free-living PA behaviour; where there was a follow-up postintervention, the primary outcome will be rate of adherence to PA behaviour improvement. A three-phase sequential explanatory synthesis of mixed studies will be employed to answer the research questions. Homogenous quantitative data will be analysed using a random-effects model of meta-analysis with results presented as relative risk for dichotomous outcomes and as weighted or standardised means for continuous outcomes. Qualitative data will be analysed using thematic synthesis. This review protocol is reported according to the Preferred Reporting Items for Systematic review and Meta-Analysis Protocols (PRISMA-P) 2015 guidelines. Trial registration number CRD42015027314. PMID:27207628

  11. Molecular typing of Japanese field isolates and live commercial vaccine strain of Mycoplasma synoviae using improved pulsed-field gel electrophoresis and vlhA gene sequencing.

    PubMed

    Harada, Kazuki; Kijima-Tanaka, Mayumi; Uchiyama, Mariko; Yamamoto, Tomoko; Oishi, Koji; Arao, Megumi; Takahashi, Toshio

    2009-12-01

    In the present study, pulsed-field gel electrophoresis (PFGE) and vlhA gene sequence analysis were applied and verified for typing the Mycoplasma synoviae live vaccine MS-H strain and field isolates from diseased chickens in Japan. The previously published PFGE protocol using SmaI digestion could not allow the discrimination of two of the 11 M. synoviae field isolates from the vaccine strain and had relatively low discrimination power (D = 0.885). On the other hand, our new PFGE protocols using BlnI and BamHI digestions as well as the vlhA sequence analysis allowed the discrimination of all 11 M. synoviae field isolates from the vaccine strain. In addition, these PFGE protocols using BlnI and BamHI digestions generated unique fragment patterns in epidemiologically unrelated isolates, including those with identical SmaI-digested patterns or vlhA gene sequences (D = 0.987 and 1.000, respectively), and generated indistinguishable or closely related patterns in epidemiologically related isolates. Therefore, we believe that they would be useful tools to determine whether M. synoviae clinical isolates from diseased chickens are derived from the vaccine strain or wild-type strain and to further elucidate the epidemiology of M. synoviae infection.

  12. Improved derivatization protocol for simultaneous determination of alkylphenol ethoxylates and brominated flame retardants followed by gas chromatography-mass spectrometry analyses.

    PubMed

    Chokwe, T B; Okonkwo, J O; Sibali, L L; Ncube, E J

    2014-01-01

    An improved derivatization protocol for the simultaneous determination of alkylphenol ethoxylates and brominated flame retardants with heptafluorobutyric anhydride under triethylamine amine base was investigated. The derivatization reaction was completed in 30 min at 50 °C using hexane as solvent. Under these conditions, it was observed that alkylphenol ethoxylates and tetrabromobisphenol A were derivatized successfully in the presence of hexabromocyclododecane, lower congeners of polybrominated biphenyls and polybrominated diphenyl ethers. The improved protocol was applied to the recover of the analytes of interest from a simulated water sample after solid phase extraction. The recoveries achieved were above 60%. The limit of detection and limit of quantification ranged from 0.01-0.20 and 0.05-0.66 μg L(-1), respectively. The improved derivatization procedure was also successfully applied to determine trace amounts of these compounds in environmental water samples. The concentrations of the targeted analytes from the environmental samples were determined from limit of quantification. The levels of the targeted compounds in the environmental samples ranged from nd-7.63 ±2.83 μg L(-1).

  13. Glutathione Ethyl Ester Supplementation during Pancreatic Islet Isolation Improves Viability and Transplant Outcomes in a Murine Marginal Islet Mass Model

    PubMed Central

    Raposo do Amaral, Alexandre S.; Pawlick, Rena L.; Rodrigues, Erika; Costal, Flavia; Pepper, Andrew; Ferreira Galvão, Flávio H.; Correa-Giannella, Maria Lucia; Shapiro, A. M.James

    2013-01-01

    Background The success of pancreatic islet transplantation still faces many challenges, mainly related to cell damage during islet isolation and early post-transplant. The increased generation of reactive oxygen species (ROS) during islet isolation and the consumption of antioxidant defenses appear to be an important pathway related to islet damage. Methodology/Principal Findings In the present study we evaluated whether supplementation of glutathione-ethyl-ester (GEE) during islet isolation could improve islet viability and transplant outcomes in a murine marginal islet mass model. We also cultured human islets for 24 hours in standard CMRL media with or without GEE supplementation. Supplementation of GEE decreased the content of ROS in isolated islets, leading to a decrease in apoptosis and maintenance of islet viability. A higher percentage of mice transplanted with a marginal mass of GEE treated islets became euglycemic after transplant. The supplementation of 20 mM GEE in cultured human islets significantly reduced the apoptosis rate in comparison to untreated islets. Conclusions/Significance GEE supplementation was able to decrease the apoptosis rate and intracellular content of ROS in isolated islets and might be considered a potential intervention to improve islet viability during the isolation process and maintenance in culture before islet transplantation. PMID:23424628

