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Sample records for improved isolation protocol

  1. 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. PMID:25355522

  2. 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.

  3. Improved isolation protocol to detect high molecular weight polysaccharide structures of Campylobacter jejuni.

    PubMed

    Kovács, Judit K; Felső, Péter; Emődy, Levente; Schneider, György; Kocsis, Béla

    2014-12-01

    Simple detection of high molecular weight, LPS-like structures of Campylobacter jejuni is still an unsolved problem. A phenol-free extraction method for the detection of HMW polysaccharide was developed without the need for Western blot. This method provides a reliable technique for large-scale screening and comparative characterization study of different isolates.

  4. Improved protocols for protein and RNA isolation from three-dimensional collagen sandwich cultures of primary hepatocytes.

    PubMed

    Heidebrecht, F; Schulz, I; Keller, M; Behrens, S-E; Bader, A

    2009-10-01

    The sandwich culture is the most widely used long-term culture system for functional primary hepatocytes. Despite its advantages, the currently available protocols for protein and RNA extraction are either time-consuming or contain steps that may skewer the results. This paper describes improved protocols for RNA and protein extraction from sandwich cultures that are easy to perform, require short working time, and use no additional enzymatic reactions that could change the expression profile of the cells. The quality of the RNA is excellent, allowing also applications requiring high purity such as microarrays. In general, the protocols are suited for any cells in 3D collagen culture. PMID:19539596

  5. 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. PMID:26874020

  6. 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

  7. REHABILITATION PROTOCOL AFTER ISOLATED POSTERIOR CRUCIATE LIGAMENT RECONSTRUCTION.

    PubMed

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

    2012-01-01

    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.

  8. 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.

  9. 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

  10. An accurate link correlation estimator for improving wireless protocol performance.

    PubMed

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

    2015-02-12

    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.

  11. 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.

  12. 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.

  13. 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. PMID:25912427

  14. 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.

  15. 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.

  16. HB - MAC: Improving the Random - HB # Authentication Protocol

    NASA Astrophysics Data System (ADS)

    Rizomiliotis, Panagiotis

    The Random - HB # protocol is a significant improvement of the HB + protocol introduced by Juels and Weis for the authentication of low-cost RFID tags. Random - HB # improves HB + in terms of both security and practicality. It is provably resistant against man-in-the-middle attacks, where the adversary can modify messages send from the reader to the tag and performs significantly better than HB + , since it reduces the transmission costs and provides more practical error rates. The only problem with Random - HB # is that the storage costs for the secret keys are insurmountable to low cost tags. The designers of the protocol have proposed also an enhanced variant which has less storage requirements, but it is not supported by a security proof. They call this variant just HB #. In this paper we propose a variant of the Random - HB #. The new proposal maintains the performance of the Random - HB #, but it requires significantly less storage for the key. To achieve that we add a lightweight message authentication code to protect the integrity of all the exchanged messages.

  17. 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.

  18. 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.

  19. A polymerase chain reaction protocol for the detection of various geographical isolates of white spot virus.

    PubMed

    Tapay, L M; Nadala, E C; Loh, P C

    1999-09-01

    Polymerase chain reaction (PCR) primers were designed based on the sequence of a cloned fragment of the white spot virus (WSV) genome and were used to detect at least four geographic isolates of WSV from both experimentally- and naturally-infected shrimp. In addition to high specificity, the one-step and two-step PCR protocols were determined to have sensitivities of 10-100 pg and 100 femtograms respectively. The two-step PCR protocol is recommended as a very sensitive and specific alternative protocol to Western blot assay for the detection of WSV.

  20. 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

  1. 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. PMID:21519906

  2. Simple algorithm for improved security in the FDDI protocol

    NASA Astrophysics Data System (ADS)

    Lundy, G. M.; Jones, Benjamin

    1993-02-01

    We propose a modification to the Fiber Distributed Data Interface (FDDI) protocol based on a simple algorithm which will improve confidential communication capability. This proposed modification provides a simple and reliable system which exploits some of the inherent security properties in a fiber optic ring network. This method differs from conventional methods in that end to end encryption can be facilitated at the media access control sublayer of the data link layer in the OSI network model. Our method is based on a variation of the bit stream cipher method. The transmitting station takes the intended confidential message and uses a simple modulo two addition operation against an initialization vector. The encrypted message is virtually unbreakable without the initialization vector. None of the stations on the ring will have access to both the encrypted message and the initialization vector except the transmitting and receiving stations. The generation of the initialization vector is unique for each confidential transmission and thus provides a unique approach to the key distribution problem. The FDDI protocol is of particular interest to the military in terms of LAN/MAN implementations. Both the Army and the Navy are considering the standard as the basis for future network systems. A simple and reliable security mechanism with the potential to support realtime communications is a necessary consideration in the implementation of these systems. The proposed method offers several advantages over traditional methods in terms of speed, reliability, and standardization.

  3. Improved 2-nitrobenzenesulfenyl method: optimization of the protocol and improved enrichment for labeled peptides.

    PubMed

    Matsuo, Ei-Ichi; Toda, Chikako; Watanabe, Makoto; Iida, Tetsuo; Masuda, Taro; Minohata, Toshikazu; Ando, Eiji; Tsunasawa, Susumu; Nishimura, Osamu

    2006-01-01

    We have developed the NBS (2-nitrobenzenesulfenyl) method, a quantitative proteome analysis method utilizing stable isotope labeling followed by mass spectrometry. The potential of this method was reported previously, and the procedure has now been further optimized. Here, we describe a procedure utilizing urea or guanidine hydrochloride as a protein denaturant, in conjunction with an improved chromatographic enrichment method for the NBS-labeled peptides using a phenyl resin column. By using this new protocol, both sample loss throughout the protocol and the elution of unwanted unlabeled peptides can be minimized, improving the efficiency of the analysis significantly.

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

    NASA Astrophysics Data System (ADS)

    Pandey, Parul; Tripathi, Maheshwari

    2016-06-01

    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.

  5. 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.

  6. 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.

  7. 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.

  8. 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

  9. MASSIVE TRANSFUSION PROTOCOL: STANDARDIZING CARE TO IMPROVE PATIENT OUTCOMES.

    PubMed

    Porteous, Joan

    2015-06-01

    Providing rapid response is a primary goal when caring for surgical patients with injuries involving massive blood loss. Massive transfusion protocols have been developed in some tertiary care health care facilities to ensure a rapid and efficient response in the provision of care to patients with a massive and uncontrolled hemorrhage. The purpose of this article is to discuss a massive transfusion protocol and to describe the process used to implement a massive transfusion protocol at Winnipeg's Health Sciences Centre (the site) as well as to describe its impact in the operating room department. PMID:26310036

  10. MASSIVE TRANSFUSION PROTOCOL: STANDARDIZING CARE TO IMPROVE PATIENT OUTCOMES.

    PubMed

    Porteous, Joan

    2015-06-01

    Providing rapid response is a primary goal when caring for surgical patients with injuries involving massive blood loss. Massive transfusion protocols have been developed in some tertiary care health care facilities to ensure a rapid and efficient response in the provision of care to patients with a massive and uncontrolled hemorrhage. The purpose of this article is to discuss a massive transfusion protocol and to describe the process used to implement a massive transfusion protocol at Winnipeg's Health Sciences Centre (the site) as well as to describe its impact in the operating room department.

  11. Improved Oocyte Isolation and Embryonic Development of Outbred Deer Mice.

    PubMed

    Choi, Jung Kyu; He, Xiaoming

    2015-01-01

    In this study, we improved the protocol for isolating cumulus-oocyte complexes (COCs) from the outbred deer mice by using only one hormone (instead of the widely used combination of two hormones) with reduced dose. Moreover, we identified that significantly more metaphase II (MII) oocytes could be obtained by supplementing epidermal growth factor (EGF) and leukemia inhibition factor (LIF) into the previously established medium for in vitro maturation (IVM) of the COCs. Furthermore, we overcame the major challenge of two-cell block during embryonic development of deer mice after either in vitro fertilization (IVF) or parthenogenetic activation (PA) of the MII oocytes, by culturing the two-cell stage embryos on the feeder layer of inactivated mouse embryonic fibroblasts (MEFs) in the medium of mouse embryonic stem cells. Collectively, this work represents a major step forward in using deer mice as an outbred animal model for biomedical research on reproduction and early embryonic development. PMID:26184014

  12. 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.

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

    PubMed

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

    2012-01-01

    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. PMID:22614273

  14. 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.

  15. 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

  16. 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.

  17. 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. PMID:26123833

  18. Comparing Breast Screening Protocols: Inserting Catch Trials Does Not Improve Sensitivity over Double Screening

    PubMed Central

    Howe, Piers D. L.

    2016-01-01

    Breast screening is an important tool for the early detection of breast cancers. However, tumours are typically present in less than 1% of mammograms. This low prevalence could cause radiologists to detect fewer tumours than they otherwise would, an issue known as the prevalence effect. The aim of our study was to investigate a novel breast screening protocol, designed to decrease the number of tumours missed by radiologists, without increasing their workload. We ran two laboratory-based experiments to assess the degree to which the novel protocol, called the catch trial (CT) protocol, resulted in greater sensitivity (d’) than the double screener protocol (DS), currently utilised in Australia. In our first experiment we found evidence that the CT protocol resulted in a criterion shift relative to the DS protocol but the evidence that sensitivity was greater in the CT protocol relative to the DS protocol was less clear. A second experiment, using more realistic stimuli that were more representative of actual tumours, also failed to find convincing evidence that sensitivity was greater in the CT protocol than in the DS protocol. This experiment instead found that both the hit rate and the false alarm rate increased in the CT protocol relative to the DS protocol. So while there was again evidence that the CT protocol induced a criterion shift, the sensitivity appeared to be approximately the same in both protocols. Our results suggest the CT protocol is unlikely to result in an improvement in sensitivity over the DS protocol, so we cannot recommend that it be trialled in a clinical setting. PMID:27723788

  19. A Simple Protocol for High Efficiency Protein Isolation After RNA Isolation from Mouse Thyroid and Other Very Small Tissue Samples.

    PubMed

    Ziros, Panos G; Chartoumpekis, Dionysios V; Sykiotis, Gerasimos P

    2016-01-01

    As a dedicated hormone-secreting organ, the thyroid gland possesses a complement of proteostatic systems, including antioxidant, unfolded protein, and autophagic responses. The vast majority of animal investigations of thyroid physiology and, more recently, proteostasis, have utilized as model the rat, rather than the mouse. This is due to the very small size of the thyroid gland in the latter, with a total weight of ~2 mg (~1 mg per thyroid lobe). However, this strategy has limited the utilization of genetic approaches, such as taking advantage of the various transgenic and knockout mouse models. Here, we describe a simple and highly efficient protocol for the simultaneous isolation of mRNA, micro-RNA and 150-200 μg of protein from as little as 1 mg of mouse thyroid tissue, the average weight of one of the two thyroid lobes, thus preserving the other lobe for immunohistochemical or other analyses. While our workflow is similar to other protocols published in the literature and/or proposed by commercial reagent providers, we have introduced a key modification that addresses efficiently the most challenging step of the protein isolation process: the solubilization of the protein pellet after RNA extraction and protein precipitation. We demonstrate the feasibility of our approach and its utility for downstream analyses (including Western blotting) that facilitate the comparative study of proteostatic pathways in the mouse thyroid. We have also successfully applied this protocol on samples from mouse liver, brown and white adipose tissue, as well as from rodent cell lines. PMID:27613051

  20. A modified protocol for RNA extraction from different peach tissues suitable for gene isolation and real-time PCR analysis.

    PubMed

    Tong, Zhaoguo; Qu, Shenchun; Zhang, Jiyu; Wang, Fei; Tao, Jianmin; Gao, Zhihong; Zhang, Zhen

    2012-03-01

    RNA extraction is the first step in the study of gene isolation and expression. However, it is difficult to extract high quantity and quality RNA from tissues containing large quantities of polysaccharides and polyphenols. Peach (Prunus persica), in addition to containing high levels of polysaccharides and polyphenols, is a challenging starting material for RNA isolation using a single method because of different amounts of those substances in diverse tissues. Based on three reported methods, we developed a modified RNA isolation protocol to solve this problem, leading to high quality and quantity of total RNA from peach mesocarp tissues of fruits which were sampled from all developmental stages and different storage periods, as well as from other tissues including flowers, leaves, stems, and roots. With our modified method, 28-650 μg of total RNA was routinely obtained from per gram of fresh material, gave at least a 1.16-fold improvement by compared with those isolated by other seven methods. The RNA extracts were successfully used in downstream applications such as RT-PCR, RACE, and real-time PCR.

  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. Full-Length gene enrichment by using an optimized RNA isolation protocol in Bixa orellana recalcitrant tissues.

    PubMed

    Rodríguez-Avila, N L; Narváez-Zapata, J A; Aguilar-Espinosa, M L; Rivera-Madrid, R

    2009-05-01

    A reliable protocol is described for isolation of large full-length cDNA from Bixa orellana mature tissues containing large quantities of pigments, phenols, and polysaccharides. This protocol involves the optimization of a commercial RNA extraction protocol in combination with a long distance reverse transcript PCR protocol. The principal advantages of this protocol are its high RNA yield and quality. The resulting RNA is suitable for RNA expression evaluation and production of large, full-length cDNA. This is the first time RNA has been isolated from all mature tissues in the tropical perennial plant B. orellana and has been proved viable for downstream applications, especially important for molecular biology studies on this economically important pigment-producing plant. PMID:19107604

  3. 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.

  4. An improved protocol for micropropagation of saltbush (Atriplex) species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Atriplex griffithsii Standl is a threatened halophytic shrub found in saline soils of isolated regions within the desert basins in Southern New Mexico and Arizona. A related species within the genus, A. canescens, has been successfully micropropagated, suggesting that clonal propagation may offer a...

  5. 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.

  6. An Optimized Protocol for Isolating Primary Epithelial Cell Chromatin for ChIP

    PubMed Central

    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. PMID:24971909

  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. Improvement in Isolation and Identification of Mouse Oogonial Stem Cells

    PubMed Central

    Lu, Zhiyong; Wu, Meng; Zhang, Jinjin; Xiong, Jiaqiang; Cheng, Jing; Shen, Wei; Luo, Aiyue; Fang, Li; Wang, Shixuan

    2016-01-01

    Female germline stem cells (FGSCs) or oogonial stem cells (OSCs) have the capacity to generate newborn oocytes and thus open a new door to fight ovarian aging and female infertility. However, the production and identification of OSCs are difficult for investigators. Rare amount of these cells in the ovary results in the failure of the acquisition of OSCs. Furthermore, the oocyte formation by OSCs in vivo was usually confirmed using tissue sections by immunofluorescence or immunohistochemistry in previous studies. STO or MEF feeder cells are derived from mouse, not human. In our study, we modified the protocol. The cells were digested from ovaries and cultured for 2-3 days and then were purified by magnetic-activated cell sorting (MACS). The ovaries and fetus of mice injected with EGFP-positive OSCs were prepared and put on the slides to directly visualize oocyte and progeny formation under microscope. Additionally, the human umbilical cord mesenchymal stem cells (hUC-MSCs) were also used as feeder cells to support the proliferation of OSCs. The results showed that all the modified procedures can significantly improve and facilitate the generation and characterization of OSCs, and hUC-MSCs as feeder will be useful for isolation and proliferation of human OSCs avoiding contamination from mouse. PMID:26635882

  10. 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.

  11. 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

  12. Improving School Climate and Reducing Teacher Isolation.

    ERIC Educational Resources Information Center

    Rothberg, Robert A.

    A survey of 196 elementary and secondary education teachers attending graduate programs in the University of Central Florida's College of Education provided data supporting previous research on teacher isolation. Over 80 percent of the teachers felt their classrooms were private worlds entered only by themselves and their students. The teachers…

  13. A novel screening protocol for the isolation of hydrogen producing Chlamydomonas reinhardtii strains

    PubMed Central

    Rühle, Thilo; Hemschemeier, Anja; Melis, Anastasios; Happe, Thomas

    2008-01-01

    Background Sealed Chlamydomonas reinhardtii cultures evolve significant amounts of hydrogen gas under conditions of sulfur depletion. However, the eukaryotic green alga goes through drastic metabolic changes during this nutritional stress resulting in cell growth inhibition and eventually cell death. This study aimed at isolating C. reinhardtii transformants which produce hydrogen under normal growth conditions to allow a continuous hydrogen metabolism without the stressful impact of nutrient deprivation. Results To achieve a steady photobiological hydrogen production, a screening protocol was designed to identify C. reinhardtii DNA insertional mutagenesis transformants with an attenuated photosynthesis to respiration capacity ratio (P/R ratio). The screening protocol entails a new and fast method for mutant strain selection altered in their oxygen production/consumption balance. Out of 9000 transformants, four strains with P/R ratios varying from virtually zero to three were isolated. Strain apr1 was found to have a slightly higher respiration rate and a significantly lower photosynthesis rate than the wild type. Sealed cultures of apr1 became anaerobic in normal growth medium (TAP) under moderate light conditions and induced [FeFe]-hydrogenase activity, yet without significant hydrogen gas evolution. However, Calvin-Benson cycle inactivation of anaerobically adapted apr1 cells in the light led to a 2-3-fold higher in vivo hydrogen production than previously reported for the sulfur-deprived C. reinhardtii wild type. Conclusion Attenuated P/R capacity ratio in microalgal mutants constitutes a platform for achieving steady state photobiological hydrogen production. Using this platform, algal hydrogen metabolism can be analyzed without applying nutritional stress. Furthermore, these strains promise to be useful for biotechnological hydrogen generation, since high in vivo hydrogen production rates are achievable under normal growth conditions, when the photosynthesis

  14. Improved Protocol For Laser Microdissection Of Human Pancreatic Islets From Surgical Specimens

    PubMed Central

    Sturm, Dorothée; Marselli, Lorella; Ehehalt, Florian; Richter, Daniela; Distler, Marius; Kersting, Stephan; Grützmann, Robert; Bokvist, Krister; Froguel, Philippe; Liechti, Robin; Jörns, Anne; Meda, Paolo; Baretton, Gustavo Bruno; Saeger, Hans-Detlev; Schulte, Anke M.; Marchetti, Piero; Solimena, Michele

    2013-01-01

    microdissection, four sections were dehydrated each time: two were placed into a foil-wrapped 50 ml tube, to protect the tissue from moisture and bleaching; the remaining two were immediately microdissected. This procedure was performed using a PALM MicroBeam instrument (Zeiss) employing the Auto Laser Pressure Catapulting (AutoLPC) mode. The completion of beta cell/islet dissection from four cryosections required no longer than 40-60 min. Cells were collected into one AdhesiveCap and lysed with 10 μl lysis buffer. Each single RNA specimen for transcriptomic analysis was obtained by combining 10 cell microdissected samples, followed by RNA extraction using the Pico Pure RNA Isolation Kit (Arcturus). This protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their rapid and accurate recognition and collection. Further improvement of this procedure could enable the dissection of phenotypically different beta cells, with possible implications for better understanding the changes associated with type 2 diabetes. PMID:23329157

  15. 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

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

    PubMed

    Fitzpatrick, Darren; Cagle, Paul; Flatow, Evan

    2016-04-01

    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.

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

    PubMed

    Fitzpatrick, Darren; Cagle, Paul; Flatow, Evan

    2016-04-01

    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

  18. 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.

  19. 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

  20. 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

  1. 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

  2. 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.

  3. 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…

  4. 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…

  5. 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…

  6. 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…

  7. 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.

  8. 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. PMID:27108331

  9. Perioperative glycemic control: use of a hospital-wide protocol to safely improve hyperglycemia.

    PubMed

    Michaelian, Nancy; Joshi, Renu; Gillman, Ed; Kratz, Ronald; Helmuth, Amy; Zimmerman, Karen; Klahre, Denise; Warner, Sandy; McBride, Vickie; Bailey, M Judy; Houseal, Linda

    2011-08-01

    Perioperative hyperglycemia impairs immunity and contributes to increased susceptibility to infection, higher incidence of multiorgan dysfunction, and greater mortality. Strict glycemic control is associated with lower infection rates, decreased length of stay (LOS), and faster recovery. A protocol that standardized preoperative education, testing, and treatment of elevated blood glucose (BG) safely improved perioperative glycemic control. Preoperative average BG improved from 191 to 155 mg/dL (P=.016); postoperative average BG decreased from 189 to 168 mg/dL (P=.094). The percentage of patients presenting with BG greater than 180 mg/dL preoperatively and achieving BG less than 180 mg/DL postoperatively increased from 21% to 43% (P = .09). Even though some results were statistically non-significant, the data showed a trend toward improvement with the new protocol. Good perioperative glycemic control, without an increased risk of hypoglycemia, is achievable. PMID:21803272

  10. A protocol for isolating insect mitochondrial genomes: a case study of NUMT in Melipona flavolineata (Hymenoptera: Apidae).

    PubMed

    Françoso, Elaine; Gomes, Fernando; Arias, Maria Cristina

    2016-07-01

    Nuclear mitochondrial DNA insertions (NUMTs) are mitochondrial DNA sequences that have been transferred into the nucleus and are recognized by the presence of indels and stop codons. Although NUMTs have been identified in a diverse range of species, their discovery was frequently accidental. Here, our initial goal was to develop and standardize a simple method for isolating NUMTs from the nuclear genome of a single bee. Subsequently, we tested our new protocol by determining whether the indels and stop codons of the cytochrome c oxidase subunit I (COI) sequence of Melipona flavolineata are of nuclear origin. The new protocol successfully demonstrated the presence of a COI NUMT. In addition to NUMT investigations, the protocol described here will also be very useful for studying mitochondrial mutations related to diseases and for sequencing complete mitochondrial genomes with high read coverage by Next-Generation technology.

  11. 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.

  12. T Lymphocyte Density and Distribution in Human Colorectal Mucosa, and Inefficiency of Current Cell Isolation Protocols

    PubMed Central

    Preza, Gloria Cuevas; Yang, Otto O.; Elliott, Julie; Anton, Peter A.; Ochoa, Maria T.

    2015-01-01

    Mucosal tissues are critical immune effector sites containing complex populations of leukocytes in a tissue microenvironment that remains incompletely understood. We identify and quantify in human distal colorectal tissue absolute mucosal CD3+ lymphocytes, including CD4+ and CD8+ subsets, by direct visualization using immunohistochemistry (IHC), immunofluorescence (IF), and an automated counting protocol (r2=0.90). Sigmoid and rectal mucosal tissues are both densely packed with T lymphocytes in the mucosal compartment. Both compartments had similar densities of CD3+ T lymphocytes with 37,400 ± 2,801 cells/mm3 and 33,700 ± 4,324 cell/mm3, respectively. Sigmoid mucosa contained 57% CD3+CD4+ and 40% CD3+CD8+ T lymphocytes which calculates to 21,300 ± 1,476/mm3 and 15,000 ± 275/mm3 T lymphocytes, respectively. Rectal mucosa had 57% CD3+CD4+ and 42% CD3+CD8+ or 21,577 ± 332, and 17,090 ± 1,206 cells/mm3, respectively. By comparison, sigmoid mucosal biopsies subjected to conventional collagenase digestion, mononuclear cell (MMC) isolation and staining for flow cytometry yielded 4,549 ± 381/mm3 and 2,708 ± 245/mm3 CD4+ and CD8+ T lymphocytes. These data suggest only ~20.7% recovery compared to IHC results for these markers. Further studies will determine if this reflects a selective bias in only CD3+, CD4+ and CD8+ T cells or can be generalized to all flow-analyzed cells from mucosal tissues for phenotyping and functional testing. PMID:25856343

  13. 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

  14. 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.

  15. 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. PMID:27218005

  16. In vitro selection and amplification protocols for isolation of aptameric sensors for small molecules.

    PubMed

    Yang, Kyung-Ae; Pei, Renjun; Stojanovic, Milan N

    2016-08-15

    We recently optimized a procedure that directly yields aptameric sensors for small molecules in so-called structure-switching format. The protocol has a high success rate, short time, and is sufficiently simple to be readily implemented in a non-specialist laboratory. We provide a stepwise guide to this selection protocol. PMID:27155227

  17. A step-by-step protocol for formaldehyde-assisted isolation of regulatory elements from Arabidopsis thaliana.

    PubMed

    Omidbakhshfard, Mohammad Amin; Winck, Flavia Vischi; Arvidsson, Samuel; Riaño-Pachón, Diego M; Mueller-Roeber, Bernd

    2014-06-01

    The control of gene expression by transcriptional regulators and other types of functionally relevant DNA transactions such as chromatin remodeling and replication underlie a vast spectrum of biological processes in all organisms. DNA transactions require the controlled interaction of proteins with DNA sequence motifs which are often located in nucleosome-depleted regions (NDRs) of the chromatin. Formaldehyde-assisted isolation of regulatory elements (FAIRE) has been established as an easy-to-implement method for the isolation of NDRs from a number of eukaryotic organisms, and it has been successfully employed for the discovery of new regulatory segments in genomic DNA from, for example, yeast, Drosophila, and humans. Until today, however, FAIRE has only rarely been employed in plant research and currently no detailed FAIRE protocol for plants has been published. Here, we provide a step-by-step FAIRE protocol for NDR discovery in Arabidopsis thaliana. We demonstrate that NDRs isolated from plant chromatin are readily amenable to quantitative polymerase chain reaction and next-generation sequencing. Only minor modification of the FAIRE protocol will be needed to adapt it to other plants, thus facilitating the global inventory of regulatory regions across species.

  18. Protocols for Biotechnological Interventions in Improvement of Vanilla (Vanilla planifolia Andrews.).

    PubMed

    Divakaran, Minoo; Babu, K Nirmal; Peter, K V

    2016-01-01

    Vanilla (Vanilla planifolia Andrews (syn. V. fragrans Salisb.), a native of Central America, is the primary source of natural vanillin and plays a major role in the global economy. The gene pool of vanilla is threatened by deforestation and overcollection that has resulted in disappearance of natural habitats and wild species. Continuous vegetative propagation and lack of natural seed set and sufficient variations in the gene pool hamper crop improvement programs. In vitro techniques, one of the key tools of plant biotechnology, can be employed for overcoming specific problems, viz. production of disease-free clones, inducing somaclonal variations, developing hybrids, gene pool conservation, incorporating desired traits by distant hybridization, genetic engineering, etc. However, realization of these objectives necessitates standardization of protocols. This chapter describes the various protocols optimized for crop improvement in Vanilla species. PMID:27108309

  19. Protocols for Biotechnological Interventions in Improvement of Vanilla (Vanilla planifolia Andrews.).

    PubMed

    Divakaran, Minoo; Babu, K Nirmal; Peter, K V

    2016-01-01

    Vanilla (Vanilla planifolia Andrews (syn. V. fragrans Salisb.), a native of Central America, is the primary source of natural vanillin and plays a major role in the global economy. The gene pool of vanilla is threatened by deforestation and overcollection that has resulted in disappearance of natural habitats and wild species. Continuous vegetative propagation and lack of natural seed set and sufficient variations in the gene pool hamper crop improvement programs. In vitro techniques, one of the key tools of plant biotechnology, can be employed for overcoming specific problems, viz. production of disease-free clones, inducing somaclonal variations, developing hybrids, gene pool conservation, incorporating desired traits by distant hybridization, genetic engineering, etc. However, realization of these objectives necessitates standardization of protocols. This chapter describes the various protocols optimized for crop improvement in Vanilla species.

  20. Comparative evaluation of a chromogenic agar medium-PCR protocol with a conventional method for isolation of Vibrio parahaemolyticus strains from environmental and clinical samples.

    PubMed

    Canizalez-Roman, Adrian; Flores-Villaseñor, Héctor; Zazueta-Beltran, Jorge; Muro-Amador, Secundino; León-Sicairos, Nidia

    2011-02-01

    Screening for pathogenic Vibrio parahaemolyticus has become routine in certain areas associated with food-borne outbreaks. This study is an evaluation of the CHROMagar Vibrio (CV) medium-PCR protocol and the conventional method (TCBS (thiosulfate-citrate-bile salts-sucrose) agar plus biochemical and Wagatsuma agar tests) for detection of V. parahaemolyticus in shrimp, water, sediment, and stool samples collected for biosurveillance in an endemic area of northwestern Mexico. A total of 131 environmental and clinical samples were evaluated. The CV medium-PCR protocol showed a significantly improved ability (P < 0.05) to isolate and detect V. parahaemolyticus, identifying isolates of this bacteria missed by the conventional method. Although some other bacteria, distinct from pathogenic V. parahaemolyticus, produced violet colonies similar to that of V. parahaemolyticus on CV medium, we were able to detect a superior number of samples of V. parahaemolyticus with the CV medium-PCR protocol than with the conventional method. The Kanagawa phenomenon is routinely determined on Wagatsuma agar for the diagnosis of V. parahaemolyticus (pathogenic) positive for thermostable direct hemolysin (TDH) in developing countries. In our results, Wagatsuma agar showed low sensitivity (65.4% at 24 h and 75.6% at 48 h) and specificity (52.4% at 48 h) for identifying V. parahaemolyticus positive for TDH. Overall, our data support the use of the CV medium-PCR protocol in place of the conventional method (TCBS-biochemical tests-Wagatsuma agar) for detection of pathogenic V. parahaemolyticus, both in terms of effectiveness and cost efficiency.

  1. An improved protocol for quantification of freshwater Actinobacteria by fluorescence in situ hybridization.

    PubMed

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

    2003-05-01

    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.

  2. An improved machine learning protocol for the identification of correct Sequest search results

    PubMed Central

    2010-01-01

    Background Mass spectrometry has become a standard method by which the proteomic profile of cell or tissue samples is characterized. To fully take advantage of tandem mass spectrometry (MS/MS) techniques in large scale protein characterization studies robust and consistent data analysis procedures are crucial. In this work we present a machine learning based protocol for the identification of correct peptide-spectrum matches from Sequest database search results, improving on previously published protocols. Results The developed model improves on published machine learning classification procedures by 6% as measured by the area under the ROC curve. Further, we show how the developed model can be presented as an interpretable tree of additive rules, thereby effectively removing the 'black-box' notion often associated with machine learning classifiers, allowing for comparison with expert rule-of-thumb. Finally, a method for extending the developed peptide identification protocol to give probabilistic estimates of the presence of a given protein is proposed and tested. Conclusions We demonstrate the construction of a high accuracy classification model for Sequest search results from MS/MS spectra obtained by using the MALDI ionization. The developed model performs well in identifying correct peptide-spectrum matches and is easily extendable to the protein identification problem. The relative ease with which additional experimental parameters can be incorporated into the classification framework, to give additional discriminatory power, allows for future tailoring of the model to take advantage of information from specific instrument set-ups. PMID:21138573

  3. 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.

  4. Cell permeable peptide of JNK inhibitor prevents islet apoptosis immediately after isolation and improves islet graft function.

    PubMed

    Noguchi, Hirofumi; Nakai, Yusuke; Matsumoto, Shinichi; Kawaguchi, Miho; Ueda, Michiko; Okitsu, Teru; Iwanaga, Yasuhiro; Yonekawa, Yukihide; Nagata, Hideo; Minami, Kohtaro; Masui, Yumi; Futaki, Shiroh; Tanaka, Koichi

    2005-08-01

    Although application of the Edmonton protocol has markedly improved outcomes for pancreatic islet transplantation, the insulin independence rate after islet transplantation from one donor pancreas has proven to remain low. During the isolation process and subsequent clinical transplantation, islets are subjected to severe adverse conditions that impair survival and ultimately contribute to graft failure. Pancreas preservation with the two-layer method (TLM) has proven to improve transplant results by protecting isolated islets against apoptosis through the mitochondrial pathway. However, pancreas storage with TLM cannot protect against activation of c-Jun NH2-terminal kinase (JNK) in isolated islets. This study investigated whether delivery of a JNK inhibitory peptide (JNKI) via the protein transduction system can prevent apoptosis of islet cells immediately after isolation. For efficient delivery of the (JNKI into isolated islets, we synthesized JNKI as a C-terminal fusion peptide with the 11-arginine protein transduction domain (11R-JNKI). 11R efficiently delivered the JNKI into isolated islets and 11R-JNKI prevented islet apoptosis immediately after isolation and improved islet graft function. These findings suggest that peptide drugs could be useful for the prevention of the impairment of islet cells and lead to improvement in the outcomes for pancreatic islet transplantation.

  5. 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

  6. 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.

  7. A universal protocol for the combined isolation of metabolites, DNA, long RNAs, small RNAs, and proteins from plants and microorganisms.

    PubMed

    Valledor, Luis; Escandón, Mónica; Meijón, Mónica; Nukarinen, Ella; Cañal, María Jesús; Weckwerth, Wolfram

    2014-07-01

    Here, we describe a method for the combined metabolomic, proteomic, transcriptomic and genomic analysis from one single sample as a major step for multilevel data integration strategies in systems biology. While extracting proteins and DNA, this protocol also allows the separation of metabolites into polar and lipid fractions, as well as RNA fractionation into long and small RNAs, thus allowing a broad range of transcriptional studies. The isolated biomolecules are suitable for analysis with different methods that range from electrophoresis and blotting to state-of-the-art procedures based on mass spectrometry (accurate metabolite profiling, shot-gun proteomics) or massive sequencing technologies (transcript analysis). The low amount of starting tissue, its cost-efficiency compared with the utilization of commercial kits, and its performance over a wide range of plant, microbial, and algal species such as Chlamydomonas, Arabidopsis, Populus, or Pinus, makes this method a universal alternative for multiple molecular isolation from plant tissues.

  8. 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

  9. 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.

  10. 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

  11. 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. PMID:25815373

  12. 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

  13. 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.

  14. Implementation of a Multidisciplinary Bleeding and Transfusion Protocol Significantly Decreases Perioperative Blood Product Utilization and Improves Some Bleeding Outcomes

    PubMed Central

    Timpa, Joseph G.; O’Meara, L. Carlisle; Goldberg, Kellen G.; Phillips, Jay P.; Crawford, Jack H.; Jackson, Kimberly W.; Alten, Jeffrey A.

    2016-01-01

    Abstract: Perioperative transfusion of blood products is associated with increased morbidity and mortality after pediatric cardiac surgery. We report the results of a quality improvement project aimed at decreasing perioperative blood product administration and bleeding after pediatric cardiopulmonary bypass (CPB) surgery. A multidisciplinary team evaluated baseline data from 99 consecutive CPB patients, focusing on the variability in transfusion management and bleeding outcomes, to create a standardized bleeding and transfusion management protocol. A total of 62 subsequent patients were evaluated after implementation of the protocol: 17 with single pass hemoconcentrated (SPHC) blood transfusion and 45 with modified ultrafiltration (MUF). Implementation of the protocol with SPHC blood led to significant decrease in transfusion of every blood product in the cardiovascular operating room and first 6 hours in cardiovascular intensive care unit ([CVICU] p < .05). Addition of MUF to the protocol led to further decrease in transfusion of all blood products compared to preprotocol. Patients <2 months old had 49% decrease in total blood product administration: 155 mL/kg preprotocol, 117 mL/kg protocol plus SPHC, and 79 mL/kg protocol plus MUF (p < .01). There were significant decreases in postoperative bleeding in the first hour after CVICU admission: 6 mL/kg preprotocol, 3.8 mL/kg protocol plus SPHC, and 2 mL/kg protocol plusMUF (p = .02). There was also significantly decreased incidence of severe postoperative bleeding (>10 mL/kg) in the first CVICU hour for protocol plus MUF patients (p < .01). Implementation of a multidisciplinary bleeding and transfusion protocol significantly decreases perioperative blood product transfusion and improves some bleeding outcomes. PMID:27134303

  15. Protocol for Optimal Quality and Quantity Pollen DNA Isolation from Honey Samples

    PubMed Central

    Lalhmangaihi, Ralte; Ghatak, Souvik; Laha, Ramachandra; Gurusubramanian, Guruswami; Kumar, Nachimuthu Senthil

    2014-01-01

    The present study illustrates an optimized sample preparation method for an efficient DNA isolation from low quantities of honey samples. A conventional PCR-based method was validated, which potentially enables characterization of plant species from as low as 3 ml bee-honey samples. In the present study, an anionic detergent was used to lyse the hard outer pollen shell, and DTT was used for isolation of thiolated DNA, as it might facilitate protein digestion and assists in releasing the DNA into solution, as well as reduce cross-links between DNA and other biomolecules. Optimization of both the quantity of honey sample and time duration for DNA isolation was done during development of this method. With the use of this method, chloroplast DNA was successfully PCR amplified and sequenced from honey DNA samples. PMID:25365793

  16. IDMA: improving the defense against malicious attack for mobile ad hoc networks based on ARIP protocol

    NASA Astrophysics Data System (ADS)

    Peng, Chaorong; Chen, Chang Wen

    2008-04-01

    Malicious nodes are mounting increasingly sophisticated attacking operations on the Mobile Ad Hoc Networks (MANETs). This is mainly because the IP-based MANETs are vulnerable to attacks by various malicious nodes. However, the defense against malicious attack can be improved when a new layer of network architecture can be developed to separate true IP address from disclosing to the malicious nodes. In this paper, we propose a new algorithm to improve the defense against malicious attack (IDMA) that is based on a recently developed Assignment Router Identify Protocol (ARIP) for the clustering-based MANET management. In the ARIP protocol, we design the ARIP architecture based on the new Identity instead of the vulnerable IP addresses to provide the required security that is embedded seamlessly into the overall network architecture. We make full use of ARIP's special property to monitor gateway forward packets by Reply Request Route Packets (RREP) without additional intrusion detection layer. We name this new algorithm IDMA because of its inherent capability to improve the defense against malicious attacks. Through IDMA, a watching algorithm can be established so as to counterattack the malicious node in the routing path when it unusually drops up packets. We provide analysis examples for IDMA for the defense against a malicious node that disrupts the route discovery by impersonating the destination, or by responding with state of corrupted routing information, or by disseminating forged control traffic. The IDMA algorithm is able to counterattack the malicious node in the cases when the node lunch DoS attack by broadcast a large number of route requests, or make Target traffic congestion by delivering huge mount of data; or spoof the IP addresses and send forge packets with a fake ID to the same Target causing traffic congestion at that destination. We have implemented IDMA algorism using the GloMoSim simulator and have demonstrated its performance under a variety of

  17. An optimized protocol for isolating lymphoid stromal cells from the intestinal lamina propria.

    PubMed

    Stzepourginski, Igor; Eberl, Gérard; Peduto, Lucie

    2015-06-01

    Mesenchymal stromal cells in lymphoid organs, also called lymphoid stromal cells (LSCs), play a pivotal role in immunity by forming specialized microenvironments that provide signals for leukocyte migration, positioning, and survival. Best characterized in lymphoid organs, LSCs are also abundant in the intestinal mucosa, which harbors a rich repertoire of immune cells. However, the lack of efficient procedures for isolation and purification of LSCs from the intestine has been a major limitation to their characterization. Here we report a new method to efficiently isolate, in addition to immune cells, viable lymphoid stromal cells and other stromal subsets from the intestinal lamina propria for subsequent phenotypic and functional analysis.

  18. Improving cardiomyocyte model fidelity and utility via dynamic electrophysiology protocols and optimization algorithms.

    PubMed

    Krogh-Madsen, Trine; Sobie, Eric A; Christini, David J

    2016-05-01

    Mathematical models of cardiac electrophysiology are instrumental in determining mechanisms of cardiac arrhythmias. However, the foundation of a realistic multiscale heart model is only as strong as the underlying cell model. While there have been myriad advances in the improvement of cellular-level models, the identification of model parameters, such as ion channel conductances and rate constants, remains a challenging problem. The primary limitations to this process include: (1) such parameters are usually estimated from data recorded using standard electrophysiology voltage-clamp protocols that have not been developed with model building in mind, and (2) model parameters are typically tuned manually to subjectively match a desired output. Over the last decade, methods aimed at overcoming these disadvantages have emerged. These approaches include the use of optimization or fitting tools for parameter estimation and incorporating more extensive data for output matching. Here, we review recent advances in parameter estimation for cardiomyocyte models, focusing on the use of more complex electrophysiology protocols and global search heuristics. We also discuss future applications of such parameter identification, including development of cell-specific and patient-specific mathematical models to investigate arrhythmia mechanisms and predict therapy strategies. PMID:26661516

  19. 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.

  20. An efficient DNA isolation protocol for filamentous cyanobacteria of the genus Arthrospira.

    PubMed

    Morin, Nicolas; Vallaeys, Tatiana; Hendrickx, Larissa; Natalie, Leys; Wilmotte, Annick

    2010-02-01

    Thanks to their photosynthetic and nutritive properties, cyanobacteria of the Arthrospira genus are of interest as food supplements, as efficient oxygen producing life support system organisms for manned space flight, and for the production of biofuels. Despite these potential valuable applications, full genome sequences and genetic information in general on Arthrospira remain scarce. This is mainly due to the difficulty to extract sufficient high molecular weight nucleic acids from these filamentous cyanobacteria. In this article, an efficient and reproducible DNA extraction procedure for cyanobacteria of the genus Arthrospira was developed. The method is based on the combination of a soft mechanical lysis with enzymatic disruption of the cell wall. The comparison with other extraction protocols clearly indicates that this optimised method allows the recovery of a larger amount of DNA. Furthermore, the extracted DNA presents a high molecular weight, a reduced degradation and an excellent overall quality. It can be directly used for molecular biology purposes such as PCR, and clone library construction.

  1. A novel protocol to accelerate dynamic combinatorial chemistry via isolation of ligand-target adducts from dynamic combinatorial libraries: a case study identifying competitive inhibitors of lysozyme.

    PubMed

    Fang, Zheng; He, Wei; Li, Xin; Li, Zhengjiang; Chen, Beining; Ouyang, Pingkai; Guo, Kai

    2013-09-15

    A novel protocol based on size-exclusion chromatography (SEC) and MS was established to accelerate dynamic combinatorial chemistry (DCC) in this study. By isolating ligand-target adducts from the dynamic combinatorial library (DCL), ligands could be identified directly by MS after denaturation. Three new inhibitors for lysozyme were discovered by this SEC-MS protocol in a case study. Km Data for these new inhibitors was also determined.

  2. 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.

  3. 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

  4. Improving patient care through implementation of nurse-driven restraint protocols.

    PubMed

    Winston, P A; Morelli, P; Bramble, J; Friday, A; Sanders, J B

    1999-08-01

    Nationally, much attention has been placed on the indiscriminate application and abuse of restraint usage. This was the impetus for health care institutions across the country to relook at the policy, practices, and procedures regarding restraints. Our health care system made changes to our restraint policy, practice guidelines, and procedures in an effort to assure protection of the patients' health and safety while preserving their dignity, rights, and well-being. The mission was to pursue a restraint-appropriate environment by restraining only those patients who were assessed as being at risk of harming self and to protect the patient or others from injury. Our overall goal was to reduce restraint usage. This article describes the current policies, practice guidelines, and procedures for identifying clinically appropriate and adequately justified situations for restraint usage. The focus is on implementation of nurse-driven restraint protocols to improve patient care. All efforts directed at improvements in restraint usage and management of a patient in restraints has reduced our overall numbers of patients in restraints as well as significantly reduced risk of incidence for patients in restraints. PMID:10476623

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

    PubMed Central

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

    2014-01-01

    Objective To assess whether an early GnRH antagonist start leads to better follicular synchronization and an improved clinical pregnancy rate (CPR). Methods 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. Results 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%). Conclusion 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. PMID:25599038

  6. 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. PMID:27091400

  7. 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

  8. 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

    2016-01-01

    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

  9. 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.

  10. A standardized protocol to reduce cerebrospinal fluid shunt infection: The Hydrocephalus Clinical Research Network Quality Improvement Initiative

    PubMed Central

    Kestle, John R. W.; Riva-Cambrin, Jay; Wellons, John C.; Kulkarni, Abhaya V.; Whitehead, William E.; Walker, Marion L.; Oakes, W. Jerry; Drake, James M.; Luerssen, Thomas G.; Simon, Tamara D.; Holubkov, Richard

    2011-01-01

    Object Quality improvement techniques are being implemented in many areas of medicine. In an effort to reduce the ventriculoperitoneal shunt infection rate, a standardized protocol was developed and implemented at 4 centers of the Hydrocephalus Clinical Research Network (HCRN). Methods The protocol was developed sequentially by HCRN members using the current literature and prior institutional experience until consensus was obtained. The protocol was prospectively applied at each HCRN center to all children undergoing a shunt insertion or revision procedure. Infections were defined on the basis of CSF, wound, or pseudocyst cultures; wound breakdown; abdominal pseudocyst; or positive blood cultures in the presence of a ventriculoatrial shunt. Procedures and infections were measured before and after protocol implementation. Results Twenty-one surgeons at 4 centers performed 1571 procedures between June 1, 2007, and February 28, 2009. The minimum follow-up was 6 months. The Network infection rate decreased from 8.8% prior to the protocol to 5.7% while using the protocol (p = 0.0028, absolute risk reduction 3.15%, relative risk reduction 36%). Three of 4 centers lowered their infection rate. Shunt surgery after external ventricular drainage (with or without prior infection) had the highest infection rate. Overall protocol compliance was 74.5% and improved over the course of the observation period. Based on logistic regression analysis, the use of BioGlide catheters (odds ratio [OR] 1.91, 95% CI 1.19–3.05; p = 0.007) and the use of antiseptic cream by any members of the surgical team (instead of a formal surgical scrub by all members of the surgical team; OR 4.53, 95% CI 1.43–14.41; p = 0.01) were associated with an increased risk of infection. Conclusions The standardized protocol for shunt surgery significantly reduced shunt infection across the HCRN. Overall protocol compliance was good. The protocol has established a common baseline within the Network, which will

  11. 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

  12. 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.

  13. 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

  14. 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.

  15. Improved (and Singular) Disinfectant Protocol for Indirectly Assessing Organic Precursor Concentrations of Trihalomethanes and Dihaloacetonitriles.

    PubMed

    Do, Thien D; Chimka, Justin R; Fairey, Julian L

    2015-08-18

    Measurements of disinfection byproduct (DBP) organic precursor concentrations (OPCs) are crucial to assess and improve DBP control processes. Typically, formation potential tests - specified in Standard Methods (SM) 5710-B/D - are used to measure OPCs. Here, we highlight several limitations of this protocol for dihaloacetonitriles and trihalomethanes and validate a novel Alternative Method (AM). The effects of pH, disinfectant type (free chlorine and monochloramine), and chlor(am)ine residual (CR) were examined on DBP formation in a suite of waters. Using the SM, DHAN decreased 43-47% as the CR increased from 3 to 5 mg L(-1) as Cl2, compromising OPC assessments. In contrast, a high monochloramine dose (250 mg L(-1) as Cl2) at pH 7.0 (the AM) accurately reflected OPCs. The two methods were compared for assessing DBP precursor removal through three granular activated carbon (GAC) columns in series. Breakthrough profiles assessed using the AM only showed DBP precursor sorption occurred in each column that decreased over time (p = 0.0001). Similarly, the AM facilitated ranking of three types of GAC compared in parallel columns, whereas the SM produced ambiguous results. Fluorescence intensity of a humic-like fluorophore (i.e., I345/425) correlated strongly to precursor removal in the GAC columns. The practical implications of the results are discussed.