  14. Simultaneous Deposition of Mass Selected Anions and Cations: Improvements in Ion Delivery for Matrix Isolation Experiments

    NASA Astrophysics Data System (ADS)

    Goodrich, Michael E.; Moore, David T.

    2016-06-01

    A focus of the research in our group has been to develop improved methods for ion delivery in matrix isolation experiments. We have previously reported a method to co-deposit low energy, mass selected metal anions and a rare gas counter cation.a A modification allowing for mass selection of both the anion and cation will be discussed. Results from preliminary experiments of mass selected, low energy Cu- and SF5+ will also be highlighted. To our knowledge, these experiments are the first time two mass selected beams of ions have been simultaneously deposited into a cryogenic matrix. Co-deposition of the ions into an argon matrix doped with 0.02% CO at 20K resulted in the observation of bands assigned to SF5+ and anionic copper carbonyl complexes, Cu(CO)n- (n=1-3). Upon irradiation of the matrix with a narrow band, blue LED, the copper carbonyl complexes are converted to the neutral analogues, while the fate of the photodetached electrons can be directly tracked, as a decrease of the SF5+ band and a growth of the neutral SF5 band are observed. aLudwig, R. M.; Moore, D. T.; J. Chem. Phys. 139, 244202 (2013).

  15. Effect of improved hydrogen supply on energy state, ureogenesis and gluconeogenesis in isolated hepatocytes.

    PubMed

    Letko, G; Halangk, W

    1986-01-01

    The cellular energy state as well as ureogenesis and gluconeogenesis as ATP-consuming processes were studied as a function of hydrogen supply of mitochondria in isolated hepatocytes. Even though the ATP turnover was quite different during ureogenesis and gluconeogenesis and in the absence of these syntheses both the safranine signals (as a probe for the mitochondrial membrane potential) and the total ATP contents of the cells did not vary to a great extent. Partial uncoupling by FCCP destroyed the mitochondrial membrane potential and decreased the cellular ATP totally. Under these conditions the presence of ornithine plus lactate maintained a measurable membrane potential and ATP level. In order to demonstrate the relevance of hydrogen supply of mitochondria to the syntheses of glucose and urea, the load on energy metabolism was additionally increased by partial uncoupling, and the cells were incubated in a parallel mode in Krebs-Henseleit buffer and an 80 mM succinate containing medium, the latter forcing succinate into the cells. After partial uncoupling, ureogenesis and gluconeogenesis as well as the ATP level were seen to decrease markedly in Krebs-Henseleit buffer, whereas in the succinate medium the improved hydrogen supply of mitochondria counteracted the drain by FCCP, and substantially higher rates of the syntheses and elevated ATP levels were maintained.

  16. A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis.

    PubMed

    Guan, Feng; Tan, Zengqi; Li, Xiang; Pang, Xingchen; Zhu, Yunlin; Li, Dongliang; Yang, Ganglong

    2015-10-30

    Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. In this study, we established the lectin-based isolation/enrichment strategy for total glycomic information. Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome.

  17. Improvement of polyvinyl alcohol properties by adding nanocrystalline cellulose isolated from banana pseudostems.

    PubMed

    Pereira, André Luís S; do Nascimento, Diego M; Souza Filho, Men de Sá M; Morais, João Paulo S; Vasconcelos, Niedja F; Feitosa, Judith P A; Brígida, Ana Iraidy S; Rosa, Morsyleide de F

    2014-11-04

    Cellulose nanocrystals (CNCs) isolated from banana pseudostems fibers (BPF) of the Pacovan variety were used as fillers in a polyvinyl alcohol (PVOH) matrix to yield a nanocomposite. The fibers from the external fractions of the BPF were alkaline bleached and hydrolyzed under acidic conditions (H2SO4 62% w/w, 70 min, 45 °C) to obtain CNCs with a length (L) of 135.0 ± 12.0 nm and a diameter (D) of 7.2 ± 1.9 nm to yield an aspect ratio (L/D) of 21.2. The CNCs were applied to PVOH films at different concentrations (0%, 1%, 3%, and 5% w/w, dry basis). With higher concentrations of CNCs, the water-vapor barrier of the films increased, while the optical properties changed very little. Increasing the concentration of the CNCs up to 3% significantly improved the mechanical properties of the nanocomposite. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Inoculation of new rhizobial isolates improve nutrient uptake and growth of bean (Phaseolus vulgaris) and arugula (Eruca sativa).