  16. Improved (and Singular) Disinfectant Protocol for Indirectly Assessing Organic Precursor Concentrations of Trihalomethanes and Dihaloacetonitriles.

    PubMed

    Do, Thien D; Chimka, Justin R; Fairey, Julian L

    2015-08-18

    Measurements of disinfection byproduct (DBP) organic precursor concentrations (OPCs) are crucial to assess and improve DBP control processes. Typically, formation potential tests - specified in Standard Methods (SM) 5710-B/D - are used to measure OPCs. Here, we highlight several limitations of this protocol for dihaloacetonitriles and trihalomethanes and validate a novel Alternative Method (AM). The effects of pH, disinfectant type (free chlorine and monochloramine), and chlor(am)ine residual (CR) were examined on DBP formation in a suite of waters. Using the SM, DHAN decreased 43-47% as the CR increased from 3 to 5 mg L(-1) as Cl2, compromising OPC assessments. In contrast, a high monochloramine dose (250 mg L(-1) as Cl2) at pH 7.0 (the AM) accurately reflected OPCs. The two methods were compared for assessing DBP precursor removal through three granular activated carbon (GAC) columns in series. Breakthrough profiles assessed using the AM only showed DBP precursor sorption occurred in each column that decreased over time (p = 0.0001). Similarly, the AM facilitated ranking of three types of GAC compared in parallel columns, whereas the SM produced ambiguous results. Fluorescence intensity of a humic-like fluorophore (i.e., I345/425) correlated strongly to precursor removal in the GAC columns. The practical implications of the results are discussed. PMID:26167626

  17. 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

  18. Cryptanalysis and improvement of three-particle deterministic secure and high bit-rate direct quantum communication protocol

    NASA Astrophysics Data System (ADS)

    Liu, Zhi-Hao; Chen, Han-Wu; Wang, Dong; Li, Wen-Qian

    2014-06-01

    The three-particle deterministic secure and high bit-rate direct quantum communication protocol and its improved version are analyzed. It shows that an eavesdropper can steal the sender's secret message by the intercept-resend attack and the entanglement attack. The original version is even fragile under denial-of-service attack. As a result, some suggestions to revise them are given.

  19. 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…

  20. 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.

  1. A Taste-intensity Visual Analog Scale: An Improved Zinc Taste-test Protocol

    PubMed Central

    Zdilla, Matthew J.; Starkey, Leah D.; Saling, Julia R.

    2015-01-01

    Context Studies employing methods for measurement of zinc sulfate taste acuity have had diverse results, and although the studies have had limitations in design and implementation, they have suggested salient findings beyond the assessment of taste acuity. These findings have included, but were not limited to, relationships between zinc sulfate taste acuity and weight, levels of serum zinc, diastolic hypertension, levels of salivary carbonic anhydrase 6, and frequency of illness. However, current protocols for zinc sulfate taste assessment are problematic and warrant improvement. Objective The current study intended to compare the ranking scale for the original Bryce-Smith and Simpson zinc taste test (BS-ZTT) with a novel, taste-intensity visual analog scale (TI-VAS) to explore the potential for improved zinc sulfate taste-acuity measurement. Design Participants were administered a 0.1% solution of zinc sulfate and assessed using the 2 scales. Setting The study took place at West Liberty University, in West Liberty, WV, USA. Participants Participants were 500 West Liberty University students who volunteered to participate in the study. Results A strong, statistically significant, positive correlation existed between the BS-ZTT and the TI-VAS scores (rs = 0.79; P < .0001; n = 491). Conclusions The study found that the BS-ZTT offered a limited number of ordinal variable ranks and depended on an examiner’s evaluation. Conversely, the TI-VAS had an expanded range of variables and was independent of the examiner. The TI-VAS can be used as a taste-acuity test for zinc sulfate and as a benchmark for future analyses of zinc sulfate taste acuity and zinc taste tests. PMID:26770137

  2. 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.

  3. 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.; Asseng, S.; Basso, B.; Ewert, F.; Wallach, D.; Baigorria, G.; Winter, J. M.

    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

  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 vascular laboratory protocol for improving and managing after-hours suspected acute deep venous thrombosis.

    PubMed

    Martin, Angela H; Eckert, George; Lemmon, Gary W; Sawchuk, Alan; Dalsing, Michael C

    2014-04-01

    This study reviews the clinical and workforce impact of a suggested protocol designed for the management of suspected acute deep venous thrombosis (DVT) in patients seen after standard vascular laboratory business hours. The protocol included the use of Wells score, D-dimer and a single dose of therapeutic anticoagulant to defer venous duplex ultrasound (VDU) testing until routine business hours unless contraindicated. Information was collected on medical history, physical exam and the timing of any diagnostic studies and treatment provided. Over 15% of studies done after-hours were deemed unnecessary by our protocol and in every individual the results were negative for an acute DVT. There were no adverse events from a one-time dose of anticoagulant. Limiting emergency VDU coverage to evaluate for acute DVT based on a management protocol can eliminate unnecessary after-hours VDU testing without having a negative impact on patient care.

  6. 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

  7. 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. PMID:25190590

  8. An Improved Harvest and in Vitro Expansion Protocol for Murine Bone Marrow-Derived Mesenchymal Stem Cells

    PubMed Central

    Xu, Song; De Becker, Ann; Van Camp, Ben; Vanderkerken, Karin; Van Riet, Ivan

    2010-01-01

    Compared to bone marrow (BM) derived mesenchymal stem cells (MSCs) from human origin or from other species, the in vitro expansion and purification of murine MSCs (mMSCs) is much more difficult because of the low MSC yield and the unwanted growth of non-MSCs in the in vitro expansion cultures. We describe a modified protocol to isolate and expand murine BM derived MSCs based on the combination of mechanical crushing and collagenase digestion at the moment of harvest, followed by an immunodepletion step using microbeads coated with CD11b, CD45 and CD34 antibodies. The number of isolated mMSCs as estimated by colony forming unit-fibroblast (CFU-F) assay showed that this modified isolation method could yield 70.0% more primary colonies. After immunodepletion, a homogenous mMSC population could already be obtained after two passages. Immunodepleted mMSCs (ID-mMSCs) are uniformly positive for stem cell antigen-1 (Sca-1), CD90, CD105 and CD73 cell surface markers, but negative for the hematopoietic surface markers CD14, CD34 and CD45. Moreover the immunodepleted cell population exhibits more differentiation potential into adipogenic, osteogenic and chondrogenic lineages. Our data illustrate the development of an efficient and reliable expansion protocol increasing the yield and purity of mMSCs and reducing the overall expansion time. PMID:21197440

  9. 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

  10. Cross Layer Adaptation of Check Intervals in Low Power Listening MAC Protocols for Lifetime Improvement in Wireless Sensor Networks

    PubMed Central

    Escolar, Soledad; Chessa, Stefano; Carretero, Jesús; Marinescu, Maria-Cristina

    2012-01-01

    Preamble sampling-based MAC protocols designed for Wireless Sensor Networks (WSN) are aimed at prolonging the lifetime of the nodes by scheduling their times of activity. This scheduling exploits node synchronization to find the right trade-off between energy consumption and delay. In this paper we consider the problem of node synchronization in preamble sampling protocols. We propose Cross Layer Adaptation of Check intervals (CLAC), a novel protocol intended to reduce the energy consumption of the nodes without significantly increasing the delay. Our protocol modifies the scheduling of the nodes based on estimating the delay experienced by a packet that travels along a multi-hop path. CLAC uses routing and MAC layer information to compute a delay that matches the packet arrival time. We have implemented CLAC on top of well-known routing and MAC protocols for WSN, and we have evaluated our implementation using the Avrora simulator. The simulation results confirm that CLAC improves the network lifetime at no additional packet loss and without affecting the end-to-end delay. PMID:23112613

  11. 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.

  12. 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. PMID:25768007

  13. 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.

  14. 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. PMID:26828226

  15. 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.

  16. 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.

  17. 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

  18. 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. PMID:27239273

  19. A Locality-Aware Cooperative Cache Management Protocol to Improve Network File System Performance

    SciTech Connect

    Jiang, Song; Petrini, Fabrizio; Ding, Xiaoning; Zhang, Xiaodong

    2006-10-10

    In a distributed environment the utilization of file buffer caches in different clients may vary greatly. Cooperative caching is used to increase cache utilization by coordinating the usage of distributed caches. Existing cooperative caching protocols mainly address organizational issues, paying little attention to exploiting locality of file access patterns. We propose a locality-aware cooperative caching protocol, called LAC, that is based on analysis and manipulation of data block reuse distance to effectively predict cache utilization and the probability of data reuse. Using a dynamically controlled synchronization technique, we make local information consistently comparable among clients. The system is highly scalable in the sense that global coordination is achieved without centralized control.

  20. 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.

  1. 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

  2. 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

  3. 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

  4. 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

  5. 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

  6. 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

  7. 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

  8. Improving the quality of perinatal mental health: a health visitor-led protocol.

    PubMed

    Lewis, Anne; Ilot, Irene; Lekka, Chrysanthi; Oluboyede, Yemi

    2011-02-01

    The mental health of mothers is of significant concern to community practitioners. This paper reports on a case study exploring the success factors of a well established, health visitor-led protocol to identify and treat women with mild to moderate depression. Data were collected through interviews with a purposive sample of 12 community practitioners, a focus group of four health visitors and observation of a multidisciplinary steering group meeting. The protocol was described as an evidence-based tool and safety net that could be used flexibly to support clinical judgments and tailored to individual needs. Success factors included frontline clinician engagement and ownership, continuity of leadership to drive development and maintain momentum, comprehensive and on-going staff training, and strategic support for the protocol as a quality indicator at a time of organisational change. Quality and clinical leadership are continuing policy priorities. The protocol enabled frontline staff to lead a service innovation, providing a standardised multiprofessional approach to women's mental health needs through effective support, advice and treatment that can be measured and quality assured. PMID:21388041

  9. Progressive Mobility Protocol Reduces Venous Thromboembolism Rate in Trauma Intensive Care Patients: A Quality Improvement Project.

    PubMed

    Booth, Kathryn; Rivet, Josh; Flici, Richelle; Harvey, Ellen; Hamill, Mark; Hundley, Douglas; Holland, Katelyn; Hubbard, Sandra; Trivedi, Apurva; Collier, Bryan

    2016-01-01

    The intensive care unit (ICU) trauma population is at high risk for complications associated with immobility. The purpose of this project was to compare ICU trauma patient outcomes before and after implementation of a structured progressive mobility (PM) protocol. Outcomes included hospital and ICU stays, ventilator days, falls, respiratory failure, pneumonia, or venous thromboembolism (VTE). In the preintervention cohort, physical therapy (PT) consults were placed 53% of the time. This rose to more than 90% during the postintervention period. PT consults seen within 24 hr rose from a baseline 23% pre- to 74%-94% in the 2 highest compliance postintervention months. On average, 40% of patients were daily determined to be too unstable for mobility per protocol guidelines-most often owing to elevated intracranial pressure. During PM sessions, there were no adverse events (i.e., extubation, hypoxia, fall). There were no significant differences in clinical outcomes between the 2 cohorts regarding hospital and ICU stays, average ventilator days, mortality, falls, respiratory failure, or pneumonia overall or within ventilated patients specifically. There was, however, a difference in the incidence of VTE between the preintervention cohort (21%) and postintervention cohort (7.5%) (p = .0004). A PM protocol for ICU trauma patients is safe and may reduce patient deconditioning and VTE complications in this high-risk population. Multidisciplinary commitment, daily protocol reinforcement, and active engagement of patients/families are the cornerstones to success in this ICU PM program. PMID:27618376

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

    DOE PAGES

    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 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

  11. 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

    2016-01-01

    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 ( 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 )). PMID:27428969

  12. Propofol improves recovery of the isolated working hypertrophic heart from ischaemia-reperfusion.

    PubMed

    King, Nicola; Al Shaama, Madj; Suleiman, M-Saadeh

    2012-11-01

    The general anaesthetic propofol shows promise in protecting normal hearts against various cardiac insults, but little is known about its cardioprotective potential in hypertrophic hearts. This study tested the hypothesis that propofol at a clinically relevant dose would enhance functional recovery in hypertrophic hearts following ischaemia. Hypertrophic hearts from spontaneously hypertensive rats and hearts from their normotensive controls, Wistar Kyoto Rats, were equilibrated in the working mode prior to global normothermic ischaemia. Reperfusion commenced with 10 min in Langendorff mode, followed by 30-min working reperfusion. Functional performance was measured throughout the working mode, whilst reperfusion damage was assessed from myocardial troponin I release during Langendorff reperfusion. Where used, 4 μg/ml propofol was added 10 min before ischaemia and was washed out 10 min into working reperfusion. An additional protocol investigated recovery of hearts protected by normothermic hyperkalaemic cardioplegic arrest. Following 20-min ischaemia, reperfusion damage was significantly worse in hypertrophic hearts compared to normal hearts, whilst addition of propofol to hypertrophic hearts significantly improved the aortic flow (31 ± 5.8 vs. 11.6 ± 2.0 ml/min, n = 6-7 ± SE, p < 0.05). Propofol also conferred significant protection following 30-min ischaemia where the recovery of cardiac output and stroke volume was similar to that for cardioplegia alone. Incubation with propofol improved the NADH/NAD(+) ratio in freshly isolated cardiomyocytes from hypertrophic hearts, suggesting possible improvements in metabolic flux. These findings suggest that propofol at the clinically relevant dose of 4 μg/ml is as effective as cardioplegic arrest in protecting hypertrophic hearts against ischaemia-reperfusion.

  13. The IMPROVE_A temperature protocol for thermal/optical carbon analysis: maintaining consistency with a long-term database.

    PubMed

    Chow, Judith C; Watson, John G; Chen, L W Antony; Chang, M C Oliver; Robinson, Norman F; Trimble, Dana; Kohl, Steven

    2007-09-01

    Thermally derived carbon fractions including organic carbon (OC) and elemental carbon (EC) have been reported for the U.S. Interagency Monitoring of PROtected Visual Environments (IMPROVE) network since 1987 and have been found useful in source apportionment studies and to evaluate quartz-fiber filter adsorption of organic vapors. The IMPROVE_A temperature protocol defines temperature plateaus for thermally derived carbon fractions of 140 degrees C for OC1, 280 degrees C for OC2, 480 degrees C for OC3, and 580 degrees C for OC4 in a helium (He) carrier gas and 580 degrees C for EC1, 740 degrees C for EC2, and 840 degrees C for EC3 in a 98% He/2% oxygen (O2) carrier gas. These temperatures differ from those used previously because new hardware used for the IMPROVE thermal/optical reflectance (IMPROVE_TOR) protocol better represents the sample temperature than did the old hardware. A newly developed temperature calibration method demonstrates that these temperatures better represent sample temperatures in the older units used to quantify IMPROVE carbon fractions from 1987 through 2004. Only the thermal fractions are affected by changes in temperature. The OC and EC by TOR are insensitive to the change in temperature protocol, and therefore the long-term consistency of the IMPROVE database is conserved. A method to detect small quantities of O2 in the pure He carrier gas shows that O2 levels above 100 ppmv also affect the comparability of thermal carbon fractions but have little effect on the IMPROVE_TOR split between OC and EC.

  14. A nurse-initiated pain protocol in the ED improves pain treatment in patients with acute musculoskeletal pain.

    PubMed

    Pierik, Jorien G J; Berben, Sivera A; IJzerman, Maarten J; Gaakeer, Menno I; van Eenennaam, Fred L; van Vugt, Arie B; Doggen, Carine J M

    2016-07-01

    While acute musculoskeletal pain is a frequent complaint, its management is often neglected. An implementation of a nurse-initiated pain protocol based on the algorithm of a Dutch pain management guideline in the emergency department might improve this. A pre-post intervention study was performed as part of the prospective PROTACT follow-up study. During the pre- (15 months, n = 504) and post-period (6 months, n = 156) patients' self-reported pain intensity and pain treatment were registered. Analgesic provision in patients with moderate to severe pain (NRS ≥4) improved from 46.8% to 68.0%. Over 10% of the patients refused analgesics, resulting into an actual analgesic administration increase from 36.3% to 46.1%. Median time to analgesic decreased from 10 to 7 min (P < 0.05), whereas time to opioids decreased from 37 to 15 min (P < 0.01). Mean pain relief significantly increased to 1.56 NRS-points, in patients who received analgesic treatment even up to 2.02 points. The protocol appeared to lead to an increase in analgesic administration, shorter time to analgesics and a higher clinically relevant pain relief. Despite improvements, suffering moderate to severe pain at ED discharge was still common. Protocol adherence needs to be studied in order to optimize pain management. PMID:26968352

  15. 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.

  16. 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.

  17. 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.

  18. Improved method for the isolation of RNA from plant tissues.

    PubMed

    Logemann, J; Schell, J; Willmitzer, L

    1987-05-15

    A fast and efficient method for the isolation of RNA from plant tissues is described. Tuber tissue is homogenized in a guanidine hydrochloride-containing buffer followed by direct extraction with phenol/chloroform. The RNA is precipitated from the aqueous phase, washed with 3 M sodium acetate and 70% ethanol, and finally dissolved in water. The yield of RNA is up to 500 micrograms/g of tissue and several tests indicate intact and nondegraded RNA. This method can be adapted to a small-scale version by the use of 1.5-ml tubes, allowing rapid isolation of RNA from a larger number of samples. Finally, this method is of particular use for isolating RNA from tissues with a high polysaccharide and nuclease content such as wounded potato tubers. PMID:2441623

  19. 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

  20. 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

  1. 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

  2. Improving the capacity of the Boström-Felbinger protocol

    NASA Astrophysics Data System (ADS)

    Cai, Qing-Yu; Li, Bai-Wen

    2004-05-01

    We show that the capacity of the Boström-Felbinger (ping-pong) protocol [

    Phys. Rev. Lett. 89, 187902 (2002)
    ] can be doubled by introducing two additional unitary operations. In message mode, Alice can use one of the four unitary operations to encode two classical bits of information. Bob can decode Alice’s information by using a Bell-basis measurement. Security against eavesdropping attack can be ensured by using two conjugate measurement bases in control mode. A message authentification method can protect this communication against a man-in-the-middle attack without eavesdropping.

  3. Improved computational neutronics methods and validation protocols for the advanced test reactor

    SciTech Connect

    Nigg, D. W.; Nielsen, J. W.; Chase, B. M.; Murray, R. K.; Steuhm, K. A.; Unruh, T.

    2012-07-01

    The Idaho National Laboratory (INL) is in the process of updating 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 purposes. 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 have been 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 flexible and repeatable ATR physics code validation protocols that are consistent with applicable national standards. (authors)

  4. 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.

  5. 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

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. New advances in dynamic cardiomyoplasty: Doppler flow wire shows improved cardiac assistance in demand protocol.

    PubMed

    Rigatelli, Gianluca; Carraro, Ugo; Barbiero, Mario; Zanchetta, Mario; Pedon, Luigi; Dimopoulos, Konstantinos; Rigatelli, Giorgio; Maiolino, Pietro; Cobelli, Franco; Riccardi, Riccardo; Volta, Sergio Dalla

    2002-01-01

    No data have been published on real cardiac assistance with demand dynamic cardiomyoplasty. We tested the utility of a Doppler flow wire in measuring beat by beat aortic flow velocity and evaluating cardiac assistance in demand cardiomyoplasty patients. The technique was tested in seven patients (M/W = 6/1; age, 57.1+/-6.2 years; atrial fibrillation/ sinus rhythm = 1/6; New York Heart Association [NYHA] classification = 1.4+/-0.5). Measurements were done using a 0.018 inch peripheral Doppler flow wire advanced through a 5 French arterial femoral sheath. Three 1 minute periods with the stimulator off, and three 1 minute periods with clinical stimulation were recorded. We measured peak aortic flow velocity in all beats. Latissimus dorsi mechanogram was simultaneously recorded. Comparison between preoperative and follow-up data showed significantly higher values of tetanic fusion frequency and ejection fraction at follow-up, whereas mean NYHA class was significantly lower. Statistical analysis showed an increase in aortic flow velocity not only in the assisted versus rest period, but also in assisted versus unassisted beats (8.42+/-6.98% and 7.55+/-3.07%). A linear correlation was found between increase in flow velocity and latissimus dorsi wrap tetanic fusion frequency (r2 = 0.53). In demand dynamic cardiomyoplasty, systolic assistance is significant and correlated to the latissimus dorsi speed of contraction; a demand stimulation protocol maintains muscle properties and increases muscle performance.

  12. 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.

  13. Combining Donor Characteristics with Immunohistological Data Improves the Prediction of Islet Isolation Success

    PubMed Central

    Girman, Peter; Zacharovova, Klara; Kriz, Jan; Fabryova, Eva; Leontovyc, Ivan; Koblas, Tomas; Kosinova, Lucie; Neskudla, Tomas; Vavrova, Ema; Habart, David; Loukotova, Sarka; Zahradnicka, Martina; Lipar, Kvetoslav; Voska, Ludek; Skibova, Jelena

    2016-01-01

    Variability of pancreatic donors may significantly impact the success of islet isolation. The aim of this study was to evaluate donor factors associated with isolation failure and to investigate whether immunohistology could contribute to organ selection. Donor characteristics were evaluated for both successful (n = 61) and failed (n = 98) islet isolations. Samples of donor pancreatic tissue (n = 78) were taken for immunohistochemical examination. Islet isolations with 250000 islet equivalents were considered successful. We confirmed that BMI of less than 25 kg/m2 (P < 0.001), cold ischemia time more than 8 hours (P < 0.01), hospitalization longer than 96 hours (P < 0.05), higher catecholamine doses (P < 0.05), and edematous pancreases (P < 0.01) all unfavorably affected isolation outcome. Subsequent immunohistochemical examination of donor pancreases confirmed significant differences in insulin-positive areas (P < 0.001). ROC analyses then established that the insulin-positive area in the pancreas could be used to predict the likely success of islet isolation (P < 0.001). At the optimal cutoff point (>1.02%), sensitivity and specificity were 89% and 76%, respectively. To conclude, while the insulin-positive area, determined preislet isolation, as a single variable, is sufficient to predict isolation outcome and helps to improve the success of this procedure, its combination with the established donor scoring system might further improve organ selection. PMID:27803935

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

    PubMed

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

    2010-01-01

    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 Rapid Serial Visual Presentation (RSVP) reading) which might plausibly produce more improvement in peripheral reading speed. Twenty-eight normally-sighted young adults were trained at 10 degrees 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.

  15. 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

  16. 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.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  18. Substance Abuse Treatment and Domestic Violence. Treatment Improvement Protocol (TIP) Series 25.

    ERIC Educational Resources Information Center

    Cook, Paddy; Gartner, Constance Grant; Markl, Lise; Henderson, Randi; Brooks, Margaret K.; Wesson, Donald; Dogoloff, Mary Lou; Vitzthum, Virginia; Hayes, Elizabeth

    The major goal of this TIP, on the best practice guidelines to improve the treatment of substance abuse, is to provide clinicians, educators, and paraprofessionals with the latest findings concerning domestic violence. The information is intended to educate providers about the needs and behaviors of batterers and survivors, and how to tailor…

  19. EQUIPping Teachers: A Protocol to Guide and Improve Inquiry-Based Instruction

    ERIC Educational Resources Information Center

    Marshall, Jeff; Horton, Robert; White, Chris

    2009-01-01

    For many years, publications such as the "National Science Education Standards" (NRC 1996) and "The Science Teacher" (TST) have encouraged teachers to focus science classes more heavily on inquiry-based instructional practice. One way to improve our teaching practice is to use a benchmark assessment to obtain a solid point of reference that…

  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. Complex social intervention for multidisciplinary teams to improve patient referrals in obstetrical care: protocol for a stepped wedge study design

    PubMed Central

    Romijn, Anita; de Bruijne, Martine C; Teunissen, Pim W; de Groot, Christianne J M; Wagner, Cordula

    2016-01-01

    Introduction In obstetrics, patients often experience referral situations between different care professionals. In these multidisciplinary teams, a focus on communication and interprofessional collaboration is needed to ensure care of high quality. Crew resource management team training is increasingly being applied in healthcare settings to improve team performance and coordination. Efforts to improve communication also include tools for standardisation such as SBAR (situation, background, assessment, recommendation). Despite the growing adoption of these interventions, evidence on their effectiveness is limited, especially on patient outcomes. This article describes a study protocol to examine the effectiveness of a crew resource management team training intervention aimed at implementing the SBAR tool for structured communication during patient referrals in obstetrical care. Methods and analysis The intervention is rolled out sequentially in five hospitals and surrounding primary care midwifery practices in the Netherlands, using a stepped wedge design. The intervention involves three phases over a period of 24 months: (1) preparation, (2) training and (3) follow-up with repeated measurements. The primary outcomes are perinatal and maternal outcomes calculated using the Adverse Outcome Index. The secondary outcomes are the reaction of participating professionals to the training programme, attitudes towards safety and teamwork (Safety Attitudes Questionnaire), cohesion (Interprofessional Collaboration Measurement Scale), use of the tool for structured communication (self-reported questionnaire) and patient experiences. These secondary outcomes from professional and patient level allow triangulation and an increased understanding of the effect of the intervention on patient outcomes. Ethics and dissemination The study was approved by the Medical Ethical Committee of the VU University Medical Centre in the Netherlands and the protocol is in accordance with Dutch

  2. 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.

  3. 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. PMID:24157882

  4. 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

  5. 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

  6. Describing methods and interventions: a protocol for the systematic analysis of the perioperative quality improvement literature

    PubMed Central

    2014-01-01

    Background Quality improvement (QI) methods are widely used in surgery in an effort to improve care, often using techniques such as Plan-Do-Study-Act cycles to implement specific interventions. Explicit definition of both the QI method and quality intervention is necessary to enable the accurate replication of effective interventions in practice, facilitate cumulative learning, reduce research waste and optimise benefits to patients. This systematic review aims to assess quality of reporting of QI methods and quality interventions in perioperative care. Methods Studies reporting on quality interventions implemented in perioperative care settings will be identified. Searches will be conducted in the Ovid SP version of Medline, Scopus, the Cochrane Central Register of Controlled Trials, the Cochrane Effective Practice and Organisation of Care database and the related articles function of PubMed. The journal BMJ Quality will be searched separately. Search strategy terms will relate to (i) surgery, (ii) QI and (iii) evaluation methods. Explicit exclusion and inclusion criteria will be applied. Data from studies will be extracted using a data extraction form. The Template for Intervention Description and Replication (TIDieR) checklist will be used to evaluate quality of reporting, together with additional items aimed at assessing QI methods specifically. Systematic review registration PROSPERO http://CRD42014012845 PMID:25193066

  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. 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

  9. Improving health-related fitness in children: the fit-4-Fun randomized controlled trial study protocol

    PubMed Central

    2011-01-01

    Background Declining levels of physical fitness in children are linked to an increased risk of developing poor physical and mental health. Physical activity programs for children that involve regular high intensity physical activity, along with muscle and bone strengthening activities, have been identified by the World Health Organisation as a key strategy to reduce the escalating burden of ill health caused by non-communicable diseases. This paper reports the rationale and methods for a school-based intervention designed to improve physical fitness and physical activity levels of Grades 5 and 6 primary school children. Methods/Design Fit-4-Fun is an 8-week multi-component school-based health-related fitness education intervention and will be evaluated using a group randomized controlled trial. Primary schools from the Hunter Region in NSW, Australia, will be invited to participate in the program in 2011 with a target sample size of 128 primary schools children (age 10-13). The Fit-4-Fun program is theoretically grounded and will be implemented applying the Health Promoting Schools framework. Students will participate in weekly curriculum-based health and physical education lessons, daily break-time physical activities during recess and lunch, and will complete an 8-week (3 × per week) home activity program with their parents and/or family members. A battery of six health-related fitness assessments, four days of pedometery-assessed physical activity and a questionnaire, will be administered at baseline, immediate post-intervention (2-months) and at 6-months (from baseline) to determine intervention effects. Details of the methodological aspects of recruitment, inclusion criteria, randomization, intervention program, assessments, process evaluation and statistical analyses are described. Discussion The Fit-4-Fun program is an innovative school-based intervention targeting fitness improvements in primary school children. The program will involve a range of evidence

  10. An Internet Intervention to Improve Asthma Management: Rationale and Protocol of a Randomized Controlled Trial

    PubMed Central

    Lau, Annie YS; Dennis, Sarah; Liaw, Siaw-Teng; Coiera, Enrico

    2013-01-01

    Background Many studies have shown the effectiveness of self-management for patients with asthma. In particular, possession and use of a written asthma action plan provided by a doctor has shown to significantly improve patients’ asthma control. Yet, uptake of a written asthma action plan and preventative asthma management is low in the community, especially amongst adults. Objective A Web-based personally controlled health management system (PCHMS) called Healthy.me will be evaluated in a 2010 CONSORT-compliant 2-group (static websites verse PCHMS) parallel randomized controlled trial (RCT) (allocation ratio 1:1). Methods The PCHMS integrates an untethered personal health record with consumer care pathways and social forums. After eligibility assessment, a sample of 300 adult patients with moderate persistent asthma will be randomly assigned to one of these arms. After 12 months of using either Healthy.me or information websites (usual care arm), a post-study assessment will be conducted. Results The primary outcome measure is possession of or revision of an asthma action plan during the study. Secondary outcome measures include: (1) adherence to the asthma action plan, (2) rate of planned and unplanned visits to healthcare providers for asthma issues, (3) usage patterns of Healthy.me and attrition rates, (4) asthma control and asthma exacerbation scores, and (5) impact of asthma on life and competing demands, and days lost from work. Conclusions This RCT will provide insights into whether access to an online PCHMS will improve uptake of a written asthma action plan and preventative asthma actions. Trial Registration Trial Registration: Australian New Zealand Clinical Trials Registry ACTRN12612000716864; https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=362714 (Archived by WebCite at http://www.webcitation.org/6IYBJGRnW). PMID:23942523

  11. Manual-Protocol Inspired Technique for Improving Automated MR Image Segmentation during Label Fusion.

    PubMed

    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:27486386

  12. 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. PMID:26719442

  13. 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.

  14. 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

  15. ASCS online fault detection and isolation based on an improved MPCA

    NASA Astrophysics Data System (ADS)

    Peng, Jianxin; Liu, Haiou; Hu, Yuhui; Xi, Junqiang; Chen, Huiyan

    2014-09-01

    Multi-way principal component analysis (MPCA) has received considerable attention and been widely used in process monitoring. A traditional MPCA algorithm unfolds multiple batches of historical data into a two-dimensional matrix and cut the matrix along the time axis to form subspaces. However, low efficiency of subspaces and difficult fault isolation are the common disadvantages for the principal component model. This paper presents a new subspace construction method based on kernel density estimation function that can effectively reduce the storage amount of the subspace information. The MPCA model and the knowledge base are built based on the new subspace. Then, fault detection and isolation with the squared prediction error (SPE) statistic and the Hotelling ( T 2) statistic are also realized in process monitoring. When a fault occurs, fault isolation based on the SPE statistic is achieved by residual contribution analysis of different variables. For fault isolation of subspace based on the T 2 statistic, the relationship between the statistic indicator and state variables is constructed, and the constraint conditions are presented to check the validity of fault isolation. Then, to improve the robustness of fault isolation to unexpected disturbances, the statistic method is adopted to set the relation between single subspace and multiple subspaces to increase the corrective rate of fault isolation. Finally fault detection and isolation based on the improved MPCA is used to monitor the automatic shift control system (ASCS) to prove the correctness and effectiveness of the algorithm. The research proposes a new subspace construction method to reduce the required storage capacity and to prove the robustness of the principal component model, and sets the relationship between the state variables and fault detection indicators for fault isolation.

  16. 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

  17. Study protocol: can a school gardening intervention improve children’s diets?

    PubMed Central

    2012-01-01

    Background The current academic literature suggests there is a potential for using gardening as a tool to improve children’s fruit and vegetable intake. This study is two parallel randomised controlled trials (RCT) devised to evaluate the school gardening programme of the Royal Horticultural Society (RHS) Campaign for School Gardening, to determine if it has an effect on children’s fruit and vegetable intake. Method/Design Trial One will consist of 26 schools; these schools will be randomised into two groups, one to receive the intensive intervention as “Partner Schools” and the other to receive the less intensive intervention as “Associate Schools”. Trial Two will consist of 32 schools; these schools will be randomised into either the less intensive intervention “Associate Schools” or a comparison group with delayed intervention. Baseline data collection will be collected using a 24-hour food diary (CADET) to collect data on dietary intake and a questionnaire exploring children’s knowledge and attitudes towards fruit and vegetables. A process measures questionnaire will be used to assess each school’s gardening activities. Discussion The results from these trials will provide information on the impact of the RHS Campaign for School Gardening on children’s fruit and vegetable intake. The evaluation will provide valuable information for designing future research in primary school children’s diets and school based interventions. Trial registration ISRCTN11396528 PMID:22537179

  18. 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. PMID:10733247

  19. 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

  20. Improving practice in community-based settings: a randomized trial of supervision – study protocol

    PubMed Central

    2013-01-01

    provide descriptive information, currently unavailable in the literature, about commonly used supervision strategies in community mental health. The Phase II randomized controlled trial of gold standard supervision strategies is, to our knowledge, the first experimental study of gold standard supervision strategies in community mental health and will yield needed information about how to leverage supervision to improve clinician fidelity and client outcomes. Trial registration ClinicalTrials.gov NCT01800266 PMID:23937766

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

    PubMed

    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 C t 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

  2. 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.

  3. 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

  4. An Improved Protocol for N-Glycosylation Analysis of Gel-Separated Sialylated Glycoproteins by MALDI-TOF/TOF

    PubMed Central

    Hao, Piliang; Ren, Yan; Xie, Yongming

    2010-01-01

    Different glycoforms of some proteins have been identified as differential spots for certain diseases in 2-DE, indicating disease-related glycosylation changes. It is routine to determine the site-specific glycosylation of nonsialylated N-glycoproteins from a single gel spot, but some obstacles still exist in analyzing sialylated glycoproteins due to the lability and higher detection limit of acid glycans in MALDI-TOF/TOF analysis. Thus, we present an improved protocol here. Tryptic glycopeptides were separated and subjected to MALDI-TOF/TOF analysis, resulting in the identification of site-specific glycosylation of high-intensity glycopeptides. Sequential deglycosylation and desialylation were used to improve the identification of glycosylation sites and desialylated glycans. The site-specific glycosylation of large glycopeptides and low-intensity glycopeptides was deduced based on the masses of glycopeptides, deglycosylated peptides and desialylated glycans. By applying it to 2-DE separated human serum, the difference of N-glycosylation was successfully determined for α1-antitrypsin between different gel spots. PMID:21124746

  5. Effects of digestion protocols on the isolation and characterization of metal-metal wear particles. I. Analysis of particle size and shape.

    PubMed

    Catelas, I; Bobyn, J D; Medley, J B; Krygier, J J; Zukor, D J; Petit, A; Huk, O L

    2001-06-01

    Isolation of metal wear particles from hip simulator lubricants or tissues surrounding implants is a challenging problem because of small particle size, their tendency to agglomerate, and their potential for chemical degradation by digestion reagents. To provide realistic measurements of size, shape, and composition of metal wear particles, it is important to optimize particle isolation and minimize particle changes due to the effects of the reagents. In this study (Part I of II), transmission electron microscopy (TEM) was used to examine and compare the effects of different isolation protocols, using enzymes or alkaline solutions, on the size and shape of three different types of cobalt-based alloy particles produced from metal-metal bearings. The effect on particle composition was examined in a subsequent study (Part II). Large particles (<1200 nm) were generated by dry abrasion of CoCrMo alloy against itself and small particles (<300 nm) were generated by hip simulator testing of a metal-metal implant pair in the presence of either distilled-deionized water or a 95% bovine serum solution. The reagents changed particle size and to a lesser extent particle shape. For both large particles and small particles generated in water, the changes in size were more extensive after alkaline than after enzymatic protocols and increased with alkaline concentration and time in solution, up to twofold at 2 h and threefold at 48 h. However, when isolating particles from 95% serum, an initial protective effect of serum proteins and/or lipids was observed. Because of this protective effect, there was no significant difference in particle size and shape for both oval and needle-shaped particles after 2 h in 2N KOH and after enzymatic treatments. However, round particles were significantly smaller after 2 h in 2N KOH than after enzymatic treatments. Particle composition may also have been affected by the 2N KOH treatment, as suggested by a difference in particle contrast under TEM

  6. Improved explant method to isolate umbilical cord-derived mesenchymal stem cells and their immunosuppressive properties.

    PubMed

    Mori, Yuka; Ohshimo, Jun; Shimazu, Takahisa; He, Haiping; Takahashi, Atsuko; Yamamoto, Yuki; Tsunoda, Hajime; Tojo, Arinobu; Nagamura-Inoue, Tokiko

    2015-04-01

    The umbilical cord (UC) has become one of the major sources of mesenchymal stem cells (MSCs). The common explant method of isolating UC-derived MSCs (UC-MSCs) involves mincing the UCs into small fragments, which are then attached to a culture dish bottom from which the MSCs migrate. However, the fragments frequently float up from the bottom of the dish, thereby reducing the cell recovery rate. To overcome this problem, we demonstrate an improved explant method for UC-MSC isolation, which involves the use of a stainless steel mesh (Cellamigo(®); Tsubakimoto Chain Co.), to protect the tissue from floating after the minced fragments are aligned at regular intervals in culture dishes. The culture medium was refreshed every 3 days and the adherent cells and tissue fragments were harvested using trypsin. The number of UC-MSCs isolated from 1 g of UC using the explant method with Cellamigo was 2.9 ± 1.4 × 10(6)/g, which was significantly higher than that obtained without Cellamigo (0.66 ± 0.53 × 10(6)/g) (n = 6, p < 0.01) when cells reached 80-90% confluence. In addition, the processing and incubation time required to reach 80-90% confluence was reduced in the improved explant method compared with the conventional method. The UC-MSCs isolated using the improved method were positive for CD105, CD73, CD90, and HLA class I expression and negative for CD45 and HLA class II expression. The isolated UC-MSCs efficiently inhibited the responder T cells induced by allogeneic dendritic cells in a mixed lymphocyte reaction. Conclusively, we demonstrated that the use of Cellamigo improves the explant method for isolating UC-MSCs.

  7. Improved explant method to isolate umbilical cord-derived mesenchymal stem cells and their immunosuppressive properties.

    PubMed

    Mori, Yuka; Ohshimo, Jun; Shimazu, Takahisa; He, Haiping; Takahashi, Atsuko; Yamamoto, Yuki; Tsunoda, Hajime; Tojo, Arinobu; Nagamura-Inoue, Tokiko

    2015-04-01

    The umbilical cord (UC) has become one of the major sources of mesenchymal stem cells (MSCs). The common explant method of isolating UC-derived MSCs (UC-MSCs) involves mincing the UCs into small fragments, which are then attached to a culture dish bottom from which the MSCs migrate. However, the fragments frequently float up from the bottom of the dish, thereby reducing the cell recovery rate. To overcome this problem, we demonstrate an improved explant method for UC-MSC isolation, which involves the use of a stainless steel mesh (Cellamigo(®); Tsubakimoto Chain Co.), to protect the tissue from floating after the minced fragments are aligned at regular intervals in culture dishes. The culture medium was refreshed every 3 days and the adherent cells and tissue fragments were harvested using trypsin. The number of UC-MSCs isolated from 1 g of UC using the explant method with Cellamigo was 2.9 ± 1.4 × 10(6)/g, which was significantly higher than that obtained without Cellamigo (0.66 ± 0.53 × 10(6)/g) (n = 6, p < 0.01) when cells reached 80-90% confluence. In addition, the processing and incubation time required to reach 80-90% confluence was reduced in the improved explant method compared with the conventional method. The UC-MSCs isolated using the improved method were positive for CD105, CD73, CD90, and HLA class I expression and negative for CD45 and HLA class II expression. The isolated UC-MSCs efficiently inhibited the responder T cells induced by allogeneic dendritic cells in a mixed lymphocyte reaction. Conclusively, we demonstrated that the use of Cellamigo improves the explant method for isolating UC-MSCs. PMID:25220032

  8. Clinical study protocol for the ARCH project - computational modeling for improvement of outcome after vascular access creation.

    PubMed

    Bode, Aron; Caroli, Anna; Huberts, Wouter; Planken, Nils; Antiga, Luca; Bosboom, Marielle; Remuzzi, Andrea; Tordoir, Jan

    2011-01-01

    Despite clinical guidelines and the possibility of diagnostic vascular imaging, creation and maintenance of a vascular access (VA) remains problematic: avoiding short- and long-term VA dysfunction is challenging. Although prognostic factors for VA dysfunction have been identified in previous studies, their potential interplay at a systemic level is disregarded. Consideration of multiple prognostic patient specific factors and their complex interaction using dedicated computational modeling tools might improve outcome after VA creation by enabling a better selection of VA configuration. These computational modeling tools are developed and validated in the ARCH project: a joint initiative of four medical centers and three industrial partners (FP7-ICT-224390). This paper reports the rationale behind computational modeling and presents the clinical study protocol designed for calibrating and validating these modeling tools. The clinical study is based on the pre-operative collection of structural and functional data at a vascular level, as well as a VA functional evaluation during the follow-up period. The strategy adopted to perform the study and for data collection is also described here. PMID:21667457

  9. Electronic and postal reminders for improving immunisation coverage in children: protocol for a systematic review and meta-analysis

    PubMed Central

    Chachou, Martel J; Mukinda, Fidele K; Motaze, Villyen; Wiysonge, Charles S

    2015-01-01

    Introduction Worldwide, suboptimal immunisation coverage causes the deaths of more than one million children under five from vaccine-preventable diseases every year. Reasons for suboptimal coverage are multifactorial, and a combination of interventions is needed to improve compliance with immunisation schedules. One intervention relies on reminders, where the health system prompts caregivers to attend immunisation appointments on time or re-engages caregivers who have defaulted on scheduled appointments. We undertake this systematic review to investigate the potential of reminders using emails, phone calls, social media, letters or postcards to improve immunisation coverage in children under five. Methods and analysis We will search for published and unpublished randomised controlled trials and non-randomised controlled trials in PubMed, Scopus, CINAHL, CENTRAL, Science Citation Index, WHOLIS, Clinicaltrials.gov and the WHO International Clinical Trials Platform. We will conduct screening of search results, study selection, data extraction and risk-of-bias assessment in duplicate, resolving disagreements by consensus. In addition, we will pool data from clinically homogeneous studies using random-effects meta-analysis; assess heterogeneity of effects using the χ2 test of homogeneity; and quantify any observed heterogeneity using the I2 statistic. Ethics and dissemination This protocol does not need approval by an ethics committee because we will use publicly available data, without directly involving human participants. The results will provide updated evidence on the effects of electronic and postal reminders on immunisation coverage, and we will discuss the applicability of the findings to low and middle-income countries. We plan to disseminate review findings through publication in a peer-reviewed journal and presentation at relevant conferences. In addition, we will prepare a policymaker-friendly summary using a validated format (eg, SUPPORT Summary) and

  10. 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.

  11. 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. PMID:26075923

  12. 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.

  13. Steps toward improving diet and exercise for cancer survivors (STRIDE): a quasi-randomised controlled trial protocol

    PubMed Central

    2014-01-01

    Background 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. Methods/Design 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. Discussion 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

  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. 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

  16. 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.

  17. Characterization of the cultivable bacterial populations associated with field grown Brassica napus L.: an evaluation of sampling and isolation protocols.

    PubMed

    Croes, Sarah; Weyens, Nele; Colpaert, Jan; Vangronsveld, Jaco

    2015-07-01

    Plant-associated bacteria are intensively investigated concerning their characteristics for plant growth promotion, biocontrol mechanisms and enhanced phytoremediation efficiency. To obtain endophytes, different sampling and isolation protocols are used although their representativeness is not always clearly demonstrated. The objective of this study was to acquire representative pictures of the cultivable bacterial root, stem and leaf communities for all Brassica napus L. individuals growing on the same field. For each plant organ, genotypic identifications of the endophytic communities were performed using three replicates. Root replicates were composed of three total root systems, whereas stem and leaf replicates needed to consist of six independent plant parts in order to be representative. Greater variations between replicates were found when considering phenotypic characteristics. Correspondence analysis revealed reliable phenotypic results for roots and even shoots, but less reliable ones for leaves. Additionally, realistic Shannon-Wiener biodiversity indices were calculated for all three organs and showed similar Evenness factors. Furthermore, it was striking that all replicates and thus the whole plant contained Pseudomonas and Bacillus strains although aboveground and belowground plant tissues differed in most dominant bacterial genera and characteristics. PMID:25367683

  18. [Improvement of the method of isolation of hydrogen-forming bacteria of Clostridium genus].

    PubMed

    Pritula, I R; Tashirev, A B

    2012-01-01

    The method of isolation and quantitative account of pure cultures of obligate anaerobic hydrogen-forming clostridia is improved. A strain of hydrogen-forming bacteria Clostridium sp. BY-11 has been isolated from the association of sporulating bacteria. Quantitative indices of hydrogen synthesis and starch fermentation have been determined when growing the strain in the liquid medium. Concentration of H2 in the gas phase was 49%, microorganisms synthesized 128 1 of H2 from 1 kg of starch, the mass of starch decreased 7 times for 6 days. The mentioned indices for hydrogen synthesis and starch fermentation and for other organic model substrates in the future are the basis for creating the industrial biotechnology for production of hydrogen as the energy carrier under disposal of ecologically dangerous solid food waste. PMID:23293828

  19. [Improvement of the method of isolation of hydrogen-forming bacteria of Clostridium genus].

    PubMed

    Pritula, I R; Tashirev, A B

    2012-01-01

    The method of isolation and quantitative account of pure cultures of obligate anaerobic hydrogen-forming clostridia is improved. A strain of hydrogen-forming bacteria Clostridium sp. BY-11 has been isolated from the association of sporulating bacteria. Quantitative indices of hydrogen synthesis and starch fermentation have been determined when growing the strain in the liquid medium. Concentration of H2 in the gas phase was 49%, microorganisms synthesized 128 1 of H2 from 1 kg of starch, the mass of starch decreased 7 times for 6 days. The mentioned indices for hydrogen synthesis and starch fermentation and for other organic model substrates in the future are the basis for creating the industrial biotechnology for production of hydrogen as the energy carrier under disposal of ecologically dangerous solid food waste.

  20. Low-dose RUTF protocol and improved service delivery lead to good programme outcomes in the treatment of uncomplicated SAM: a programme report from Myanmar.

    PubMed

    James, Philip T; Van den Briel, Natalie; Rozet, Aurélie; Israël, Anne-Dominique; Fenn, Bridget; Navarro-Colorado, Carlos

    2015-10-01

    The treatment of uncomplicated severe acute malnutrition (SAM) requires substantial amounts of ready-to-use therapeutic food (RUTF). In 2009, Action Contre la Faim anticipated a shortfall of RUTF for their nutrition programme in Myanmar. A low-dose RUTF protocol to treat children with uncomplicated SAM was adopted. In this protocol, RUTF was dosed according to beneficiary's body weight, until the child reached a Weight-for-Height z-score of ≥-3 and mid-upper arm circumference ≥110 mm. From this point, the child received a fixed quantity of RUTF per day, independent of body weight until discharge. Specific measures were implemented as part of this low-dose RUTF protocol in order to improve service quality and beneficiary support. We analysed individual records of 3083 children treated from July 2009 to January 2010. Up to 90.2% of children recovered, 2.0% defaulted and 0.9% were classified as non-responders. No deaths were recorded. Among children who recovered, median [IQR] length of stay and weight gain were 42 days [28; 56] and 4.0 g kg(-1) day(-1) [3.0; 5.7], respectively. Multivariable logistic regression showed that children older than 48 months had higher odds of non-response to treatment than younger children (adjusted odds ratio: 3.51, 95% CI: 1.67-7.42). Our results indicate that a low-dose RUTF protocol, combined with specific measures to ensure good service quality and beneficiary support, was successful in treating uncomplicated SAM in this setting. This programmatic experience should be validated by randomised studies aiming to test, quantify and attribute the effect of the protocol adaptation and programme improvements presented here.

  1. Low–dose RUTF protocol and improved service delivery lead to good programme outcomes in the treatment of uncomplicated SAM: a programme report from Myanmar

    PubMed Central

    James, Philip T; Van den Briel, Natalie; Rozet, Aurélie; Israël, Anne-Dominique; Fenn, Bridget; Navarro-Colorado, Carlos

    2015-01-01

    The treatment of uncomplicated severe acute malnutrition (SAM) requires substantial amounts of ready-to-use therapeutic food (RUTF). In 2009, Action Contre la Faim anticipated a shortfall of RUTF for their nutrition programme in Myanmar. A low-dose RUTF protocol to treat children with uncomplicated SAM was adopted. In this protocol, RUTF was dosed according to beneficiary's body weight, until the child reached a Weight-for-Height z-score of ≥−3 and mid-upper arm circumference ≥110 mm. From this point, the child received a fixed quantity of RUTF per day, independent of body weight until discharge. Specific measures were implemented as part of this low-dose RUTF protocol in order to improve service quality and beneficiary support. We analysed individual records of 3083 children treated from July 2009 to January 2010. Up to 90.2% of children recovered, 2.0% defaulted and 0.9% were classified as non-responders. No deaths were recorded. Among children who recovered, median [IQR] length of stay and weight gain were 42 days [28; 56] and 4.0 g kg–1 day–1 [3.0; 5.7], respectively. Multivariable logistic regression showed that children older than 48 months had higher odds of non-response to treatment than younger children (adjusted odds ratio: 3.51, 95% CI: 1.67–7.42). Our results indicate that a low-dose RUTF protocol, combined with specific measures to ensure good service quality and beneficiary support, was successful in treating uncomplicated SAM in this setting. This programmatic experience should be validated by randomised studies aiming to test, quantify and attribute the effect of the protocol adaptation and programme improvements presented here. PMID:25850698

  2. Low-dose RUTF protocol and improved service delivery lead to good programme outcomes in the treatment of uncomplicated SAM: a programme report from Myanmar.

    PubMed

    James, Philip T; Van den Briel, Natalie; Rozet, Aurélie; Israël, Anne-Dominique; Fenn, Bridget; Navarro-Colorado, Carlos

    2015-10-01

    The treatment of uncomplicated severe acute malnutrition (SAM) requires substantial amounts of ready-to-use therapeutic food (RUTF). In 2009, Action Contre la Faim anticipated a shortfall of RUTF for their nutrition programme in Myanmar. A low-dose RUTF protocol to treat children with uncomplicated SAM was adopted. In this protocol, RUTF was dosed according to beneficiary's body weight, until the child reached a Weight-for-Height z-score of ≥-3 and mid-upper arm circumference ≥110 mm. From this point, the child received a fixed quantity of RUTF per day, independent of body weight until discharge. Specific measures were implemented as part of this low-dose RUTF protocol in order to improve service quality and beneficiary support. We analysed individual records of 3083 children treated from July 2009 to January 2010. Up to 90.2% of children recovered, 2.0% defaulted and 0.9% were classified as non-responders. No deaths were recorded. Among children who recovered, median [IQR] length of stay and weight gain were 42 days [28; 56] and 4.0 g kg(-1) day(-1) [3.0; 5.7], respectively. Multivariable logistic regression showed that children older than 48 months had higher odds of non-response to treatment than younger children (adjusted odds ratio: 3.51, 95% CI: 1.67-7.42). Our results indicate that a low-dose RUTF protocol, combined with specific measures to ensure good service quality and beneficiary support, was successful in treating uncomplicated SAM in this setting. This programmatic experience should be validated by randomised studies aiming to test, quantify and attribute the effect of the protocol adaptation and programme improvements presented here. PMID:25850698

  3. 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…

  4. Membrane Protein Production in Escherichia coli: Protocols and Rules.

    PubMed

    Angius, Federica; Ilioaia, Oana; Uzan, Marc; Miroux, Bruno

    2016-01-01

    Functional and structural studies on membrane proteins are limited by the difficulty to produce them in large amount and in a functional state. In this review, we provide protocols to achieve high-level expression of membrane proteins in Escherichia coli. The T7 RNA polymerase-based expression system is presented in detail and protocols to assess and improve its efficiency are discussed. Protocols to isolate either membrane or inclusion bodies and to perform an initial qualitative test to assess the solubility of the recombinant protein are also included. PMID:27485328

  5. Membrane Protein Production in Escherichia coli: Protocols and Rules.

    PubMed

    Angius, Federica; Ilioaia, Oana; Uzan, Marc; Miroux, Bruno

    2016-01-01

    Functional and structural studies on membrane proteins are limited by the difficulty to produce them in large amount and in a functional state. In this review, we provide protocols to achieve high-level expression of membrane proteins in Escherichia coli. The T7 RNA polymerase-based expression system is presented in detail and protocols to assess and improve its efficiency are discussed. Protocols to isolate either membrane or inclusion bodies and to perform an initial qualitative test to assess the solubility of the recombinant protein are also included.

  6. 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

    2012-01-01

    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. PMID:22653603

  7. An improved isolated working rabbit heart preparation using red cell enhanced perfusate.

    PubMed Central

    Chen, V.; Chen, Y. H.; Downing, S. E.

    1987-01-01

    The performance of isolated working rabbit hearts perfused with Krebs-Henseleit (KH) buffer was compared with those in which the buffer was supplemented with washed human red blood cells (KH + RBC) at a hematocrit of 15 percent. When perfused with KH alone at 70 cm H2O afterload and paced at 240 beats/minute, coronary flow was more than double, whereas aortic flow was 40-60 percent of that in hearts perfused with KH + RBC, regardless of left atrial filling pressures (LAFP). Peak systolic pressure reached a plateau at 120 mm Hg in KH + RBC, but at 95 mm Hg in the KH group. Stroke work, however, was similar in the two groups. Despite the high coronary flow, oxygen uptake by hearts perfused with KH was substantially less and did not respond to increases in LAFP as in those perfused with KH + RBC. There was a 20 percent drop in ATP and glycogen content after 90 minutes' perfusion. In contrast, isolated hearts perfused with RBC-enriched buffer remained stable for at least 150 minutes. Irrespective of the perfusate, triacylglycerol content of the muscle remained at similar levels throughout the course of study. Increasing RBC in the perfusate from 15 percent to 25 percent had no additional effect on cardiac performance or oxygen consumption. Our findings demonstrate that in the isolated working rabbit heart inclusion of RBC in the perfusate improves mechanical and metabolic stability by providing an adequate oxygen supply. PMID:3604287

  8. 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.

  9. 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.

  10. Optical mapping analysis of the spatiotemporal pattern of experimental tachyarrhythmia in improved isolated rat atrium preparation.

    PubMed

    Sakai, Tetsuro

    2008-04-01

    We have studied the experimental tachyarrhythmia in an improved isolated rat atrial preparation for the optical mapping of excitation spread. The atrial preparation, including the right or left auricle, was dissected from the adult rat heart, and an artificial hole was made in the center of the preparation. The preparation was then stained with a fast merocyanine-rhodanine voltage-sensitive dye (NK2761). Using a multi-element (16 x 16) photodiode array, the spread of excitation was assessed optically by timing the initiation of the action potential-related extrinsic absorption changes. In comparison with the intact isolated right atrial preparation, which we used previously, the mapping of the excitation spread was much easier and more precise because of the simple structure of the preparation. The electrical stimulation applied by a bipolar electrode evoked the sustained excitation with a fast rhythm, which we termed "experimental tachyarrhythmia" (ET). We optically mapped the spatiotemporal patterns of the spread of excitation during the initiation and the maintenance phases of ET. In most cases, a rotation of the excitatory wave around the artificial hole, i.e., a circus movement of the excitatory wave, was observed. These maps suggest that this circus movement resembles the basic mechanism of the tachycardia-like excitation observed in the intact isolated right atrial preparation. On the other hand, the appearance of an ectopic pacemaker with a fast rhythm was also observed. In some examples, two ectopic pacemakers appeared simultaneously. We consider that the experiment using the improved preparation is a superb in vitro model of atrial arrhythmia.

  11. Isolation of an oxalate-resistant Ashbya gossypii strain and its improved riboflavin production.

    PubMed

    Sugimoto, Takashi; Morimoto, Aki; Nariyama, Masashi; Kato, Tatsuya; Park, Enoch Y

    2010-01-01

    An oxalate-resistant strain of Ashbya gossypii was naturally isolated from spores grown on an oxalate-containing medium, and its medium was optimized to improve riboflavin production. Riboflavin production by the resistant strain was three-fold higher than that by the wild-type organism when grown in flask cultures. Medium optimization increased the riboflavin production by the resistant strain to 5 g l(-1), which was five-fold higher than that obtained by the wild-type strain. The productivity was reproduced in a 3-l bioreactor. During the early growth phase, the specific activity of isocitrate lyase in the oxalate-resistant strain was slightly higher than that in the wild-type strain. Proteomic analysis of the oxalate-resistant strain revealed that the expression of aldose reductase and cobalamin-independent methionine synthase decreased significantly. This is the first report that describes the natural isolation of a riboflavin producer using an antimetabolite-containing medium to enhance the riboflavin production level. This method should also be useful for improving the productivity of other bioproducts since it does not require any mutations or genetic modifications of the microorganism. PMID:19826846

  12. 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. PMID:26679437

  13. 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

  14. 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

  15. Common pitfalls of stem cell differentiation: a guide to improving protocols for neurodegenerative disease models and research.

    PubMed

    Engel, Martin; Do-Ha, Dzung; Muñoz, Sonia Sanz; Ooi, Lezanne

    2016-10-01

    Induced pluripotent stem cells and embryonic stem cells have revolutionized cellular neuroscience, providing the opportunity to model neurological diseases and test potential therapeutics in a pre-clinical setting. The power of these models has been widely discussed, but the potential pitfalls of stem cell differentiation in this research are less well described. We have analyzed the literature that describes differentiation of human pluripotent stem cells into three neural cell types that are commonly used to study diseases, including forebrain cholinergic neurons for Alzheimer's disease, midbrain dopaminergic neurons for Parkinson's disease and cortical astrocytes for neurodegenerative and psychiatric disorders. Published protocols for differentiation vary widely in the reported efficiency of target cell generation. Additionally, characterization of the cells by expression profile and functionality differs between studies and is often insufficient, leading to highly variable protocol outcomes. We have synthesized this information into a simple methodology that can be followed when performing or assessing differentiation techniques. Finally we propose three considerations for future research, including the use of physiological O2 conditions, three-dimensional co-culture systems and microfluidics to control feeding cycles and growth factor gradients. Following these guidelines will help researchers to ensure that robust and meaningful data is generated, enabling the full potential of stem cell differentiation for disease modeling and regenerative medicine. PMID:27154043

  16. Common pitfalls of stem cell differentiation: a guide to improving protocols for neurodegenerative disease models and research.

    PubMed

    Engel, Martin; Do-Ha, Dzung; Muñoz, Sonia Sanz; Ooi, Lezanne

    2016-10-01

    Induced pluripotent stem cells and embryonic stem cells have revolutionized cellular neuroscience, providing the opportunity to model neurological diseases and test potential therapeutics in a pre-clinical setting. The power of these models has been widely discussed, but the potential pitfalls of stem cell differentiation in this research are less well described. We have analyzed the literature that describes differentiation of human pluripotent stem cells into three neural cell types that are commonly used to study diseases, including forebrain cholinergic neurons for Alzheimer's disease, midbrain dopaminergic neurons for Parkinson's disease and cortical astrocytes for neurodegenerative and psychiatric disorders. Published protocols for differentiation vary widely in the reported efficiency of target cell generation. Additionally, characterization of the cells by expression profile and functionality differs between studies and is often insufficient, leading to highly variable protocol outcomes. We have synthesized this information into a simple methodology that can be followed when performing or assessing differentiation techniques. Finally we propose three considerations for future research, including the use of physiological O2 conditions, three-dimensional co-culture systems and microfluidics to control feeding cycles and growth factor gradients. Following these guidelines will help researchers to ensure that robust and meaningful data is generated, enabling the full potential of stem cell differentiation for disease modeling and regenerative medicine.

  17. 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. PMID:25946328

  18. 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.

  19. Isolation and characterization of an Ashbya gossypii mutant for improved riboflavin production.

    PubMed

    Wei, Shiping; Hurley, James; Jiang, Zhenglong; Wang, Siwen; Wang, Yuanyuan

    2012-04-01

    The use of the filamentous fungus, Ashbya gossypii, to improve riboflavin production at an industrial scale is described in this paper. A riboflavin overproducing strain was isolated by ultraviolet irradiation. Ten minutes after spore suspensions of A. gossypii were irradiated by ultraviolet light, a survival rate of 5.5% spores was observed, with 10% of the surviving spores giving rise to riboflavin-overproducing mutants. At this time point, a stable mutant of the wild strain was isolated. Riboflavin production of the mutant was two fold higher than that of the wild strain in flask culture. When the mutant was growing on the optimized medium, maximum riboflavin production could reach 6.38 g/l. It has even greater promise to increase its riboflavin production through dynamic analysis of its growth phase parameters, and riboflavin production could reach 8.12 g/l with pH was adjusted to the range of 6.0-7.0 using KH2PO4 in the later growth phase. This mutant has the potential to be used for industrial scale riboflavin production. PMID:24031850

  20. Isolation and characterization of an Ashbya gossypii mutant for improved riboflavin production

    PubMed Central

    Wei, Shiping; Hurley, James; Jiang, Zhenglong; Wang, Siwen; Wang, Yuanyuan

    2012-01-01

    The use of the filamentous fungus, Ashbya gossypii, to improve riboflavin production at an industrial scale is described in this paper. A riboflavin overproducing strain was isolated by ultraviolet irradiation. Ten minutes after spore suspensions of A. gossypii were irradiated by ultraviolet light, a survival rate of 5.5% spores was observed, with 10% of the surviving spores giving rise to riboflavin-overproducing mutants. At this time point, a stable mutant of the wild strain was isolated. Riboflavin production of the mutant was two fold higher than that of the wild strain in flask culture. When the mutant was growing on the optimized medium, maximum riboflavin production could reach 6.38 g/l. It has even greater promise to increase its riboflavin production through dynamic analysis of its growth phase parameters, and riboflavin production could reach 8.12 g/l with pH was adjusted to the range of 6.0-7.0 using KH2PO4 in the later growth phase. This mutant has the potential to be used for industrial scale riboflavin production. PMID:24031850

  1. Improvement of Isolated Myoclonus Phenotype in Myoclonus Dystonia after Pallidal Deep Brain Stimulation

    PubMed Central

    Ramdhani, Ritesh A.; Frucht, Steven J.; Behnegar, Anousheh; Kopell, Brian H.

    2016-01-01

    Background Myoclonus–dystonia is a condition that manifests predominantly as myoclonic jerks with focal dystonia. It is genetically heterogeneous with most mutations in the epsilon sarcoglycan gene (SGCE). In medically refractory cases, deep brain stimulation (DBS) has been shown to provide marked sustainable clinical improvement, especially in SGCE-positive patients. We present two patients with myoclonus–dystonia (one SGCE positive and the other SGCE negative) who have the isolated myoclonus phenotype and had DBS leads implanted in the bilateral globus pallidus internus (GPi). Methods We review their longitudinal Unified Myoclonus Rating Scale scores along with their DBS programming parameters and compare them with published cases in the literature. Results Both patients demonstrated complete amelioration of all aspects of myoclonus within 6–12 months after surgery. The patient with the SGCE-negative mutation responded just as well as the patient who was SGCE positive. High-frequency stimulation (130 Hz) with amplitudes greater than 2.5 V provided therapeutic benefit. Discussion This case series demonstrates that high frequency GPi-DBS is effective in treating isolated myoclonus in myoclonus–dystonia, regardless of the presence of SGCE mutation. PMID:26989574

  2. 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

  3. Down-regulation of hippocampal BDNF and Arc associated with improvement in aversive spatial memory performance in socially isolated rats.

    PubMed

    Pisu, Maria Giuseppina; Dore, Riccardo; Mostallino, Maria Cristina; Loi, Manila; Pibiri, Francesca; Mameli, Roberta; Cadeddu, Roberto; Secci, Pietro Paolo; Serra, Mariangela

    2011-09-12

    Rats deprived of social contact with other rats at a young age experience a form of prolonged stress that leads to long-lasting changes in behavioral profile. Such isolation is thought to be anxiogenic for these normally gregarious animals, and the abnormal reactivity of isolated rats to environmental stimuli is thought to be a product of prolonged stress. We now show that isolation of rats at weaning reduced immobility time in the forced swim test, decreased sucrose intake and preference, and down-regulated both brain-derived neurotrophic factor (BDNF) and activity-regulated cytoskeletal associated protein (Arc) in the hippocampus. In the Morris water maze, isolated rats showed a reduced latency to reach the hidden platform during training, indicative of an improved learning performance, compared with group-housed rats. The cumulative search error during place training trials indicated a reliable difference between isolated and group-housed rats on days 4 and 5. The probe trial revealed a significant decrease of the average proximity to the target location in the isolated rats suggesting an improvement in spatial memory. Isolated rats also showed an increase in the plasma level of corticosterone on the 5th day of training and increased expression of BDNF and Arc in the hippocampus on both days 1 and 5. These results show that social isolation from weaning in rats results in development of depressive-like behavior but has a positive effect on spatial learning, supporting the existence of a facilitating effect of stress on cognitive function.

  4. 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

    2015-10-01

    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.

  5. Improved method for isolation of lycopsamine from roots of comfrey (Symphytum officinale).

    PubMed

    Janes, Damjan; Kalamar, Bostjan; Kreft, Samo

    2012-07-01

    An improved method for the isolation and purification of pyrrolizidine alkaloids from comfrey (Symphytum officinale L.) roots was developed, introducing very fast, selective and ion residue-free reduction of N-oxides followed by ion-exchange chromatography giving a non-aqueous solution of alkaloids, from which solvents can be easily removed. With this procedure the use of large volumes of organic solvents, very slow reduction of N-oxides and input of additional impurities was avoided. Lycopsamine, which proved to be the major alkaloid, was additionally purified by preparative layer chromatography (PLC) and high performance liquid chromatography (HPLC). The identity of the alkaloid was confirmed by (I)H NMR spectroscopy and mass spectrometry. PMID:22908565

  6. Isolation of lipase producing Bacillus sp. from olive mill wastewater and improving its enzyme activity.

    PubMed

    Ertuğrul, Sevgi; Dönmez, Gönül; Takaç, Serpil

    2007-11-19

    The bacteria that could grow on media containing olive mill wastewater (OMW) were isolated and their lipase production capacities were investigated. The strain possessing the highest lipase activity among 17 strains grown on tributyrin agar medium was identified as Bacillus sp. The effect of initial pH on the lipase activity was investigated in tributyrin medium and pH 6 was found to be the optimal. The liquid medium composition was improved by replacing tributyrin with various carbon sources. Among the media containing different compositions of triolein, trimyristin, trilaurin, tricaprin, tricaprylin, tributyrin, triacetin, Tween 80, OMW, glucose, and whey; the medium contained 20% whey +1% triolein was found to give the highest lipase activity. Cultivation of Bacillus sp. in the optimal medium at pH 6 and 30 degrees C for 64h resulted in the extracellular and intracellular lipase activities of 15 and 168U/ml, respectively.

  7. 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

    2014-11-01

    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.

  8. 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.

  9. A comparative evaluation of culture conditions for short-term maintenance (<24 hr) of human islets isolated using the Edmonton protocol.

    PubMed

    Matsumoto, Shinichi; Goel, Shilpa; Qualley, Sabrina; Strong, D. Michael; Reems, Jo Anna

    2003-01-01

    Once human islets are isolated, they are typically transplanted into type 1 diabetic recipients within 2 h of isolation. This time restriction makes it difficult for patients to travel from distant locations to receive an islet transplant and it also makes it difficult to complete pre-release quality control assessments (i.e., endotoxin and gram stain) before the expiration of the islet product. Therefore, there were two goals for this study. The first was to measure the stability of islets after a 24 h culture period using CMRL media 1066 (CMRL) supplemented with either fetal bovine serum (FBS); albumin or insulin transferrin and selenium (ITS). The second was to determine the impact of cell concentration and media depth on islet stability. The results of the study indicated that culture recoveries at 37 degrees C with CMRL + ITS (also known as Memphis media) were higher (64.1 +/- 8.3%) than with CMRL supplemented with FBS (38.7 +/- 9.7%) or albumin (47.6 +/- 8.2%) and that post-culture islet viabilities, post-culture purities and stimulation indexes (SIs) were comparable. In the second series of experiments, the results showed that islets recoveries and SIs in cultures with low islet concentrations (300 IE/ml) were significantly better than cultures at high islet concentrations (1500 IE/ml). Additionally, at a shallow media depth (1.4 vs. 7 mm of media) the SI of the islets improved, and this effect was independent of the additive (i.e., FBS, albumin and ITS).

  10. 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

  11. 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.

  12. 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)

  13. Improving the communication reliability of body sensor networks based on the IEEE 802.15.4 protocol.

    PubMed

    Gomes, Diogo; Afonso, José A

    2014-03-01

    Body sensor networks (BSNs) enable continuous monitoring of patients anywhere, with minimum constraints to daily life activities. Although the IEEE 802.15.4 and ZigBee(®) (ZigBee Alliance, San Ramon, CA) standards were mainly developed for use in wireless sensors network (WSN) applications, they are also widely used in BSN applications because of device characteristics such as low power, low cost, and small form factor. However, compared with WSNs, BSNs present some very distinctive characteristics in terms of traffic and mobility patterns, heterogeneity of the nodes, and quality of service requirements. This article evaluates the suitability of the carrier sense multiple access-collision avoidance protocol, used by the IEEE 802.15.4 and ZigBee standards, for data-intensive BSN applications, through the execution of experimental tests in different evaluation scenarios, in order to take into account the effects of contention, clock drift, and hidden nodes on the communication reliability. Results show that the delivery ratio may decrease substantially during transitory periods, which can last for several minutes, to a minimum of 90% with retransmissions and 13% without retransmissions. This article also proposes and evaluates the performance of the BSN contention avoidance mechanism, which was designed to solve the identified reliability problems. This mechanism was able to restore the delivery ratio to 100% even in the scenario without retransmissions.

  14. Robust Optimization of Biological Protocols

    PubMed Central

    Flaherty, Patrick; Davis, Ronald W.

    2015-01-01

    When conducting high-throughput biological experiments, it is often necessary to develop a protocol that is both inexpensive and robust. Standard approaches are either not cost-effective or arrive at an optimized protocol that is sensitive to experimental variations. We show here a novel approach that directly minimizes the cost of the protocol while ensuring the protocol is robust to experimental variation. Our approach uses a risk-averse conditional value-at-risk criterion in a robust parameter design framework. We demonstrate this approach on a polymerase chain reaction protocol and show that our improved protocol is less expensive than the standard protocol and more robust than a protocol optimized without consideration of experimental variation. PMID:26417115

  15. Optimising text messaging to improve adherence to web-based smoking cessation treatment: a randomised control trial protocol

    PubMed Central

    Graham, Amanda L; Jacobs, Megan A; Cohn, Amy M; Cha, Sarah; Abroms, Lorien C; Papandonatos, George D; Whittaker, Robyn

    2016-01-01

    Introduction Millions of smokers use the Internet for smoking cessation assistance each year; however, most smokers engage minimally with even the best designed websites. The ubiquity of mobile devices and their effectiveness in promoting adherence in other areas of health behaviour change make them a promising tool to address adherence in Internet smoking cessation interventions. Text messaging is used by most adults, and messages can proactively encourage use of a web-based intervention. Text messaging can also be integrated with an Internet intervention to facilitate the use of core Internet intervention components. Methods and analysis We identified four aspects of a text message intervention that may enhance its effectiveness in promoting adherence to a web-based smoking cessation programme: personalisation, integration, dynamic tailoring and message intensity. Phase I will use a two-level full factorial design to test the impact of these four experimental features on adherence to a web-based intervention. The primary outcome is a composite metric of adherence that incorporates general utilisation metrics (eg, logins, page views) and specific feature utilisation shown to predict abstinence. Participants will be N=860 adult smokers who register on an established Internet cessation programme and enrol in its text message programme. Phase II will be a two-arm randomised trial to compare the efficacy of the web-based cessation programme alone and in conjunction with the optimised text messaging intervention on 30-day point prevalence abstinence at 9 months. Phase II participants will be N=600 adult smokers who register to use an established Internet cessation programme and enrol in text messaging. Secondary analyses will explore whether adherence mediates the effect of treatment condition on outcome. Ethics and dissemination This protocol was approved by Chesapeake IRB. We will disseminate study results through peer-reviewed manuscripts and conference

  16. Various irrigation protocols for final rinse to improve bond strengths of fiber posts inside the root canal.

    PubMed

    Bitter, Kerstin; Hambarayan, Ani; Neumann, Konrad; Blunck, Uwe; Sterzenbach, Guido

    2013-08-01

    Irrigation after post space preparation and its effects on the bond strength of different adhesive strategies are still an issue of interest. The aim of the present study was to investigate the effects of various irrigation protocols (IPs) on the push-out bond strengths of fiber posts. One-hundred and fifty extracted human anterior teeth were divided into three groups (n = 50 each) and endodontically treated. The post space was irrigated before post placement using the following five IPs in each group: IP1 (control): 5 ml of distilled water; IP2: 5.25% sodium hypochlorite (NaOCl) applied with passive ultrasonic irrigation (PUI); IP3: 1% NaOCl applied with PUI; IP4: 18% ethylenediaminetetracetic acid (EDTA) followed by 5.25% NaOCl; and IP5: 2% chlorhexidine (CHX). Fiber posts placement in each group was performed using three different adhesive strategies. A self-etch adhesive system revealed higher bond strength (16.2 ± 6.9 MPa) compared with an etch-and-rinse adhesive (8.5 ± 4.2 MPa) and a self-adhesive resin cement (9.2 ± 4.7 MPa). The use of 18% EDTA/5.25% NaOCl increased the bond strength of the self-adhesive resin cement. Conversely, this IP decreased the bond strength for the etch-and-rinse adhesive system, whilst 1% NaOCl enhanced the bond strength for the same system. Consequently, each adhesive strategy may need to be adapted to a specific IP.

  17. Microwave irradiation is a useful tool for improving isolation of actinomycetes from soil.

    PubMed

    Wang, D S; Xue, Q H; Zhu, W J; Zhao, J; Duan, J L; Shen, G H

    2013-01-01

    Actinomycetes are an important source of novel, biologically active compounds. New methods need to be developed for isolating previously unknown actinomycetes from soil. The objective of this experiment was to study microwave irradiation of soil as a means for isolating previously unknown actinomycetes. Soil samples were collected at ten elevations between 800 and 3670 m on Taibai Mountain, Shaanxi Province, China. Moistened soil samples were irradiated at 120 W heating power (2450 MHz) for 3 min using a household microwave oven. Irradiation increased total actinomycete, streptomycete, and antagonistic actinomycete counts on three types of culture media. Irradiation also increased the number of culturable actinomycete isolates. Some actinomycete isolates were culturable only after the soil was irradiated, whereas other isolates could not be cultured after irradiation. Irradiation of soil from elevations > 3000 m increased actinomycete counts significantly but had little effect on the number of culturable actinomycete isolates. In contrast, irradiation of samples from elevations < 3000 m had relatively little effect on actinomycete counts, but significantly increased the number of culturable actinomycete isolates. We used 16S rDNA sequence analysis to identify 14 actinomycete isolates that were only culturable after irradiation. Microwave irradiation of soil was helpful for isolating Streptomyces spp., Nocardia spp., Streptosporangium spp., and Lentzea spp. Slightly more than 90% of the identified actinomycete species were biologically active. In conclusion, microwave irradiation is a useful tool for isolating biologically active actinomycetes from soil. PMID:23718054

  18. Microwave irradiation is a useful tool for improving isolation of actinomycetes from soil.

    PubMed

    Wang, D S; Xue, Q H; Zhu, W J; Zhao, J; Duan, J L; Shen, G H

    2013-01-01

    Actinomycetes are an important source of novel, biologically active compounds. New methods need to be developed for isolating previously unknown actinomycetes from soil. The objective of this experiment was to study microwave irradiation of soil as a means for isolating previously unknown actinomycetes. Soil samples were collected at ten elevations between 800 and 3670 m on Taibai Mountain, Shaanxi Province, China. Moistened soil samples were irradiated at 120 W heating power (2450 MHz) for 3 min using a household microwave oven. Irradiation increased total actinomycete, streptomycete, and antagonistic actinomycete counts on three types of culture media. Irradiation also increased the number of culturable actinomycete isolates. Some actinomycete isolates were culturable only after the soil was irradiated, whereas other isolates could not be cultured after irradiation. Irradiation of soil from elevations > 3000 m increased actinomycete counts significantly but had little effect on the number of culturable actinomycete isolates. In contrast, irradiation of samples from elevations < 3000 m had relatively little effect on actinomycete counts, but significantly increased the number of culturable actinomycete isolates. We used 16S rDNA sequence analysis to identify 14 actinomycete isolates that were only culturable after irradiation. Microwave irradiation of soil was helpful for isolating Streptomyces spp., Nocardia spp., Streptosporangium spp., and Lentzea spp. Slightly more than 90% of the identified actinomycete species were biologically active. In conclusion, microwave irradiation is a useful tool for isolating biologically active actinomycetes from soil.

  19. Investigating the organisational impacts of quality improvement: a protocol for a realist evaluation of improvement approaches drawing on the Resource Based View of the Firm

    PubMed Central

    Burton, Christopher R; Rycroft Malone, Jo; Robert, Glenn; Willson, Alan; Hopkins, Angela

    2014-01-01

    Introduction Little is understood about the role of quality improvement in enabling health organisations to survive and thrive in the contemporary context of financial and economic challenges. We will draw on the theoretical foundations of the ‘Resource Based View of the Firm’ (RBV) to develop insights into why health organisations engage in improvement work, how impacts are conceptualised, and ‘what works’ in delivering these impacts. Specifically, RBV theorises that the mix and use of resources across different organisations may explain differences in performance. Whether improvement work influences these resources is unclear. Methods and analysis Case study research will be conducted across health organisations participating in four approaches to improvement, including: a national improvement programme; a multiorganisational partnership around implementation; an organisational strategy for quality improvement; and a coproduction project designed to enhance the experience of a clinical service from the perspective of patients. Data will comprise in-depth interviews with key informants, observation of key events and documents; analysed within and then across cases. Adopting a realist perspective, the core tenets of RBV will be evaluated as a programme theory, focusing on the interplay between organisational conditions and behavioural or resource responses that are reported through engagement in improvement. Ethics and dissemination The study has been approved by Bangor University Ethics Committee. The investigation will not judge the relative merits of different approaches to healthcare quality improvement. Rather, we will develop unique insights into the organisational consequences, and dependencies of quality improvement, providing an opportunity to add to the explanatory potential of RBV in this and other contexts. In addition to scientific and lay reports of the study findings, research outputs will include a framework for constructing the economic

  20. 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).

  1. Spatial and temporal variability of compound-specific stable isotope (CSSI) biomarkers in soil and sediment tracing: towards improved sampling protocols

    NASA Astrophysics Data System (ADS)

    Reiffarth, Dominic; Petticrew, Ellen; Owens, Philip; Lobb, David

    2016-04-01

    The use of CSSI in biomarkers, specifically fatty acids and derivatives thereof, has recently been investigated as a potential tracer in soil and sediment fingerprinting. The use of CSSIs is of interest because of the potential to discern sediment providence based on land use, which is often difficult or not possible with other tracing techniques alone, such as geochemistry and fallout radionuclides. However, challenges exist in producing a representative sample of potential source materials. This presentation focuses on the development of improved protocols for sample collection. The data presented here are part of a larger investigation into using CSSIs as tracers in an agricultural watershed (South Tobacco Creek) in southern Manitoba, Canada. Extensive sampling was performed throughout the 2012 and 2013 growing seasons in several locations within the watershed, with a focus on capturing within and between field spatial and temporal variability in one particular sub-watershed (the "Stepler" watershed). The Stepler watershed provided a unique opportunity to perform sampling in a natural environment where agricultural crops were hydrologically separated, thereby allowing for a sampling regime of transects strategically placed with little influence from nearby crops. A portion of the data which has been analyzed, showing temporal and spatial variability in terms of carbon stable isotope signal, biomarker concentrations and soil organic carbon, is presented. As CSSI protocols for tracing are still in development, these data aid in determining the robustness of the technique as well as helping to inform sampling approaches.

  2. The Internal Coherence Assessment Protocol & Developmental Framework: Building the Organizational Capacity for Instructional Improvement in Schools. Research Paper

    ERIC Educational Resources Information Center

    Elmore, Richard F.; Forman, Michelle L.; Stosich, Elizabeth L.; Bocala, Candice

    2014-01-01

    Purpose: In this paper we describe the Internal Coherence (IC) model of assessment and professional development, a set of clinical tools and practices designed to help practitioners foster the organizational conditions required for whole-school instructional improvement. Proposed Conceptual Argument: We argue that the data captured by the IC…

  3. Improved Picture Naming in Chronic Aphasia after Tms to Part of Right Broca's Area: An Open-Protocol Study

    ERIC Educational Resources Information Center

    Naeser, M.A.; Martin, P.I.; Nicholas, M.; Baker, E.H.; Seekins, H.; Kobayashi, M.; Theoret, H.; Fregni, F.; Maria-Tormos, J.; Kurland, J.; Doron, K.W.; Pascual-Leone, A.

    2005-01-01

    Functional imaging studies with nonfluent aphasia patients have observed ''over-activation'' in right (R) language homologues. This may represent a maladaptive strategy; suppression may result in language improvement. We applied slow, 1 Hz repetitive transcranial magnetic stimulation (rTMS) to an anterior portion of R Broca's homologue daily, for…

  4. Determining Specificity of Motor Imagery Training for Upper Limb Improvement in Chronic Stroke Patients: A Training Protocol and Pilot Results

    ERIC Educational Resources Information Center

    Craje, Celine

    2010-01-01

    Motor imagery (MI) refers to the mental rehearsal of a movement without actual motor output. MI training has positive effects on upper limb recovery after stroke. However, until now it is unclear whether this effect is specific to the trained task or a more general motor skill improvement. This study was set up to advance our insights into the…

  5. Effects of different Brush Border Membrane Vesicle isolation protocols on proteomic analysis of Cry1Ac binding proteins from the midgut of Helicoverpa armigera

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The brush border membrane vesicles (BBMV) isolated from midgut cells of insect have been widely used for studying of the binding receptors and action mode of Cry proteins produced by Bacillus thuringiensis (Bt). There are several methods for isolating insect BBMV used in one-dimension electrophoresi...

  6. A multi-level system quality improvement intervention to reduce racial disparities in hypertension care and control: study protocol

    PubMed Central

    2013-01-01

    Background Racial disparities in blood pressure control have been well documented in the United States. Research suggests that many factors contribute to this disparity, including barriers to care at patient, clinician, healthcare system, and community levels. To date, few interventions aimed at reducing hypertension disparities have addressed factors at all of these levels. This paper describes the design of Project ReD CHiP (Reducing Disparities and Controlling Hypertension in Primary Care), a multi-level system quality improvement project. By intervening on multiple levels, this project aims to reduce disparities in blood pressure control and improve guideline concordant hypertension care. Methods Using a pragmatic trial design, we are implementing three complementary multi-level interventions designed to improve blood pressure measurement, provide patient care management services and offer expanded provider education resources in six primary care clinics in Baltimore, Maryland. We are staggering the introduction of the interventions and will use Statistical Process Control (SPC) charting to determine if there are changes in outcomes at each clinic after implementation of each intervention. The main hypothesis is that each intervention will have an additive effect on improvements in guideline concordant care and reductions in hypertension disparities, but the combination of all three interventions will result in the greatest impact, followed by blood pressure measurement with care management support, blood pressure measurement with provider education, and blood pressure measurement only. This study also examines how organizational functioning and cultural competence affect the success of the interventions. Discussion As a quality improvement project, Project ReD CHiP employs a novel study design that specifically targets multi-level factors known to contribute to hypertension disparities. To facilitate its implementation and improve its sustainability, we have

  7. Evidence-based improvement of the National Trauma Triage Protocol: The Glasgow Coma Scale versus Glasgow Coma Scale motor subscale

    PubMed Central

    Brown, Joshua B.; Forsythe, Raquel M.; Stassen, Nicole A.; Peitzman, Andrew B.; Billiar, Timothy R.; Sperry, Jason L.; Gestring, Mark L.

    2015-01-01

    BACKGROUND Ideal triage uses simple criteria to identify severely injured patients. Glasgow Coma Scale motor (GCSm) may be easier for field use and was considered for the National Trauma Triage Protocol (NTTP). This study evaluated performance of the NTTP if GCSm is substituted for the current GCS score ≤ 13 criterion. METHODS Subjects in the National Trauma Data Bank undergoing scene transport were included. Presence of NTTP physiologic (Step 1) and anatomic (Step 2) criteria was determined. GCSm score ≤ 5 was defined as a positive criterion. Trauma center need (TCN) was defined as Injury Severity Score (ISS) > 15, intensive care unit admission, urgent operation, or emergency department death. Test characteristics were calculated to predict TCN. Area under the curve was compared between GCSm and GCS scores, individually and within the NTTP. Logistic regression was used to determine the association of GCSm score ≤ 5 and GCS score ≤ 13 with TCN after adjusting for other triage criteria. Predicted versus actual TCN was compared. RESULTS There were 811,143 subjects. Sensitivity was lower (26.7% vs. 30.3%), specificity was higher (95.1% vs. 93.1%), and accuracy was similar (66.1% vs. 66.3%) for GCSm score ≤ 5 compared with GCS score ≤ 13. Incorporated into the NTTP Steps 1 + 2, GCSm score ≤ 5 traded sensitivity (60.4% vs. 62.1%) for specificity (67.1% vs. 65.7%) with similar accuracy (64.2% vs. 64.2%) to GCS score ≤ 13. There was no difference in the area under the curve between GCSm score ≤ 5 and GCS score ≤ 13 when incorporated into the NTTP Steps 1 + 2 (p = 0.10). GCSm score ≤ 5 had a stronger association with TCN (odds ratio, 3.37; 95% confidence interval, 3.27–3.48; p < 0.01) than GCS score ≤ 13 (odds ratio, 3.03; 95% confidence interval, 2.94–3.13; p < 0.01). GCSm had a better fit of predicted versus actual TCN than GCS at the lower end of the scales. CONCLUSION GCSm score ≤ 5 increases specificity at the expense of sensitivity

  8. Communication complexity protocols for qutrits

    SciTech Connect

    Tamir, Boaz

    2007-03-15

    Consider a function where its entries are distributed among many parties. Suppose each party is allowed to send only a limited amount of information to a referee. The referee can use a classical protocol to compute the value of the global function. Is there a quantum protocol improving the results of all classical protocols? In a recent work Brukner et al. showed the deep connection between such problems and the theory of Bell inequalities. Here we generalize the theory to trits. There, the best classical protocol fails whereas the quantum protocol yields the correct answer.

  9. 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.

  10. Optimization protocols and improved strategies of cross-linked enzyme aggregates technology: current development and future challenges.

    PubMed

    Cui, Jian Dong; Jia, Shi Ru

    2015-03-01

    Cross-linked enzyme aggregate (CLEA) technology has been regarded as an effective carrier-free immobilization method. This method is very attractive due to its simplicity and robustness, as well as for the possibility of using the crude enzyme extract and the opportunity to co-immobilize multiple different enzymes. The resulting CLEAs generally exhibit high catalyst productivities, improved storage and operational stability and are easy to recycle. Nowadays, although the technology has been applied to various enzymes, some undesirable properties have limited its further application. To overcome these limitations, novel strategies have been developing in recent years. This mini-review focuses on process optimization, new improved strategies and the latest advances on CLEAs technology.

  11. 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.

  12. 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. PMID:24519013

  13. REFLEX, a social-cognitive group treatment to improve insight in schizophrenia: study protocol of a multi-center RCT

    PubMed Central

    2011-01-01

    Background Insight is impaired in a majority of people with schizophrenia. Impaired insight is associated with poorer outcomes of the disorder. Based on existing literature, we developed a model that explains which processes may possibly play a role in impaired insight. This model was the starting point of the development of REFLEX: a brief psychosocial intervention to improve insight in schizophrenia. REFLEX is a 12-sessions group training, consisting of three modules of four sessions each. Modules in this intervention are: "coping with stigma", "you and your personal narrative", and "you in the present". Methods/Design REFLEX is currently evaluated in a multicenter randomized controlled trial. Eight mental health institutions in the Netherlands participate in this evaluation. Patients are randomly assigned to either REFLEX or an active control condition, existing of cognitive remediation exercises in a group. In a subgroup of patients, fMRI scans are made before and after training in order to assess potential haemodynamic changes associated with the effects of the training. Discussion REFLEX is one of the few interventions aiming specifically to improving insight in schizophrenia and has potential value for improving insight. Targeting insight in schizophrenia is a complex task, that comes with several methodological issues. These issues are addressed in the discussion of this paper. Trial registration Current Controlled Trials: ISRCTN50247539 PMID:21975132

  14. Assessing the effectiveness and cost effectiveness of adaptive e-Learning to improve dietary behaviour: protocol for a systematic review

    PubMed Central

    2010-01-01

    Background The composition of habitual diets is associated with adverse or protective effects on aspects of health. Consequently, UK public health policy strongly advocates dietary change for the improvement of population health and emphasises the importance of individual empowerment to improve health. A new and evolving area in the promotion of dietary behavioural change is e-Learning, the use of interactive electronic media to facilitate teaching and learning on a range of issues, including diet and health. The aims of this systematic review are to determine the effectiveness and cost-effectiveness of adaptive e-Learning for improving dietary behaviours. Methods/Design The research will consist of a systematic review and a cost-effectiveness analysis. Studies will be considered for the review if they are randomised controlled trials, involving participants aged 13 or over, which evaluate the effectiveness or efficacy of interactive software programmes for improving dietary behaviour. Primary outcome measures will be those related to dietary behaviours, including estimated intakes of energy, nutrients and dietary fibre, or the estimated number of servings per day of foods or food groups. Secondary outcome measures will be objective clinical measures that are likely to respond to changes in dietary behaviours, such as anthropometry or blood biochemistry. Knowledge, self-efficacy, intention and emotion will be examined as mediators of dietary behaviour change in order to explore potential mechanisms of action. Databases will be searched using a comprehensive four-part search strategy, and the results exported to a bibliographic database. Two review authors will independently screen results to identify potentially eligible studies, and will independently extract data from included studies, with any discrepancies at each stage settled by a third author. Standardised forms and criteria will be used. A descriptive analysis of included studies will describe study design

  15. Study protocol: using the Q-STEPS to assess and improve the quality of physical activity programmes for the elderly

    PubMed Central

    2012-01-01

    Background Aging is one of the most important and obvious phenomenon observed in our society. In the past years, there has been a growing concern in designing physical activity (PA) programmes for elderly people, because evidence suggests that such health promotion interventions may reduce the deleterious effects of the ageing process. Accordingly, a growing body of literature points to the importance of a sound approach to planning and evaluation in order to improve the quality of PA programmes. However, while numerous PA programmes have been designed for the elderly in recent years, their evaluation has been scarce. Quality management processes and tools provide a practical way for organisations to assess, identify and shed light on the areas requiring improvement. The Quality Self-assessment Tool for Exercise Programmes for Seniors (Q-STEPS) seems to provide a framework tailored to evaluate PA programmes for the elderly. Findings The primary purpose of this study is 1) to determine feasibility, acceptability and usability of the Q-STEPS. Secondary purposes of the study are: 2) to examine the quality of the PA programmes for elderly people developed by the Portuguese Local Administration over a three-year period of self-assessments in terms of: a) Enabler domains (Leadership, Policy and Strategy, People, Partnership and Resources, Processes); b) Result domains (Customer Results, People Results, Society Results and Key Performance Results); 3) to estimate the association between the use of Q-STEPS and some indicators relating to the elderly participants, during the three self-assessments, such as: attendance rates, physical fitness, health-related quality of life and the elderly’s perceived quality of the programme. The study will be conducted in PA programmes for elderly adults from mainland Portuguese municipalities over a three-year period. The project will adopt a participative quality improvement approach that features annual learning cycles of: 1) self

  16. Ethics in public health research: a research protocol to evaluate the effectiveness of public-private partnerships as a means to improve health and welfare systems worldwide.