    PubMed

    de Souza, Eduardo M; Bassani, Victor L; Sperotto, Raul A; Granada, Camille E

    2016-08-01

    In the current agricultural model, the massive use of chemical fertilizer causes environmental and economic losses. Inoculation of plant-growth-promoting (PGP) nitrogen-fixing bacteria is an alternative to the use of nitrogen, phosphorus and potassium fertilizers. In this study, rhizobia strains isolated from common bean (Phaseolus vulgaris) root nodules were evaluated in an effort to identify an efficient nitrogen-fixing rhizobia strain able to improve bean germination and growth. Common bean plants were collected from seven sites in southern Brazil, and 210 native rhizobia isolates were obtained. Evaluation of PGP traits showed that most of the rhizobia isolates were non-siderophore producers and weak indolic compounds producers. Under laboratory conditions, rhizobia isolates E15 (Rhizobium leguminosarum) and L5 (Rhizobium radiobacter) increase germination percentage, length, and dry weight of common bean and arugula (Eruca sativa) seedlings. Under greenhouse conditions, common bean plants inoculated with the rhizobia isolates VC28 and L15 (both Rhizobium fabae) presented the highest nodule number and shoot dry matter, while VC28 also presented the highest values of shoot nitrogen and potassium. Isolate L17 presented highly effective N fixation, even with reduced nodulation. These new rhizobia isolates are attractive PGP alternatives to chemical fertilizers. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  19. Optimizing isolation culture and freezing methods to preserve Wharton's jelly's mesenchymal stem cell (MSC) properties: an MSC banking protocol validation for the Hellenic Cord Blood Bank.

    PubMed

    Chatzistamatiou, Theofanis K; Papassavas, Andreas C; Michalopoulos, Efstathios; Gamaloutsos, Christos; Mallis, Panagiotis; Gontika, Ioanna; Panagouli, Effrosyni; Koussoulakos, Stauros L; Stavropoulos-Giokas, Catherine

    2014-12-01

    Mesenchymal stem or stromal cells (MSCs) are a heterogeneous population that can be isolated from many tissues including umbilical cord Wharton's jelly (UC-WJ). Although initially limited in studies such as a hematopoietic stem cell transplantation adjuvant, an increasing number of clinical trials consider MSCs as a potential anti-inflammatory or a regenerative medicine agent. It has been proposed that creating a repository of MSCs would increase their availability for clinical applications. The aim of this study was to assess the optimal isolation and cryopreservation procedures to facilitate WJ MSC banking. Cells were isolated from UC-WJ using enzymatic digestion or plastic adhesion methods. Their isolation efficacy, growth kinetics, immunophenotype, and differentiation potential were studied, as well as the effects of freezing. Flow cytometry for common MSC markers was performed on all cases and differentiation was shown with histocytochemical staining. Finally, the isolation efficacy on cryopreserved WJ tissue fragments was tested. MSC isolation was successful using both isolation methods on fresh UC-WJ tissue. However, UC-WJ MSC isolation from frozen tissue fragments was impossible. Flow cytometry analysis revealed that only MSC markers were expressed on the surface of the isolated cells while differentiation assays showed that they were capable of trilinear differentiation. All the above characteristics were also preserved in isolated UC-WJ MSCs over the cryopreservation study period. These data showed that viable MSCs can only be isolated from fresh UC-WJ tissue, setting the foundation for clinical-grade banking. © 2014 AABB.

  20. Improved production of isomaltulose by a newly isolated mutant of Serratia sp. cells immobilized in calcium alginate.

    PubMed

    Kim, Yonghwan; Koo, Bong-Seong; Lee, Hyeon-Cheol; Yoon, Youngdae

    2015-03-01

    Isomaltulose, also known as palatinose, is produced by sucrose isomerase and has been highlighted as a sugar substitute due to a number of advantageous properties. For the massive production of isomaltulose, high resistance to sucrose and stability of sucrose isomerase as well as sucrose conversion yields would be critical factors. We describe a series of screening procedures to isolate the mutant strain of Serratia sp. possessing enhanced isomaltulose production with improved stability. The new Serratia sp. isolated from a series of screening procedures allowed us to produce isomaltulose from 60% sucrose solution, with over 90% conversion yield. Moreover, when this strain was immobilized in calcium alginate beads and placed in a medium containing 60% sucrose, it showed over 70% sucrose conversion yields for 30 cycles of repeated-batch reactions. Thus, improved conversion activity and stability of the newly isolated Serratia sp. strain in the present study would be highly valuable for industries related to isomaltulose production.