    PubMed

    Barr, Donald A

    2007-01-01

    Public-private partnerships have become a common approach to health care problems worldwide. Many public-private partnerships were created during the late 1990s, but most were focused on specific diseases such as HIV/AIDS, tuberculosis, and malaria. Recently there has been enthusiasm for using public-private partnerships to improve the delivery of health and welfare services for a wider range of health problems, especially in developing countries. The success of public-private partnerships in this context appears to be mixed, and few data are available to evaluate their effectiveness. This analysis provides an overview of the history of health-related public-private partnerships during the past 20 years and describes a research protocol commissioned by the World Health Organization to evaluate the effectiveness of public-private partnerships in a research context.

  17. Effectiveness of a smartphone application for improving healthy lifestyles, a randomized clinical trial (EVIDENT II): study protocol

    PubMed Central

    2014-01-01

    Background New technologies could facilitate changes in lifestyle and improve public health. However, no large randomized, controlled studies providing scientific evidence of the benefits of their use have been made. The aims of this study are to develop and validate a smartphone application, and to evaluate the effect of adding this tool to a standardized intervention designed to improve adherence to the Mediterranean diet and to physical activity. An evaluation is also made of the effect of modifying habits upon vascular structure and function, and therefore on arterial aging. Methods/Design A randomized, double-blind, multicenter, parallel group clinical trial will be carried out. A total of 1215 subjects under 70 years of age from the EVIDENT trial will be included. Counseling common to both groups (control and intervention) will be provided on adaptation to the Mediterranean diet and on physical activity. The intervention group moreover will receive training on the use of a smartphone application designed to promote a healthy diet and increased physical activity, and will use the application for three months. The main study endpoints will be the changes in physical activity, assessed by accelerometer and the 7-day Physical Activity Recall (PAR) interview, and adaptation to the Mediterranean diet, as evaluated by an adherence questionnaire and a food frequency questionnaire (FFQ). Evaluation also will be made of vascular structure and function based on central arterial pressure, the radial augmentation index, pulse velocity, the cardio-ankle vascular index, and carotid intima-media thickness. Discussion Confirmation that the new technologies are useful for promoting healthier lifestyles and that their effects are beneficial in terms of arterial aging will have important clinical implications, and may contribute to generalize their application in favor of improved population health. Trial registration Clinical Trials.gov Identifier: NCT02016014 PMID:24628961

  18. Effectiveness of implementation strategies in improving physician adherence to guideline recommendations in heart failure: a systematic review protocol

    PubMed Central

    Van Spall, Harriette G C; Shanbhag, Deepti; Gabizon, Itzhak; Ibrahim, Quazi; Graham, Ian D; Harlos, Karen; Haynes, R Brian; Connolly, Stuart J

    2016-01-01

    Introduction The uptake of Clinical Practice Guideline (CPG) recommendations that improve outcomes in heart failure (HF) remains suboptimal. We will conduct a systematic review to identify implementation strategies that improve physician adherence to class I recommendations, those with clear evidence that benefits outweigh the risks. We will use American, Canadian and European HF guidelines as our reference. Methods and analysis We will conduct a literature search in the databases of MEDLINE, EMBASE, HEALTHSTAR, CINAHL, Cochrane Library, Campbell Collaboration, Joanna Briggs Institute Evidence Based Practice, Centre for Reviews and Dissemination and Evidence Based Practice Centres. We will include prospective studies evaluating implementation interventions aimed at improving uptake of class I CPG recommendations in HF. We will extract data in duplicate. We will classify interventions according to their level of application (ie, provider, organisation, systems level) and common underlying characteristics (eg, education, decision-support, financial incentives) using the Cochrane Effective Practice and Organisation of Care Taxonomy. We will assess the impact of the intervention on adherence to the CPGs. Outcomes will include proportion of eligible patients who were: prescribed a CPG-recommended pharmacological treatment; referred for device consideration; provided self-care education at discharge; and provided left ventricular function assessment. We will include clinical outcomes such as hospitalisations, readmissions and mortality, if data is available. We will identify the common elements of successful and failing interventions, and examine the context in which they were applied, using the Process Redesign contextual framework. We will synthesise the results narratively and, if appropriate, will pool results for meta-analysis. Discussion and dissemination In this review, we will assess the impact of implementation strategies and contextual factors on physician

  19. Treating Adult Asthma Exacerbations With a 2-Day Course of Dexamethasone in the Emergency Department: New Protocols to Improve Compliance.

    PubMed

    Evans, Dian Dowling; Clinton Shedd, Glenn

    2016-01-01

    The Research to Practice column is intended to improve the research critique skills of the advanced practice registered nurse and emergency nurse (RN) and to assist with the translation of research into practice. For each column, a topic and a research study are selected. The stage is set with a case presentation. The research article is then reviewed and critiqued, and the findings are discussed in relation to the case presented. In the current column, we examine the findings of from their article, titled "Two Days of Dexamethasone Versus 5 Days of Prednisone in the Treatment of Acute Asthma: A Randomized Controlled Trial." PMID:27482988

  20. Home based exercise to improve turning and mobility performance among community dwelling older adults: protocol for a randomized controlled trial

    PubMed Central

    2014-01-01

    Background Turning is a common activity for older people, and is one of the activities commonly associated with falls during walking. Falls that occur while walking and turning have also been associated with an increased risk of hip fracture in older people. Despite the importance of stability during turning, there has been little focus on identifying this impairment in at risk older people, or in evaluating interventions aiming to improve this outcome. This study will evaluate the effectiveness of a 16 week tailored home based exercise program in older adults aged (50 years and above) who were identified as having unsteadiness during turning. Methods/Design A single blind randomized controlled trial will be conducted, with assessors blind to group allocation. Study participants will be aged 50 years and above, living in the community and have been identified as having impaired turning ability [outside of age and gender normal limits on the Step Quick Turn (180 degree turn) task on the Neurocom® Balance Master with long plate]. After a comprehensive baseline assessment, those classified as having balance impairment while turning will be randomized to intervention or control group. The intervention group will receive a 16 week individualized balance and strength home exercise program, based on the Otago Exercise Program with additional exercises focused on improving turning ability. Intervention group will attend four visit to the assessment centre over 16 weeks period, for provision, monitoring, modification of the exercise and encourage ongoing participation. Participants in the control group will continue with their usual activities. All participants will be re-assessed on completion of the 16 week program. Primary outcome measures will be the Step Quick Turn Test and Timed-Up and Go test. Secondary outcomes will include other clinical measures of balance, psychological aspects of falls, incidence of falls and falls risk factors. Discussion Results of this study

  1. Study protocol for a randomized controlled trial to assess the feasibility of an open label intervention to improve hydroxyurea adherence in youth with sickle cell disease

    PubMed Central

    Smaldone, Arlene; Findley, Sally; Bakken, Suzanne; Matiz, L. Adriana; Rosenthal, Susan L.; Jia, Haomiao; Matos, Sergio; Manwani, Deepa; Green, Nancy S.

    2016-01-01

    Background Community health workers (CHW) are increasingly recognized as a strategy to improve health outcomes for the underserved with chronic diseases but has not been formally explored in adolescents with sickle cell disease (SCD). SCD primarily affects African American, Hispanic and other traditionally underserved populations. Hydroxyurea (HU), an oral, once-daily medication, is the only approved therapeutic drug for sickle cell disease and markedly reduces symptoms, morbidity and mortality and improves quality of life largely by increasing hemoglobin F blood levels. This paper presents the rationale, study design and protocol for an open label randomized controlled trial to improve parent-youth partnerships in self-management and medication adherence to HU in adolescents with SCD. Methods/Design A CHW intervention augmented by text messaging was designed for adolescents with SCD ages 10–18 years and their parents to improve daily HU adherence. Thirty adolescent parent dyads will be randomized with 2:1 intervention group allocation. Intervention dyads will establish a relationship with a culturally aligned CHW to identify barriers to HU use, identify cues to build a habit, and develop a dyad partnership to improve daily HU adherence and achieve their individualized “personal best” hemoglobin F target. Intervention feasibility, acceptability and efficacy will be assessed via a 2-site trial. Outcomes of interest are HU adherence, dyad self-management communication, quality of life, and resource use. Discussion Despite known benefits, poor HU adherence is common. If feasible and acceptable, the proposed intervention may improve health of underserved adolescents with SCD by enhancing long-term HU adherence. PMID:27327779

  2. Improved online δ18O measurements of nitrogen- and sulfur-bearing organic materials and a proposed analytical protocol

    USGS Publications Warehouse

    Qi, H.; Coplen, T.B.; Wassenaar, L.I.

    2011-01-01

    It is well known that N2 in the ion source of a mass spectrometer interferes with the CO background during the δ18O measurement of carbon monoxide. A similar problem arises with the high-temperature conversion (HTC) analysis of nitrogenous O-bearing samples (e.g. nitrates and keratins) to CO for δ18O measurement, where the sample introduces a significant N2 peak before the CO peak, making determination of accurate oxygen isotope ratios difficult. Although using a gas chromatography (GC) column longer than that commonly provided by manufacturers (0.6 m) can improve the efficiency of separation of CO and N2 and using a valve to divert nitrogen and prevent it from entering the ion source of a mass spectrometer improved measurement results, biased δ18O values could still be obtained. A careful evaluation of the performance of the GC separation column was carried out. With optimal GC columns, the δ18O reproducibility of human hair keratins and other keratin materials was better than ±0.15 ‰ (n = 5; for the internal analytical reproducibility), and better than ±0.10 ‰ (n = 4; for the external analytical reproducibility).

  3. Randomised controlled trial of an in-home monitoring intervention to improve health outcomes for type 2 diabetes: study protocol.

    PubMed

    Carlisle, Karen; Warren, Robin; Scuffham, Paul; Cheffins, Tracy

    2012-01-01

    Type 2 diabetes is a leading cause of death and morbidity and is a health priority in Australia. This randomised controlled trial will explore whether remote access to clinical care, supported by telehealth technologies over high speed broadband, leads to improved diabetes control in a way that benefits patients, carers and clinicians and improves the overall health system. People in the intervention arm of the trial will receive additional diabetes care from a care coordinator nurse via an in-home broadband communication device that can capture clinical measures, provide regular health assessments and videoconference with other health professionals when required. Patients in the control arm of the trial will receive usual care from their GP and participate in the clinical measurement and quality of life components of the evaluation. The trial evaluation will include biomedical, psychological, self-management and quality of life measures. Data on utilisation rates and satisfaction with the technology will be collected and cost -effectiveness analyses undertaken. The role of this technology in health care reform will be explored. PMID:23138078

  4. Step-by-step protocol to perfuse and dissect the mouse parotid gland and isolation of high-quality RNA from murine and human parotid tissue.

    PubMed

    Watermann, Christoph; Valerius, Klaus Peter; Wagner, Steffen; Wittekindt, Claus; Klussmann, Jens Peter; Baumgart-Vogt, Eveline; Karnati, Srikanth

    2016-04-01

    Macroscopic identification and surgical removal of the mouse parotid gland is demanding because of its anatomic location and size. Moreover, the mouse parotid gland contains high concentrations of RNases, making it difficult to isolate high-quality RNA. So far, appropriate methods for optimal perfusion-fixation and dissection of mouse parotid glands, as well as the isolation of high quality RNA from this tissue, are not available. Here we present a simple, optimized, step-by-step surgical method to perfuse and isolate murine parotid glands. We also compared two common RNA extraction methods (RNeasy Mini Kit versus TRIzol) for their yields of high-quality, intact RNA from human and murine parotid gland tissues that were either snap-frozen or immersed in RNAlater stabilization solution. Mouse parotid tissue that was perfused and immersed in RNAlater and human samples immersed in RNAlater exhibited the best RNA quality, independent of the isolation method. PMID:27071609

  5. Normalization of coagulopathy is associated with improved outcome after isolated traumatic brain injury.

    PubMed

    Epstein, Daniel S; Mitra, Biswadev; Cameron, Peter A; Fitzgerald, Mark; Rosenfeld, Jeffrey V

    2016-07-01

    Acute traumatic coagulopathy (ATC) has been reported in the setting of isolated traumatic brain injury (iTBI) and is associated with poor outcomes. We aimed to evaluate the effectiveness of procoagulant agents administered to patients with ATC and iTBI during resuscitation, hypothesizing that timely normalization of coagulopathy may be associated with a decrease in mortality. A retrospective review of the Alfred Hospital trauma registry, Australia, was conducted and patients with iTBI (head Abbreviated Injury Score [AIS] ⩾3 and all other body AIS <3) and coagulopathy (international normalized ratio ⩾1.3) were selected for analysis. Data on procoagulant agents used (fresh frozen plasma, platelets, cryoprecipitate, prothrombin complex concentrates, tranexamic acid, vitamin K) were extracted. Among patients who had achieved normalization of INR or survived beyond 24hours and were not taking oral anticoagulants, the association of normalization of INR and death at hospital discharge was analyzed using multivariable logistic regression analysis. There were 157 patients with ATC of whom 68 (43.3%) received procoagulant products within 24hours of presentation. The median time to delivery of first products was 182.5 (interquartile range [IQR] 115-375) minutes, and following administration of coagulants, time to normalization of INR was 605 (IQR 274-1146) minutes. Normalization of INR was independently associated with significantly lower mortality (adjusted odds ratio 0.10; 95% confidence interval 0.03-0.38). Normalization of INR was associated with improved mortality in patients with ATC in the setting of iTBI. As there was a substantial time lag between delivery of products and eventual normalization of coagulation, specific management of coagulopathy should be implemented as early as possible. PMID:26947341

  6. Improvement of device isolation using field implantation for GaN MOSFETs

    NASA Astrophysics Data System (ADS)

    Jiang, Ying; Wang, Qingpeng; Zhang, Fuzhe; Li, Liuan; Shinkai, Satoko; Wang, Dejun; Ao, Jin-Ping

    2016-03-01

    Gallium nitride (GaN) metal-oxide-semiconductor field-effect transistors (MOSFETs) with boron field implantation isolation and mesa isolation were fabricated and characterized. The process of boron field implantation was altered and subsequently conducted after performing high-temperature ohmic annealing and gate oxide thermal treatment. Implanted regions with high resistivity were achieved. The circular MOSFET fabricated in the implanted region showed an extremely low current of 6.5 × 10-12 A under a gate voltage value up to 10 V, thus demonstrating that the parasitic MOSFET in the isolation region was eliminated by boron field implantation. The off-state drain current of the rectangular MOSFET with boron field implantation was 5.5 × 10-11 A, which was only one order of magnitude higher than the 6.6 × 10-12 A of the circular device. By contrast, the rectangular MOSFET with mesa isolation presented an off-state drain current of 3.2 × 10-9 A. The field isolation for GaN MOSFETs was achieved by using boron field implantation. The implantation did not reduce the field-effect mobility. The isolation structure of both mesa and implantation did not influence the subthreshold swing, whereas the isolation structure of only the implantation increased the subthreshold swing. The breakdown voltage of the implanted region with 5 μm spacing was up to 901.5 V.

  7. Periodontal treatment to improve glycaemic control in diabetic patients: study protocol of the randomized, controlled DIAPERIO trial

    PubMed Central

    Vergnes, Jean-Noel; Arrivé, Elise; Gourdy, Pierre; Hanaire, Hélène; Rigalleau, Vincent; Gin, Henri; Sédarat, Cyril; Dorignac, Georges; Bou, Christophe; Sixou, Michel; Nabet, Cathy

    2009-01-01

    Background Periodontitis is a common, chronic inflammatory disease caused by gram-negative bacteria leading to destruction of tissues supporting the teeth. Epidemiological studies have consistently shown increased frequency, extent and severity of periodontitis among diabetic adults. More recently, some controlled clinical trials have also suggested that periodontal treatment could improve glycaemic control in diabetic patients. However current evidence does not provide sufficient information on which to confidently base any clinical recommendations. The main objective of this clinical trial is to assess whether periodontal treatment could lead to a decrease in glycated haemoglobin levels in metabolically unbalanced diabetic patients suffering from chronic periodontitis. Methods The DIAPERIO trial is an open-label, 13-week follow-up, randomized, controlled trial. The total target sample size is planned at 150 participants, with a balanced (1:1) treatment allocation (immediate treatment vs delayed treatment). Periodontal treatment will include full mouth non-surgical scaling and root planing, systemic antibiotherapy, local antiseptics (chlorhexidine 0.12%) and oral health instructions. The primary outcome will be the difference in change of HbA1c between the two groups after the 13-weeks' follow-up. Secondary outcomes will be the difference in change of fructosamine levels and quality of life between the two groups. Discussion The DIAPERIO trial will provide insight into the question of whether periodontal treatment could lead to an improvement in glycaemic control in metabolically unbalanced diabetic patients suffering from periodontitis. The results of this trial will help to provide evidence-based recommendations for clinicians and a draft framework for designing national health policies. Trial registration Current Controlled Trials ISRCTN15334496 PMID:19646281

  8. 3PFDB+: improved search protocol and update for the identification of representatives of protein sequence domain families.

    PubMed

    Joseph, Agnel P; Shingate, Prashant; Upadhyay, Atul K; Sowdhamini, R

    2014-01-01

    Protein domain families are usually classified on the basis of similarity of amino acid sequences. Selection of a single representative sequence for each family provides targets for structure determination or modeling and also enables fast sequence searches to associate new members to a family. Such a selection could be challenging since some of these domain families exhibit huge variation depending on the number of members in the family, the average family sequence length or the extent of sequence divergence within a family. We had earlier created 3PFDB database as a repository of best representative sequences, selected from each PFAM domain family on the basis of high coverage. In this study, we have improved the database using more efficient strategies for the initial generation of sequence profiles and implement two independent methods, FASSM and HMMER, for identifying family members. HMMER employs a global sequence similarity search, while FASSM relies on motif identification and matching. This improved and updated database, 3PFDB+ generated in this study, provides representative sequences and profiles for PFAM families, with 13 519 family representatives having more than 90% family coverage. The representative sequence is also highlighted in a two-dimensional plot, which reflects the relative divergence between family members. Representatives belonging to small families with short sequences are mainly associated with low coverage. The set of sequences not recognized by the family representative profiles, highlight several potential false or weak family associations in PFAM. Partial domains and fragments dominate such cases, along with sequences that are highly diverged or different from other family members. Some of these outliers were also predicted to have different secondary structure contents, which reflect different putative structure or functional roles for these domain sequences. Database URL: http://caps.ncbs.res.in/3pfdbplus/.

  9. Improving the management of multimorbidity in general practice: protocol of a cluster randomised controlled trial (The 3D Study)

    PubMed Central

    Chaplin, Katherine; Bower, Peter; Brookes, Sara; Fitzpatrick, Bridie; Guthrie, Bruce; Shaw, Alison; Mercer, Stewart; Rafi, Imran; Thorn, Joanna

    2016-01-01

    Introduction An increasing number of people are living with multimorbidity. The evidence base for how best to manage these patients is weak. Current clinical guidelines generally focus on single conditions, which may not reflect the needs of patients with multimorbidity. The aim of the 3D study is to develop, implement and evaluate an intervention to improve the management of patients with multimorbidity in general practice. Methods and analysis This is a pragmatic two-arm cluster randomised controlled trial. 32 general practices around Bristol, Greater Manchester and Glasgow will be randomised to receive either the ‘3D intervention’ or usual care. 3D is a complex intervention including components affecting practice organisation, the conduct of patient reviews, integration with secondary care and measures to promote change in practice organisation. Changes include improving continuity of care and replacing reviews of each disease with patient-centred reviews with a focus on patients' quality of life, mental health and polypharmacy. We aim to recruit 1383 patients who have 3 or more chronic conditions. This provides 90% power at 5% significance level to detect an effect size of 0.27 SDs in the primary outcome, which is health-related quality of life at 15 months using the EQ-5D-5L. Secondary outcome measures assess patient centredness, illness burden and treatment burden. The primary analysis will be a multilevel regression model adjusted for baseline, stratification/minimisation, clustering and important co-variables. Nested process evaluation will assess implementation, mechanisms of effectiveness and interaction of the intervention with local context. Economic analysis of cost-consequences and cost-effectiveness will be based on quality-adjusted life years. Ethics and dissemination This study has approval from South-West (Frenchay) National Health Service (NHS) Research Ethics Committee (14/SW/0011). Findings will be disseminated via final report, peer

  10. Trunk muscle exercises as a means of improving postural stability in people with Parkinson's disease: a protocol for a randomised controlled trial

    PubMed Central

    Hubble, Ryan P; Naughton, Geraldine A; Silburn, Peter A; Cole, Michael H

    2014-01-01

    Introduction Exercise has been shown to improve clinical measures of strength, balance and mobility, and in some cases, has improved symptoms of tremor and rigidity in people with Parkinson's disease (PD). However, to date, no research has examined whether improvements in trunk control can remedy deficits in dynamic postural stability in this population. The proposed randomised controlled trial aims to establish whether a 12-week exercise programme aimed at improving dynamic postural stability in people with PD; (1) is more effective than education; (2) is more effective when training frequency is increased; and (3) provides greater long-term benefits than education. Methods/design Forty-five community-dwelling individuals diagnosed with idiopathic PD with a falls history will be recruited. Participants will complete baseline assessments including tests of cognition, vision, disease severity, fear of falling, mobility and quality of life. Additionally, participants will complete a series of standing balance tasks to evaluate static postural stability, while dynamic postural control will be measured during walking using head and trunk-mounted three-dimensional accelerometers. Following baseline testing, participants will be randomly-assigned to one of three intervention groups, who will receive either exercise once per week, exercise 3 days/week, or education. Participants will repeat the same battery of tests conducted at baseline after the 12-week intervention and again following a further 12-week sustainability period. Discussion This study has the potential to show that low-intensity and progressive trunk exercises can provide a non-invasive and effective means for maintaining or improving postural stability for people with PD. Importantly, if the programme is noted to be effective, it could be easily performed by patients within their home environment or under the guidance of available allied health professionals. Trial registration number The protocol for

  11. Improving outcomes of preschool language delay in the community: protocol for the Language for Learning randomised controlled trial

    PubMed Central

    2012-01-01

    Background Early language delay is a high-prevalence condition of concern to parents and professionals. It may result in lifelong deficits not only in language function, but also in social, emotional/behavioural, academic and economic well-being. Such delays can lead to considerable costs to the individual, the family and to society more widely. The Language for Learning trial tests a population-based intervention in 4 year olds with measured language delay, to determine (1) if it improves language and associated outcomes at ages 5 and 6 years and (2) its cost-effectiveness for families and the health care system. Methods/Design A large-scale randomised trial of a year-long intervention targeting preschoolers with language delay, nested within a well-documented, prospective, population-based cohort of 1464 children in Melbourne, Australia. All children received a 1.25-1.5 hour formal language assessment at their 4th birthday. The 200 children with expressive and/or receptive language scores more than 1.25 standard deviations below the mean were randomised into intervention or ‘usual care’ control arms. The 20-session intervention program comprises 18 one-hour home-based therapeutic sessions in three 6-week blocks, an outcome assessment, and a final feed-back/forward planning session. The therapy utilises a ‘step up-step down’ therapeutic approach depending on the child’s language profile, severity and progress, with standardised, manualised activities covering the four language development domains of: vocabulary and grammar; narrative skills; comprehension monitoring; and phonological awareness/pre-literacy skills. Blinded follow-up assessments at ages 5 and 6 years measure the primary outcome of receptive and expressive language, and secondary outcomes of vocabulary, narrative, and phonological skills. Discussion A key strength of this robust study is the implementation of a therapeutic framework that provides a standardised yet tailored approach for

  12. Improving the normalization of complex interventions: measure development based on normalization process theory (NoMAD): study protocol

    PubMed Central

    2013-01-01

    Background Understanding implementation processes is key to ensuring that complex interventions in healthcare are taken up in practice and thus maximize intended benefits for service provision and (ultimately) care to patients. Normalization Process Theory (NPT) provides a framework for understanding how a new intervention becomes part of normal practice. This study aims to develop and validate simple generic tools derived from NPT, to be used to improve the implementation of complex healthcare interventions. Objectives The objectives of this study are to: develop a set of NPT-based measures and formatively evaluate their use for identifying implementation problems and monitoring progress; conduct preliminary evaluation of these measures across a range of interventions and contexts, and identify factors that affect this process; explore the utility of these measures for predicting outcomes; and develop an online users’ manual for the measures. Methods A combination of qualitative (workshops, item development, user feedback, cognitive interviews) and quantitative (survey) methods will be used to develop NPT measures, and test the utility of the measures in six healthcare intervention settings. Discussion The measures developed in the study will be available for use by those involved in planning, implementing, and evaluating complex interventions in healthcare and have the potential to enhance the chances of their implementation, leading to sustained changes in working practices. PMID:23578304

  13. A study protocol: a community pharmacy-based intervention for improving the management of sleep disorders in the community settings

    PubMed Central

    2014-01-01

    Background Sleep disorders are very common in the community and are estimated to affect up to 45% of the world’s population. Pharmacists are in a position to give advice and provide appropriate services to individuals who are unable to easily access medical care. The purpose of this study is to develop an intervention to improve the management of sleep disorders in the community. The aims are– (1) to evaluate the effectiveness of a community pharmacy-based intervention in managing sleep disorders, (2) to evaluate the role of actigraph as an objective measure in monitoring certain sleep disorders and (3) to evaluate the extended role of community pharmacists in managing sleep disorders. This intervention is developed to monitor individuals undergoing treatment and overcome the difficulties in validating self-reported feedback. Method/design This is a community-based intervention, prospective, controlled trial, with one intervention group and one control group, comparing individuals receiving a structured intervention with those receiving usual care for sleep-related disorders at community pharmacies. Discussion This study will demonstrate the utilisation and efficacy of community pharmacy-based intervention to manage sleep disorders in the community, and will assess the possibility of implementing this intervention into the community pharmacy workflow. Trial registration Australian New Zealand Clinical Trial Registry: ACTRN12612000825853 PMID:24533916

  14. Continuous quality improvement (CQI) in addiction treatment settings: design and intervention protocol of a group randomized pilot study

    PubMed Central

    2014-01-01

    Background Few studies have designed and tested the use of continuous quality improvement approaches in community based substance use treatment settings. Little is known about the feasibility, costs, efficacy, and sustainment of such approaches in these settings. Methods/Design A group-randomized trial using a modified stepped wedge design is being used. In the first phase of the study, eight programs, stratified by modality (residential, outpatient) are being randomly assigned to the intervention or control condition. In the second phase, the initially assigned control programs are receiving the intervention to gain additional information about feasibility while sustainment is being studied among the programs initially assigned to the intervention. Discussion By using this design in a pilot study, we help inform the field about the feasibility, costs, efficacy and sustainment of the intervention. Determining information at the pilot stage about costs and sustainment provides value for designing future studies and implementation strategies with the goal to reduce the time between intervention development and translation to real world practice settings. PMID:24467770

  15. An improved representation of geographically isolated wetlands in a watershed-scale hydrologic model

    EPA Science Inventory

    Geographically isolated wetlands (GIWs), defined as wetlands surrounded by uplands, provide an array of ecosystem goods and services. Within the United States, federal regulatory protections for GIWs are contingent, in part, on the quantification of their singular or aggregate ef...

  16. Improving Post-Hospitalization Transition Outcomes through Accessible Health Information Technology and Caregiver Support: Protocol for a Randomized Controlled Trial

    PubMed Central

    Piette, John D; Striplin, Dana; Marinec, Nicolle; Chen, Jenny; Gregory, Lynn A; Sumerlin, Denise L; DeSantis, Angela M; Gibson, Carolyn; Crause, Ingrid; Rouse, Marylena; Aikens, James E

    2015-01-01

    Objective The goal of this trial is to evaluate a novel intervention designed to improve post-hospitalization support for older adults with chronic conditions via: (a) direct tailored communication to patients using regular automated calls post discharge, (b) support for informal caregivers outside of the patient’s household via structured automated feedback about the patient’s status plus advice about how caregivers can help, and (c) support for care management including a web-based disease management tool and alerts about potential problems. Methods 846 older adults with common chronic conditions are being identified upon hospital admission. Patients are asked to identify a “CarePartner” (CP) living outside their household, i.e., an adult child or other social network member willing to play an active role in their post-discharge transition support. Patient-CP pairs are randomized to the intervention or usual care. Intervention patients receive automated assessment and behavior change calls, and their CPs receives structured feedback and advice via email and automated calls following each assessment. Clinical teams have access to assessment results via the web and receive automated reports about urgent health problems. Patients complete surveys at baseline, 30 days, and 90 days post discharge; utilization data is obtained from hospital records. CPs, other caregivers, and clinicians are interviewed to evaluate intervention effects on processes of self-care support, caregiver stress and communication, and the intervention’s potential for broader implementation. The primary outcome is 30-day readmission rates; other outcomes measured at 30 days and 90 days include functional status, self-care behaviors, and mortality risk. Conclusion This trial uses accessible health technologies and coordinated communication among informal caregivers and clinicians to fill the growing gap between what discharged patients need and available resources. A unique feature of

  17. Can virtual nature improve patient experiences and memories of dental treatment? A study protocol for a randomized controlled trial

    PubMed Central

    2014-01-01

    Background Dental anxiety and anxiety-related avoidance of dental care create significant problems for patients and the dental profession. Distraction interventions are used in daily medical practice to help patients cope with unpleasant procedures. There is evidence that exposure to natural scenery is beneficial for patients and that the use of virtual reality (VR) distraction is more effective than other distraction interventions, such as watching television. The main aim of this randomized controlled trial is to determine whether the use of VR during dental treatment can improve the overall dental experience and recollections of treatment for patients, breaking the negative cycle of memories of anxiety leading to further anxiety, and avoidance of future dental appointments. Additionally, the aim is to test whether VR benefits dental patients with all levels of dental anxiety or whether it could be especially beneficial for patients suffering from higher levels of dental anxiety. The third aim is to test whether the content of the VR distraction can make a difference for its effectiveness by comparing two types of virtual environments, a natural environment and an urban environment. Methods/design The effectiveness of VR distraction will be examined in patients 18 years or older who are scheduled to undergo dental treatment for fillings and/or extractions, with a maximum length of 30 minutes. Patients will be randomly allocated into one of three groups. The first group will be exposed to a VR of a natural environment. The second group will be exposed to a VR of an urban environment. A third group consists of patients who receive standard care (control group). Primary outcomes relate to patients’ memories of the dental treatment one week after treatment: (a) remembered pain, (b) intrusive thoughts and (c) vividness of memories. Other measures of interest are the dental experience, the treatment experience and the VR experience. Trial registration Current

  18. Tephrochronology of last termination sequences in Europe: a protocol for improved analytical precision and robust correlation procedures (a joint SCOTAV-INTIMATE proposal)

    NASA Astrophysics Data System (ADS)

    Turney, Chris S. M.; Lowe, J. John; Davies, Siwan M.; Hall, Valerie; Lowe, David J.; Wastegård, Stefan; Hoek, Wim Z.; Alloway, Brent

    2004-02-01

    The precise sequence of events during the Last Termination (18 000-9000 ka 14C yr BP), and the extent to which major environmental changes were synchronous, are difficult to establish using the radiocarbon method alone because of serious distortions of the radiocarbon time-scale, as well as the influences of site-specific errors that can affect the materials dated. Attention has therefore turned to other methods that can provide independent tests of the chronology and correlation of events during the Last Termination. With emphasis on European sequences, we summarise here the potential of tephrostratigraphy and tephrochronology to fulfil this role. Recent advances in the detection and analysis of hidden tephra layers (cryptotephra) indicate that some tephras of Last Termination age are much more widespread in Europe than appreciated hitherto, and a number of new tephra deposits have also been identified. There is much potential for developing an integrated tephrochronological framework for Europe, which can help to underpin the overall chronology of events during the Last Termination. For that potential to be realised, however, there needs to be a more systematic and robust analysis of tephra layers than has been the practice in the past. We propose a protocol for improving analytical and reporting procedures, as well as the establishment of a centralised data base of the results, which will provide an important geochronological tool to support a diverse range of stratigraphical studies, including opportunities to reassess volcanic hazards. Although aimed primarily at Europe, the protocol proposed here is of equal relevance to other regions and periods of interest. Copyright

  19. Significant Improvements in Cognitive Performance Post-Transcranial, Red/Near-Infrared Light-Emitting Diode Treatments in Chronic, Mild Traumatic Brain Injury: Open-Protocol Study

    PubMed Central

    Zafonte, Ross; Krengel, Maxine H.; Martin, Paula I.; Frazier, Judith; Hamblin, Michael R.; Knight, Jeffrey A.; Meehan, William P.; Baker, Errol H.

    2014-01-01

    Abstract This pilot, open-protocol study examined whether scalp application of red and near-infrared (NIR) light-emitting diodes (LED) could improve cognition in patients with chronic, mild traumatic brain injury (mTBI). Application of red/NIR light improves mitochondrial function (especially in hypoxic/compromised cells) promoting increased adenosine triphosphate (ATP) important for cellular metabolism. Nitric oxide is released locally, increasing regional cerebral blood flow. LED therapy is noninvasive, painless, and non-thermal (cleared by the United States Food and Drug Administration [FDA], an insignificant risk device). Eleven chronic, mTBI participants (26–62 years of age, 6 males) with nonpenetrating brain injury and persistent cognitive dysfunction were treated for 18 outpatient sessions (Monday, Wednesday, Friday, for 6 weeks), starting at 10 months to 8 years post- mTBI (motor vehicle accident [MVA] or sports-related; and one participant, improvised explosive device [IED] blast injury). Four had a history of multiple concussions. Each LED cluster head (5.35 cm diameter, 500 mW, 22.2 mW/cm2) was applied for 10 min to each of 11 scalp placements (13 J/cm2). LEDs were placed on the midline from front-to-back hairline; and bilaterally on frontal, parietal, and temporal areas. Neuropsychological testing was performed pre-LED, and at 1 week, and 1 and 2 months after the 18th treatment. A significant linear trend was observed for the effect of LED treatment over time for the Stroop test for Executive Function, Trial 3 inhibition (p=0.004); Stroop, Trial 4 inhibition switching (p=0.003); California Verbal Learning Test (CVLT)-II, Total Trials 1–5 (p=0.003); and CVLT-II, Long Delay Free Recall (p=0.006). Participants reported improved sleep, and fewer post-traumatic stress disorder (PTSD) symptoms, if present. Participants and family reported better ability to perform social, interpersonal, and occupational functions. These open-protocol data suggest

  20. Significant improvements in cognitive performance post-transcranial, red/near-infrared light-emitting diode treatments in chronic, mild traumatic brain injury: open-protocol study.

    PubMed

    Naeser, Margaret A; Zafonte, Ross; Krengel, Maxine H; Martin, Paula I; Frazier, Judith; Hamblin, Michael R; Knight, Jeffrey A; Meehan, William P; Baker, Errol H

    2014-06-01

    This pilot, open-protocol study examined whether scalp application of red and near-infrared (NIR) light-emitting diodes (LED) could improve cognition in patients with chronic, mild traumatic brain injury (mTBI). Application of red/NIR light improves mitochondrial function (especially in hypoxic/compromised cells) promoting increased adenosine triphosphate (ATP) important for cellular metabolism. Nitric oxide is released locally, increasing regional cerebral blood flow. LED therapy is noninvasive, painless, and non-thermal (cleared by the United States Food and Drug Administration [FDA], an insignificant risk device). Eleven chronic, mTBI participants (26-62 years of age, 6 males) with nonpenetrating brain injury and persistent cognitive dysfunction were treated for 18 outpatient sessions (Monday, Wednesday, Friday, for 6 weeks), starting at 10 months to 8 years post- mTBI (motor vehicle accident [MVA] or sports-related; and one participant, improvised explosive device [IED] blast injury). Four had a history of multiple concussions. Each LED cluster head (5.35 cm diameter, 500 mW, 22.2 mW/cm(2)) was applied for 10 min to each of 11 scalp placements (13 J/cm(2)). LEDs were placed on the midline from front-to-back hairline; and bilaterally on frontal, parietal, and temporal areas. Neuropsychological testing was performed pre-LED, and at 1 week, and 1 and 2 months after the 18th treatment. A significant linear trend was observed for the effect of LED treatment over time for the Stroop test for Executive Function, Trial 3 inhibition (p=0.004); Stroop, Trial 4 inhibition switching (p=0.003); California Verbal Learning Test (CVLT)-II, Total Trials 1-5 (p=0.003); and CVLT-II, Long Delay Free Recall (p=0.006). Participants reported improved sleep, and fewer post-traumatic stress disorder (PTSD) symptoms, if present. Participants and family reported better ability to perform social, interpersonal, and occupational functions. These open-protocol data suggest that placebo

  1. Significant improvements in cognitive performance post-transcranial, red/near-infrared light-emitting diode treatments in chronic, mild traumatic brain injury: open-protocol study.

    PubMed

    Naeser, Margaret A; Zafonte, Ross; Krengel, Maxine H; Martin, Paula I; Frazier, Judith; Hamblin, Michael R; Knight, Jeffrey A; Meehan, William P; Baker, Errol H

    2014-06-01

    This pilot, open-protocol study examined whether scalp application of red and near-infrared (NIR) light-emitting diodes (LED) could improve cognition in patients with chronic, mild traumatic brain injury (mTBI). Application of red/NIR light improves mitochondrial function (especially in hypoxic/compromised cells) promoting increased adenosine triphosphate (ATP) important for cellular metabolism. Nitric oxide is released locally, increasing regional cerebral blood flow. LED therapy is noninvasive, painless, and non-thermal (cleared by the United States Food and Drug Administration [FDA], an insignificant risk device). Eleven chronic, mTBI participants (26-62 years of age, 6 males) with nonpenetrating brain injury and persistent cognitive dysfunction were treated for 18 outpatient sessions (Monday, Wednesday, Friday, for 6 weeks), starting at 10 months to 8 years post- mTBI (motor vehicle accident [MVA] or sports-related; and one participant, improvised explosive device [IED] blast injury). Four had a history of multiple concussions. Each LED cluster head (5.35 cm diameter, 500 mW, 22.2 mW/cm(2)) was applied for 10 min to each of 11 scalp placements (13 J/cm(2)). LEDs were placed on the midline from front-to-back hairline; and bilaterally on frontal, parietal, and temporal areas. Neuropsychological testing was performed pre-LED, and at 1 week, and 1 and 2 months after the 18th treatment. A significant linear trend was observed for the effect of LED treatment over time for the Stroop test for Executive Function, Trial 3 inhibition (p=0.004); Stroop, Trial 4 inhibition switching (p=0.003); California Verbal Learning Test (CVLT)-II, Total Trials 1-5 (p=0.003); and CVLT-II, Long Delay Free Recall (p=0.006). Participants reported improved sleep, and fewer post-traumatic stress disorder (PTSD) symptoms, if present. Participants and family reported better ability to perform social, interpersonal, and occupational functions. These open-protocol data suggest that placebo

  2. Supporting the improvement and management of prescribing for urinary tract infections (SIMPle): protocol for a cluster randomized trial

    PubMed Central

    2013-01-01

    Background The overuse of antimicrobials is recognized as the main selective pressure driving the emergence and spread of antimicrobial resistance in human bacterial pathogens. Urinary tract infections (UTIs) are among the most common infections presented in primary care and empirical antimicrobial treatment is currently recommended. Previous research has identified that a substantial proportion of Irish general practitioners (GPs) prescribe antimicrobials for UTIs that are not in accordance with the Guidelines for Antimicrobial Prescribing in Primary Care in Ireland. The aim of this trial is to design, implement and evaluate the effectiveness of a complex intervention on GP antimicrobial prescribing and adult (18 years of age and over) patients’ antimicrobial consumption when presenting with a suspected UTI. Methods/design The Supporting the Improvement and Management of Prescribing for urinary tract infections (SIMPle) study is a three-armed intervention with practice-level randomization. Adult patients presenting with suspected UTIs in primary care will be included in the study. The intervention integrates components for both GPs and patients. For GPs the intervention includes interactive workshops, audit and feedback reports and automated electronic prompts summarizing recommended first-line antimicrobial treatment and, for one intervention arm, a recommendation to consider delayed antimicrobial treatment. For patients, multimedia applications and information leaflets are included. Thirty practices will be recruited to the study; laboratory data indicate that 2,038 patients will be prescribed an antimicrobial in the study. The primary outcome is a change in prescribing of first-line antimicrobials for UTIs in accordance with the Guidelines for Antimicrobial Prescribing in Primary Care in Ireland. The study will take place over 15 months with a six-month intervention period. Data will be collected through a remote electronic anonymized data-extraction system

  3. Improvements on Restricted Insecticide Application Protocol for Control of Human and Animal African Trypanosomiasis in Eastern Uganda

    PubMed Central

    Muhanguzi, Dennis; Picozzi, Kim; Hatendorf, Jan; Thrusfield, Michael; Welburn, Susan Christina; Kabasa, John David; Waiswa, Charles

    2014-01-01

    Background African trypanosomes constrain livestock and human health in Sub-Saharan Africa, and aggravate poverty and hunger of these otherwise largely livestock-keeping communities. To solve this, there is need to develop and use effective and cheap tsetse control methods. To this end, we aimed at determining the smallest proportion of a cattle herd that needs to be sprayed on the legs, bellies and ears (RAP) for effective Human and Animal African Trypanosomiasis (HAT/AAT) control. Methodology/Principal finding Cattle in 20 villages were ear-tagged and injected with two doses of diminazene diaceturate (DA) forty days apart, and randomly allocated to one of five treatment regimens namely; no treatment, 25%, 50%, 75% monthly RAP and every 3 month Albendazole drench. Cattle trypanosome re-infection rate was determined by molecular techniques. ArcMap V10.3 was used to map apparent tsetse density (FTD) from trap catches. The effect of graded RAP on incidence risk ratios and trypanosome prevalence was determined using Poisson and logistic random effect models in R and STATA V12.1 respectively. Incidence was estimated at 9.8/100 years in RAP regimens, significantly lower compared to 25.7/100 years in the non-RAP regimens (incidence rate ratio: 0.37; 95% CI: 0.22–0.65; P<0.001). Likewise, trypanosome prevalence after one year of follow up was significantly lower in RAP animals than in non-RAP animals (4% vs 15%, OR: 0.20, 95% CI: 0.08–0.44; P<0.001). Contrary to our expectation, level of protection did not increase with increasing proportion of animals treated. Conclusions/significance Reduction in RAP coverage did not significantly affect efficacy of treatment. This is envisaged to improve RAP adaptability to low income livestock keepers but needs further evaluation in different tsetse challenge, HAT/AAT transmission rates and management systems before adopting it for routine tsetse control programs. PMID:25356758

  4. The implementation of a translational study involving a primary care based behavioral program to improve blood pressure control: The HTN-IMPROVE study protocol (01295)

    PubMed Central

    2010-01-01

    Background Despite the impact of hypertension and widely accepted target values for blood pressure (BP), interventions to improve BP control have had limited success. Objectives We describe the design of a 'translational' study that examines the implementation, impact, sustainability, and cost of an evidence-based nurse-delivered tailored behavioral self-management intervention to improve BP control as it moves from a research context to healthcare delivery. The study addresses four specific aims: assess the implementation of an evidence-based behavioral self-management intervention to improve BP levels; evaluate the clinical impact of the intervention as it is implemented; assess organizational factors associated with the sustainability of the intervention; and assess the cost of implementing and sustaining the intervention. Methods The project involves three geographically diverse VA intervention facilities and nine control sites. We first conduct an evaluation of barriers and facilitators for implementing the intervention at intervention sites. We examine the impact of the intervention by comparing 12-month pre/post changes in BP control between patients in intervention sites versus patients in the matched control sites. Next, we examine the sustainability of the intervention and organizational factors facilitating or hindering the sustained implementation. Finally, we examine the costs of intervention implementation. Key outcomes are acceptability and costs of the program, as well as changes in BP. Outcomes will be assessed using mixed methods (e.g., qualitative analyses--pattern matching; quantitative methods--linear mixed models). Discussion The study results will provide information about the challenges and costs to implement and sustain the intervention, and what clinical impact can be expected. PMID:20637095

  5. ICSI improves fertilization in isolated teratozoospermic men: a study with strictly controlled external factors and WHO-5 standard.

    PubMed

    Zhu, Yuan; Wu, Qiong-fang; Zhou, Xiao-Jun; Xin, Cai-lin; Ling, Ge; Lu, Guang-xiu

    2013-02-01

    A clear clinical management pathway (conventional in vitro fertilization, IVF or intracytoplasmic sperm injection, ICSI) for treating patients with teratozoospermia is lacking. Here we conducted a retrospective study of fertility indices in 2,178 IVF/ICSI cycles in order to reevaluate clinical management of couples with isolated teratozoospermia (< 4% morphologically normal sperms and normal sperm concentration and motility with the standard of WHO-5).We strictly controlled external factors that could affect oocyte quality or endometrial receptivity to minimize the impact of confounders. Fertilization, total fertilization failure, embryo quality, blastocyst formation rate, and pregnancy rate were studied. Retrospectively, in conventional IVF cycles a significantly lower fertilization rate and higher total fertilization failure rate were observed in couples with isolated teratozoospermia as compared to couples with a normal semen profile. Furthermore, when ICSI was used to treat these teratozoospermic couples, improvement in fertilization was noted. However, the embryo quality, blastocyst formation rate, and pregnancy of couples with isolated teratozoospermia were not enhanced by ICSI. Multiple variable analysis showed that many factors including percentage of morphologically normal sperm are statistically correlated with fertilization rate and total fertilization failure in conventional IVF cycles. In addition the insemination method was correlated with fertilization rate in cases with isolated teratozoospermia. Further studies are warranted to compare outcomes of conventional IVF and ICSI in cases of isolated teratozoospermia, where less than 5 oocytes are retrieved.

  6. A cluster randomized controlled trial aimed at implementation of local quality improvement collaboratives to improve prescribing and test ordering performance of general practitioners: Study Protocol

    PubMed Central

    Trietsch, Jasper; van der Weijden, Trudy; Verstappen, Wim; Janknegt, Rob; Muijrers, Paul; Winkens, Ron; van Steenkiste, Ben; Grol, Richard; Metsemakers, Job

    2009-01-01

    Background The use of guidelines in general practice is not optimal. Although evidence-based methods to improve guideline adherence are available, variation in physician adherence to general practice guidelines remains relatively high. The objective for this study is to transfer a quality improvement strategy based on audit, feedback, educational materials, and peer group discussion moderated by local opinion leaders to the field. The research questions are: is the multifaceted strategy implemented on a large scale as planned?; what is the effect on general practitioners' (GPs) test ordering and prescribing behaviour?; and what are the costs of implementing the strategy? Methods In order to evaluate the effects, costs and feasibility of this new strategy we plan a multi-centre cluster randomized controlled trial (RCT) with a balanced incomplete block design. Local GP groups in the south of the Netherlands already taking part in pharmacotherapeutic audit meeting groups, will be recruited by regional health officers. Approximately 50 groups of GPs will be randomly allocated to two arms. These GPs will be offered two different balanced sets of clinical topics. Each GP within a group will receive comparative feedback on test ordering and prescribing performance. The feedback will be discussed in the group and working agreements will be created after discussion of the guidelines and barriers to change. The data for the feedback will be collected from existing and newly formed databases, both at baseline and after one year. Discussion We are not aware of published studies on successes and failures of attempts to transfer to the stakeholders in the field a multifaceted strategy aimed at GPs' test ordering and prescribing behaviour. This pragmatic study will focus on compatibility with existing infrastructure, while permitting a certain degree of adaptation to local needs and routines. Trial registration Nederlands Trial Register ISRCTN40008171 PMID:19222840

  7. Paramagnetic cellulose DNA isolation improves DNA yield and quality among diverse plant taxa1

    PubMed Central

    Moeller, Jackson R.; Moehn, Nicholas R.; Waller, Donald M.; Givnish, Thomas J.

    2014-01-01

    • Premise of the study: The chemical diversity of land plants ensures that no single DNA isolation method results in high yield and purity with little effort for all species. Here we evaluate a new technique originally developed for forensic science, based on MagnaCel paramagnetic cellulose particles (PMC), to determine its efficacy in extracting DNA from 25 plant species representing 21 families and 15 orders. • Methods and Results: Yield and purity of DNA isolated by PMC, DNeasy Plant Mini Kit (silica column), and cetyltrimethylammonium bromide (CTAB) methods were compared among four individuals for each of 25 plant species. PMC gave a twofold advantage in average yield, and the relative advantage of the PMC method was greatest for samples with the lowest DNA yields. PMC also produced more consistent sample purity based on absorbance ratios at 260:280 and 260:230 nm. • Conclusions: PMC technology is a promising alternative for plant DNA isolation. PMID:25309836

  8. Xenogenic transfer of isolated murine mitochondria into human rho0 cells can improve respiratory function.

    PubMed

    Katrangi, Eyad; D'Souza, Gerard; Boddapati, Sarathi V; Kulawiec, Mariola; Singh, Keshav K; Bigger, Brian; Weissig, Volkmar

    2007-12-01

    Mitochondrial DNA mutations are the direct cause of several physiological disorders and are also associated with the aging process. The modest progress made over the past two decades towards manipulating the mitochondrial genome and understanding its function within living mammalian cells means that cures for mitochondrial DNA mutations are still elusive. Here, we report that transformed mammalian cells internalize exogenous isolated mitochondria upon simple co-incubation. We first demonstrate the physical presence of internalized mitochondria within recipient cells using fluorescence microscopy. Second, we show that xenogenic transfer of murine mitochondria into human cells lacking functional mitochondria can functionally restore respiration in cells lacking mtDNA. Third, utilizing the natural competence of isolated mitochondria to take up linear DNA molecules, we demonstrate the feasibility of using cellular internalization of isolated exogenous mitochondria as a potential tool for studying mitochondrial genetics in living mammalian cells. PMID:18069915

  9. Evaluation of an internet-based aftercare program to improve vocational reintegration after inpatient medical rehabilitation: study protocol for a cluster-randomized controlled trial

    PubMed Central

    2013-01-01

    Background Mental disorders are the main reasons for rising proportions of premature pension in most high-income countries. Although inpatient medical rehabilitation has increasingly targeted work-related stress, there is still a lack of studies on the transfer of work-specific interventions into work contexts. Therefore, we plan to evaluate an online aftercare program aiming to improve vocational reintegration after medical rehabilitation. Methods Vocationally strained patients (n = 800) aged between 18 and 59 years with private internet access are recruited in psychosomatic, orthopedic and cardiovascular rehabilitation clinics in Germany. During inpatient rehabilitation, participants in stress management group training are cluster-randomized to the intervention or control group. The intervention group (n = 400) is offered an internet-based aftercare with weekly writing tasks and therapeutic feedback, a patient forum, a self-test and relaxation exercises. The control group (n = 400) obtains regular e-mail reminders with links to publicly accessible information about stress management and coping. Assessments are conducted at the beginning of inpatient rehabilitation, the end of inpatient rehabilitation, the end of aftercare, and 9 months later. The primary outcome is a risk score for premature pension, measured by a screening questionnaire at follow-up. Secondary outcome measures include level of vocational stress, physical and mental health, and work capacity at follow-up. Discussion We expect the intervention group to stabilize the improvements achieved during inpatient rehabilitation concerning stress management and coping, resulting in an improved vocational reintegration. The study protocol demonstrates the features of internet-based aftercare in rehabilitation. Trial registration International Standard Randomised Controlled Trial Number Register (ISRCTN:ISRCTN33957202) PMID:23351836

  10. Improving physical functional and quality of life in older adults with multiple sclerosis via a DVD-delivered exercise intervention: a study protocol

    PubMed Central

    Wójcicki, Thomas R; Roberts, Sarah A; Learmonth, Yvonne C; Hubbard, Elizabeth A; Kinnett-Hopkins, Dominque; Motl, Robert W; McAuley, Edward

    2014-01-01

    Introduction There is a need to identify innovative, low-cost and broad-reaching strategies for promoting exercise and improving physical function in older adults with multiple sclerosis (MS). Methods and analysis This randomised controlled pilot trial will test the efficacy of a 6-month, DVD-delivered exercise intervention to improve functional performance and quality of life in older adults with MS. Participants will be randomised either into a DVD-delivered exercise condition or an attentional control condition. This novel approach to programme delivery provides participants with detailed exercise instructions which are presented in a progressive manner and includes a variety of modifications to better meet varying levels of physical abilities. The targeted exercises focus on three critical elements of functional fitness: flexibility, strength and balance. It is hypothesised that participants who are randomised to the exercise DVD condition will demonstrate improvements in physical function compared with participants assigned to the attentional control condition. Data analysis will include a 2 (condition)×2 (time) mixed factor analysis of variance (ANOVA) that follows intent-to-treat principles, as well as an examination of effect sizes. Participants will take part in qualitative interviews about perspectives on physical activity and programme participation. Ethics and dissemination The study protocol was approved by a university institutional review board and registered with a federal database. Participants will be asked to read and sign a detailed informed consent document and will be required to provide a physician's approval to participate in the study. The exercise DVDs include an overview of safety-related concerns and recommendations relative to exercise participation, as well as detailed instructions highlighting the proper execution of each exercise presented on screen. Following completion of this trial, data will be immediately analysed and results

  11. A Comparative experimental study of media access protocols for wireless radio networks

    SciTech Connect

    Barrett, C. L.; Drozda, M.; Marathe, M. V.

    2001-05-24

    ) Performance analysis of protocols at a given level in the protocol stack need to be studied not locally in isolation but as a part of the complete protocol stack. The results suggest that in order to improve the performance of a communication network, it will be important to study the entire protocol stuck as a single algorithmic construct; optimizing individual layers in the 7 layer OSI stack will not yield performance improvements beyond a point.

  12. Rational Protocols

    NASA Astrophysics Data System (ADS)

    Cachin, Christian

    Security research continues to provide a plethora of new protocols and mechanisms; these solutions patch either existing vulnerabilities found in practical systems or solve hypothetical security problems in the sense that the problem is often conceived at the same time when the first solution is proposed. Yet only a very small fraction of this research is relevant to ordinary users in the sense that they are willing to actually deploy the technology.

  13. Improvement of lightweight floating ceiling design with optimum stiffener and isolator locations

    NASA Astrophysics Data System (ADS)

    Hui, C. K.; Ng, C. F.

    2009-11-01

    In this paper, a new design concept of a lightweight floating ceiling with a special arrangement of stiffener beams and isolators is proposed to enhance vibration isolation performance. The key design parameters of resonant frequency of bending and mode shape factor on vibration transmission are determined with some simple formulae. Structural vibrations and noise radiation are evaluated with finite element models (FEM) for various designs. The optimum ceiling designs are applied in a studio, and field measurements with reliable frequency range of 30-400 Hz are conducted to confirm the theoretical results. The analyses ascertain that four design features ensure the optimum vibration isolation performance: the stiffener beams must be installed at the nodal line of fundamental bending resonance of the plate; smaller panels should be applied instead of a large panel covering the whole area of the ceiling, and joints should be free; the isolators should be placed at the nodal point of the bending mode of the plate; and the fundamental bending resonance frequency must not match the modal critical frequency. The proposed stiffened wooden panel design achieved a noise and vibration reduction of 20 dB in the frequency range of 40-100 Hz, and was better than the performance of a concrete floating floor.

  14. Engagement, Alignment, and Rigor as Vital Signs of High-Quality Instruction: A Classroom Visit Protocol for Instructional Improvement and Research

    ERIC Educational Resources Information Center

    Early, Diane M.; Rogge, Ronald D.; Deci, Edward L.

    2014-01-01

    This paper investigates engagement (E), alignment (A), and rigor (R) as vital signs of high-quality teacher instruction as measured by the EAR Classroom Visit Protocol, designed by the Institute for Research and Reform in Education (IRRE). Findings indicated that both school leaders and outside raters could learn to score the protocol with…

  15. How to Improve the Peer Review Method: Free-Selection vs Assigned-Pair Protocol Evaluated in a Computer Networking Course

    ERIC Educational Resources Information Center

    Papadopoulos, Pantelis M.; Lagkas, Thomas D.; Demetriadis, Stavros N.

    2012-01-01

    This study provides field research evidence on the efficiency of a "free-selection" peer review assignment protocol as compared to the typically implemented "assigned-pair" protocol. The study employed 54 sophomore students who were randomly assigned into three groups: Assigned-Pair (AP) (the teacher assigns student works for review to student…

  16. Ease-of-use protocol for the rapid detection of third-generation cephalosporin resistance in Enterobacteriaceae isolated from blood cultures using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry.

    PubMed

    Foschi, C; Compri, M; Smirnova, V; Denicolò, A; Nardini, P; Tamburini, M V; Lombardo, D; Landini, M P; Ambretti, S

    2016-06-01

    An ease-of-use protocol for the identification of resistance against third-generation cephalosporins in Enterobacteriaceae isolated from blood culture bottles was evaluated using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. A cefotaxime hydrolysis assay from chocolate agar subcultures using antibiotic discs and without inoculum standardization was developed for routine work flow, with minimal hands-on time. This assay showed good performance in distinguishing between cefotaxime-susceptible and cefotaxime-resistant strains, with excellent results for Escherichia coli (sensitivity 94.7%, specificity 100%). However, cefotaxime resistance was not detected reliably in Enterobacteriaceae expressing AmpC genes or carbapenemase-producing Klebsiella pneumoniae. PMID:27105753

  17. Patients With Isolated PCL Injuries Improve From Surgery as Much as Patients With ACL Injuries After 2 Years

    PubMed Central

    Owesen, Christian; Sivertsen, Einar Andreas; Engebretsen, Lars; Granan, Lars-Petter; Årøen, Asbjørn

    2015-01-01

    Background: Reports on outcome after posterior cruciate ligament (PCL) reconstruction often contain both isolated PCL and combined knee ligament injuries. This makes it difficult to conclude on the outcome after reconstruction of isolated PCL injuries. Purpose: To investigate the outcome after PCL reconstruction in patients with an isolated PCL injury and to compare this with the outcome of patients treated with reconstruction after isolated anterior cruciate ligament (ACL) injuries. Study Design: Cohort study; Level of evidence, 3. Methods: Seventy-one patients with an isolated PCL injury that was reconstructed surgically and who had registered in the Norwegian Knee Ligament Registry between 2004 and 2010 were included in this study. Patients with isolated ACL reconstructions (n = 9661) who had registered in the same period were included for comparison. Knee Injury and Osteoarthritis Outcome Score (KOOS) was used as the patient-reported outcome measure. Preoperative and 2-year postoperative KOOS scores were compared. Changes in KOOS score reported by the PCL patients were compared with changes reported by the ACL patients. Results: At the 2-year postoperative follow-up of the PCL-reconstructed patients, the patient-reported outcome was improved, measured by KOOS as follows: pain, 15.1 (95% CI, 8.5-21.8; P < .001); symptoms, 0.9 (95% CI, –6.6 to 8.3; P = .82); activities of daily living, 13.2 (95% CI, 6.6-13.9; P < .001); sports, 20.7 (95% CI, 11.8-29.4; P < .001); and quality of life, 26.6 (95% CI, 18.9-34.2; P < .001). According to the KOOS, the incremental improvements were similar for PCL and ACL patients. Time from injury to surgery was longer for the PCL patients compared with ACL patients (median, 21.5 vs 8.0 months; P < .001). Conclusion: Patients undergoing PCL reconstruction can expect the same improvements in KOOS score as patients undergoing ACL reconstruction. However, PCL patients start out with an inferior score on average and consequently end up

  18. Portrait of trauma care in Quebec's rural emergency departments and identification of priority intervention needs to improve the quality of care: a study protocol

    PubMed Central

    Fleet, Richard; Tounkara, Fatoumata Korika; Ouimet, Mathieu; Dupuis, Gilles; Poitras, Julien; Tanguay, Alain; Fortin, Jean Paul; Trottier, Jean-Guy; Ouellet, Jean; Lortie, Gilles; Plant, Jeff; Morris, Judy; Chauny, Jean Marc; Lauzier, François; Légaré, France

    2016-01-01

    Introduction Trauma remains the primary cause of death in individuals under 40 years of age in Canada. In Quebec, the Trauma Care Continuum (TCC) has been demonstrated to be effective in decreasing the mortality rate among trauma victims. Although rural citizens are at greater risk for trauma and trauma death, no empirical data concerning the effectiveness of the TCC for the rural population in Quebec are available. The emergency departments (EDs) are important safety nets for rural citizens. However, our data indicate that access to diagnostic support services, such as intensive care units and CT is limited in rural areas. The objectives are to (1) draw a portrait of trauma services in rural EDs; (2) explore geographical variations in trauma care in Quebec; (3) identify adaptable factors that could reduce variation; and (4) establish consensus solutions for improving the quality of care. Methods and analysis The study will take place from November 2015 to November 2018. A mixed methodology (qualitative and quantitative) will be used. We will include data (2009–2013) from all trauma victims treated in the 26 rural EDs and tertiary/secondary care centres in Quebec. To meet objectives 1 and 2, data will be gathered from the Ministry's Database of the Quebec Trauma Registry Information System. For objectives 3 and 4, the project will use the Delphi method to develop consensus solutions for improving the quality of trauma care in rural areas. Data will be analysed using a Poisson regression to compare mortality rate during hospital stay or death on ED arrival (objectives 1 and 2). Average scores and 95% CI will be calculated for the Delphi questionnaire (objectives 3 and 4). Ethics and dissemination This protocol has been approved by CSSS Alphonse-Desjardins research ethics committee (Project MP-HDL-2016-003). The results will be published in peer-reviewed journals. PMID:27098826

  19. Improved protocol to purify untagged amelogenin - Application to murine amelogenin containing the equivalent P70→T point mutation observed in human amelogenesis imperfecta.

    PubMed

    Buchko, Garry W; Shaw, Wendy J

    2015-01-01

    Amelogenin is the predominant extracellular protein responsible for converting carbonated hydroxyapatite into dental enamel, the hardest and most heavily mineralized tissue in vertebrates. Despite much effort, the precise mechanism by which amelogenin regulates enamel formation is not fully understood. To assist efforts aimed at understanding the biochemical mechanism of enamel formation, more facile protocols to purify recombinantly expressed amelogenin, ideally without any tag to assist affinity purification, are advantageous. Here we describe an improved method to purify milligram quantities of amelogenin that exploits its high solubility in 2% glacial acetic acid under conditions of low ionic strength. The method involves heating the frozen cell pellet for two 15min periods at ∼70°C with 2min of sonication in between, dialysis twice in 2% acetic acid (1:250 v/v), and reverse phase chromatography. A further improvement in yield is obtained by resuspending the frozen cell pellet in 6M guanidine hydrochloride in the first step. The acetic acid heating method is illustrated with a murine amelogenin containing the corresponding P70→T point mutation observed in an human amelogenin associated with amelogenesis imperfecta (P71T), while the guanidine hydrochloride heating method is illustrated with wild type murine amelogenin (M180). The self-assembly properties of P71T were probed by NMR chemical shift perturbation studies as a function of protein (0.1-1.8mM) and NaCl (0-367mM) concentration. Relative to similar studies with wild type murine amelogenin, P71T self-associates at lower protein or salt concentrations with the interactions initiated near the N-terminus.

  20. Improved protocol to purify untagged amelogenin – Application to murine amelogenin containing the equivalent P70→T point mutation observed in human amelogenesis imperfecta

    SciTech Connect

    Buchko, Garry W.; Shaw, Wendy J.

    2014-10-13

    Amelogenin is the predominant extracellular protein responsible for converting carbonated hydroxyapatite into dental enamel, the hardest and most heavily mineralized tissue in vertebrates. Despite much effort, the precise mechanism by which amelogenin regulates enamel formation is not fully understood. To assist efforts aimed at understanding the biochemical mechanism of enamel formation, more facile protocols to purify recombinantly expressed amelogenin, ideally without any tag to assist affinity purification, are advantageous. Here we describe an improved method to purify milligram quantities of amelogenin that exploits its high solubility in 2% glacial acetic acid under conditions of low ionic strength. The method involves heating the frozen cell pellet for two 15 min periods at ~70 ºC with two minutes of sonication in between, dialysis twice in 2% acetic acid (1:250 v/v), and reverse phase chromatography. A further improvement in yield is obtained by resuspending the frozen cell pellet in 6 M guanidine hydrochloride in the first step. The acetic acid heating method is illustrated with a murine amelogenin containing the corresponding P70→T point mutation observed in an human amelogenin associated with amelogenesis imperfecta (P71T), while the guanidine hydrochloride heating method is illustrated with wild type murine amelogenin (M180). The self-assembly properties of P71T were probed by NMR chemical shift perturbation studies as a function of protein (0.1 to 1.8 mM) and NaCl (0 to 367 mM) concentration. In conclusion, relative to similar studies with wild type murine amelogenin, P71T self-associates at lower protein or salt concentrations with the interactions initiated near the N-terminus.

  1. Improved protocol to purify untagged amelogenin – Application to murine amelogenin containing the equivalent P70→T point mutation observed in human amelogenesis imperfecta

    DOE PAGES

    Buchko, Garry W.; Shaw, Wendy J.

    2014-10-13

    Amelogenin is the predominant extracellular protein responsible for converting carbonated hydroxyapatite into dental enamel, the hardest and most heavily mineralized tissue in vertebrates. Despite much effort, the precise mechanism by which amelogenin regulates enamel formation is not fully understood. To assist efforts aimed at understanding the biochemical mechanism of enamel formation, more facile protocols to purify recombinantly expressed amelogenin, ideally without any tag to assist affinity purification, are advantageous. Here we describe an improved method to purify milligram quantities of amelogenin that exploits its high solubility in 2% glacial acetic acid under conditions of low ionic strength. The method involvesmore » heating the frozen cell pellet for two 15 min periods at ~70 ºC with two minutes of sonication in between, dialysis twice in 2% acetic acid (1:250 v/v), and reverse phase chromatography. A further improvement in yield is obtained by resuspending the frozen cell pellet in 6 M guanidine hydrochloride in the first step. The acetic acid heating method is illustrated with a murine amelogenin containing the corresponding P70→T point mutation observed in an human amelogenin associated with amelogenesis imperfecta (P71T), while the guanidine hydrochloride heating method is illustrated with wild type murine amelogenin (M180). The self-assembly properties of P71T were probed by NMR chemical shift perturbation studies as a function of protein (0.1 to 1.8 mM) and NaCl (0 to 367 mM) concentration. In conclusion, relative to similar studies with wild type murine amelogenin, P71T self-associates at lower protein or salt concentrations with the interactions initiated near the N-terminus.« less

  2. Line-of-sight performance improvement with reaction-wheel isolation

    NASA Technical Reports Server (NTRS)

    Rodden, J. J.; Dougherty, H. J.; Reschke, L. F.; Hasha, M. D.; Davis, L. P.

    1986-01-01

    Design techniques used to isolate and damp vibrations from the reaction control wheels assembly (RCA) from the structural dynamics of the Hubble Space Telescope are described. The four RCAs can generate sufficient torque for large maneuvers and precision control during fine pointing. The RCAs are to operate continuously during observations and thereby pose the potential of inducing jitter. The image stability must be constrained to less than 0.007 arcsec rms per 24 hr and pointing accuracy must be 0.01 arcsec for 100 hr. Vibration-producing characteristics of the RCAs are identified, and test results are provided from measurements of the magnitudes of the associated disturbances. Design features and test results for the vibration isolators employed with the RCAs to effect viscous damping of RCA axial perturbations are presented. The dampers are expected to reduce jitter effects of RCA operation by a half.

  3. Development of an improved selective agar medium for isolation of Yersinia pestis.

    PubMed

    Ber, Raphael; Mamroud, Emanuelle; Aftalion, Moshe; Tidhar, Avital; Gur, David; Flashner, Yehuda; Cohen, Sara

    2003-10-01

    Existing media designed for selective isolation of clinically important members of the genus Yersinia were found to be unsatisfactory for the growth and isolation of Yersinia pestis. We report the development of a new selective agar medium (termed BIN) that supports the growth of Y. pestis. The development of the formulation of this medium was based on a fluorescence screening system designed for monitoring bacterial growth on semisolid media, using a green fluorescent protein-expressing strain. High-throughput combinatorial experiments can be conducted for the quantitative evaluation of the effect of different medium components on growth. Generation of fluorescence plots in this system, using microplates, allowed the quantitative evaluation of the growth rate of Y. pestis EV76 cultures in different agar compositions. The final BIN formulation is based on brain heart infusion agar, to which the selective agents irgasan, cholate salts, crystal violet, and nystatin were introduced. It was found that BIN agar is more efficient in supporting colony formation and recovery of Y. pestis than are the conventional semisolid media MacConkey agar and Yersinia-selective agar (cefsulodin-irgasan-novobiocin agar). The advantage of BIN over other media has been also demonstrated in recovering virulent Y. pestis from the mixed bacterial populations found in decaying carcasses of infected mice. The BIN medium is suggested as a selective medium for isolation and recovery of Y. pestis from various backgrounds.

  4. Memantine improves memory for taste-avoidance learning in day-old chicks exposed to isolation stress.

    PubMed

    Barber, Teresa A; Meyers, Ryan A; McGettigan, Brian F

    2010-04-01

    Activation of NMDA receptors by glutamate is particularly important in the initial stages of memory consolidation. Memantine, a noncompetitive NMDA receptor antagonist, ameliorates memory impairment under certain circumstances, despite blocking the activation of NMDA receptors. The present experiments tested the hypothesis that memantine can improve memory deficits induced by isolation stress in day-old chicks (Gallus gallus domesticus) trained in a one-trial taste-avoidance task. Three experiments assessed the effects of memantine at different concentrations and in combination with isolation stress. The results of Experiment 1 indicate that, under normal, non-stressed conditions, memory in control animals is strong and 15.0 mM memantine impairs memory, similar to that seen in many studies of the effects of NMDA receptor antagonists on learning. However, the results of Experiments 2 and 3 showed that, when chicks were exposed to isolation stress during the pre-training period, memory formation for saline-injected control animals was impaired and 5.0 mM memantine significantly improved memory in an inverted U-shaped dose response function. The current results extend the findings that memantine can ameliorate memory impairment and supports the hypothesis that memantine, despite its action to reduce NMDA receptor activity, can facilitate normalized memory acquisition.

  5. A novel modified ultra-long agonist protocol improves the outcome of high body mass index women with polycystic ovary syndrome undergoing IVF/ICSI.

    PubMed

    Tu, Jiongjiong; Lin, Ge; Lu, Changfu; Gong, Fei

    2014-03-01

    In an attempt to evaluate the effectiveness of a novel modified ultra-long agonist (ULA) protocol on polycystic ovary syndrome (PCOS) patients undergoing in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI), a retrospective study of 499 women employed with either ULA or conventional long agonist (LA) protocol was analyzed. In high BMI group (>25 kg/m²), the ULA protocol yielded significant higher clinic pregnancy rate (PR) (70.2% versus 50.8%, p < 0.05), implantation rate (52.7% versus 35.7%, p < 0.05) and live birth rate (63.8% versus 39.0%, p < 0.05) when compared with LA protocol. In low BMI group (≤25 kg/m²), the ULA protocol also demonstrated a higher clinic PR (70.8% versus 59.5%, p < 0.05) whereas implantation rate and live birth rate are comparable. Within ULA protocol, the clinic PR, implantation rate and live birth rate are similar between high and low BMI patients. Similarly, the clinic PR and live birth rate demonstrated no significant difference within LA group but there is a significant lower implantation rate (35.7% versus 63.9%, p < 0.05) observed in high BMI patients. No difference in miscarriage rate and severe OHSS rate was found among all groups. In conclusion, ULA protocol benefits the IVF outcomes of PCOS patients with high BMI status.

  6. ISOLATION OF MOUSE NEUTROPHILS

    PubMed Central

    Swamydas, Muthulekha; Luo, Yi; Dorf, Martin E.; Lionakis, Michail S.

    2015-01-01

    Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited and acquired qualitative and quantitative neutrophil defects are at high risk for developing bacterial and fungal infections and suffering adverse outcomes from these infections. Therefore, research aiming at defining the molecular factors that modulate neutrophil effector function under homeostatic conditions and during infection is essential for devising strategies to augment neutrophil function and improve the outcome of infected individuals. This unit describes a reproducible density gradient centrifugation-based protocol that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice both at the steady state and following infection with Candida albicans as described in UNIT 19.6. In another protocol, we also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or Fluorescence-activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver or the spleen. Finally, methods for isolating neutrophils from mouse peritoneal fluid and peripheral blood are included. Mouse neutrophils isolated by these protocols can be used for examining several aspects of cellular function ex vivo including pathogen binding, phagocytosis and killing, neutrophil chemotaxis, oxidative burst, degranulation and cytokine production, and for performing neutrophil adoptive transfer experiments. PMID:26237011

  7. Very small embryonic-like stem-cell optimization of isolation protocols: an update of molecular signatures and a review of current in vivo applications

    PubMed Central

    Shin, Dong-Myung; Suszynska, Malwina; Mierzejewska, Kasia; Ratajczak, Janina; Ratajczak, Mariusz Z

    2013-01-01

    As the theory of stem cell plasticity was first proposed, we have explored an alternative hypothesis for this phenomenon: namely that adult bone marrow (BM) and umbilical cord blood (UCB) contain more developmentally primitive cells than hematopoietic stem cells (HSCs). In support of this notion, using multiparameter sorting we were able to isolate small Sca1+Lin−CD45− cells and CD133+Lin−CD45− cells from murine BM and human UCB, respectively, which were further enriched for the detection of various early developmental markers such as the SSEA antigen on the surface and the Oct4 and Nanog transcription factors in the nucleus. Similar populations of cells have been found in various organs by our team and others, including the heart, brain and gonads. Owing to their primitive cellular features, such as the high nuclear/cytoplasm ratio and the presence of euchromatin, they are called very small embryonic-like stem cells (VSELs). In the appropriate in vivo models, VSELs differentiate into long-term repopulating HSCs, mesenchymal stem cells (MSCs), lung epithelial cells, cardiomyocytes and gametes. In this review, we discuss the most recent data from our laboratory and other groups regarding the optimal isolation procedures and describe the updated molecular characteristics of VSELs. PMID:24232255

  8. The Use of Metal Filled Via Holes for Improving Isolation in LTCC RF and Wireless Multichip Packages

    NASA Technical Reports Server (NTRS)

    Ponchak, George E.; Chun, Donghoon; Yook, Jong-Gwan; Katehi, Linda P. B.

    1999-01-01

    LTCC MCMs (Low Temperature Cofired Ceramic MultiChip Module) for RF and wireless systems often use metal filled via holes to improve isolation between the stripline and microstrip interconnects. In this paper, results from a 3D-FEM electromagnetic characterization of microstrip and stripline interconnects with metal filled via fences for isolation are presented. It is shown that placement of a via hole fence closer than three times the substrate height to the transmission lines increases radiation and coupling. Radiation loss and reflections are increased when a short via fence is used in areas suspected of having high radiation. Also, via posts should not be separated by more than three times the substrate height for low radiation loss, coupling, and suppression of higher order modes in a package.

  9. Thermophilic and cellulolytic consortium isolated from composting plants improves anaerobic digestion of cellulosic biomass: Toward a microbial resource management approach.

    PubMed

    Kinet, R; Destain, J; Hiligsmann, S; Thonart, P; Delhalle, L; Taminiau, B; Daube, G; Delvigne, F

    2015-01-01

    A cellulolytic consortium was isolated from a composting plant in order to boost the initial hydrolysis step encountered in anaerobic digestion. Improvement of the cellulose degradation, as well as biogas production, was observed for the cultures inoculated with the exogenous consortium. Metagenomics analyses pointed out a weak richness (related to the number of OTUs) of the exogenous consortium induced by the selective pressure (cellulose as sole carbon source) met during the initial isolation steps. Main microbial strains determined were strictly anaerobic and belong to the Clostridia class. During cellulose anaerobic degradation, pH drop induced a strong modification of the microbial population. Despite the fact that richness and evenness were very weak, the exogenous consortium was able to adapt and to maintain the cellulolytic degradation potential. This important result point out the fact that simplified microbial communities could be used in order to increase the robustness of mixed cultures involved in environmental biotechnology. PMID:25879181

  10. Thermophilic and cellulolytic consortium isolated from composting plants improves anaerobic digestion of cellulosic biomass: Toward a microbial resource management approach.

    PubMed

    Kinet, R; Destain, J; Hiligsmann, S; Thonart, P; Delhalle, L; Taminiau, B; Daube, G; Delvigne, F

    2015-01-01

    A cellulolytic consortium was isolated from a composting plant in order to boost the initial hydrolysis step encountered in anaerobic digestion. Improvement of the cellulose degradation, as well as biogas production, was observed for the cultures inoculated with the exogenous consortium. Metagenomics analyses pointed out a weak richness (related to the number of OTUs) of the exogenous consortium induced by the selective pressure (cellulose as sole carbon source) met during the initial isolation steps. Main microbial strains determined were strictly anaerobic and belong to the Clostridia class. During cellulose anaerobic degradation, pH drop induced a strong modification of the microbial population. Despite the fact that richness and evenness were very weak, the exogenous consortium was able to adapt and to maintain the cellulolytic degradation potential. This important result point out the fact that simplified microbial communities could be used in order to increase the robustness of mixed cultures involved in environmental biotechnology.

  11. Improved Method for Isolation of Microbial RNA from Biofuel Feedstock for Metatranscriptomics

    SciTech Connect

    Piao, Hailan; Markillie, Lye Meng; Culley, David E.; Mackie, Roderick I.; Hess, Matthias

    2013-03-28

    Metatranscriptomics—gene express profiling via DNA sequencing—is a powerful tool to identify genes that are ac- tively expressed and might contribute to the phenotype of individual organisms or the phenome (the sum of several phenotypes) of a microbial community. Furthermore, metatranscriptome studies can result in extensive catalogues of genes that encode for enzymes of industrial relevance. In both cases, a major challenge for generating a high quality metatranscriptome is the extreme lability of RNA and its susceptibility to ubiquitous RNAses. The microbial commu- nity (the microbiome) of the cow rumen efficiently degrades lignocelullosic biomass, generates significant amounts of methane, a greenhouse gas twenty times more potent than carbon dioxide, and is of general importance for the physio- logical wellbeing of the host animal. Metatranscriptomes of the rumen microbiome from animals kept under different conditions and from various types of rumen-incubated biomass can be expected to provide new insights into these highly interesting phenotypes and subsequently provide the framework for an enhanced understanding of this socio- economically important ecosystem. The ability to isolate large amounts of intact RNA will significantly facilitate accu- rate transcript annotation and expression profiling. Here we report a method that combines mechanical disruption with chemical homogenization of the sample material and consistently yields 1 mg of intact RNA from 1 g of rumen-in- cubated biofuel feedstock. The yield of total RNA obtained with our method exceeds the RNA yield achieved with pre- viously reported isolation techniques, which renders RNA isolated with the method presented here as an ideal starting material for metatranscriptomic analyses and other molecular biology applications that require significant amounts of starting material.

  12. Improved quality and yield of islets isolated from human pancreata using a two-step digestion method.

    PubMed

    Kenmochi, T; Miyamoto, M; Une, S; Nakagawa, Y; Moldovan, S; Navarro, R A; Benhamou, P Y; Brunicardi, F C; Mullen, Y

    2000-03-01

    A new approach, involving a two-step digestion process and Los Angeles preservation solution #1 (LAP-1), a cold storage solution, was developed for isolation of high-quality islets from human pancreata for transplantation. This approach markedly improves the islet yield, purity and viability, and the isolation success rate. In this method, the pancreas was digested first in warm collagenase solution for up to 20 minutes. After decanting the enzyme solution, partially digested tissue was dissociated by gentle agitation in cold LAP-1 solution without additional collagenase. The digested tissues were stored in cold LAP-1 solution until islet purification on Euro-Ficoll. Forty-six islet isolations were performed consecutively by the new method (group 1). These results were compared to those obtained earlier with 46 consecutive isolations, using our previous method that had been used before development of the new method (group 2). Our old method was a modification of Ricordi's method involving only warm collagenase digestion and the storage of digested tissues in cold Hanks balanced salt solution. All pancreata were partial, containing the body and tail. There were no significant differences in both groups with regard to the donor age, cold ischemic time, harvesting conditions, and pancreatic weight. Pancreas digestion was completed in approximately 1 hour in both groups. The isolation success rate as determined by viable islets after 2 days in culture was 93.5% (43 of 46 cases) in group 1, and 56.5% (26 of the 46) in group 2. Immediately after isolation, the new method yielded a total of 335,739 +/- 36,244 islets equivalent to 150 microm (IEQ) and 6,233 +/- 681 IEQ/g of pancreas with 83 +/- 2.5% purity, whereas the old method yielded a total of 195,587 +/- 25,242 IEQ and 3,763 +/- 5,509 IEQ/g with 69.2 +/- 4.7% purity. Isolated islets in group 1 maintained a good three-dimensional structure, displayed normal insulin release to high glucose stimulation in vitro, and

  13. Improved dose calculation accuracy for low energy brachytherapy by optimizing dual energy CT imaging protocols for noise reduction using sinogram affirmed iterative reconstruction.

    PubMed

    Landry, Guillaume; Gaudreault, Mathieu; van Elmpt, Wouter; Wildberger, Joachim E; Verhaegen, Frank

    2016-03-01

    The goal of this study was to evaluate the noise reduction achievable from dual energy computed tomography (CT) imaging (DECT) using filtered backprojection (FBP) and iterative image reconstruction algorithms combined with increased imaging exposure. We evaluated the data in the context of imaging for brachytherapy dose calculation, where accurate quantification of electron density ρe and effective atomic number Zeff is beneficial. A dual source CT scanner was used to scan a phantom containing tissue mimicking inserts. DECT scans were acquired at 80 kVp/140Sn kVp (where Sn stands for tin filtration) and 100 kVp/140Sn kVp, using the same values of the CT dose index CTDIvol for both settings as a measure for the radiation imaging exposure. Four CTDIvol levels were investigated. Images were reconstructed using FBP and sinogram affirmed iterative reconstruction (SAFIRE) with strength 1,3 and 5. From DECT scans two material quantities were derived, Zeff and ρe. DECT images were used to assign material types and the amount of improperly assigned voxels was quantified for each protocol. The dosimetric impact of improperly assigned voxels was evaluated with Geant4 Monte Carlo (MC) dose calculations for an (125)I source in numerical phantoms. Standard deviations for Zeff and ρe were reduced up to a factor ∼2 when using SAFIRE with strength 5 compared to FBP. Standard deviations on Zeff and ρe as low as 0.15 and 0.006 were achieved for the muscle insert representing typical soft tissue using a CTDIvol of 40 mGy and 3mm slice thickness. Dose calculation accuracy was generally improved when using SAFIRE. Mean (maximum absolute) dose errors of up to 1.3% (21%) with FBP were reduced to less than 1% (6%) with SAFIRE at a CTDIvol of 10 mGy. Using a CTDIvol of 40mGy and SAFIRE yielded mean dose calculation errors of the order of 0.6% which was the MC dose calculation precision in this study and no error was larger than ±2.5% as opposed to errors of up to -4% with FPB. This

  14. A study protocol of a randomised controlled trial to investigate if a community based strength training programme improves work task performance in young adults with Down syndrome

    PubMed Central

    2010-01-01

    Background Muscle strength is important for young people with Down syndrome as they make the transition to adulthood, because their workplace activities typically emphasise physical rather than cognitive skills. Muscle strength is reduced up to 50% in people with Down syndrome compared to their peers without disability. Progressive resistance training improves muscle strength and endurance in people with Down syndrome. However, there is no evidence on whether it has an effect on work task performance or physical activity levels. The aim of this study is to investigate if a student-led community-based progressive resistance training programme can improve these outcomes in adolescents and young adults with Down syndrome. Methods A randomised controlled trial will compare progressive resistance training with a control group undertaking a social programme. Seventy adolescents and young adults with Down syndrome aged 14-22 years and mild to moderate intellectual disability will be randomly allocated to the intervention or control group using a concealed method. The intervention group will complete a 10-week, twice a week, student-led progressive resistance training programme at a local community gymnasium. The student mentors will be undergraduate physiotherapy students. The control group will complete an arts/social programme with a student mentor once a week for 90 minutes also for 10 weeks to control for the social aspect of the intervention. Work task performance (box stacking, pail carry), muscle strength (1 repetition maximum for chest and leg press) and physical activity (frequency, duration, intensity over 7-days) will be assessed at baseline (Week 0), following the intervention (Week 11), and at 3 months post intervention (Week 24) by an assessor blind to group allocation. Data will be analysed using ANCOVA with baseline measures as covariates. Discussion This paper outlines the study protocol for a randomised controlled trial on the effects of progressive

  15. A water-based training program that include perturbation exercises to improve stepping responses in older adults: study protocol for a randomized controlled cross-over trial

    PubMed Central

    Melzer, Itshak; Elbar, Ori; Tsedek, Irit; Oddsson, Lars IE

    2008-01-01

    Background Gait and balance impairments may increase the risk of falls, the leading cause of accidental death in the elderly population. Fall-related injuries constitute a serious public health problem associated with high costs for society as well as human suffering. A rapid step is the most important protective postural strategy, acting to recover equilibrium and prevent a fall from initiating. It can arise from large perturbations, but also frequently as a consequence of volitional movements. We propose to use a novel water-based training program which includes specific perturbation exercises that will target the stepping responses that could potentially have a profound effect in reducing risk of falling. We describe the water-based balance training program and a study protocol to evaluate its efficacy (Trial registration number #NCT00708136). Methods/Design The proposed water-based training program involves use of unpredictable, multi-directional perturbations in a group setting to evoke compensatory and volitional stepping responses. Perturbations are made by pushing slightly the subjects and by water turbulence, in 24 training sessions conducted over 12 weeks. Concurrent cognitive tasks during movement tasks are included. Principles of physical training and exercise including awareness, continuity, motivation, overload, periodicity, progression and specificity were used in the development of this novel program. Specific goals are to increase the speed of stepping responses and improve the postural control mechanism and physical functioning. A prospective, randomized, cross-over trial with concealed allocation, assessor blinding and intention-to-treat analysis will be performed to evaluate the efficacy of the water-based training program. A total of 36 community-dwelling adults (age 65–88) with no recent history of instability or falling will be assigned to either the perturbation-based training or a control group (no training). Voluntary step reaction times

  16. Effectiveness of Motivational Interviewing in improving lipid level in patients with dyslipidemia assisted by general practitioners: Dislip-EM study protocol

    PubMed Central

    2011-01-01

    Background The non-pharmacological approach to cholesterol control in patients with hyperlipidemia is based on the promotion of a healthy diet and physical activity. Thus, to help patients change their habits, it is essential to identify the most effective approach. Many efforts have been devoted to explain changes in or adherence to specific health behaviors. Such efforts have resulted in the development of theories that have been applied in prevention campaigns, and that include brief advice and counseling services. Within this context, Motivational Interviewing has proven to be effective in changing health behaviors in specific cases. However, more robust evidence is needed on the effectiveness of Motivational Interviewing in treating chronic pathologies -such as dyslipidemia- in patients assisted by general practitioners. This article describes a protocol to assess the effectiveness of MI as compared with general practice (brief advice), with the aim of improving lipid level control in patients with dyslipidemia assisted by a general practitioner. Methods/Design An open, two-arm parallel, multicentre, cluster, controlled, randomized, clinical trial will be performed. A total of 48-50 general practitioners from 35 public primary care centers in Spain will be randomized and will recruit 436 patients with dyslipidemia. They will perform an intervention based either on Motivational Interviewing or on the usual brief advice. After an initial assessment, follow-ups will be performed at 2, 4, 8 and 12 months. Primary outcomes are lipid levels (total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides) and cardiovascular risk. The study will assess the degree of dietary and physical activity improvement, weight loss in overweight patients, and adherence to treatment guidelines. Discussion Motivational interview skills constitute the primary strategies GPs use to treat their patients. Having economical, simple, effective and applicable techniques is essential

  17. Study protocol of a randomised controlled trial of intranasal ketamine compared with intranasal fentanyl for analgesia in children with suspected, isolated extremity fractures in the paediatric emergency department

    PubMed Central

    Reynolds, Stacy L; Studnek, Jonathan R; Bryant, Kathleen; VanderHave, Kelly; Grossman, Eric; Moore, Charity G; Young, James; Hogg, Melanie; Runyon, Michael S

    2016-01-01

    Introduction Fentanyl is the most widely studied intranasal (IN) analgesic in children. IN subdissociative (INSD) ketamine may offer a safe and efficacious alternative to IN fentanyl and may decrease overall opioid use during the emergency department (ED) stay. This study examines the feasibility of a larger, multicentre clinical trial comparing the safety and efficacy of INSD ketamine to IN fentanyl and the potential role for INSD ketamine in reducing total opioid medication usage. Methods and analysis This double-blind, randomised controlled, pilot trial will compare INSD ketamine (1 mg/kg) to IN fentanyl (1.5 μg/kg) for analgesia in 80 children aged 4–17 years with acute pain from a suspected, single extremity fracture. The primary safety outcome for this pilot trial will be the frequency of cumulative side effects and adverse events at 60 min after drug administration. The primary efficacy outcome will be exploratory and will be the mean reduction of pain scale scores at 20 min. The study is not powered to examine efficacy. Secondary outcome measures will include the total dose of opioid pain medication in morphine equivalents/kg/hour (excluding study drug) required during the ED stay, number and reason for screen failures, time to consent, and the number and type of protocol deviations. Patients may receive up to 2 doses of study drug. Ethics and dissemination This study was approved by the US Food and Drug Administration, the local institutional review board and the study data safety monitoring board. This study data will be submitted for publication regardless of results and will be used to establish feasibility for a multicentre, non-inferiority trial. Trial registration number NCT02521415. PMID:27609854

  18. Trichuris suis ova in relapsing-remitting multiple sclerosis and clinically isolated syndrome (TRIOMS): study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background Trichuris suis ova is a probiotic treatment based on the hygiene hypothesis. It has been demonstrated as safe and effective in autoimmune inflammatory bowel diseases and clinical trials indicate that helminth infections also have an immunomodulatory effect in multiple sclerosis. We hypothesize that administering 2,500 Trichuris suis ova eggs orally every two weeks for 12 months is - due to its immunomodulatory and anti-inflammatory effect - significantly more effective than oral placebo in preventing new T2 and Gd+ lesions, as quantified by cerebral MRI and clinical examination, in relapsing-remitting multiple sclerosis and clinically isolated syndrome. Methods/Design Fifty patients with relapsing-remitting multiple sclerosis or clinically isolated syndrome with clinical activity, not undergoing any standard therapies, will be randomized 1:1 to Trichuris suis ova 2,500 eggs every two weeks or matching placebo. The safety, tolerability and effect on disease activity and in vivo mechanisms of action of Trichuris suis ova in MS will be assessed by neurological, laboratory and immunological exams and magnetic resonance imaging throughout the 12-month treatment period and over a follow-up period of 6 months. Various immunological analyses will be used to assess the overall patient immune response prior to and at varying time points following treatment with Trichuris suis ova. Discussion We anticipate that Trichuris suis ova will be well tolerated and more effective than the placebo in preventing new T2 and Gd+ lesions, as quantified by MRI. We also expect the Th1/Th17 proinflammatory response to shift towards the more anti-inflammatory Th2 response. This study has important clinical implications and will involve extensive research on the immunology of helminth therapy. Trial registration ClinicalTrials.gov: NCT01413243 PMID:23782752

  19. The efficiency of using a seismic base isolation system for a 2D concrete frame founded upon improved soft soil with rigid inclusions

    NASA Astrophysics Data System (ADS)

    Awwad, Talal; Donia, Modar

    2016-03-01

    2D finite element models were developed to analyze the effect of improved soft-soil foundation on the efficiency of a base-isolated concrete frame. Static and dynamic analyses were performed for a frame on raft foundation. Non-improved and improved soft-soil foundation using rigid inclusions were considered, as well as the use of high damping rubber bearing as base isolation. Results show that the use of rigid inclusions increases the efficiency of base isolation; base shear is reduced by 38% and maximum acceleration at the top of the frame by 30%.

  20. Gallic acid isolated from Spirogyra sp. improves cardiovascular disease through a vasorelaxant and antihypertensive effect.