  1. Anethole improves the in vitro development of isolated caprine secondary follicles.

    PubMed

    Sá, N A R; Araújo, V R; Correia, H H V; Ferreira, A C A; Guerreiro, D D; Sampaio, A M; Escobar, E; Santos, F W; Moura, A A; Lôbo, C H; Ceccatto, V M; Campello, C C; Rodrigues, A P R; Leal-Cardoso, J H; Figueiredo, J R

    2017-02-01

    The aim of this study was to investigate the effect of three concentrations of anethole (30, 300, and 2000 μg/mL) on survival, antrum formation, follicular diameter, and oocyte maturation in the caprine species. The study also evaluated the effects of anethole on transcripts of ICAM-1, CAV-1, TIMP-2, and PAI-1 genes and levels of reactive oxygen species (ROS) in isolated goat preantral ovarian follicles before and after in vitro culture for 18 days. Preantral follicles were isolated from goat ovaries and individually cultured in alpha minimum essential medium modified (α-MEM(+)), defined as the control treatment, α-MEM(+) supplemented with ascorbic acid at a concentration of 100 μg/mL (AA), or α-MEM(+) supplemented with three different concentrations of anethole (30, 300, 2000 μg/mL) for a period of 18 days. Treatments were named as α-MEM(+), AA, AN30, AN300, and AN2000, respectively. After culture, the follicles were opened, the cumulus oocytes complex (COCs) were removed and matured in vitro. The walls of the follicles were used for the quantitation of mRNA by quantitative real-time polymerase chain reaction. Finally, the medium collected at the end of culture was used for the measurements of ROS. After 18 days of culture, the AA treatment showed the percentage of intact follicles and follicular diameter significantly higher compared with the other treatments. However, daily growth rate, antrum formation, and also oocyte diameter were similar among the treatments. In addition, compared with AA, the rate of oocytes for in vitro maturation (diameter ≥ 110 μm) and the meiosis resumption rate were significantly higher in the treatments AN30 and AN2000, respectively. When assessing gene related to remodeling of the basement membrane, significant differences in mRNA levels for ICAM-1, CAV-1, TIMP-2, and PAI-1 were observed in comparison with Day 0, i.e., in the noncultured control. In addition, the ROS from Day 12, all treatments with the addition

  2. Application of a standardised protocol for hepatic venous pressure gradient measurement improves quality of readings and facilitates reduction of variceal bleeding in cirrhotics

    PubMed Central

    Tey, Tze Tong; Gogna, Apoorva; Irani, Farah Gillan; Too, Chow Wei; Lo, Hoau Gong Richard; Tan, Bien Soo; Tay, Kiang Hiong; Lui, Hock Foong; Chang, Pik Eu Jason

    2016-01-01

    INTRODUCTION Hepatic venous pressure gradient (HVPG) measurement is recommended for prognostic and therapeutic indications in centres with adequate resources and expertise. Our study aimed to evaluate the quality of HVPG measurements at our centre before and after introduction of a standardised protocol, and the clinical relevance of the HVPG to variceal bleeding in cirrhotics. METHODS HVPG measurements performed at Singapore General Hospital from 2005–2013 were retrospectively reviewed. Criteria for quality HVPG readings were triplicate readings, absence of negative pressure values and variability of ≤ 2 mmHg. The rate of variceal bleeding was compared in cirrhotics who achieved a HVPG response to pharmacotherapy (reduction of the HVPG to < 12 mmHg or by ≥ 20% of baseline) and those who did not. RESULTS 126 HVPG measurements were performed in 105 patients (mean age 54.7 ± 11.4 years; 55.2% men). 80% had liver cirrhosis and 20% had non-cirrhotic portal hypertension (NCPH). The mean overall HVPG was 13.5 ± 7.2 mmHg, with a significant difference between the cirrhosis and NCPH groups (p < 0.001). The proportion of quality readings significantly improved after the protocol was introduced. HVPG response was achieved in 28 (33.3%, n = 84) cirrhotics. Nine had variceal bleeding over a median follow-up of 29 months. The rate of variceal bleeding was significantly lower in HVPG responders compared to nonresponders (p = 0.025). CONCLUSION The quality of HVPG measurements in our centre improved after the introduction of a standardised protocol. A HVPG response can prognosticate the risk of variceal bleeding in cirrhotics. PMID:26996384

  3. Improved technique for fluorescence in situ hybridisation analysis of isolated nuclei from archival, B5 or formalin fixed, paraffin wax embedded tissue.