    PubMed

    Kang, Nalae; Lee, Ji-Hyeok; Lee, WonWoo; Ko, Ju-Young; Kim, Eun-A; Kim, Jin-Soo; Heu, Min-Soo; Kim, Gwang Hoon; Jeon, You-Jin

    2015-03-01

    In this study, we investigated the vasorelaxant and antihypertensive effects of gallic acid (GA), a polyphenol isolated from the green alga Spirogyra sp., to assess its suitability as a therapeutic for cardiovascular diseases (CVDs). We examined the effect of GA on endothelium-dependent vasorelaxation in human umbilical vein endothelial cells (HUVECs). GA increased nitric oxide (NO) levels by increasing phosphorylation of endothelial nitric oxide synthase (eNOS), and its effect on NO production was attenuated by pretreatment with the eNOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME). We also investigated its antihypertensive effect by examining GA-mediated inhibition of angiotensin-I converting enzyme (ACE). GA inhibited ACE with a half-maximal inhibitory concentration (IC50) value of 37.38 ± 0.39 μg/ml. In silico simulations revealed that GA binds to the active site of ACE (PDB: 1O86) with a binding energy of -270.487 kcal/mol. Furthermore, GA clearly reduced blood pressure in spontaneously hypertensive rats (SHR) to an extent comparable to captopril. These results suggest that GA isolated from Spirogyra sp. exerts multiple therapeutic effects and has potential as a CVD treatment. PMID:25727171

  1. Gallic acid isolated from Spirogyra sp. improves cardiovascular disease through a vasorelaxant and antihypertensive effect.

    PubMed

    Kang, Nalae; Lee, Ji-Hyeok; Lee, WonWoo; Ko, Ju-Young; Kim, Eun-A; Kim, Jin-Soo; Heu, Min-Soo; Kim, Gwang Hoon; Jeon, You-Jin

    2015-03-01

    In this study, we investigated the vasorelaxant and antihypertensive effects of gallic acid (GA), a polyphenol isolated from the green alga Spirogyra sp., to assess its suitability as a therapeutic for cardiovascular diseases (CVDs). We examined the effect of GA on endothelium-dependent vasorelaxation in human umbilical vein endothelial cells (HUVECs). GA increased nitric oxide (NO) levels by increasing phosphorylation of endothelial nitric oxide synthase (eNOS), and its effect on NO production was attenuated by pretreatment with the eNOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME). We also investigated its antihypertensive effect by examining GA-mediated inhibition of angiotensin-I converting enzyme (ACE). GA inhibited ACE with a half-maximal inhibitory concentration (IC50) value of 37.38 ± 0.39 μg/ml. In silico simulations revealed that GA binds to the active site of ACE (PDB: 1O86) with a binding energy of -270.487 kcal/mol. Furthermore, GA clearly reduced blood pressure in spontaneously hypertensive rats (SHR) to an extent comparable to captopril. These results suggest that GA isolated from Spirogyra sp. exerts multiple therapeutic effects and has potential as a CVD treatment.

  2. A best practice fall prevention exercise program to improve balance, strength / power, and psychosocial health in older adults: study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background With increasing age neuromuscular deficits (e.g., sarcopenia) may result in impaired physical performance and an increased risk for falls. Prominent intrinsic fall-risk factors are age-related decreases in balance and strength / power performance as well as cognitive decline. Additional studies are needed to develop specifically tailored exercise programs for older adults that can easily be implemented into clinical practice. Thus, the objective of the present trial is to assess the effects of a fall prevention program that was developed by an interdisciplinary expert panel on measures of balance, strength / power, body composition, cognition, psychosocial well-being, and falls self-efficacy in healthy older adults. Additionally, the time-related effects of detraining are tested. Methods/Design Healthy old people (n = 54) between the age of 65 to 80 years will participate in this trial. The testing protocol comprises tests for the assessment of static / dynamic steady-state balance (i.e., Sharpened Romberg Test, instrumented gait analysis), proactive balance (i.e., Functional Reach Test; Timed Up and Go Test), reactive balance (i.e., perturbation test during bipedal stance; Push and Release Test), strength (i.e., hand grip strength test; Chair Stand Test), and power (i.e., Stair Climb Power Test; countermovement jump). Further, body composition will be analysed using a bioelectrical impedance analysis system. In addition, questionnaires for the assessment of psychosocial (i.e., World Health Organisation Quality of Life Assessment-Bref), cognitive (i.e., Mini Mental State Examination), and fall risk determinants (i.e., Fall Efficacy Scale – International) will be included in the study protocol. Participants will be randomized into two intervention groups or the control / waiting group. After baseline measures, participants in the intervention groups will conduct a 12-week balance and strength / power exercise intervention 3 times per week, with

  3. Analyzing the effect of routing protocols on media access control protocols in radio networks

    SciTech Connect

    Barrett, C. L.; Drozda, M.; Marathe, A.; Marathe, M. V.

    2002-01-01

    We study the effect of routing protocols on the performance of media access control (MAC) protocols in wireless radio networks. Three well known MAC protocols: 802.11, CSMA, and MACA are considered. Similarly three recently proposed routing protocols: AODV, DSR and LAR scheme 1 are considered. The experimental analysis was carried out using GloMoSim: a tool for simulating wireless networks. The main focus of our experiments was to study how the routing protocols affect the performance of the MAC protocols when the underlying network and traffic parameters are varied. The performance of the protocols was measured w.r.t. five important parameters: (i) number of received packets, (ii) average latency of each packet, (iii) throughput (iv) long term fairness and (v) number of control packets at the MAC layer level. Our results show that combinations of routing and MAC protocols yield varying performance under varying network topology and traffic situations. The result has an important implication; no combination of routing protocol and MAC protocol is the best over all situations. Also, the performance analysis of protocols at a given level in the protocol stack needs to be studied not locally in isolation but as a part of the complete protocol stack. A novel aspect of our work is the use of statistical technique, ANOVA (Analysis of Variance) to characterize the effect of routing protocols on MAC protocols. This technique is of independent interest and can be utilized in several other simulation and empirical studies.

  4. A new improved protocol for in vitro intratubular dentinal bacterial contamination for antimicrobial endodontic tests: standardization and validation by confocal laser scanning microscopy

    PubMed Central

    de ANDRADE, Flaviana Bombarda; ARIAS, Marcela Paola Castro; MALIZA, Amanda Garcia Alves; DUARTE, Marco Antonio Hungaro; GRAEFF, Márcia Sirlene Zardin; AMOROSO-SILVA, Pablo Andrés; MIDENA, Raquel Zanin; de MORAES, Ivaldo Gomes

    2015-01-01

    Objectives To compare three methods of intratubular contamination that simulate endodontic infections using confocal laser scanning microscopy (CLSM). Material and Methods Two pre-existing models of dentinal contamination were used to induce intratubular infection (groups A and B). These methods were modified in an attempt to improve the model (group C). Among the modifications it may be included: specimen contamination for five days, ultrasonic bath with BHI broth after specimen sterilization, use of E. faecalis during the exponential growth phase, greater concentration of inoculum, and two cycles of centrifugation on alternate days with changes of culture media. All specimens were longitudinally sectioned and stained with of LIVE/DEAD® for 20 min. Specimens were assessed using CLSM, which provided images of the depth of viable bacterial proliferation inside the dentinal tubules. Additionally, three examiners used scores to classify the CLSM images according to the following parameters: homogeneity, density, and depth of the bacterial contamination inside the dentinal tubules. Kruskal-Wallis and Dunn’s tests were used to evaluate the live and dead cells rates, and the scores obtained. Results The contamination scores revealed higher contamination levels in group C when compared with groups A and B (p<0.05). No differences were observed between group A and B (p>0.05). The volume of live cells in group C was higher than in groups A and B (p<0.05). Conclusion The new protocol for intratubular infection resulted in high and uniform patterns of bacterial contamination and higher cell viability in all specimens when compared with the current methods. PMID:26200524

  5. Improved outcome in high-risk childhood acute lymphoblastic leukemia defined by prednisone-poor response treated with double Berlin-Frankfurt-Muenster protocol II.

    PubMed

    Aricò, Maurizio; Valsecchi, Maria Grazia; Conter, Valentino; Rizzari, Carmelo; Pession, Andrea; Messina, Chiara; Barisone, Elena; Poggi, Vincenzo; De Rossi, Giulio; Locatelli, Franco; Micalizzi, Maria Concetta; Basso, Giuseppe; Masera, Giuseppe

    2002-07-15

    One hundred ninety-eight children and adolescents were entered in the Associazione Italiana di Ematologia ed Oncologia Pediatrica (AIEOP)-ALL95 study for high-risk acute lymphoblastic leukemia (ALL). Inclusion criteria were poor response to initial prednisone/intrathecal methotrexate (prednisone-poor response [PPR]), resistance to induction therapy, translocation t(9;22), infants with the t(4;11), or CD10(-) ALL. The event-free survival (EFS) rate at 4 years was 56.5% (SE, 3.9%) for the entire group. The overall EFS rate in the current study was significantly better (P =.002) than that obtained in a comparable group of patients treated in the early 1990s in the AIEOP-ALL91 study. In particular, patients with PPR had a 4-year EFS of 61.1% (SE, 4.4%) versus 42.8% (SE, 5.4%) in the ALL 91 study (P =.008). Among PPR patients, those who were PPR-only (60.1%)-that is, they achieved CR and were negative for t(9;22) and t(4;11) translocations-had the best outcomes with this intensive treatment, even when additional adverse features (hyperleukocytosis, T phenotype) were present (4-year EFS, 70.1%; SE, 4.7%). We attribute this improvement to the replacement of 6 alternating blocks of non-cross-resistant drugs with an 8-drug reinduction regimen (Berlin-Frankfurt-Muenster [BFM] protocol II), repeated twice, in the context of a standard BFM-type intensive chemotherapy for high-risk ALL. This modified therapy may lead to high cure rates for patients defined as at high risk for intrinsic resistance to corticosteroids only.

  6. Novel enabling technologies of gene isolation and plant transformation for improved crop protection

    SciTech Connect

    Torok, Tamas

    2013-02-04

    Meeting the needs of agricultural producers requires the continued development of improved transgenic crop protection products. The completed project focused on developing novel enabling technologies of gene discovery and plant transformation to facilitate the generation of such products.

  7. Application of In Situ Fiberization for fabrication of improved strain isolation pads and graphite epoxy composites

    NASA Technical Reports Server (NTRS)

    Rosser, R. W.; Seibold, R. W.; Basiulis, D. I.

    1982-01-01

    The feasibility of applying the in situ fiberization process to the fabrication of strain isolation pads (SIP) for the Space Shuttle and to the fabrication of graphite-epoxy composites was evaluated. The ISF process involves the formation of interconnected polymer fiber networks by agitation of dilute polymer solutions under controlled conditions. High temperature polymers suitable for SIP use were fiberized and a successful fiberization of polychloro trifluoroethylene, a relatively high melting polymer, was achieved. Attempts to fiberize polymers with greater thermal stability were unsuccessful, apparently due to characteristics caused by the presence of aromaticity in the backbone of such materials. Graphite-epoxy composites were fabricated by interconnecting two dimensional arrays of graphite fiber with polypropylene IS fibers with subsequent epoxy resin impregnation. Mechanical property tests were performed on laminated panels of this material to evaluate intralaminar and interlaminar shear strength, and thus fracture toughness. Test results were generally unpromising.

  8. Dichloromethane dehalogenase with improved catalytic activity isolated from a fast-growing dichloromethane-utilizing bacterium

    SciTech Connect

    Scholtz, R.; Egli, C.; Cook, A.M.; Leisinger, T. ); Wackett, L.P. )

    1988-12-01

    A methylotrophic bacterium, denoted strain DM11, was isolated from groundwater and shown to utilize dichloromethane or dibromomethane as the sole carbon and energy source. The new isolate grew at the high rate of 0.22 h{sup {minus}1} compared with 11 previously characterized dichloromethane-utilizing bacteria ({mu}{sub max}, 0.08 h{sup {minus}1}). The dichloromethane dehalogenase from strain DM11 (group B enzyme) was purified by anion-exchange chromatography. It was shown to be substantially different from the set of dichloromethane dehalogenases from the 11 slow-growing strains (group A enzymes) that had previously been demonstrated to be identical. The V{sub max} for the group B enzyme was 97 mkat/kg of protein, some 5.6-fold higher than that of the group A enzymes. The group A dehalogenases showed hyperbolic saturation with the cosubstrate glutathione, whereas the group B enzyme showed positive cooperativity in glutathione binding. Only 1 of 15 amino acids occupied common positions at the N termini, and amino acid contents were substantially different in group A and group B dehalogenases. Immunological assays demonstrated weak cross-reactivity between the two enzymes. Despite the observed structural and kinetic differences, there is potentially evolutionary relatedness between group A and group B enzymes, as indicated by (i) hybridization of DM11 DNA with a gene probe of the group A enzyme, (ii) a common requirement for glutathione in catalysis, and (iii) similar subunit molecular weights of about 34,000.

  9. Isolation from Animal Tissue and Genetic Transformation of Coxiella burnetii Are Facilitated by an Improved Axenic Growth Medium▿

    PubMed Central

    Omsland, Anders; Beare, Paul A.; Hill, Joshua; Cockrell, Diane C.; Howe, Dale; Hansen, Bryan; Samuel, James E.; Heinzen, Robert A.

    2011-01-01

    We recently described acidified citrate cysteine medium (ACCM), which supports host cell-free (axenic) growth of Coxiella burnetii. After 6 days of incubation, greater than 3 logs of growth was achieved with the avirulent Nine Mile phase II (NMII) strain. Here, we describe modified ACCM and culture conditions that support improved growth of C. burnetii and their use in genetic transformation and pathogen isolation from tissue samples. ACCM was modified by replacing fetal bovine serum with methyl-β-cyclodextrin to generate ACCM-2. Cultivation of NMII in ACCM-2 with moderate shaking and in 2.5% oxygen yielded 4 to 5 logs of growth over 7 days. Similar growth was achieved with the virulent Nine Mile phase I and G isolates of C. burnetii. Colonies that developed after 6 days of growth in ACCM-2 agarose were approximately 0.5 mm in diameter, roughly 5-fold larger than those formed in ACCM agarose. By electron microscopy, colonies consisted primarily of the C. burnetii small cell variant morphological form. NMII was successfully cultured in ACCM-2 when medium was inoculated with as little as 10 genome equivalents contained in tissue homogenates from infected SCID mice. A completely axenic C. burnetii genetic transformation system was developed using ACCM-2 that allowed isolation of transformants in about 2 1/2 weeks. Transformation experiments demonstrated clonal populations in colonies and a transformation frequency of approximately 5 × 10−5. Cultivation in ACCM-2 will accelerate development of C. burnetii genetic tools and provide a sensitive means of primary isolation of the pathogen from Q fever patients. PMID:21478315

  10. Using electronic technology to improve clinical care – results from a before-after cluster trial to evaluate assessment and classification of sick children according to Integrated Management of Childhood Illness (IMCI) protocol in Tanzania

    PubMed Central

    2013-01-01

    Background Poor adherence to the Integrated Management of Childhood Illness (IMCI) protocol reduces the potential impact on under-five morbidity and mortality. Electronic technology could improve adherence; however there are few studies demonstrating the benefits of such technology in a resource-poor settings. This study estimates the impact of electronic technology on adherence to the IMCI protocols as compared to the current paper-based protocols in Tanzania. Methods In four districts in Tanzania, 18 clinics were randomly selected for inclusion. At each site, observers documented critical parts of the clinical assessment of children aged 2 months to 5 years. The first set of observations occurred during examination of children using paper-based IMCI (pIMCI) and the next set of observations occurred during examination using the electronic IMCI (eIMCI). Children were re-examined by an IMCI expert and the diagnoses were compared. A total of 1221 children (671 paper, 550 electronic) were observed. Results For all ten critical IMCI items included in both systems, adherence to the protocol was greater for eIMCI than for pIMCI. The proportion assessed under pIMCI ranged from 61% to 98% compared to 92% to 100% under eIMCI (p < 0.05 for each of the ten assessment items). Conclusions Use of electronic systems improved the completeness of assessment of children with acute illness in Tanzania. With the before-after nature of the design, potential for temporal confounding is the primary limitation. However, the data collection for both phases occurred over a short period (one month) and so temporal confounding was expected to be minimal. The results suggest that the use of electronic IMCI protocols can improve the completeness and consistency of clinical assessments and future studies will examine the long-term health and health systems impact of eIMCI. PMID:23981292

  11. Oral contraceptive pretreatment does not improve outcome in microdose gonadotrophin-releasing hormone agonist protocol among poor responder intracytoplasmic sperm injection patients

    PubMed Central

    Berker, Bulent; Turhan, Nilgun Ozturk; Satiroglu, Hakan

    2008-01-01

    Purpose To compare oral contraceptive (OC) pretreatment plus microdose GnRH-a in flare-up protocol and non-OC microdose GnRH-a in flare-up protocol among poor responder ICSI patients. Methods A retrospective analysis of poor responder ICSI patients. Patients were divided into two groups according to used microdose protocol. Precycle treatment with OC followed by follicular phase administration of 40 μg sc leuprolide acetate (LA) every 12 h beginning on after 2 day pill-free period and rFSH administration was begun on the third day of LA administration (OC-Group, n = 26). Alternatively on day 2 after menses, patients were administered similar stimulation regime (non-OC Group, n = 27). Results There were no significant differences between groups in the number of oocytes, peak estradiol levels, endometrial thickness, fertilization rates and embryo quality. Implantations and pregnancy rates per embryo transfer were similar. Conclusion OC pretreatment plus microdose GnRHa in flare-up protocol does not offer advantages over non-OC microdose GnRHa in flare-up protocol among poor responder ICSI patients. PMID:18253823

  12. Resolution improvement of isolated line pattern in quarter-micrometer level by layout-optimized assistant pattern method

    NASA Astrophysics Data System (ADS)

    Tounai, Keiichiro; Aizaki, Naoaki

    1996-06-01

    In the previous report, we showed the optimized results of off-axis illumination (OAI) for 0.25 micrometers resist patterning. Wide DOF was obtained for dense patterns like 1:1 L and S pattern, however, DOF was small for the sparse patterns. In order to widen the DOF for isolated line patterns under the OAI, we have examined the assistant pattern method, which needs the only unresolved additional patterns beside the isolated pattern. Optimization has been carried out by light intensity simulation under the following criteria; CD variation within plus or minus 10 percent, resist thickness loss of 10 percent and unresolved assistant patterns. CD value has been defined by the threshold method of aerial images. The evaluation method by aerial image is useful for rough estimate because the calculation is very rapid. However, the difference from the experimental results cannot be ignored in some cases, especially, in important CD- focus characteristics. For more accurate evaluation, we also investigated the result difference using a simplified resist development model. This model is useful for rapid calculation as light intensity calculation. From the evaluation of the exposure dose and focus latitude (EFL), the optimal layout must be decided considering exposure dose latitude decrease by resist development process. We have additionally investigated the combination effect of assistant pattern and halftone phase-shifting under OAI. EFL was improved a little by halftone phase- shifting mask without assistant patterns, on the other hand, EFL was improved by combination of HPSM and assistant patterns.

  13. Improved isolation strategies allowed the phenotypic differentiation of two Nitrospira strains from widespread phylogenetic lineages.

    PubMed

    Nowka, Boris; Off, Sandra; Daims, Holger; Spieck, Eva

    2015-03-01

    The second step of nitrification, the oxidation of nitrite to nitrate, is vital for the functioning of the nitrogen cycle, but our understanding of the ecological roles of the involved microorganisms is still limited. The known diversity of Nitrospira, the most widely distributed nitrite-oxidizing bacteria, has increased remarkably by analyses of 16S rRNA and functional gene sequences. However, only few representatives could be brought into laboratory cultures so far. In this study, two Nitrospira from activated sludge were isolated using novel approaches together with established methods. Highly enriched 'Candidatus Nitrospira defluvii' was separated from concomitant heterotrophs by taking advantage of its resistance against ampicillin and acriflavine. Beside this member of lineage I, a novel species of lineage II, named N. lenta, was initially enriched at 10°C and finally purified by using optical tweezers. The tolerance to elevated nitrite levels was much higher in N. defluvii than in the more fastidious N. lenta and was accompanied by pronounced biofilm formation. Phylogenetic classification of 12 additional enrichments indicated that Nitrospira lineage I is common in extreme and moderate ecosystems like lineage II. The new cultures will help to explore physiological and genomic differences affecting niche separation between members of this highly diverse genus.

  14. Improved isolation strategies allowed the phenotypic differentiation of two Nitrospira strains from widespread phylogenetic lineages.

    PubMed

    Nowka, Boris; Off, Sandra; Daims, Holger; Spieck, Eva

    2015-03-01

    The second step of nitrification, the oxidation of nitrite to nitrate, is vital for the functioning of the nitrogen cycle, but our understanding of the ecological roles of the involved microorganisms is still limited. The known diversity of Nitrospira, the most widely distributed nitrite-oxidizing bacteria, has increased remarkably by analyses of 16S rRNA and functional gene sequences. However, only few representatives could be brought into laboratory cultures so far. In this study, two Nitrospira from activated sludge were isolated using novel approaches together with established methods. Highly enriched 'Candidatus Nitrospira defluvii' was separated from concomitant heterotrophs by taking advantage of its resistance against ampicillin and acriflavine. Beside this member of lineage I, a novel species of lineage II, named N. lenta, was initially enriched at 10°C and finally purified by using optical tweezers. The tolerance to elevated nitrite levels was much higher in N. defluvii than in the more fastidious N. lenta and was accompanied by pronounced biofilm formation. Phylogenetic classification of 12 additional enrichments indicated that Nitrospira lineage I is common in extreme and moderate ecosystems like lineage II. The new cultures will help to explore physiological and genomic differences affecting niche separation between members of this highly diverse genus. PMID:25764560

  15. Isolating the perceptual from the social: tapping in shared space results in improved synchrony.

    PubMed

    Wu, David W-L; Chapman, Craig S; Walker, Esther; Bischof, Walter F; Kingstone, Alan

    2013-10-01

    Current theory suggests that interpersonal synchrony is an important social behavior in that it not only serves as a form of "social glue," but it also arises automatically in a social context. Theorists suggest potential mechanisms for interpersonal synchrony, ranging from a "low-level" social-perceptual system account to a "high-level" social-motivational explanation. Past studies that suggest synchrony can be influenced by social factors do not discriminate between these accounts. The current investigation seeks to isolate the effect of the high-level social system on interpersonal synchrony by investigating the effects of spatial proximity on unintentional coordinated tapping between two naïve participants. Dyads performed a synchronization-continuation task either in the same room, in different rooms, or in different rooms but with the ability to hear each other tap. Participant taps were represented by a box that flashed on the monitor to control visual information across all three conditions. Same-room dyads had increased coordination over different-room dyads, whereas dyads that shared audio but were in different rooms showed an intermediate level of coordination. The present study demonstrates that shared space, independent of perceptual differences in stimuli, can increase unintentional coordinated tapping. PMID:23750971

  16. Evaluation of selected white-rot fungal isolates for improving the sugar yield from wheat straw.

    PubMed

    Cianchetta, Stefano; Di Maggio, Barbara; Burzi, Pier Luigi; Galletti, Stefania

    2014-05-01

    Biological pretreatment of lignocellulosic biomass by fungi can represent a low-cost and eco-friendly alternative to physicochemical methods to facilitate enzymatic hydrolysis. However, fungal metabolism can cause cellulose loss and it is therefore necessary to use the appropriate fungal strain-biomass type combination. In this work, the effects of biological pretreatments carried out by five different fungi on enzymatic hydrolysis of wheat straw were investigated. The best results were obtained with a Ceriporiopsis subvermispora strain, which minimized weight and cellulose losses and gave the highest net sugar yield (calculated with respect to the holocellulose content of the untreated straw), up to 44 % after a 10-week pretreatment, more than doubling the yields obtained with the other isolates. Moreover, prolonging the pretreatment from 4 up to 10 weeks produced a 2-fold increase, up to 60 %, in digestibility (sugar yield, calculated considering the holocellulose content of the pretreated material). The hemicellulose content of the pretreated material resulted inversely correlated with digestibility, and it could thus be utilized as an index of the pretreatment efficacy. Finally, a correlation was also found between digestibility and the difference between the absorbance values at 290 and 320 nm of pretreated wheat straw extracts.

  17. Choosing GnRH Antagonist Protocol Shows Improved Oocyte and Embryo Quality, Coherent with the Perifollicular Vascularity (PFV) in Assisted Reproductive Techniques

    PubMed Central

    Ramachandran, Amar; Kumar, Pratap

    2015-01-01

    Introduction The parent oocyte from which the embryo is derived, determines its quality and the perifollicular vascularity (PFV) determines the micro-environment of the developing ovum. The PFV correlates well with the follicular oxygenation, oocyte maturation and embryo viability. PFV is imaged with Power Doppler Ultrasound. Aim To study and compare the association of the PFV of follicles with the quality of the oocytes and embryos in agonist and antagonist protocol, employed in Assisted Reproductive techniques (ART). Study Design A prospective observational study was conducted on 75 patients, who were recruited for ART cycles, out of which 25 were given the Agonist protocol and the remaining 50 received the Antagonist protocol. Materials and Methods The patients underwent the stimulation protocol. The PFV of preovulatory follicles were studied with Transvaginal Power Doppler and graded. Each oocyte retrieved carried the same label of its parent follicle. Embryos were cultured. The embryologist was blinded. The oocyte and embryo quality were assessed and compared with the PFV of parent follicle. Results Follicles with grade 1 and 2 PFV were predominantly observed. The yield of oocytes was independent of PFV. The mean yield of good quality embryos in conjunction with the PFV of the parent follicle was found to be highly significant in both the groups. The antagonist group had statistically significant yield of mature oocytes and embryos, compared to the agonist group. Conclusion Antagonist protocol had favourable outcomes compared with the agonist protocol. The retrieval of oocytes, even the mature ones and the yield of high grade embryos were found higher. As the PFV increased, the yield and overall pregnancy rates were higher. PFV as assessed by Power Doppler is a useful non-invasive biomarker of embryo quality and can be employed in conjunction with other biomarkers in ART to predict successful outcome. PMID:26674932

  18. Dosimetry Formalism and Implementation of a Homogenous Irradiation Protocol to Improve the Accuracy of Small Animal Whole-Body Irradiation Using a 137Cs Irradiator.

    PubMed

    Brodin, N Patrik; Chen, Yong; Yaparpalvi, Ravindra; Guha, Chandan; Tomé, Wolfgang A

    2016-02-01

    Shielded Cs irradiators are routinely used in pre-clinical radiation research to perform in vitro or in vivo investigations. Without appropriate dosimetry and irradiation protocols in place, there can be large uncertainty in the delivered dose of radiation between irradiated subjects that could lead to inaccurate and possibly misleading results. Here, a dosimetric evaluation of the JL Shepard Mark I-68A Cs irradiator and an irradiation technique for whole-body irradiation of small animals that allows one to limit the between subject variation in delivered dose to ±3% are provided. Mathematical simulation techniques and Gafchromic EBT film were used to describe the region within the irradiation cavity with homogeneous dose distribution (100% ± 5%), the dosimetric impact of varying source-to-subject distance, and the variation in attenuation thickness due to turntable rotation. Furthermore, an irradiation protocol and dosimetry formalism that allows calculation of irradiation time for whole-body irradiation of small animals is proposed that is designed to ensure a more consistent dose delivery between irradiated subjects. To compare this protocol with the conventional irradiation protocol suggested by the vendor, high-resolution film dosimetry measurements evaluating the dose difference between irradiation subjects and the dose distribution throughout subjects was performed using phantoms resembling small animals. Based on these results, there can be considerable variation in the delivered dose of > ± 5% using the conventional irradiation protocol for whole-body irradiation doses below 5 Gy. Using the proposed irradiation protocol this variability can be reduced to within ±3% and the dosimetry formalism allows for more accurate calculation of the irradiation time in relation to the intended prescription dose. PMID:26710162

  19. Publishing protocols for partnered research.

    PubMed

    Hysong, Sylvia J; Woodard, LeChauncy; Garvin, Jennifer H; Murawsky, Jeffrey; Petersen, Laura A

    2014-12-01

    Published scientific protocols are advocated as a means of controlling bias in research reporting. Indeed, many journals require a study protocol with manuscript submission. However, publishing protocols of partnered research (PPR) can be challenging in light of the research model's dynamic nature, especially as no current reporting standards exist. Nevertheless, as these protocols become more prevalent, a priori documentation of methods in partnered research studies becomes increasingly important. Using as illustration a suite of studies aimed at improving coordination and communication in the primary care setting, we sought to identify challenges in publishing PPR relative to traditional designs, present alternative solutions to PPR publication, and propose an initial checklist of content to be included in protocols of partnered research. Challenges to publishing PPR include reporting details of research components intended to be co-created with operational partners, changes to sampling and entry strategy, and alignment of scientific and operational goals. Proposed solutions include emulating reporting standards of qualitative research, participatory action research, and adaptive trial designs, as well as embracing technological tools that facilitate publishing adaptive protocols, with version histories that are able to be updated as major protocol changes occur. Finally, we present a proposed checklist of reporting elements for partnered research protocols. PMID:25355092

  20. Publishing protocols for partnered research.

    PubMed

    Hysong, Sylvia J; Woodard, LeChauncy; Garvin, Jennifer H; Murawsky, Jeffrey; Petersen, Laura A

    2014-12-01

    Published scientific protocols are advocated as a means of controlling bias in research reporting. Indeed, many journals require a study protocol with manuscript submission. However, publishing protocols of partnered research (PPR) can be challenging in light of the research model's dynamic nature, especially as no current reporting standards exist. Nevertheless, as these protocols become more prevalent, a priori documentation of methods in partnered research studies becomes increasingly important. Using as illustration a suite of studies aimed at improving coordination and communication in the primary care setting, we sought to identify challenges in publishing PPR relative to traditional designs, present alternative solutions to PPR publication, and propose an initial checklist of content to be included in protocols of partnered research. Challenges to publishing PPR include reporting details of research components intended to be co-created with operational partners, changes to sampling and entry strategy, and alignment of scientific and operational goals. Proposed solutions include emulating reporting standards of qualitative research, participatory action research, and adaptive trial designs, as well as embracing technological tools that facilitate publishing adaptive protocols, with version histories that are able to be updated as major protocol changes occur. Finally, we present a proposed checklist of reporting elements for partnered research protocols.

  1. Improvement of signal-to-noise ratio using iterative reconstruction in a 99m Tc-ECD split-dose injection protocol.

    PubMed

    Yokoi, Takashi; Shinohara, Hiroyuki; Takaki, Akihiro

    2003-08-01

    optimal injection dose ratio. In a clinical study, the addition method provided better quality images than the subtraction method. The ROI values of rest images estimated by the subtraction method were close to the results obtained with the addition method (ROI(sub)=1.01 ROI(add)-0.312, r=0.999). The ROI values of the ACZ images estimated by the subtraction method also agreed with the results obtained using the addition method, but the correlation was slightly worse (ROI(sub)=1.03 ROI(add)-2.23, r=0.995). Quantitative ROI values were quite similar between the methods. Our results demonstrated that the quality of reconstructed rest and ACZ-loaded images were significantly better with the addition method than with the conventional subtraction method. We conclude that the proposed method will be useful as a practical reconstruction algorithm to improve the S/N in an equal-volume split-dose injection protocol using (99m)Tc-ECD.

  2. Does Interpersonal Psychotherapy improve clinical care for adolescents with depression attending a rural child and adolescent mental health service? Study protocol for a cluster randomised feasibility trial

    PubMed Central

    Bearsley-Smith, Cate; Browne, Mark Oakley; Sellick, Ken; Villanueva, Elmer V; Chesters, Janice; Francis, Karen; Reddy, Prasuna

    2007-01-01

    Background Depression amongst adolescents is a costly societal problem. Little research documents the effectiveness of public mental health services in mapping this problem. Further, it is not clear whether usual care in such services can be improved via clinician training in a relevant evidence based intervention. One such intervention, found to be effective and easily learned amongst novice clinicians, is Interpersonal Psychotherapy (IPT). The study described in the current paper has two main objectives. First, it aims to investigate the impact on clinical care of implementing Interpersonal Psychotherapy for Adolescents for the treatment of adolescent depression within a rural mental health service compared with Treatment as Usual (TAU). The second objective is to record the process and challenges (i.e. feasibility, acceptability, sustainability) associated with implementing and evaluating an evidence-based intervention within a community service. This paper outlines the study rationale and design for this community based research trial. Methods/design The study involves a cluster randomisation trial to be conducted within a Child and Adolescent Mental Health Service in rural Australia. All clinicians in the service will be invited to participate. Participating clinicians will be randomised via block design at each of four sites to (a) training and delivery of IPT, or (b) TAU. The primary measure of impact on care will be a clinically significant change in depressive symptomatology, with secondary outcomes involving treatment satisfaction and changes in other symptomatology. Participating adolescents with significant depressive symptomatology, aged 12 to 18 years, will complete assessment measures at Weeks 0, 12 and 24 of treatment. They will also complete a depression inventory once a month during that period. This study aims to recruit 60 adolescent participants and their parent/guardian/s. A power analysis is not indicated as an intra-class correlation

  3. Primary care Identification and Referral to Improve Safety of women experiencing domestic violence (IRIS): protocol for a pragmatic cluster randomised controlled trial

    PubMed Central

    2010-01-01

    Background Domestic violence, which may be psychological, physical, sexual, financial or emotional, is a major public health problem due to the long-term health consequences for women who have experienced it and for their children who witness it. In populations of women attending general practice, the prevalence of physical or sexual abuse in the past year from a partner or ex-partner ranges from 6 to 23%, and lifetime prevalence from 21 to 55%. Domestic violence is particularly important in general practice because women have many contacts with primary care clinicians and because women experiencing abuse identify doctors and nurses as professionals from whom they would like to get support. Yet health professionals rarely ask about domestic violence and have little or no training in how to respond to disclosure of abuse. Methods/Design This protocol describes IRIS, a pragmatic cluster randomised controlled trial with the general practice as unit of randomisation. Our trial tests the effectiveness and cost-effectiveness of a training and support programme targeted at general practice teams. The primary outcome is referral of women to specialist domestic violence agencies. Forty-eight practices in two UK cities (Bristol and London) are randomly allocated, using minimisation, into intervention and control groups. The intervention, based on an adult learning model in an educational outreach framework, has been designed to address barriers to asking women about domestic violence and to encourage appropriate responses to disclosure and referral to specialist domestic violence agencies. Multidisciplinary training sessions are held with clinicians and administrative staff in each of the intervention practices, with periodic feedback of identification and referral data to practice teams. Intervention practices have a prompt to ask about abuse integrated in the electronic medical record system. Other components of the intervention include an IRIS champion in each practice

  4. Characterizing the Interaction Between Routing and MAC Protocols in Ad-Hoc Networks

    SciTech Connect

    Barrett, C. L.; Drozda, M.; Marathe, A.; Marathe, M. V.

    2002-01-01

    We empirically study the effect of mobility on the performance of protocols designed for wireless ad-hoc networks. An important objective is to study the interaction of the Routing and MAC layer protocols under different mobility parameters. We use three basic mobility models: grid mobility model, random waypoint model, and exponential correlated random model. The performance of protocols is measured in terms of (i) latency, (ii) throughput, (iii) number of packets received, (iv) long term fairness and (v) number of control packets at the MAC and routing layer level. Three different commonly studied routing protocols are used: AODV, DSR and LAR1. Similarly three well known MAC protocols are used: MACA, 802.1 1 and CSMA. Our main contribution is simulation based experiments coupled with rigorous statistical analysis to characterize the interaction of MAC layer protocols with routing layer protocols in ad-hoc networks. From the results, we can conclude the following: e No single MAC or Routing protocol dominated the other protocols in their class. Probably more interestingly, no MAURouting protocol combination was better than other combinations over all scenarios and response variables. 0 In general, it is not meaningful to speak about a MAC or a routing protocol in isolation. Presence of interaction leads to trade-offs between the amount of control packets generated by each layer. The results raise the possibility of improving the performance of a particular MAC layer protocol by using a cleverly designed routing protocol or vice-versa. Thus in order to improve the performanceof a communication network, it is important to study the entire protocol stack as a single algorithmic construct; optimizing individual layers in the seven layer OS1 stack will not yield performance improvements beyond a point. A methodological contribution of this paper is the use of statistical methods such as analysis of variance (ANOVA), to characterize the interaction between the protocols

  5. New opportunities for improved ribotyping of C. difficile clinical isolates by exploring their genomes.

    PubMed

    Gürtler, Volker; Grando, Danilla

    2013-06-01

    Clostridium difficile causes outbreaks of infectious diarrhoea, most commonly occurring in healthcare institutions. Recently, concern has been raised with reports of C. difficile disease in those traditionally thought to be at low risk i.e. community acquired rather than healthcare acquired. This has increased awareness for the need to track outbreaks and PCR-ribotyping has found widespread use to elucidate epidemiologically linked isolates. PCR-ribotyping uses conserved regions of the 16S rRNA gene and 23S rRNA gene as primer binding sites to produce varying PCR products due to the intergenic spacer (ITS1) regions of the multiple operons. With the explosion of whole genome sequence data it became possible to analyse the start of the 23S rRNA gene for a more accurate selection of regions closer to the end of the ITS1. However the following questions must still be asked: (i) Does the chromosomal organisation of the rrn operon vary between C. difficile strains? and (ii) just how conserved are the primer binding regions? Eight published C. difficile genomes have been aligned to produce a detailed database of indels of the ITS1's from the rrn operon sets. An iPad Filemaker Go App has been constructed and named RiboTyping (RT). It contains detail such as sequences, ribotypes, strain numbers, GenBank numbers and genome position numbers. Access to various levels of the database is provided so that details can be printed. There are three main regions of the rrn operon that have been analysed by the database and related to each other by strain, ribotype and operon: (1) 16S gene (2) ITS1 indels (3) 23S gene. This has enabled direct intra- and inter-genomic comparisons at the strain, ribotype and operon (allele) levels in each of the three genomic regions. This is the first time that such an analysis has been done. By using the RT App with search criteria it will be possible to select probe combinations for specific strains/ribotypes/rrn operons for experiments to do with

  6. Reprint of New opportunities for improved ribotyping of C. difficile clinical isolates by exploring their genomes.

    PubMed

    Gürtler, Volker; Grando, Danilla

    2013-12-01

    Clostridium difficile causes outbreaks of infectious diarrhoea, most commonly occurring in healthcare institutions. Recently, concern has been raised with reports of C. difficile disease in those traditionally thought to be at low risk i.e. community acquired rather than healthcare acquired. This has increased awareness for the need to track outbreaks and PCR-ribotyping has found widespread use to elucidate epidemiologically linked isolates. PCR-ribotyping uses conserved regions of the 16S rRNA gene and 23S rRNA gene as primer binding sites to produce varying PCR products due to the intergenic spacer (ITS1) regions of the multiple operons. With the explosion of whole genome sequence data it became possible to analyse the start of the 23S rRNA gene for a more accurate selection of regions closer to the end of the ITS1. However the following questions must still be asked: (i) Does the chromosomal organisation of the rrn operon vary between C. difficile strains? and (ii) just how conserved are the primer binding regions? Eight published C. difficile genomes have been aligned to produce a detailed database of indels of the ITS1's from the rrn operon sets. An iPad Filemaker Go App has been constructed and named RiboTyping (RT). It contains detail such as sequences, ribotypes, strain numbers, GenBank numbers and genome position numbers. Access to various levels of the database is provided so that details can be printed. There are three main regions of the rrn operon that have been analysed by the database and related to each other by strain, ribotype and operon: (1) 16S gene (2) ITS1 indels (3) 23S gene. This has enabled direct intra- and inter-genomic comparisons at the strain, ribotype and operon (allele) levels in each of the three genomic regions. This is the first time that such an analysis has been done. By using the RT App with search criteria it will be possible to select probe combinations for specific strains/ribotypes/rrn operons for experiments to do with

  7. Sequence tagged microsatellite profiling (STMP): improved isolation of DNA sequence flanking target SSRs

    PubMed Central

    Hayden, M. J.; Good, G.; Sharp, P. J.

    2002-01-01

    Sequence tagged microsatellite profiling (STMP) enables the rapid development of large numbers of co-dominant DNA markers, known as sequence tagged microsatellites (STMs). Each STM is amplified by PCR using a single primer specific to the conserved DNA sequence flanking the microsatellite repeat in combination with a universal primer that anchors to the 5′-ends of the microsatellites. It is also possible to convert STMs into conventional microsatellite, or simple sequence repeat (SSR), markers that are amplified using a pair of primers flanking the repeat sequence. Here, we describe a modification of the STMP procedure to significantly improve the capacity to convert STMs into conventional SSRs and, therefore, facilitate the development of highly specific DNA markers for purposes such as marker-assisted breeding. The usefulness of this technique was demonstrated in bread wheat. PMID:12466561

  8. Sequence tagged microsatellite profiling (STMP): improved isolation of DNA sequence flanking target SSRs.

    PubMed

    Hayden, M J; Good, G; Sharp, P J

    2002-12-01

    Sequence tagged microsatellite profiling (STMP) enables the rapid development of large numbers of co-dominant DNA markers, known as sequence tagged microsatellites (STMs). Each STM is amplified by PCR using a single primer specific to the conserved DNA sequence flanking the microsatellite repeat in combination with a universal primer that anchors to the 5'-ends of the microsatellites. It is also possible to convert STMs into conventional microsatellite, or simple sequence repeat (SSR), markers that are amplified using a pair of primers flanking the repeat sequence. Here, we describe a modification of the STMP procedure to significantly improve the capacity to convert STMs into conventional SSRs and, therefore, facilitate the development of highly specific DNA markers for purposes such as marker-assisted breeding. The usefulness of this technique was demonstrated in bread wheat. PMID:12466561

  9. An environmental disinfection odyssey: evaluation of sequential interventions to improve disinfection of Clostridium difficile isolation rooms.

    PubMed

    Sitzlar, Brett; Deshpande, Abhishek; Fertelli, Dennis; Kundrapu, Sirisha; Sethi, Ajay K; Donskey, Curtis J

    2013-05-01

    OBJECTIVE. Effective disinfection of hospital rooms after discharge of patients with Clostridium difficile infection (CDI) is necessary to prevent transmission. We evaluated the impact of sequential cleaning and disinfection interventions by culturing high-touch surfaces in CDI rooms after cleaning. DESIGN. Prospective intervention. SETTING. A Veterans Affairs hospital. INTERVENTIONS. During a 21-month period, 3 sequential tiered interventions were implemented: (1) fluorescent markers to provide monitoring and feedback on thoroughness of cleaning facility-wide, (2) addition of an automated ultraviolet radiation device for adjunctive disinfection of CDI rooms, and (3) enhanced standard disinfection of CDI rooms, including a dedicated daily disinfection team and implementation of a process requiring supervisory assessment and clearance of terminally cleaned CDI rooms. To determine the impact of the interventions, cultures were obtained from CDI rooms after cleaning and disinfection. RESULTS. The fluorescent marker intervention improved the thoroughness of cleaning of high-touch surfaces (from 47% to 81% marker removal; P < .0001). Relative to the baseline period, the prevalence of positive cultures from CDI rooms was reduced by 14% (P=.024), 48% (P <.001), and 89% (P=.006) with interventions 1, 2, and 3, respectively. During the baseline period, 67% of CDI rooms had positive cultures after disinfection, whereas during interventions periods 1, 2, and 3 the percentages of CDI rooms with positive cultures after disinfection were reduced to 57%, 35%, and 7%, respectively. CONCLUSIONS. An intervention that included formation of a dedicated daily disinfection team and implementation of a standardized process for clearing CDI rooms achieved consistent CDI room disinfection. Culturing of CDI rooms provides a valuable tool to drive improvements in environmental disinfection.

  10. An environmental disinfection odyssey: evaluation of sequential interventions to improve disinfection of Clostridium difficile isolation rooms.

    PubMed

    Sitzlar, Brett; Deshpande, Abhishek; Fertelli, Dennis; Kundrapu, Sirisha; Sethi, Ajay K; Donskey, Curtis J

    2013-05-01

    OBJECTIVE. Effective disinfection of hospital rooms after discharge of patients with Clostridium difficile infection (CDI) is necessary to prevent transmission. We evaluated the impact of sequential cleaning and disinfection interventions by culturing high-touch surfaces in CDI rooms after cleaning. DESIGN. Prospective intervention. SETTING. A Veterans Affairs hospital. INTERVENTIONS. During a 21-month period, 3 sequential tiered interventions were implemented: (1) fluorescent markers to provide monitoring and feedback on thoroughness of cleaning facility-wide, (2) addition of an automated ultraviolet radiation device for adjunctive disinfection of CDI rooms, and (3) enhanced standard disinfection of CDI rooms, including a dedicated daily disinfection team and implementation of a process requiring supervisory assessment and clearance of terminally cleaned CDI rooms. To determine the impact of the interventions, cultures were obtained from CDI rooms after cleaning and disinfection. RESULTS. The fluorescent marker intervention improved the thoroughness of cleaning of high-touch surfaces (from 47% to 81% marker removal; P < .0001). Relative to the baseline period, the prevalence of positive cultures from CDI rooms was reduced by 14% (P=.024), 48% (P <.001), and 89% (P=.006) with interventions 1, 2, and 3, respectively. During the baseline period, 67% of CDI rooms had positive cultures after disinfection, whereas during interventions periods 1, 2, and 3 the percentages of CDI rooms with positive cultures after disinfection were reduced to 57%, 35%, and 7%, respectively. CONCLUSIONS. An intervention that included formation of a dedicated daily disinfection team and implementation of a standardized process for clearing CDI rooms achieved consistent CDI room disinfection. Culturing of CDI rooms provides a valuable tool to drive improvements in environmental disinfection. PMID:23571361

  11. Improvement in The Function of Isolated Rat Pancreatic Islets through Reduction of Oxidative Stress Using Traditional Iranian Medicine

    PubMed Central

    Mahroui, Neda; Mirzaei, Sanaz; Siahpoosh, Zahra; D.4, Pharm.; Nili-Ahmadabadi, Amir; Mohammadirad, Azadeh; Baeeri, Maryam; Hajiaghaie, Reza; Abdollahi, Mohammad

    2014-01-01

    Objective Pancreatic islets have fewer antioxidant enzymes than other tissues and thus are vulnerable to oxidative stress. In the present study, the effects of nine specifically selected Iranian medical plants on the mitochondria function and survival of isolated rat islets were examined. Materials and Methods In this experimental study, following laparotomy, pancreases of rats were removed and the islets isolated and incubated in vitro for 24 hours. Logarithmic doses of plant materials were added to the islets and incubated for an additional 24 hours after which the viability of the cells and production of reactive oxygen species (ROS) were measured. Levels of insulin production in relation to static and stimulated glucose concen- trations were also determined. Results The tested compounds markedly increased survival of the islet cells, their mi- tochondrial activity, and insulin levels at the same time as reducing production of ROS. Greatest effects were observed in the following order: Peganum harmala, Glycyrrhiza glabra, Satureja hortensis, Rosmarinus officinalis, Teucrium scordium, Aloe vera, Zingiber officinale, Silybum marianum, and Hypericum perforatum at doses of 10, 103, 104, 10, 102, 102, 10-1, 10 and 103μgmL-1, respectively. Conclusion Based on these results, we suggest that pretreatment with these select- ed Iranian medical plants can improve the outcomes of pancreas transplants and grafts through the control of oxidative stress damage. PMID:24567945

  12. Simple method for isolation of glyceraldehyde 3-phosphate dehydrogenase and the improvement of myofibril gel properties.