    PubMed

    Schurter, M J; LeBrun, D P; Harrison, K J

    2002-04-01

    Fluorescence in situ hybridisation (FISH) is an effective method to detect chromosomal alterations in a variety of tissue types, including archived paraffin wax embedded specimens fixed in B5 or formalin. However, precipitating fixatives such as B5 have been known to produce unsatisfactory results in comparison with formalin when used for FISH. This study describes an effective nuclear isolation and FISH procedure for B5 and formalin fixed tissue, optimising the nuclear isolation step and nuclei pretreatments using tonsil and mantle cell lymphoma specimens. The protocol presented can be used to isolate nuclei and perform FISH on B5 or formalin fixed, paraffin wax embedded samples from a variety of tissue types.

  4. Improving the efficiency of isolated microspore culture in six-row spring barley: II-exploring novel growth regulators to maximize embryogenesis and reduce albinism.

    PubMed

    Esteves, Patricio; Clermont, Isabelle; Marchand, Suzanne; Belzile, François

    2014-06-01

    Two alternative cytokinins, thidiazuron and meta-topoline, were tested in isolated microspore culture on recalcitrant barley genotypes (six-row, spring), and green plant regeneration was improved substantially. Doubled-haploid (DH) plants are coveted in plant breeding and in genetic studies, since they are rapidly obtained and perfectly homozygous. In barley, DHs are produced mainly via androgenesis, and isolated microspore culture (IMC) constitutes the method offering the greatest potential efficiency. However, IMC can often be challenging in some genotypes because of low yield of microspores, low regeneration and high incidence of albinism. Six-row spring-type barleys, the predominant type grown in Eastern Canada, are considered recalcitrant in this regard. Our general objective was to optimize an IMC protocol for DH production in six-row spring barley. In particular, we explored the use of alternative hormones in the induction medium (thidiazuron and dicamba), and in the regeneration medium (meta-topoline). This optimization was performed on two typical six-row spring (ACCA and Léger), a two-row spring (Gobernadora) and a two-row winter (Igri) barley cultivar. When 6-benzyl-aminopurine (BAP) was replaced by a combination of thidiazuron and dicamba in the induction medium, a 5.1-fold increase (P < 0.01) in the production of green plants resulted. This increase was mainly achieved by a reduction of albinism. Moreover, a 2.9-fold increase (P < 0.01) in embryo differentiation into green plants was obtained using meta-topoline instead of BAP in the regeneration medium. Together, these innovations allowed us to achieve a substantial improvement in the efficiency of IMC in this recalcitrant type of barley. These results were later successfully validated using sets of F1s from a six-row spring barley breeding program.

  5. Early Intervention with a Parent-Delivered Massage Protocol Directed at Tactile Abnormalities Decreases Severity of Autism and Improves Child-to-Parent Interactions: A Replication Study

    PubMed Central

    Silva, Louisa M. T.; Gabrielsen, Kristen R.; Budden, Sarojini S.; Buenrostro, Martha; Horton, Gretchen

    2015-01-01

    Tactile abnormalities are severe and universal in preschool children with autism. They respond well to treatment with a daily massage protocol directed at tactile abnormalities (QST massage for autism). Treatment is based on a model for autism proposing that tactile impairment poses a barrier to development. Two previous randomized controlled trials evaluating five months of massage treatment reported improvement of behavior, social/communication skills, and tactile and other sensory symptoms. This is the first report from a two-year replication study evaluating the protocol in 103 preschool children with autism. Parents gave daily treatment; trained staff gave weekly treatment and parent support. Five-month outcomes replicated earlier studies and showed normalization of receptive language (18%, P = .03), autistic behavior (32%, P = .006), total sensory abnormalities (38%, P = .0000005), tactile abnormalities (49%, P = .0002), and decreased autism severity (medium to large effect size, P = .008). In addition, parents reported improved child-to-parent interactions, bonding, and decreased parenting stress (44%, P = .00008). Early childhood special education programs are tasked with addressing sensory abnormalities and engaging parents in effective home programs. Until now, they have lacked research-based methods to do so. This program fulfills the need. It is recommended to parents and ECSE programs (ages 3–5) at autism diagnosis. PMID:25878901

  6. Normalization of coagulopathy is associated with