    PubMed

    Miyaguchi, Yuji; Sakamoto, Taro; Sasaki, Shun; Nakade, Koji; Tanabe, Manabu; Ichinoseki, Satoko; Numata, Masahiro; Kosai, Kiichi

    2011-02-01

    Porcine glycolytic enzyme, glyceraldehyde 3-phosphate dehydrogenase (G3PD) was prepared effectively by a combination of ethylene diamine tetra-acetate (EDTA) pretreatment and affinity purification. After salting out of porcine sarcoplasmic proteins (SP) with ammonium sulfate at 75% saturation, the obtained supernatant (SP-f3) was treated with EDTA, leaving G3PD in the supernatant (G3PD-E) and most other SPs in the precipitate. At that time, the separation of G3PD-E required more than 20 mmol/L EDTA. G3PD-E was then subjected to affinity purification by batchwise method using blue-sepharose CL-6B, and purified G3PD (G3PD-AP) was obtained using 2 mol/L potassium chloride (KCl) as an eluent. Texture analysis showed that the hardness, adhesiveness and gumminess of the myofibril gel at 0.2-mol/L NaCl increased with the addition of G3PD-AP. Scanning electron microscopy revealed that the G3PD-AP reinforced the gel network of the myofibril. However, scanning electron micrograph analysis showed that the network-structure of the gel by the addition of G3PD-AP developed in a different manner from that by adding 0.6 mol/L NaCl. These results showed that glycolytic enzyme, G3PD, contributes to the improvement of the rheological properties of meat products.

  13. Improving central line infection rates in the neonatal intensive care unit: Effect of hospital location, site of insertion, and implementation of catheter-associated bloodstream infection protocols

    PubMed Central

    Freeman, Jennifer J.; Gadepalli, Samir K.; Siddiqui, Sabina M.; Jarboe, Marcus D.; Hirschl, Ronald B.

    2016-01-01

    Introduction Catheter associated blood stream infections (CABSI) are morbid and expensive for all ages, including neonates. Thus far, the impact of CABSI prevention protocols, such as insertion and maintenance bundles, in the neonatal intensive care unit (NICU) is largely unknown. We hypothesized that lines placed in the operating room (OR) would have a lower infection rate due to established insertion protocols and a more sterile environment. Methods Retrospective chart review of NICU patients who received a percutaneous or tunneled central venous catheter between 2005 and 2012. ECMO cannulas, PICC and umbilical catheters were excluded. Variables of interest included demographics, anatomical site, hospital location, line days, and line infection. Line infection was defined as a positive blood culture drawn through the catheter. Results A total of 368 catheters were placed in 285 NICU patients. Majority of catheters (65.5%) were placed in OR. Saphenous and femoral veins were most common anatomical sites (50.8%). Twenty-eight catheters were infected (7.6%). After adjusting for pre-operative antibiotics, anatomical site, and SNAPPE-II scores, lines placed in OR were three times less likely to become infected (Odds Ratio=0.32, p=0.038). Although implementation of CABSI prevention protocols resulted in statistically significant reductions in infection (Odds Ratio=0.4, p=0.043), lines placed in the OR remained less likely to become infected. Conclusions NICU line infection rates decreased with implementation of CABSI prevention protocols. Despite this implementation, catheters placed in the NICU continued to have higher infection rates. As a result, when patient status allows it, we recommend that central lines in newborns be placed in the operating room. PMID:25783394

  14. Equal Improvement in Men and Women in the Treatment of Urologic Chronic Pelvic Pain Syndrome Using a Multi-modal Protocol with an Internal Myofascial Trigger Point Wand.

    PubMed

    Anderson, Rodney U; Wise, David; Sawyer, Tim; Nathanson, Brian H; Nevin Smith, J

    2016-06-01

    Both men and women require treatment for urologic chronic pelvic pain syndromes (UCPPS), which includes interstitial cystitis/painful bladder syndrome, pelvic floor dysfunction, and chronic prostatitis/chronic pelvic pain syndrome. However, it is unknown if men and women respond differently to a protocol that includes specific physical therapy self-treatment using an internal trigger point wand and training in paradoxical relaxation. We performed a retrospective analysis by gender in a single arm, open label, single center clinical trial designed to evaluate the safety and effectiveness of a protocol for the treatment of UCPPS from October, 2008 to May, 2011. 314 adult men (79.9 %) and 79 (20.1 %) women met inclusion criteria. The median duration of symptoms was 60 months. The protocol required an initial 6-day clinic for training followed by a 6-month self-treatment period. The treatment included self-administered pelvic floor trigger point release with an internal trigger point device for physical therapy along with paradoxical relaxation training. Notable gender differences in prior treatments were observed. Men had a lower median [Interquartile Range] NIH-CPSI score at baseline than women (27 [21, 31] vs. 29 [22, 33], p = 0.04). Using a 1-10 scale with 10 = Most Severe, the median reduction in trigger point sensitivity was 3 units for both men and women after 6 months therapy (p = 0.74). A modified Intention to Treat analysis and a multivariate regression analysis found similar results. We conclude that men and women have similar, significant reductions in trigger point sensitivity with this protocol. PMID:26721470

  15. Equal Improvement in Men and Women in the Treatment of Urologic Chronic Pelvic Pain Syndrome Using a Multi-modal Protocol with an Internal Myofascial Trigger Point Wand.

    PubMed

    Anderson, Rodney U; Wise, David; Sawyer, Tim; Nathanson, Brian H; Nevin Smith, J

    2016-06-01

    Both men and women require treatment for urologic chronic pelvic pain syndromes (UCPPS), which includes interstitial cystitis/painful bladder syndrome, pelvic floor dysfunction, and chronic prostatitis/chronic pelvic pain syndrome. However, it is unknown if men and women respond differently to a protocol that includes specific physical therapy self-treatment using an internal trigger point wand and training in paradoxical relaxation. We performed a retrospective analysis by gender in a single arm, open label, single center clinical trial designed to evaluate the safety and effectiveness of a protocol for the treatment of UCPPS from October, 2008 to May, 2011. 314 adult men (79.9 %) and 79 (20.1 %) women met inclusion criteria. The median duration of symptoms was 60 months. The protocol required an initial 6-day clinic for training followed by a 6-month self-treatment period. The treatment included self-administered pelvic floor trigger point release with an internal trigger point device for physical therapy along with paradoxical relaxation training. Notable gender differences in prior treatments were observed. Men had a lower median [Interquartile Range] NIH-CPSI score at baseline than women (27 [21, 31] vs. 29 [22, 33], p = 0.04). Using a 1-10 scale with 10 = Most Severe, the median reduction in trigger point sensitivity was 3 units for both men and women after 6 months therapy (p = 0.74). A modified Intention to Treat analysis and a multivariate regression analysis found similar results. We conclude that men and women have similar, significant reductions in trigger point sensitivity with this protocol.

  16. Improvement of forage production in Calliandra calothyrsus: methodology for the identification of an effective inoculum containing Rhizobium strains and arbuscular mycorrhizal isolates.

    PubMed

    Lesueur, D; Ingleby, K; Odee, D; Chamberlain, J; Wilson, J; Tiki Manga, T; Sarrailh, J M; Pottinger, A

    2001-10-01

    The overall aim of this paper is to describe the selection of effective rhizobia and arbuscular mycorrhizas (AM), which after inoculation, will significantly improve the forage production of Calliandra calothyrsus under field conditions. To achieve this objective, the following activities were carried out: (i) establishment from both nodules and soil samples of a collection of microsymbionts (rhizobium and AM) of C. calothyrsus from Central America (Mexico, Honduras, Guatemala, Nicaragua and Costa Rica), also from outside its native range in Cameroon, Kenya and New Caledonia; (ii) identification under glasshouse conditions of the most effective rhizobia and AM isolates; (iii) production of a solid selected inoculum for field trials; (iv) examination of the impact of the inoculation on the growth of C. calothyrsus monitored under nursery conditions. We have screened 446 rhizobia strains in the nursery and identified six as being very effective at nodulating the host plant. They originated from Costa Rica (CCCR15 and CCCR1), from New Caledonia (CCNC26), from Cameroon (CCC22) and from Kenya (KWN35 and KCC6). In relation to AM, five isolates have been selected for the ability to infect and promote growth of the host plant--two isolates of Gigaspora albida isolated from Kenya (GA1b and GA2); one isolate of Scutellospora verrucosa isolated from Kenya (SV2c); one isolate of Scutellospora calospora isolated from Guatemala (SC2) and one isolate of Glomus etunicatum isolated from Honduras (GE1). Further experiments will test these selected inocula, singly and in mixtures, in order to obtain an inoculant which significantly improves the growth of C. calothyrsus and to enable its distribution to farmers who use this woody legume for forage production on their farms. PMID:11566397

  17. Improvement of levitation force characteristics in magnetic levitation type seismic isolation device composed of HTS bulk and permanent magnet

    NASA Astrophysics Data System (ADS)

    Tsuda, M.; Kawasaki, T.; Yagai, T.; Hamajima, T.

    2008-02-01

    Magnetic levitation type seismic isolation device composed of HTS bulks and permanent magnets can theoretically remove horizontal vibration completely. It is, however, not easy to generate the large levitation force by using only the levitation system composed of HTS bulk and permanent magnet (HTS-PM system). We focused on a hybrid levitation system composed of the HTS-PM system and the PM-PM system composed of only permanent magnets and investigated the suitable arranging method of the hybrid system for improving levitation force and obtaining stable levitation. In order to clarify the most suitable permanent magnet arrangement in the PM-PM system for the levitation force improvement, repulsive force between permanent magnets was measured in various kinds of the PM-PM system. The maximum repulsive force per unit area in the PM-PM system was at least three times larger than the levitation force per unit area in the HTS-PM system, so that the levitation force in the hybrid system was larger than that of the HTS-PM system. Stable levitation was also achieved in the hybrid system. This is because repulsive force in the PM-PM system against horizontal displacement was much smaller than restoring force in the HTS-PM system.

  18. Study on biodegradation of Mazut by newly isolated strain Enterobacter cloacae BBRC10061: improving and kinetic investigation

    PubMed Central

    2013-01-01

    Mazut as a source content of various hydrocarbons is hard to be degraded and its cracking could turn mazut into useful materials. Nevertheless degradation of mazut by routine methods is too expensive but application of indigenous microorganisms as biocatalysts could be effective and important to lower the costs and expand its consumption. Mazut biodegradation can be improved using various strategies; Therefore in this study newly isolated strain Enterobacter cloacae BBRC 10061 was used in a method of gradual addition of mazut into medium and its results were compared with simple addition method. To investigate degradation of mazut by BBRC 10061, influence of increase of mazut concentration was assayed based on gradual addition method. Also different kinetic models were used to evaluate kinetics of the process. Results showed that gradual addition method has been a beneficial technique for improvement of mazut degradation because bacterial induction to produce biosurfactant and essential enzymes for cracking mazut was higher during process. Although addition of more mazut increased the rate of biodegradation but percentage of degradation decreased. pH of medium decreased during biodegradation period while electric potential increased. Also the biodegradation kinetics was not fitted with the biokinetic models; therefore kinetics of biodegradation of mazut has to be studied by new models. PMID:23369455

  19. Effect of oral nitrates on pulse pressure and arterial elasticity in patients aged over 65 years with refractory isolated systolic hypertension: study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background Isolated systolic hypertension is a highly prevalent disease among the elderly. The little available evidence on the efficacy of nitrates for treating the disease is based on small experimental studies. Methods/design We performed a multicenter, randomized, double-blind, phase III, placebo-controlled trial in 154 patients aged over 65 years with refractory isolated systolic hypertension. Patients were randomized to placebo or 40 mg/day of extended-release isosorbide mononitrate added to standard therapy and titrated to 60 mg/day at week 6 if blood pressure exceeded 140/90 mmHg. The primary objective was to assess the effect on clinical pulse pressure of extended-release isosorbide mononitrate added to standard therapy in patients aged over 65 years with refractory isolated systolic hypertension after 3 months of treatment. The secondary objectives were as follows: to quantify the effect of adding the study drug on central blood pressure and vascular compliance using the augmentation index and pulse wave velocity; to evaluate the safety profile by recording adverse effects (frequency, type, severity) and the percentage of patients who had to withdraw from the trial because of adverse events; to quantify the percentage of patients who reach a clinical systolic blood pressure <140 mmHg or <130 mmHg measured by ambulatory blood pressure monitoring; and to quantify the change in pulse pressure measured by ambulatory blood pressure monitoring. Discussion Few clinical trials have been carried out to test the effect of oral nitrates on isolated systolic hypertension, even though these agents seem to be effective. Treatment with extended-release isosorbide mononitrate could improve control of systolic blood pressure without severe side effects, thus helping to reduce the morbidity and mortality of the disease. Trial registration EUDRACT Number: 2012-002988-10 PMID:24228894

  20. Development of an improved isolation approach and simple sequence repeat markers to characterize Phytophthora capsici populations in irrigation ponds in southern Georgia.

    PubMed

    Wang, Ziying; Langston, David B; Csinos, Alexander S; Gitaitis, Ronald D; Walcott, Ronald R; Ji, Pingsheng

    2009-09-01

    Phytophthora capsici, the causal agent of Phytophthora blight, is a major concern in vegetable production in Georgia and many other states in the United States. Contamination of irrigation water sources by P. capsici may be an important source of inoculum for the pathogen. A simple method was developed in this study to improve the efficiency of recovering P. capsici from fruits used as baits in irrigation ponds. In contrast to direct isolation on agar plates, infected fruit tissues were used to inoculate stems of pepper seedlings, and the infected pepper stems were used for isolation on agar plates. With isolation through inoculation of pepper stems, the frequency of recovering P. capsici from infected eggplant and pear fruits increased from 13.9% to 77.7% and 8.1% to 53.5%, respectively, compared with direct isolation on agar plates. P. capsici was isolated from seven out of nine irrigation ponds evaluated, with most of the ponds containing both A1 and A2 mating types and a 4:5 ratio of A1 to A2 when isolates from all ponds were calculated. All P. capsici isolates were pathogenic on squash plants, and only a small proportion (8.2%) of the isolates were resistant or intermediately sensitive to mefenoxam. Simple sequence repeats (SSRs) were identified through bioinformatics mining of 55,848 publicly available expressed sequence tags of P. capsici in dbEST GenBank. Thirty-one pairs of SSR primers were designed, and SSR analysis indicated that the 61 P. capsici isolates from irrigation ponds were genetically distinct. Cluster analysis separated the isolates into five genetic clusters with no more than two genetic groups in one pond, indicating relatively low P. capsici genetic diversity in each pond. The isolation method and SSR markers developed for P. capsici in this study could contribute to a more comprehensive understanding of the genetic diversity of this important pathogen. PMID:19581483

  1. EFFECT OF MOBILITY ON PERFORMANCE OF WIRELESS AD-HOC NETWORK PROTOCOLS.

    SciTech Connect

    Barrett, C. L.; Drozda, M.; Marathe, M. V.; Marathe, A.

    2001-01-01

    We empirically study the effect of mobility on the performance of protocols designed for wireless adhoc networks. An important ohjective is to study the interaction of the Routing and MAC layer protocols under different mobility parameters. We use three basic mobility models: grid mobility model, random waypoint model, and exponential correlated random model. The performance of protocols was measured in terms of (i) latency, (ii) throughput, (iii) number of packels received, (iv) long term fairness and (v) number of control packets at the MAC layer level. Three different commonly studied routing protocols were used: AODV, DSR and LAR1. Similarly three well known MAC protocols were used: MACA, 802.1 1 and CSMA. The inair1 conclusion of our study include the following: 1. 'I'he performance of the: network varies widely with varying mobility models, packet injection rates and speeds; and can ba in fact characterized as fair to poor depending on the specific situation. Nevertheless, in general, it appears that the combination of AODV and 802.1 I is far better than other combination of routing and MAC protocols. 2. MAC layer protocols interact with routing layer protocols. This concept which is formalized using statistics implies that in general it is not meaningful to speak about a MAC or a routing protocol in isolation. Such an interaction leads to trade-offs between the amount of control packets generated by each layer. More interestingly, the results wise the possibility of improving the performance of a particular MAC layer protocol by using a cleverly designed routing protocol or vice-versa. 3. Routing prolocols with distributed knowledge about routes are more suitable for networks with mobility. This is seen by comparing the performance of AODV with DSR or LAR scheme 1. In DSli and IAR scheme 1, information about a computed path is being stored in the route query control packct. 4. MAC layer protocols have varying performance with varying mobility models. It is

  2. Isolation, Culture, and Staining of Single Myofibers

    PubMed Central

    Gallot, Yann Simon; Hindi, Sajedah M.; Mann, Aman K.; Kumar, Ashok

    2016-01-01

    Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate ex vivo satellite cells activation through isolation of single myofibers from extensor digitorum longus (EDL) muscle of mice and culturing and staining of myofiber-associated satellite cells with the markers of self-renewal, proliferation, and differentiation.

  3. Prior to Conception: The Role of an Acupuncture Protocol in Improving Women's Reproductive Functioning Assessed by a Pilot Pragmatic Randomised Controlled Trial

    PubMed Central

    Cochrane, Suzanne; Smith, Caroline A.; Possamai-Inesedy, Alphia; Bensoussan, Alan

    2016-01-01

    The global average of couples with fertility problems is 9%. Assisted reproductive technologies are often inaccessible. Evidence points to acupuncture offering an opportunity to promote natural fertility. This study asked whether providing a multiphasic fertility acupuncture protocol to women with sub/infertility would increase their awareness of fertility and achieve normalisation of their menstrual cycle compared with a lifestyle control. In a pragmatic randomised controlled trial sub/infertile women were offered an intervention of acupuncture and lifestyle modification or lifestyle modification only. There was a statistically significant increase in fertility awareness in the acupuncture group (86.4%, 19) compared to 40% (n = 8) of the lifestyle only participants (Relative Risk (RR) 2.38, 95% confidence interval (CI) of 1.25, 4.50), with an adjusted p value of 0.011. Changes in menstrual regularity were not statistically significant. There was no statistical difference in the pregnancy rate with seven women (adjusted p = 0.992) achieving pregnancy during the course of the study intervention. Those receiving the acupuncture conceived within an average of 5.5 weeks compared to 10.67 weeks for the lifestyle only group (p = 0.422). The acupuncture protocol tested influenced women who received it compared to women who used lifestyle modification alone: their fertility awareness and wellbeing increased, and those who conceived did so in half the time. PMID:27242910

  4. Clinical Protocol Information System

    PubMed Central

    Wirtschafter, David D.; Gams, Richard; Ferguson, Carol; Blackwell, William; Boackle, Paul

    1980-01-01

    The Clinical Protocol Information System (CPIS) supports the clinical research and patient care objectives of the SouthEastern Cancer Study Group (SEG). The information system goals are to improve the evaluability of clinical trials, decrease the frequency of adverse patient events, implement drug toxicity surveillance, improve the availability of study data and demonstrate the criteria for computer networks that can impact on the general medical care of the community. Nodes in the network consist of Data General MicroNova MP-100 minicomputers that drive the interactive data dialogue and communicate with the network concentrator (another DG MicroNova) in Birmingham. Functions supported include: source data editing, care “advice,” care “audit,” care “explanation,” and treatment note printing. The complete database is updated nightly and resides on UAB's IBM 370/158-AP.

  5. Electrophysiological Characterization of Ts6 and Ts7, K+ Channel Toxins Isolated through an Improved Tityus serrulatus Venom Purification Procedure

    PubMed Central

    Cerni, Felipe A.; Pucca, Manuela B.; Peigneur, Steve; Cremonez, Caroline M.; Bordon, Karla C. F.; Tytgat, Jan; Arantes, Eliane C.

    2014-01-01

    In Brazil, Tityus serrulatus (Ts) is the species responsible for most of the scorpion related accidents. Among the Ts toxins, the neurotoxins with action on potassium channels (α-KTx) present high interest, due to their effect in the envenoming process and the ion channel specificity they display. The α-KTx toxins family is the most relevant because its toxins can be used as therapeutic tools for specific target cells. The improved isolation method provided toxins with high resolution, obtaining pure Ts6 and Ts7 in two chromatographic steps. The effects of Ts6 and Ts7 toxins were evaluated in 14 different types of potassium channels using the voltage-clamp technique with two-microelectrodes. Ts6 toxin shows high affinity for Kv1.2, Kv1.3 and Shaker IR, blocking these channels in low concentrations. Moreover, Ts6 blocks the Kv1.3 channel in picomolar concentrations with an IC50 of 0.55 nM and therefore could be of valuable assistance to further designing immunosuppressive therapeutics. Ts7 toxin blocks multiple subtypes channels, showing low selectivity among the channels analyzed. This work also stands out in its attempt to elucidate the residues important for interacting with each channel and, in the near future, to model a desired drug. PMID:24590385

  6. Outlining a selection procedure for Saccharomyces cerevisiae isolated from grape marc to improve fermentation process and distillate quality.

    PubMed

    Bovo, Barbara; Carlot, Milena; Fontana, Federico; Lombardi, Angiolella; Soligo, Stefano; Giacomini, Alessio; Corich, Viviana

    2015-04-01

    Nowadays grape marc represents one of the main by-product of winemaking. Many South Europe countries valorize this ligno-cellulosic waste through fermentation and distillation for industrial alcoholic beverage production. The storage of marcs is a crucial phase in the distillation process, due to the physicochemical transformations ascribed to microbial activity. Among the methods adopted by distillers to improve the quality of spirits, the use of selected yeasts has not been explored so far, therefore in this work we evaluated the selection criteria of Saccharomyces cerevisiae strains for grape marc fermentation. The proposed selection procedure included three steps: characterization of phenotypical traits, evaluation of selected strains on pasteurised grape marc at lab-scale (100 g) and pilot-scale fermentation (350 kg). This selection process was applied on 104 strains isolated from grape marcs of different origins and technological treatment. Among physiological traits, β-glucosidase activity level as quality trait seems to be only partially involved in increasing varietal flavour. More effective in describing yeast impact on distillate quality is the ratio higher alcohols/esters that indicates strain ability to increase positive flavours. Finally, evaluating grape marc as source of selected yeasts, industrial treatment rather than varietal origin seems to shape strain technological and quality traits. PMID:25475330

  7. Method for microRNA isolation from clinical serum samples.

    PubMed

    Li, Yu; Kowdley, Kris V

    2012-12-01

    MicroRNAs are a group of intracellular noncoding RNA molecules that have been implicated in a variety of human diseases. Because of their high stability in blood, microRNAs released into circulation could be potentially utilized as noninvasive biomarkers for diagnosis or prognosis. Current microRNA isolation protocols are specifically designed for solid tissues and are impractical for biomarker development utilizing small-volume serum samples on a large scale. Thus, a protocol for microRNA isolation from serum is needed to accommodate these conditions in biomarker development. To establish such a protocol, we developed a simplified approach to normalize sample input by using single synthetic spike-in microRNA. We evaluated three commonly used commercial microRNA isolation kits for the best performance by comparing RNA quality and yield. The manufacturer's protocol was further modified to improve the microRNA yield from 200μl of human serum. MicroRNAs isolated from a large set of clinical serum samples were tested on the miRCURY LNA real-time PCR panel and confirmed to be suitable for high-throughput microRNA profiling. In conclusion, we have established a proven method for microRNA isolation from clinical serum samples suitable for microRNA biomarker development.

  8. Interpolation of recurrence and hashing entanglement distillation protocols

    SciTech Connect

    Vollbrecht, Karl Gerd H.; Verstraete, Frank

    2005-06-15

    We construct interesting entanglement distillation protocols by interpolating between the recurrence and hashing protocols. This leads to asymptotic two-way distillation protocols, resulting in an improvement of the distillation rate for all mixed Bell diagonal entangled states, even for the ones with very high fidelity. We also present a method for how entanglement-assisted distillation protocol can be converted into nonentanglement-assisted protocols with the same yield.

  9. Isolation of Mouse Pancreatic Islets of Langerhans.

    PubMed

    Ramírez-Domínguez, Miriam

    2016-01-01

    The aim of any pancreatic islet isolation is obtaining pure, viable and functional pancreatic islets, either for in vitro or in vivo purposes. The islets of Langerhans are complex microorgans with the important role of regulating glucose homeostasis. Imbalances in glucose homeostasis lead to diabetes, which is defined by the American Diabetes Association as a "group of metabolic diseases characterized by hyperglycemia resulting from defects in insulin secretion, insulin action or both" (American Diabetes Association 2011). Currently, the rising demand of human islets is provoking a shortage of this tissue, limiting research and clinical practice on this field. In this scenario, it is essential to investigate and improve islet isolation procedures in animal models, while keeping in mind the anatomical and functional differences between species. This chapter discusses the main aspects of mouse islet isolation research, highlighting the critical factors and shortcomings to take into account for the selection and/or optimization of a mouse islet isolation protocol. PMID:27586420

  10. Can the Isolated-Elements Strategy Be Improved by Targeting Points of High Cognitive Load for Additional Practice?

    ERIC Educational Resources Information Center

    Ayres, Paul

    2013-01-01

    Reducing problem complexity by isolating elements has been shown to be an effective instructional strategy. Novices, in particular, benefit from learning from worked examples that contain partially interacting elements rather than worked examples that provide full interacting elements. This study investigated whether the isolating-elements…

  11. Massive transfusion and massive transfusion protocol

    PubMed Central

    Patil, Vijaya; Shetmahajan, Madhavi

    2014-01-01

    Haemorrhage remains a major cause of potentially preventable deaths. Rapid transfusion of large volumes of blood products is required in patients with haemorrhagic shock which may lead to a unique set of complications. Recently, protocol based management of these patients using massive transfusion protocol have shown improved outcomes. This section discusses in detail both management and complications of massive blood transfusion. PMID:25535421

  12. Massive transfusion and massive transfusion protocol.

    PubMed

    Patil, Vijaya; Shetmahajan, Madhavi

    2014-09-01

    Haemorrhage remains a major cause of potentially preventable deaths. Rapid transfusion of large volumes of blood products is required in patients with haemorrhagic shock which may lead to a unique set of complications. Recently, protocol based management of these patients using massive transfusion protocol have shown improved outcomes. This section discusses in detail both management and complications of massive blood transfusion.

  13. An improved method for the isolation of rat alveolar type II lung cells: Use in the Comet assay to determine DNA damage induced by cigarette smoke.

    PubMed

    Dalrymple, Annette; Ordoñez, Patricia; Thorne, David; Dillon, Debbie; Meredith, Clive

    2015-06-01

    Smoking is a cause of serious diseases, including lung cancer, emphysema, chronic bronchitis and heart disease. DNA damage is thought to be one of the mechanisms by which cigarette smoke (CS) initiates disease in the lung. Indeed, CS induced DNA damage can be measured in vitro and in vivo. The potential of the Comet assay to measure DNA damage in isolated rat lung alveolar type II epithelial cells (AEC II) was explored as a means to include a genotoxicity end-point in rodent sub-chronic inhalation studies. In this study, published AEC II isolation methods were improved to yield viable cells suitable for use in the Comet assay. The improved method reduced the level of basal DNA damage and DNA repair in isolated AEC II. CS induced DNA damage could also be quantified in isolated cells following a single or 5 days CS exposure. In conclusion, the Comet assay has the potential to determine CS or other aerosol induced DNA damage in AEC II isolated from rodents used in sub-chronic inhalation studies.

  14. Improvement of methyl orange dye biotreatment by a novel isolated strain, Aeromonas veronii GRI, by SPB1 biosurfactant addition.

    PubMed

    Mnif, Inès; Maktouf, Sameh; Fendri, Raouia; Kriaa, Mouna; Ellouze, Semia; Ghribi, Dhouha

    2016-01-01

    Aeromonas veronii GRI (KF964486), isolated from acclimated textile effluent after selective enrichment on azo dye, was assessed for methyl orange biodegradation potency. Results suggested the potential of this bacterium for use in effective treatment of azo-dye-contaminated wastewaters under static conditions at neutral and alkaline pH value, characteristic of typical textile effluents. The strain could tolerate higher doses of dyes as it was able to decolorize up to 1000 mg/l. When used as microbial surfactant to enhance methyl orange biodecolorization, Bacillus subtilis SPB1-derived lipopeptide accelerated the decolorization rate and maximized slightly the decolorization efficiency at an optimal concentration of about 0.025%. In order to enhance the process efficiency, a Taguchi design was conducted. Phytotoxicity bioassay using sesame and radish seeds were carried out to assess the biotreatment effectiveness. The bacterium was able to effectively decolorize the azo dye when inoculated with an initial optical density of about 0.5 with 0.25% sucrose, 0.125% yeast extract, 0.01% SPB1 biosurfactant, and when conducting an agitation phase of about 24 h after static incubation. Germination potency showed an increase toward the nonoptimized conditions indicating an improvement of the biotreatment. When comparing with synthetic surfactants, a drastic decrease and an inhibition of orange methyl decolorization were observed in the presence of CTAB and SDS. The nonionic surfactant Tween 80 had a positive effect on methyl orange biodecolorization. Also, studies ensured that methyl orange removal by this strain could be due to endocellular enzymatic activities. To conclude, the addition of SPB1 bioemulsifier reduced energy costs by reducing effective decolorization period, biosurfactant stimulated bacterial decolorization method may provide highly efficient, inexpensive, and time-saving procedure in treatment of textile effluents. PMID:26396008

  15. Improvement of methyl orange dye biotreatment by a novel isolated strain, Aeromonas veronii GRI, by SPB1 biosurfactant addition.

    PubMed

    Mnif, Inès; Maktouf, Sameh; Fendri, Raouia; Kriaa, Mouna; Ellouze, Semia; Ghribi, Dhouha

    2016-01-01

    Aeromonas veronii GRI (KF964486), isolated from acclimated textile effluent after selective enrichment on azo dye, was assessed for methyl orange biodegradation potency. Results suggested the potential of this bacterium for use in effective treatment of azo-dye-contaminated wastewaters under static conditions at neutral and alkaline pH value, characteristic of typical textile effluents. The strain could tolerate higher doses of dyes as it was able to decolorize up to 1000 mg/l. When used as microbial surfactant to enhance methyl orange biodecolorization, Bacillus subtilis SPB1-derived lipopeptide accelerated the decolorization rate and maximized slightly the decolorization efficiency at an optimal concentration of about 0.025%. In order to enhance the process efficiency, a Taguchi design was conducted. Phytotoxicity bioassay using sesame and radish seeds were carried out to assess the biotreatment effectiveness. The bacterium was able to effectively decolorize the azo dye when inoculated with an initial optical density of about 0.5 with 0.25% sucrose, 0.125% yeast extract, 0.01% SPB1 biosurfactant, and when conducting an agitation phase of about 24 h after static incubation. Germination potency showed an increase toward the nonoptimized conditions indicating an improvement of the biotreatment. When comparing with synthetic surfactants, a drastic decrease and an inhibition of orange methyl decolorization were observed in the presence of CTAB and SDS. The nonionic surfactant Tween 80 had a positive effect on methyl orange biodecolorization. Also, studies ensured that methyl orange removal by this strain could be due to endocellular enzymatic activities. To conclude, the addition of SPB1 bioemulsifier reduced energy costs by reducing effective decolorization period, biosurfactant stimulated bacterial decolorization method may provide highly efficient, inexpensive, and time-saving procedure in treatment of textile effluents.

  16. An innovative telemedicine knowledge translation program to improve quality of care in intensive care units: protocol for a cluster randomized pragmatic trial

    PubMed Central

    Scales, Damon C; Dainty, Katie; Hales, Brigette; Pinto, Ruxandra; Fowler, Robert A; Adhikari, Neill KJ; Zwarenstein, Merrick

    2009-01-01

    Background There are challenges to timely adoption of, and ongoing adherence to, evidence-based practices known to improve patient care in the intensive care unit (ICU). Quality improvement initiatives using a collaborative network approach may increase the use of such practices. Our objective is to evaluate the effectiveness of a novel knowledge translation program for increasing the proportion of patients who appropriately receive the following six evidence-based care practices: venous thromboembolism prophylaxis; ventilator-associated pneumonia prevention; spontaneous breathing trials; catheter-related bloodstream infection prevention; decubitus ulcer prevention; and early enteral nutrition. Methods and design We will conduct a pragmatic cluster randomized active control trial in 15 community ICUs and one academic ICU in Ontario, Canada. The intervention is a multifaceted videoconferenced educational and problem-solving forum to organize knowledge translation strategies, including comparative audit and feedback, educational sessions from content experts, and dissemination of algorithms. Fifteen individual ICUs (clusters) will be randomized to receive quality improvement interventions targeting one of the best practices during each of six study phases. Each phase lasts four months during the first study year and three months during the second. At the end of each study phase, ICUs are assigned to an intervention for a best practice not yet received according to a random schedule. The primary analysis will use patient-level process-of-care data to measure the intervention's effect on rates of adoption and adherence of each best practice in the targeted ICU clusters versus controls. Discussion This study design evaluates a new system for knowledge translation and quality improvement across six common ICU problems. All participating ICUs receive quality improvement initiatives during every study phase, improving buy-in. This study design could be considered for other

  17. Alternative parallel ring protocols

    NASA Technical Reports Server (NTRS)

    Mukkamala, R.; Foudriat, E. C.; Maly, Kurt J.; Kale, V.

    1990-01-01

    Communication protocols are know to influence the utilization and performance of communication network. The effect of two token ring protocols on a gigabit network with multiple ring structure is investigated. In the first protocol, a mode sends at most one message on receiving a token. In the second protocol, a mode sends all the waiting messages when a token is received. The behavior of these protocols is shown to be highly dependent on the number of rings as well as the load in the network.

  18. Comparison of seven protocols to identify fecal contamination sources using Escherichia coli

    USGS Publications Warehouse

    Stoeckel, D.M.; Mathes, M.V.; Hyer, K.E.; Hagedorn, C.; Kator, H.; Lukasik, J.; O'Brien, T. L.; Fenger, T.W.; Samadpour, M.; Strickler, K.M.; Wiggins, B.A.

    2004-01-01

    Microbial source tracking (MST) uses various approaches to classify fecal-indicator microorganisms to source hosts. Reproducibility, accuracy, and robustness of seven phenotypic and genotypic MST protocols were evaluated by use of Escherichia coli from an eight-host library of known-source isolates and a separate, blinded challenge library. In reproducibility tests, measuring each protocol's ability to reclassify blinded replicates, only one (pulsed-field gel electrophoresis; PFGE) correctly classified all test replicates to host species; three protocols classified 48-62% correctly, and the remaining three classified fewer than 25% correctly. In accuracy tests, measuring each protocol's ability to correctly classify new isolates, ribotyping with EcoRI and PvuII approached 100% correct classification but only 6% of isolates were classified; four of the other six protocols (antibiotic resistance analysis, PFGE, and two repetitive-element PCR protocols) achieved better than random accuracy rates when 30-100% of challenge isolates were classified. In robustness tests, measuring each protocol's ability to recognize isolates from nonlibrary hosts, three protocols correctly classified 33-100% of isolates as "unknown origin," whereas four protocols classified all isolates to a source category. A relevance test, summarizing interpretations for a hypothetical water sample containing 30 challenge isolates, indicated that false-positive classifications would hinder interpretations for most protocols. Study results indicate that more representation in known-source libraries and better classification accuracy would be needed before field application. Thorough reliability assessment of classification results is crucial before and during application of MST protocols.

  19. Duration of luteal support (DOLS) with progesterone pessaries to improve the success rates in assisted conception: study protocol for a randomized controlled trial

    PubMed Central

    2012-01-01

    Background Luteal support with progesterone is necessary for successful implantation of the embryo following egg collection and embryo transfer in an in-vitro fertilization (IVF) cycle. Progesterone has been used for as little as 2 weeks and for as long as 12 weeks of gestation. The optimal length of treatment is unresolved at present and it remains unclear how long to treat women receiving luteal supplementation. Design The trial is a prospective, randomized, double-blind, placebo-controlled trial to investigate the effect of the duration of luteal support with progesterone in IVF cycles. Following 2 weeks standard treatment and a positive biochemical pregnancy test, this randomized control trial will allocate women to a supplementary 8 weeks treatment with vaginal progesterone or 8 weeks placebo. Further studies would be required to investigate whether additional supplementation with progesterone is beneficial in early pregnancy. Discussion Currently at the Hewitt Centre, approximately 32.5% of women have a positive biochemical pregnancy test 2 weeks after embryo transfer. It is this population that is eligible for trial entry and randomization. Once the patient has confirmed a positive urinary pregnancy test they will be invited to join the trial. Once the consent form has been completed by the patient a trial prescription sheet will be sent to pharmacy with a stated collection time. The patient can then be randomized and the drugs dispensed according to pharmacy protocol. A blood sample will then be drawn for measurement of baseline hormone levels (progesterone, estradiol, free beta-human chorionic gonadotrophin, pregnancy-associated plasma protein-A, Activin A, Inhibin A and Inhibin B). The primary outcome measure is the proportion of all randomized women that continue successfully to a viable pregnancy (at least one fetus with fetal heart rate >100 beats/minute) on transabdominal/transvaginal ultrasound at 10 weeks post embryo transfer/12 weeks gestation

  20. Using patients’ experiences of adverse events to improve health service delivery and practice: protocol of a data linkage study of Australian adults age 45 and above

    PubMed Central

    Walton, Merrilyn; Smith-Merry, Jennifer; Harrison, Reema; Manias, Elizabeth; Iedema, Rick; Kelly, Patrick

    2014-01-01

    Introduction Evidence of patients’ experiences is fundamental to creating effective health policy and service responses, yet is missing from our knowledge of adverse events. This protocol describes explorative research redressing this significant deficit; investigating the experiences of a large cohort of recently hospitalised patients aged 45 years and above in hospitals in New South Wales (NSW), Australia. Methods and analysis The 45 and Up Study is a cohort of 265 000 adults aged 45 years and above in NSW. Patients who were hospitalised between 1 January and 30 June 2014 will be identified from this cohort using data linkage and a random sample of 20 000 invited to participate. A cross-sectional survey (including qualitative and quantitative components) will capture patients’ experiences in hospital and specifically of adverse events. Approximately 25% of respondents are likely to report experiencing an adverse event. Quantitative components will capture the nature and type of events as well as common features of patients’ experiences. Qualitative data provide contextual knowledge of their condition and care and the impact of the event on individuals. Respondents who do not report an adverse event will report their experience in hospital and be the control group. Statistical and thematic analysis will be used to present a patient perspective of their experiences in hospital; the characteristics of patients experiencing an adverse event; experiences of information sharing after an event (open disclosure) and the other avenues of redress pursued. Interviews with key policymakers and a document analysis will be used to create a map of the current practice. Ethics and dissemination Dissemination via a one-day workshop, peer-reviewed publications and conference presentations will enable effective clinical responses and service provision and policy responses to adverse events to be developed. PMID:25311039

  1. Using two simple methods of Ar-Ar(F) self-assembly and isolation chromophores to further improve the comprehensive performance of NLO dendrimers.

    PubMed

    Wu, Wenbo; Yu, Gui; Liu, Yunqi; Ye, Cheng; Qin, Jingui; Li, Zhen

    2013-01-01

    Herein, through a combination of divergent and convergent approaches, coupled with the utilization of the powerful Sharpless "click chemistry" reaction, two series of high-generation nonlinear optical (NLO) dendrimers have been conveniently prepared in high purity and satisfactory yields. Perfluoroaromatic rings and isolation chromophores were introduced to further improve their comprehensive performance. Thanks to the effects of Ar-Ar(F) self-assembly and the isolation chromophores, coupled with perfect 3D spatial isolation from the highly branched structure of the dendrimer, G5-PFPh-NS displayed very large NLO efficiency (up to 257 pm V(-1)), which is, to the best of our knowledge, the new record highest value reported so far for simple azo chromophore moieties. High-quality wide optical transparency and good stability were also achieved.

  2. Supplemented base medium containing Amburana cearensis associated with FSH improves in vitro development of isolated goat preantral follicles.

    PubMed

    Gouveia, B B; Macedo, T J S; Santos, J M S; Barberino, R S; Menezes, V G; Müller, M C; Almeida, J R G S; Figueiredo, J R; Matos, M H T

    2016-09-15

    The effects of Amburana cearensis ethanolic extract, with or without addition of a mix of supplements associated or not with FSH, on in vitro morphology and development of caprine secondary follicles were evaluated. In experiment 1, isolated follicles (250 μm in diameter) were cultured for 12 days in alpha-modified minimal essential medium (α-MEM) alone (control) or in medium composed of different concentrations of A. cearensis extract (Amb 0.1; 0.2, or 0.4 mg/mL). In experiment 2, culture media were α-MEM or Amb 0.2 mg/mL (both without supplements), or these same media supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred as α-MEM(+) and Amb 0.2(+), respectively), or these last groups also supplemented with sequential FSH (100 ng/mL from Day 0 to Day 6; 500 ng/mL from Day 6 to Day 12), constituting groups α-MEM(+) + FSH and Amb 0.2(+) + FSH. At the end of culture in experiment 1, control medium (α-MEM) and Amb 0.2 mg/mL had higher percentages (P < 0.05) of morphologically normal follicles and percentage of fully grown oocytes, i.e., oocyte greater than 110 μm, compared to the other A. cearensis extract concentrations. In experiment 2, all supplemented media had higher percentages (P < 0.05) of normal follicles and antrum formation than nonsupplemented media. In addition, follicles cultured in Amb 0.2(+) + FSH showed an average increase in diameter higher (P < 0.05) than the other treatments. Oocytes cultured in both treatments supplemented with FSH showed greater glutathione and active mitochondria levels than nonsupplemented media but similar to the other treatments. In conclusion, A. cearensis extract (0.2 mg/mL) added by supplements and FSH improves follicular growth. Therefore, it can be an alternative culture medium for goat preantral follicle development. PMID:27287468

  3. Improving outcomes for severe sepsis and septic shock: tools for early identification of at-risk patients and treatment protocol implementation.

    PubMed

    Rivers, Emanuel P; Ahrens, Tom

    2008-07-01

    Sepsis is a significant problem, and septicemia is the 10th leading cause of death in the United States. Sepsis incidence is increasing, and the mortality rate is 20% to 50% for patients with severe sepsis. This article identifies methods for improving outcomes of severe sepsis and septic shock. Included are recommendations for diagnosis and treatment. Case studies are included.

  4. A multi-centre randomised controlled trial of rehabilitation aimed at improving outdoor mobility for people after stroke: Study protocol for a randomised controlled trial

    PubMed Central

    2012-01-01

    . Discussion This study protocol describes a pragmatic randomised controlled trial that will hopefully provide robust evidence of the benefit of outdoor mobility interventions after stroke for clinicians working in the community. The results will be available towards the end of 2012. Trial registration ISRCTN58683841 PMID:22721452

  5. A protocol for improving mapping and assessing of seagrass abundance along the West Central Coast of Florida using Landsat TM and EO-1 ALI/Hyperion images

    NASA Astrophysics Data System (ADS)

    Pu, Ruiliang; Bell, Susan

    2013-09-01

    Seagrass habitats are characteristic features of shallow waters worldwide and provide a variety of ecosystem functions. Remote sensing techniques can help collect spatial and temporal information about seagrass resources. In this study, we evaluate a protocol that utilizes image optimization algorithms followed by atmospheric and sunglint corrections to the three satellite sensors [Landsat 5 Thematic Mapper (TM), Earth Observing-1 (EO-1) Advanced Land Imager (ALI) and Hyperion (HYP)] and a fuzzy synthetic evaluation technique to map and assess seagrass abundance in Pinellas County, FL, USA. After image preprocessed with image optimization algorithms and atmospheric and sunglint correction approaches, the three sensors' data were used to classify the submerged aquatic vegetation cover (%SAV cover) into 5 classes with a maximum likelihood classifier. Based on three biological metrics [%SAV, leaf area index (LAI), and Biomass] measured from the field, nine multiple regression models were developed for estimating the three biometrics with spectral variables derived from the three sensors' data. Then, five membership maps were created with the three biometrics along with two environmental factors (water depth and distance-to-shoreline). Finally, seagrass abundance maps were produced by using a fuzzy synthetic evaluation technique and five membership maps. The experimental results indicate that the HYP sensor produced the best results of the 5-class classification of %SAV cover (overall accuracy = 87% and Kappa = 0.83 vs. 82% and 0.77 by ALI and 79% and 0.73 by TM) and better multiple regression models for estimating the three biometrics (R2 = 0.66, 0.62 and 0.61 for %SAV, LAI and Biomass vs. 0.62, 0.61 and 0.55 by ALI and 0.58, 0.56 and 0.52 by TM) for creating seagrass abundance maps along with two environmental factors. Combined our results demonstrate that the image optimization algorithms and the fuzzy synthetic evaluation technique were effective in mapping

  6. Improving advance care planning for English-speaking and Spanish-speaking older adults: study protocol for the PREPARE randomised controlled trial

    PubMed Central

    Sudore, Rebecca L; Barnes, Deborah E; Le, Gem M; Ramos, Roberto; Osua, Stacy J; Richardson, Sarah A; Boscardin, John; Schillinger, Dean

    2016-01-01

    Introduction Advance care planning (ACP) is a process that allows patients to identify their goals for medical care. Traditionally, ACP has focused on completing advance directives; however, we have expanded the ACP paradigm to also prepare patients to communicate their wishes and make informed decisions. To this end, we created an ACP website called PREPARE (http://www.prepareforyourcare.org) to prepare diverse English-speaking and Spanish-speaking older adults for medical decision-making. Here, we describe the study protocol for a randomised controlled efficacy trial of PREPARE in a safety-net setting. The goal is to determine the efficacy of PREPARE to engage diverse English-speaking and Spanish-speaking older adults in a full spectrum of ACP behaviours. Methods and analysis We include English-speaking and Spanish-speaking adults from an urban public hospital who are ≥55 years old, have ≥2 chronic medical conditions and have seen a primary care physician ≥2 times in the last year. Participants are randomised to the PREPARE intervention (review PREPARE and an easy-to-read advance directive) or the control arm (only the easy-to-read advance directive). The primary outcome is documentation of an advance directive and/or ACP discussion. Secondary outcomes include ACP behaviour change processes measured with validated surveys (eg, self-efficacy, readiness) and a broad range of ACP actions (eg, choosing a surrogate, identifying goals for care, discussing ACP with clinicians and/or surrogates). Using blinded outcome ascertainment, outcomes will be measured at 1 week and at 3, 6 and 12 months, and compared between study arms using mixed-effects logistic regression and mixed-effects linear, Poisson or negative binomial regression. Ethics and dissemination This study has been approved by the appropriate Institutional Review Boards and is guided by input from patient and clinical advisory boards and a data safety monitoring board. The results of this study will

  7. Improving implementation of evidence-based practice in mental health service delivery: protocol for a cluster randomised quasi-experimental investigation of staff-focused values interventions

    PubMed Central

    2013-01-01

    files and staff work samples are reviewed for each condition to determine the impact of implementation. Self-determination theory and theories of organisational change are used to interpret the data. Discussion The research adds to the current knowledge base related to worker motivation and uptake of workplace practice. It describes a structured protocol that aims to enhance worker autonomy for imposed workplace practices. The research will inform how best to measure and conceptualise transfer. These findings will apply particularly to contexts where individuals are not ‘volunteers’ in requisite change processes. Trial registration ACTRN: ACTRN12613000353796. PMID:23819816

  8. Isolation and characterisation of selected lactic acid bacteria for improved processing of Nem chua, a traditional fermented meat from Vietnam.

    PubMed

    Nguyen, H T H; Elegado, F B; Librojo-Basilio, N T; Mabesa, R C; Dizon, E I

    2010-03-01

    Nem chua is a traditional lactic acid fermented meat of Vietnam that is consumed raw. During fermentation, the number of acid-producing bacteria increased significantly and by the final day (4th), it had reached approximately 8.4 log cfu/g, contributing to 94% of the total microbiota of the product. Out of 85 isolates obtained from the product, only 44 were confirmed Gram-positive and catalase-negative. These putative LAB isolates were purified and screened for antimicrobial and proteolytic activity. None of the isolates showed bacteriocin activity using the 'spot-on lawn' method, but showed antimicrobial activity against Lactobacillus sakei and Enterococcus faecium using deferred assay. Four isolates showed good proteolytic activities at pH 7.0. These isolates were identified as Lactobacillus plantarum using an API 50 CHL i.d. kit. Sequencing of the partial 16S rRNA gene amplified by primers 1101F (5'-AACGAGCGCAACCC-3') and 1407R (5'-GACGGGCGGTGTGTAC-3') showed 98% homology to Lactobacillus plantarum WCFS1.

  9. Licklider Transmission Protocol Implementation

    NASA Technical Reports Server (NTRS)

    Burleigh, Scott C.; Krupiarz, Chris

    2011-01-01

    This software is an implementation of the Licklider Transmission Protocol (LTP), a communications protocol intended to support the Bundle Protocol in Delay-Tolerant Network (DTN) operations. LTP is designed to provide retransmission-based reliability over links characterized by extremely long message round-trip times and/or frequent interruptions in connectivity. Communication in interplanetary space is the most prominent example of this sort of environment, and LTP is principally aimed at supporting long-haul reliable transmission over deep-space RF links. Like any reliable transport service employing ARQ (Automatic Repeat re-Quests), LTP is stateful. In order to assure the reception of a block of data it has sent, LTP must retain for possible retransmission all portions of that block which might not have been received yet. In order to do so, it must keep track of which portions of the block are known to have been received so far, and which are not, together with any additional information needed for purposes of retransmitting part, or all, of the block. Long round-trip times mean substantial delay between the transmission of a block of data and the reception of an acknowledgement from the block s destination, signaling arrival of the block. If LTP postponed transmission of additional blocks of data until it received acknowledgement of the arrival of all prior blocks, valuable opportunities to use what little deep space transmission bandwidth is available would be forever lost. For this reason, LTP is based in part on a notion of massive state retention. Any number of requested transmission conversations (sessions) may be concurrently in flight at various displacements along the link between two LTP engines, and the LTP engines must necessarily retain transmission status and retransmission resources for all of them. Moreover, if any of the data of a given block are lost en route, it will be necessary to retain the state of that transmission during an additional

  10. Isolation of clonal cultures of endosymbiotic green algae from their ciliate hosts.

    PubMed

    Achilles-Day, Undine E M; Day, John G

    2013-03-01

    Using Paramecium bursaria as a model organism improved protocols have been developed to isolate clonal endosymbiotic algae. This involved micromanipulation of individual protists, rupturing to release endosymbionts followed by enrichment on complex media and a series of plating steps, under low light (PAR ~10μmol photons m(-2)s(-1)).

  11. Improvement of hospital care for patients with non-Hodgkin’s lymphoma: protocol for a cluster randomized controlled trial (PEARL study)

    PubMed Central

    2013-01-01

    Background Malignant lymphomas constitute a diverse group of cancers of lymphocytes. One well-known disease is Hodgkin’s lymphoma; the others are classified as non-Hodgkin’s lymphoma (NHL). NHLs are the most common hematologic neoplasms in adults worldwide, and in 2012 over 170,000 new cases were estimated in the United States and Europe. In previous studies, several practice gaps in hospital care for patients with NHL have been identified. To decrease this variation in care, the present study aims to perform a problem analysis in which barriers to and facilitators for optimal NHL care will be identified and, based on these findings, to develop (tailored) improvement strategies. Subsequently, we will assess the effectiveness, feasibility and costs of the improvement strategies. Methods/design Barriers and facilitators will be explored using the literature, using interviews and questionnaires among physicians involved in NHL care, and patients diagnosed with NHL. The results will be used to develop a tailored improvement strategy. A cluster randomized controlled trial involving 19 Dutch hospitals will be conducted. Hospitals will be randomized to receive either an improvement strategy tailored to the barriers and facilitators found or, a standard strategy of audit and feedback. The effects of both strategies will be evaluated using previously developed quality indicators. Adherence to the indicators will be measured before and after the intervention period based on medical records from newly diagnosed NHL patients. To study the feasibility of both strategies, a process evaluation will be additionally performed. Data about exposure to the different elements of the strategies will be collected using questionnaires. Economic evaluation from a healthcare perspective will compare the two implementation strategies, where the costs of the implementation strategy and changes in healthcare consumption will be assessed. Discussion The presence of variation in the use of

  12. Improving physical health and reducing substance use in psychosis – randomised control trial (IMPACT RCT): study protocol for a cluster randomised controlled trial

    PubMed Central

    2013-01-01

    Background Cardiovascular morbidity and mortality is increased in individuals with severe mental illnesses. We set out to establish a multicentre, two arm, parallel cluster randomized controlled trial (RCT) of a health promotion intervention (HPI), IMPACT Therapy. The patient-tailored IMPACT Therapy aims to target one or more health behaviours from a pre-defined list that includes cannabis use; alcohol use; other substance use; cigarette smoking; exercise; diet and diabetic control, prioritising those identified as problematic by the patient, taking a motivational interviewing and CBT approach. Methods Impact therapy will be delivered by care coordinators in the community to the treatment group and will be compared to treatment as usual (TAU). The main hypothesis is that the addition of IMPACT Therapy (HPI) to TAU will be more effective than TAU alone in improving patients’ quality of life as measured by the Short Form-36, including mental health and physical health subscales on completion of the intervention at 12 months post randomisation. A subsidiary hypothesis will be that addition of IMPACT Therapy (HPI) will be more cost-effective than TAU alone in improving health in people with SMI 12 months from baseline. The IMPACT therapy patient groups’ improvement in quality of life, as well as its cost effectiveness, is hypothesised to be maintained at 15 months. Outcomes will be analyzed on an intention-to-treat (ITT) basis. Discussion The results of the trial will provide information about the effectiveness of the IMPACT therapy programme in supporting community mental health teams to address physical comorbidity in severe mental illness. Trial registration ISRCTN58667926. PMID:24131496

  13. Investigating a TELEmedicine solution to improve MEDication adherence in chronic Heart Failure (TELEMED-HF): study protocol for a randomized controlled trial

    PubMed Central

    2011-01-01

    Background Frequent rehospitalisations and poorer survival chances in heart failure (HF) patients may partly be explained by poor medication adherence. There are multiple medication-related reasons for suboptimal adherence, but psychological reasons may also be important. A novel TELEmonitoring device may improve MEDication adherence in HF patients (TELEMED-HF). TELEMED-HF is a randomized, controlled clinical intervention trial designed to examine (1) the efficacy and cost-efficiency of an electronic medication adherence support system in improving and monitoring HF patients' medication adherence; (2) the effect of medication adherence on hospitalizations and health care consumption; as well as on (3) clinical characteristics, and Quality of Life (QoL); and (4) clinical, sociodemographic, and psychological determinants of medication adherence. Methods/Design Consecutive patients with chronic, systolic HF presenting to the outpatient clinic of the TweeSteden Hospital, The Netherlands, will be approached for study participation and randomly assigned (1:1) following blocked randomization procedures to the intervention (n = 200) or usual care arm (n = 200). Patients in the intervention arm use the medication support device for six months in addition to usual care. Post-intervention, patients return to usual care only and all patients participate in four follow-up occasions over 12 months. Primary endpoints comprise objective and subjective medication adherence, healthcare consumption, number of hospitalizations, and cost-effectiveness. Secondary endpoints include disease severity, physical functioning, and QoL. Discussion The TELEMED-HF study will provide us a comprehensive understanding of medication adherence in HF patients, and will show whether telemonitoring is effective and cost-efficient in improving adherence and preventing hospitalization in HF patients. Trial registration number NCT01347528. PMID:21999637

  14. Study protocol for a randomized, controlled trial comparing the efficacy of two educational interventions to improve inhalation techniques in patients with chronic obstructive pulmonary disease (COPD): TIEPOC Study

    PubMed Central

    Leiva-Fernández, José; Leiva-Fernández, Francisca; Vázquez-Alarcón, Rubén L; García-Ruiz, Antonio; Prados-Torres, Daniel; Barnestein-Fonseca, Pilar

    2014-01-01

    Background: An appropriate inhalation technique and adherence to treatment are both critical determinants of the success of chronic obstructive pulmonary disease (COPD) management. We have observed that up to 75% of patients do not use a successful inhalation technique. Knowledge evaluation and frequent reassessment of inhaler use, together with education of patients and healthcare professionals, can significantly improve the benefits that patients with COPD will derive from inhaler therapy. The objective of this study is to test the efficacy of two educational interventions to improve inhalation techniques in patients with COPD. Methods: Multicenter randomized controlled trial with 296 patients diagnosed with COPD selected by a non-probabilistic method of sampling from seven Spanish Primary Care Centers. The patients will be divided into three groups by block randomization. The three groups are: 1) control; 2) Intervention A; and 3) Intervention B. The control group will comprise patients with no explanations or written information; the Intervention A group will comprise patients to whom we give written information only; and the Intervention B group will comprise patients to whom we give written information plus instructor training. Every patient in each group will be visited four times during the year of the study at the health centers. Discussion: Our hypothesis is that the application of educational interventions (A or B) in patients with COPD who use inhaler therapy will increase the number of patients who perform a correct inhalation technique by at least 25%. We will evaluate the effectiveness of these interventions on patient inhalation technique improvement, where feasible within the context of clinical practice. PMID:24991223

  15. A cluster randomized trial to improve adherence to evidence-based guidelines on diabetes and reduce clinical inertia in primary care physicians in Belgium: study protocol [NTR 1369

    PubMed Central

    Borgermans, Liesbeth; Goderis, Geert; Broeke, Carine Van Den; Mathieu, Chantal; Aertgeerts, Bert; Verbeke, Geert; Carbonez, An; Ivanova, Anna; Grol, Richard; Heyrman, Jan

    2008-01-01

    Background Most quality improvement programs in diabetes care incorporate aspects of clinician education, performance feedback, patient education, care management, and diabetes care teams to support primary care physicians. Few studies have applied all of these dimensions to address clinical inertia. Aim To evaluate interventions to improve adherence to evidence-based guidelines for diabetes and reduce clinical inertia in primary care physicians. Design Two-arm cluster randomized controlled trial. Participants Primary care physicians in Belgium. Interventions Primary care physicians will be randomly allocated to 'Usual' (UQIP) or 'Advanced' (AQIP) Quality Improvement Programs. Physicians in the UQIP will receive interventions addressing the main physician, patient, and office system factors that contribute to clinical inertia. Physicians in the AQIP will receive additional interventions that focus on sustainable behavior changes in patients and providers. Outcomes Primary endpoints are the proportions of patients within targets for three clinical outcomes: 1) glycosylated hemoglobin < 7%; 2) systolic blood pressure differences ≤130 mmHg; and 3) low density lipoprotein/cholesterol < 100 mg/dl. Secondary endpoints are individual improvements in 12 validated parameters: glycosylated hemoglobin, low and high density lipoprotein/cholesterol, total cholesterol, systolic blood pressure, diastolic blood pressure, weight, physical exercise, healthy diet, smoking status, and statin and anti-platelet therapy. Primary and secondary analysis Statistical analyses will be performed using an intent-to-treat approach with a multilevel model. Linear and generalized linear mixed models will be used to account for the clustered nature of the data, i.e., patients clustered withinimary care physicians, and repeated assessments clustered within patients. To compare patient characteristics at baseline and between the intervention arms, the generalized estimating equations (GEE) approach

  16. Feasibility intervention trial of two types of improved cookstoves in three resource-limited settings: study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background Exposure to biomass fuel smoke is one of the leading risk factors for disease burden worldwide. International campaigns are currently promoting the widespread adoption of improved cookstoves in resource-limited settings, yet little is known about the cultural and social barriers to successful improved cookstove adoption and how these barriers affect environmental exposures and health outcomes. Design We plan to conduct a one-year crossover, feasibility intervention trial in three resource-limited settings (Kenya, Nepal and Peru). We will enroll 40 to 46 female primary cooks aged 20 to 49 years in each site (total 120 to 138). Methods At baseline, we will collect information on sociodemographic characteristics and cooking practices, and measure respiratory health and blood pressure for all participating women. An initial observational period of four months while households use their traditional, open-fire design cookstoves will take place prior to randomization. All participants will then be randomized to receive one of two types of improved, ventilated cookstoves with a chimney: a commercially-constructed cookstove (Envirofit G3300/G3355) or a locally-constructed cookstove. After four months of observation, participants will crossover and receive the other improved cookstove design and be followed for another four months. During each of the three four-month study periods, we will collect monthly information on self-reported respiratory symptoms, cooking practices, compliance with cookstove use (intervention periods only), and measure peak expiratory flow, forced expiratory volume at 1 second, exhaled carbon monoxide and blood pressure. We will also measure pulmonary function testing in the women participants and 24-hour kitchen particulate matter and carbon monoxide levels at least once per period. Discussion Findings from this study will help us better understand the behavioral, biological, and environmental changes that occur with a cookstove

  17. The braided single-stage protocol for quantum secure communication

    NASA Astrophysics Data System (ADS)

    Darunkar, Bhagyashri; Verma, Pramode K.

    2014-05-01

    This paper presents the concept and implementation of a Braided Single-stage Protocol for quantum secure communication. The braided single-stage protocol is a multi-photon tolerant secure protocol. This multi-photon tolerant protocol has been implemented in the laboratory using free-space optics technology. The proposed protocol capitalizes on strengths of the three-stage protocol and extends it with a new concept of braiding. This protocol overcomes the limitations associated with the three-stage protocol in the following ways: It uses the transmission channel only once as opposed to three times in the three-stage protocol, and it is invulnerable to man-in-the-middle attack. This paper also presents the error analysis resulting from the misalignment of the devices in the implementation. The experimental results validate the efficient use of transmission resources and improvement in the data transfer rate.

  18. Evaluations and modifications of semi-selective media for improved isolation of Agrobacterium tumefaciens biovar 1 from cultivated walnut

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Agrobacterium tumefaciens, the causal agent of crown gall of walnut, is an aerobic, Gram negative bacterium belonging to the family Rhizobiaceae. Like many in this group, A. tumefaciens is a common inhabitant of soil and plant host tissue. Isolation from these complex environments is difficult even ...

  19. MDCK-SIAT1 cells show improved isolation rates for recent human influenza viruses compared to conventional MDCK cells.

    PubMed

    Oh, Ding Yuan; Barr, Ian G; Mosse, Jenny A; Laurie, Karen L

    2008-07-01

    The ability to isolate and propagate influenza virus is an essential tool for the yearly surveillance of circulating virus strains and to ensure accurate clinical diagnosis for appropriate treatment. The suitability of MDCK-SIAT1 cells, engineered to express increased levels of alpha-2,6-linked sialic acid receptors, as an alternative to conventional MDCK cells for isolation of circulating influenza virus was assessed. A greater number of influenza A (H1N1 and H3N2) and B viruses from stored human clinical specimens collected between 2005 and 2007 were isolated following inoculation in MDCK-SIAT1 cells than in MDCK cells. In addition, a higher titer of virus was recovered following culture in MDCK-SIAT1 cells. All A(H1N1) viruses recovered from MDCK-SIAT1 cells were able to agglutinate both turkey and guinea pig red blood cells (RBC), while half of the A(H3N2) viruses recovered after passage in MDCK-SIAT1 cells lost the ability to agglutinate turkey RBC. Importantly, the HA-1 domain of the hemagglutinin gene was genetically stable after passaging in MDCK-SIAT1 cells, a feature not always seen following MDCK cell or embryonated chicken egg passage of human influenza virus. These data indicate that the MDCK-SIAT1 cell line is superior to conventional MDCK cells for isolation of human influenza virus from clinical specimens and may be used routinely for the isolation and propagation of current human influenza viruses for surveillance, diagnostic, and research purposes.

  20. The RTS2 protocol

    NASA Astrophysics Data System (ADS)

    Kubánek, Petr; Jelínek, Martin; French, John; Prouza, Michal; Vítek, Stanislav; Castro-Tirado, Alberto J.; Reglero, Victor

    2008-07-01

    Remote Telescope System 2nd version (RTS2) is an open source project aimed at developing a software environment to control a fully robotic observatory. RTS2 consists of various components, which communicate via an ASCII based protocol. As the protocol was from the beginning designed as an observatory control system, it provides some unique features, which are hard to find in the other communication systems. These features include advanced synchronisation mechanisms and strategies for setting variables. This presentation describes the protocol and its unique features. It also assesses protocol performance, and provides examples how the RTS2 library can be used to quickly build an observatory control system.

  1. Improving access to preparatory information for children undergoing general anaesthesia for tooth extraction and their families: study protocol for a Phase III randomized controlled trial

    PubMed Central

    2014-01-01

    Background Children can find anaesthesia induction especially distressing and postoperative psychological and physical morbidity are common. Preparation programmes for general anaesthesia (GA) are highly effective in reducing this distress. A Phase II study has already verified the effectiveness of a prototype preoperative GA-coping computer game to help children cope with induction in a dental GA setting. The biggest patient users of pediatric GA services in the UK are children who need to have teeth removed (estimated to be 100,000 yearly). Tooth decay is the most common disease in children worldwide. This study is a Phase III randomized controlled trial (RCT) and will evaluate the effectiveness of the new internet version of this game. Methods/design The Phase III RCT will use a double-blind three-armed design. The clinical trial will recruit up to 210 children and will compare the web-based game against standard care and another non-medical game. At least 53 patients in each group will be required for 90% statistical power. Distress will be assessed through an evaluation of the child’s behaviour during the visit and later parental reports of physical and psychological morbidity. The satisfaction of parents and children will be measured; the mode of usage of the web-based game will be automatically recorded and the impact on the service (for example, recovery time and throughput) will be reported. The Phase III study primary outcome will measure: (1) patient experience: acceptance of anaesthetic induction, child cooperation and distress, reduction of peri- and postoperative morbidity, child and family satisfaction, and (2) service improvement: anaesthetic time and improvement in throughput. Measures will be administered at baseline, at the time of the GA treatment visit, and at 48 hours and one week postoperatively. Discussion This study aims to determine the effectiveness of an online GA-coping game for children and families undergoing tooth extraction under

  2. An e-Health Decision Support System for Improving Compliance of Health Workers to the Maternity Care Protocols in South Africa

    PubMed Central

    Horner, V.; Rautenbach, P.; Mbananga, N.; Mashamba, T.; Kwinda, H.

    2013-01-01

    Background There are problems of quality in maternity services at primary health care level in South Africa. Many of these problems can be traced to non-adherence to the maternity care guidelines and lack of tools to support clinic staff and managers in their roles. Objective: The aim of this research was to investigate the utility of e-health (computerized) decision support systems at addressing the problem of compliance of health workers to the maternity care guidelines at primary health care in South Africa. At present there are no documented studies on e-health clinical decision support systems for primary health care in South Africa, though clinical decision support systems for primary health care are listed as part of the e-health strategy of the National Department of Health. Methods: An e-health decision support system named the Bacis (Basic Antenatal Care Information System) Program was developed, then implemented and evaluated at a primary health care clinic. The duration of the study was two years: this includes development, implementation and evaluation. Results and Conclusion There was an overall improvement in compliance from 85.1 % to 89.3%. This result was not statistically significant. However when results were stratified into specific categories, the Bacis Program showed statistically significant improvement in compliance over the checklist system in three out of nine important categories. These are compliance at booking, patients younger than 18 years and patients booking after week 20. Further, insights and experience were also gained on development and implementation of clinical information systems at resource strained environments such as primary health care in South Africa. These results, insights and experience are invaluable for the implementation of the proposed e-health strategy in South Africa. PMID:23650485

  3. Multistrategy childcare-based intervention to improve compliance with nutrition guidelines versus usual care in long day care services: a study protocol for a randomised controlled trial

    PubMed Central

    Seward, Kirsty; Finch, Meghan; Wiggers, John; Wyse, Rebecca; Jones, Jannah; Gillham, Karen; Yoong, Sze Lin

    2016-01-01

    Introduction Interventions to improve child diet are recommended as dietary patterns developed in childhood track into adulthood and influence the risk of chronic disease. For child health, childcare services are required to provide foods to children consistent with nutrition guidelines. Research suggests that foods and beverages provided by services to children are often inconsistent with nutrition guidelines. The primary aim of this study is to assess, relative to a usual care control group, the effectiveness of a multistrategy childcare-based intervention in improving compliance with nutrition guidelines in long day care services. Methods and analysis The study will employ a parallel group randomised controlled trial design. A sample of 58 long day care services that provide all meals (typically includes 1 main and 2 mid-meals) to children while they are in care, in the Hunter New England region of New South Wales, Australia, will be randomly allocated to a 6-month intervention to support implementation of nutrition guidelines or a usual care control group in a 1:1 ratio. The intervention was designed to overcome barriers to the implementation of nutrition guidelines assessed using the theoretical domains framework. Intervention strategies will include the provision of staff training and resources, audit and feedback, ongoing support and securing executive support. The primary outcome of the trial will be the change in the proportion of long day care services that have a 2-week menu compliant with childcare nutrition guidelines, measured by comprehensive menu assessments. As a secondary outcome, child dietary intake while in care will also be assessed. To assess the effectiveness of the intervention, the measures will be undertaken at baseline and ∼6 months postbaseline. Ethics and dissemination The study was approved by the Hunter New England Human Research Ethics Committee. Study findings will be disseminated widely through peer-reviewed publications. PMID

  4. Efficacy and safety of a multifactor intervention to improve therapeutic adherence in patients with chronic obstructive pulmonary disease (COPD): protocol for the ICEPOC study

    PubMed Central

    2011-01-01

    Background Low therapeutic adherence to medication is very common. Clinical effectiveness is related to dose rate and route of administration and so poor therapeutic adherence can reduce the clinical benefit of treatment. The therapeutic adherence of patients with chronic obstructive pulmonary disease (COPD) is extremely poor according to most studies. The research about COPD adherence has mainly focussed on quantifying its effect, and few studies have researched factors that affect non-adherence. Our study will evaluate the effectiveness of a multifactor intervention to improve the therapeutic adherence of COPD patients. Methods/Design A randomized controlled clinical trial with 140 COPD diagnosed patients selected by a non-probabilistic method of sampling. Subjects will be randomly allocated into two groups, using the block randomization technique. Every patient in each group will be visited four times during the year of the study. Intervention: Motivational aspects related to adherence (beliefs and behaviour): group and individual interviews; cognitive aspects: information about illness; skills: inhaled technique training. Reinforcement of the cognitive-emotional aspects and inhaled technique training will be carried out in all visits of the intervention group. Discussion Adherence to a prescribed treatment involves a behavioural change. Cognitive, emotional and motivational aspects influence this change and so we consider the best intervention procedure to improve adherence would be a cognitive and emotional strategy which could be applied in daily clinical practice. Our hypothesis is that the application of a multifactor intervention (COPD information, dose reminders and reinforcing audiovisual material, motivational aspects and inhalation technique training) to COPD patients taking inhaled treatment will give a 25% increase in the number of patients showing therapeutic adherence in this group compared to the control group. We will evaluate the effectiveness

  5. Study protocol: Improving patient choice in treating low back pain (IMPACT - LBP): A randomised controlled trial of a decision support package for use in physical therapy

    PubMed Central

    2011-01-01

    Background Low back pain is a common and costly condition. There are several treatment options for people suffering from back pain, but there are few data on how to improve patients' treatment choices. This study will test the effects of a decision support package (DSP), designed to help patients seeking care for back pain to make better, more informed choices about their treatment within a physiotherapy department. The package will be designed to assist both therapist and patient. Methods/Design Firstly, in collaboration with physiotherapists, patients and experts in the field of decision support and decision aids, we will develop the DSP. The work will include: a literature and evidence review; secondary analysis of existing qualitative data; exploration of patients' perspectives through focus groups and exploration of experts' perspectives using a nominal group technique and a Delphi study. Secondly, we will carry out a pilot single centre randomised controlled trial within NHS Coventry Community Physiotherapy. We will randomise physiotherapists to receive either training for the DSP or not. We will randomly allocate patients seeking treatment for non specific low back pain to either a physiotherapist trained in decision support or to receive usual care. Our primary outcome measure will be patient satisfaction with treatment at three month follow-up. We will also estimate the cost-effectiveness of the intervention, and assess the value of conducting further research. Discussion Informed shared decision-making should be an important part of any clinical consultation, particularly when there are several treatments, which potentially have moderate effects. The results of this pilot will help us determine the benefits of improving the decision-making process in clinical practice on patient satisfaction. Trial registration Current Controlled Trials ISRCTN46035546 PMID:21352528

  6. Cryptanalysis of the arbitrated quantum signature protocols

    SciTech Connect

    Gao Fei; Qin Sujuan; Guo Fenzhuo; Wen Qiaoyan

    2011-08-15

    As a new model for signing quantum messages, arbitrated quantum signature (AQS) has recently received a lot of attention. In this paper we study the cryptanalysis of previous AQS protocols from the aspects of forgery and disavowal. We show that in these protocols the receiver, Bob, can realize existential forgery of the sender's signature under known message attack. Bob can even achieve universal forgery when the protocols are used to sign a classical message. Furthermore, the sender, Alice, can successfully disavow any of her signatures by simple attack. The attack strategies are described in detail and some discussions about the potential improvements of the protocols are given. Finally we also present several interesting topics on AQS protocols that can be studied in future.

  7. Accuracy of NHANES periodontal examination protocols.

    PubMed

    Eke, P I; Thornton-Evans, G O; Wei, L; Borgnakke, W S; Dye, B A

    2010-11-01

    This study evaluates the accuracy of periodontitis prevalence determined by the National Health and Nutrition Examination Survey (NHANES) partial-mouth periodontal examination protocols. True periodontitis prevalence was determined in a new convenience sample of 454 adults ≥ 35 years old, by a full-mouth "gold standard" periodontal examination. This actual prevalence was compared with prevalence resulting from analysis of the data according to the protocols of NHANES III and NHANES 2001-2004, respectively. Both NHANES protocols substantially underestimated the prevalence of periodontitis by 50% or more, depending on the periodontitis case definition used, and thus performed below threshold levels for moderate-to-high levels of validity for surveillance. Adding measurements from lingual or interproximal sites to the NHANES 2001-2004 protocol did not improve the accuracy sufficiently to reach acceptable sensitivity thresholds. These findings suggest that NHANES protocols produce high levels of misclassification of periodontitis cases and thus have low validity for surveillance and research.

  8. Security Weaknesses in Arbitrated Quantum Signature Protocols

    NASA Astrophysics Data System (ADS)

    Liu, Feng; Zhang, Kejia; Cao, Tianqing

    2014-01-01

    Arbitrated quantum signature (AQS) is a cryptographic scenario in which the sender (signer), Alice, generates the signature of a message and then a receiver (verifier), Bob, can verify the signature with the help of a trusted arbitrator, Trent. In this paper, we point out there exist some security weaknesses in two AQS protocols. Our analysis shows Alice can successfully disavow any of her signatures by a simple attack in the first protocol. Furthermore, we study the security weaknesses of the second protocol from the aspects of forgery and disavowal. Some potential improvements of this kind of protocols are given. We also design a new method to authenticate a signature or a message, which makes AQS protocols immune to Alice's disavowal attack and Bob's forgery attack effectively.

  9. Cryptanalysis of the arbitrated quantum signature protocols

    NASA Astrophysics Data System (ADS)

    Gao, Fei; Qin, Su-Juan; Guo, Fen-Zhuo; Wen, Qiao-Yan

    2011-08-01

    As a new model for signing quantum messages, arbitrated quantum signature (AQS) has recently received a lot of attention. In this paper we study the cryptanalysis of previous AQS protocols from the aspects of forgery and disavowal. We show that in these protocols the receiver, Bob, can realize existential forgery of the sender's signature under known message attack. Bob can even achieve universal forgery when the protocols are used to sign a classical message. Furthermore, the sender, Alice, can successfully disavow any of her signatures by simple attack. The attack strategies are described in detail and some discussions about the potential improvements of the protocols are given. Finally we also present several interesting topics on AQS protocols that can be studied in future.

  10. Telehealth system (e-CUIDATE) to improve quality of life in breast cancer survivors: rationale and study protocol for a randomized clinical trial

    PubMed Central

    2013-01-01

    Background Breast cancer survivors suffer physical impairment after oncology treatment. This impairment reduces quality of life (QoL) and increase the prevalence of handicaps associated to unhealthy lifestyle (for example, decreased aerobic capacity and strength, weight gain, and fatigue). Recent work has shown that exercise adapted to individual characteristics of patients is related to improved overall and disease-free survival. Nowadays, technological support using telerehabilitation systems is a promising strategy with great advantage of a quick and efficient contact with the health professional. It is not known the role of telerehabilitation through therapeutic exercise as a support tool to implement an active lifestyle which has been shown as an effective resource to improve fitness and reduce musculoskeletal disorders of these women. Methods / Design This study will use a two-arm, assessor blinded, parallel randomized controlled trial design. People will be eligible if: their diagnosis is of stages I, II, or IIIA breast cancer; they are without chronic disease or orthopedic issues that would interfere with ability to participate in a physical activity program; they had access to the Internet and basic knowledge of computer use or living with a relative who has this knowledge; they had completed adjuvant therapy except for hormone therapy and not have a history of cancer recurrence; and they have an interest in improving lifestyle. Participants will be randomized into e-CUIDATE or usual care groups. E-CUIDATE give participants access to a range of contents: planning exercise arranged in series with breathing exercises, mobility, strength, and stretching. All of these exercises will be assigned to women in the telerehabilitation group according to perceived needs. The control group will be asked to maintain their usual routine. Study endpoints will be assessed after 8 weeks (immediate effects) and after 6 months. The primary outcome will be QoL measured by The

  11. Clinician-led improvement in cancer care (CLICC) - testing a multifaceted implementation strategy to increase evidence-based prostate cancer care: phased randomised controlled trial - study protocol

    PubMed Central

    2014-01-01

    Background Clinical practice guidelines have been widely developed and disseminated with the aim of improving healthcare processes and patient outcomes but the uptake of evidence-based practice remains haphazard. There is a need to develop effective implementation methods to achieve large-scale adoption of proven innovations and recommended care. Clinical networks are increasingly being viewed as a vehicle through which evidence-based care can be embedded into healthcare systems using a collegial approach to agree on and implement a range of strategies within hospitals. In Australia, the provision of evidence-based care for men with prostate cancer has been identified as a high priority. Clinical audits have shown that fewer than 10% of patients in New South Wales (NSW) Australia at high risk of recurrence after radical prostatectomy receive guideline recommended radiation treatment following surgery. This trial will test a clinical network-based intervention to improve uptake of guideline recommended care for men with high-risk prostate cancer. Methods/Design In Phase I, a phased randomised cluster trial will test a multifaceted intervention that harnesses the NSW Agency for Clinical Innovation (ACI) Urology Clinical Network to increase evidence-based care for men with high-risk prostate cancer following surgery. The intervention will be introduced in nine NSW hospitals over 10 months using a stepped wedge design. Outcome data (referral to radiation oncology for discussion of adjuvant radiotherapy in line with guideline recommended care or referral to a clinical trial of adjuvant versus salvage radiotherapy) will be collected through review of patient medical records. In Phase II, mixed methods will be used to identify mechanisms of provider and organisational change. Clinicians’ knowledge and attitudes will be assessed through surveys. Process outcome measures will be assessed through document review. Semi-structured interviews will be conducted to elucidate

  12. Microsatellite Instability Predicts Improved Response to Adjuvant Therapy With Irinotecan, Fluorouracil, and Leucovorin in Stage III Colon Cancer: Cancer and Leukemia Group B Protocol 89803

    PubMed Central

    Bertagnolli, Monica M.; Niedzwiecki, Donna; Compton, Carolyn C.; Hahn, Hejin P.; Hall, Margaret; Damas, Beatrice; Jewell, Scott D.; Mayer, Robert J.; Goldberg, Richard M.; Saltz, Leonard B.; Warren, Robert S.; Redston, Mark

    2009-01-01

    Purpose Colon cancers exhibiting DNA mismatch repair (MMR) defects demonstrate distinct clinical and pathologic features, including better prognosis and reduced response to fluorouracil (FU) –based chemotherapy. This prospective study investigated adjuvant chemotherapy containing FU and irinotecan in patients with MMR deficient (MMR-D) colon cancers. Patients and Methods Cancer and Leukemia Group B 89803 randomly assigned 1,264 patients with stage III colon cancer to postoperative weekly bolus FU/leucovorin (LV) or weekly bolus irinotecan, FU, and LV (IFL). The primary end point was overall survival; disease-free survival (DFS) was a secondary end point. Tumor expression of the MMR proteins, MLH1 and MSH2, was determined by immunohistochemistry (IHC). DNA microsatellite instability was also assessed using a panel of mono- and dinucleotide markers. Tumors with MMR defects were those demonstrating loss of MMR protein expression (MMR-D) and/or microsatellite instability high (MSI-H) genotype. Results Of 723 tumor cases examined by genotyping and IHC, 96 (13.3%) were MMR-D/MSI-H. Genotyping results were consistent with IHC in 702 cases (97.1%). IFL-treated patients with MMR-D/MSI-H tumors showed improved 5-year DFS as compared with those with mismatch repair intact tumors (0.76; 95% CI, 0.64 to 0.88 v 0.59; 95% CI, 0.53 to 0.64; P = .03). This relationship was not observed among patients treated with FU/LV. A trend toward longer DFS was observed in IFL-treated patients with MMR-D/MSI-H tumors as compared with those receiving FU/LV (0.57; 95% CI, 0.42 to 0.71 v 0.76; 95% CI, 0.64 to 0.88; P = .07; hazard ratio interaction between tumor status and treatment, 0.51; likelihood ratio P = .117). Conclusion Loss of tumor MMR function may predict improved outcome in patients treated with the IFL regimen as compared with those receiving FU/LV. PMID:19273709

  13. Effectiveness of a smart phone app on improving immunization of children in rural Sichuan Province, China: study protocol for a paired cluster randomized controlled trial

    PubMed Central

    2014-01-01

    Background Although good progress has been achieved in expanding immunization of children in China, disparities exist across different provinces. Information gaps both from the service supply and demand sides hinder timely vaccination of children in rural areas. The rapid development of mobile health technology (mHealth) provides unprecedented opportunities for improving health services and reaching underserved populations. However, there is a lack of literature that rigorously evaluates the impact of mHealth interventions on immunization coverage as well as the usability and feasibility of smart phone applications (apps). This study aims to assess the effectiveness of a smart phone-based app (Expanded Program on Immunization app, or EPI app) on improving the coverage of children’s immunization. Methods/Design This cluster randomized trial will take place in Xuanhan County, Sichuan Province, China. Functionalities of the app include the following: to make appointments automatically, record and update children’s immunization information, generate a list of children who missed their vaccination appointments, and send health education information to village doctors. After pairing, 36 villages will be randomly allocated to the intervention arm (n = 18) and control arm (n = 18). The village doctors in the intervention arm will use the app while the village doctors in the control arm will record and manage immunization in the usual way in their catchment areas. A household survey will be used at baseline and at endline (8 months of implementation). The primary outcome is full-dose coverage and the secondary outcome is immunization coverage of the five vaccines that are included in the national Expanded Program on Immunization program as well as Hib vaccine, Rotavirus vaccine and Pneumococcal conjugate vaccine. Multidimensional evaluation of the app will also be conducted to assess usability and feasibility. Discussion This study is the first to evaluate the

  14. Happy@Work: protocol for a web-based randomized controlled trial to improve mental well-being among an Asian working population

    PubMed Central

    2014-01-01

    Background Mental health issues pose a serious concern in the workplace for the huge productivity loss and financial burden associated with it. Unlike the traditional ‘fixing-what-is-wrong’ approach, positive psychology offers a less-stigmatized way to promote mental health. Psychological capital, a concept originated from positive psychology, has been proven effective in improving mental well-being and work performance. However, little evidence exists for its implementation among Asian working population or its cost-benefit for organizations adopting such promotion strategy. The current study is designed to assess the protective effects of a web-based psychology capital intervention among Hong Kong working population on individuals’ mental health and work performance, as well as organizations’ return-on-investment. Methods/Design A two-arm randomized controlled trial design will be adopted. Eligible working adults will be randomly allocated to either the intervention group or the waiting-list control group, with 177 participants in each arm. The intervention, which consists of four web-based training sessions, each targeting one of the psychological capital components (hope, efficacy, optimism and resilience), will be implemented over a 4-week period. On-line surveys will assess the participants in each group at baseline, intervention completion, 1 and 3 months after the completion. The primary outcome is individuals’ psychological capital level; secondary outcomes include individuals’ well-being, depressive symptoms, work engagement and productivity. Return-on-investment will be calculated from the employers’ perspective based on productivity gain, savings in medical expenditure, as well as operation and time costs. Analysis will follow the intention-to-treat principle. Discussion This is the first experimental study that explores the applicability of psychological capital development among Asian population. Through investigating changes in

  15. Encapsulation of carvacrol, a monoterpene present in the essential oil of oregano, with β-cyclodextrin, improves the pharmacological response on cancer pain experimental protocols.

    PubMed

    Guimarães, Adriana Gibara; Oliveira, Marlange Almeida; Alves, Rafael dos Santos; Menezes, Paula dos Passos; Serafini, Mairim Russo; Araújo, Adriano Antunes de Souza; Bezerra, Daniel Pereira; Quintans Júnior, Lucindo José

    2015-02-01

    Cancer pain is a major public health problem worldwide due to the strong impact on the quality of life of patients and side effects of the existing therapeutic options. Monoterpenes, as carvacrol (CARV), have been extensively studied about their therapeutic properties, especially their importance in the control of painful conditions and inflammation, which can be improved through the use of inclusion complexes of β-cyclodextrin (β-CD). We evaluated the effect of encapsulation of CARV in β-CD (CARV/β-CD) on the nociception induced by tumor cells (Sarcoma 180) in rodents. Inclusion complexes were prepared in two different procedures and characterized through thermal analysis and scanning electron microscopy. CARV/β-CD complex was administered (50 mg/kg, p.o.) in mice with tumor on the hind paw and was able to reduce the hyperalgesia (von Frey) during 24 h, unlike the free CARV (100 mg/kg, p.o.), which promoted effects until 9 h. Administration on alternate days of complex of CARV/β-CD (12.5-50 mg/kg, p.o.) reduced hyperalgesia, as well as spontaneous and palpation-induced nociception. However, pure CARV (50 mg/kg) did not cause significant changes in nociceptive responses. Together, these results produced evidence that the encapsulation of carvacrol in β-cyclodextrin can be useful for the development of new options for pain management. PMID:25557507

  16. Improving the care of people with traumatic brain injury through the Neurotrauma Evidence Translation (NET) program: protocol for a program of research.

    PubMed

    Green, Sally E; Bosch, Marije; McKenzie, Joanne E; O'Connor, Denise A; Tavender, Emma J; Bragge, Peter; Chau, Marisa; Pitt, Veronica; Rosenfeld, Jeffrey V; Gruen, Russell L

    2012-01-01

    The Neurotrauma Evidence Translation (NET) program was funded in 2009 to increase the uptake of research evidence in the clinical care of patients who have sustained traumatic brain injury. This paper reports the rationale and plan for this five-year knowledge translation research program. The overarching aims of the program are threefold: to improve outcomes for people with traumatic brain injury; to create a network of neurotrauma clinicians and researchers with expertise in knowledge translation and evidence-based practice; and to contribute knowledge to the field of knowledge translation research. The program comprises a series of interlinked projects spanning varying clinical environments and disciplines relevant to neurotrauma, anchored within four themes representing core knowledge translation activities: reviewing research evidence; understanding practice; developing and testing interventions for practice change; and building capacity for knowledge translation in neurotrauma. The program uses a range of different methods and study designs, including: an evidence fellowship program; conduct of and training in systematic reviews; mixed method study designs to describe and understand factors that influence current practices (e.g., semi-structured interviews and surveys); theory-based methods to develop targeted interventions aiming to change practice; a cluster randomised trial to test the effectiveness of a targeted theory-informed intervention; stakeholder involvement activities; and knowledge translation events such as consensus conferences.

  17. Study to Improve Cardiovascular Outcomes in high-risk older patieNts (ICON1) with acute coronary syndrome: study design and protocol of a prospective observational study

    PubMed Central

    Kunadian, Vijay; Neely, R Dermot G; Sinclair, Hannah; Batty, Jonathan A; Veerasamy, Murugapathy; Ford, Gary A; Qiu, Weiliang

    2016-01-01

    Introduction The ICON1 study (a study to Improve Cardiovascular Outcomes in high-risk older patieNts with acute coronary syndrome) is a prospective observational study of older patients (≥75 years old) with non-ST-elevation acute coronary syndrome managed by contemporary treatment (pharmacological and invasive). The aim of the study was to determine the predictors of poor cardiovascular outcomes in this age group and to generate a risk prediction tool. Methods and analysis Participants are recruited from 2 tertiary hospitals in the UK. Baseline evaluation includes frailty, comorbidity, cognition and quality-of-life measures, inflammatory status assessed by a biomarker panel, including microRNAs, senescence assessed by telomere length and telomerase activity, cardiovascular status assessed by arterial stiffness, endothelial function, carotid intima media thickness and left ventricular systolic and diastolic function, and coronary plaque assessed by virtual histology intravascular ultrasound and optical coherence tomography. The patients are followed-up at 30 days and at 1 year for primary outcome measures of death, myocardial infarction, stroke, unplanned revascularisation, bleeding and rehospitalisation. Ethics and dissemination The study has been approved by the regional ethics committee (REC 12/NE/016). Findings of the study will be presented in scientific sessions and will be published in peer-reviewed journals. Trial registration number NCT01933581: Pre-results. PMID:27554105

  18. Improving the care of people with traumatic brain injury through the Neurotrauma Evidence Translation (NET) program: protocol for a program of research.

    PubMed

    Green, Sally E; Bosch, Marije; McKenzie, Joanne E; O'Connor, Denise A; Tavender, Emma J; Bragge, Peter; Chau, Marisa; Pitt, Veronica; Rosenfeld, Jeffrey V; Gruen, Russell L

    2012-01-01

    The Neurotrauma Evidence Translation (NET) program was funded in 2009 to increase the uptake of research evidence in the clinical care of patients who have sustained traumatic brain injury. This paper reports the rationale and plan for this five-year knowledge translation research program. The overarching aims of the program are threefold: to improve outcomes for people with traumatic brain injury; to create a network of neurotrauma clinicians and researchers with expertise in knowledge translation and evidence-based practice; and to contribute knowledge to the field of knowledge translation research. The program comprises a series of interlinked projects spanning varying clinical environments and disciplines relevant to neurotrauma, anchored within four themes representing core knowledge translation activities: reviewing research evidence; understanding practice; developing and testing interventions for practice change; and building capacity for knowledge translation in neurotrauma. The program uses a range of different methods and study designs, including: an evidence fellowship program; conduct of and training in systematic reviews; mixed method study designs to describe and understand factors that influence current practices (e.g., semi-structured interviews and surveys); theory-based methods to develop targeted interventions aiming to change practice; a cluster randomised trial to test the effectiveness of a targeted theory-informed intervention; stakeholder involvement activities; and knowledge translation events such as consensus conferences. PMID:22866892

  19. Encapsulation of carvacrol, a monoterpene present in the essential oil of oregano, with β-cyclodextrin, improves the pharmacological response on cancer pain experimental protocols.

    PubMed

    Guimarães, Adriana Gibara; Oliveira, Marlange Almeida; Alves, Rafael dos Santos; Menezes, Paula dos Passos; Serafini, Mairim Russo; Araújo, Adriano Antunes de Souza; Bezerra, Daniel Pereira; Quintans Júnior, Lucindo José

    2015-02-01

    Cancer pain is a major public health problem worldwide due to the strong impact on the quality of life of patients and side effects of the existing therapeutic options. Monoterpenes, as carvacrol (CARV), have been extensively studied about their therapeutic properties, especially their importance in the control of painful conditions and inflammation, which can be improved through the use of inclusion complexes of β-cyclodextrin (β-CD). We evaluated the effect of encapsulation of CARV in β-CD (CARV/β-CD) on the nociception induced by tumor cells (Sarcoma 180) in rodents. Inclusion complexes were prepared in two different procedures and characterized through thermal analysis and scanning electron microscopy. CARV/β-CD complex was administered (50 mg/kg, p.o.) in mice with tumor on the hind paw and was able to reduce the hyperalgesia (von Frey) during 24 h, unlike the free CARV (100 mg/kg, p.o.), which promoted effects until 9 h. Administration on alternate days of complex of CARV/β-CD (12.5-50 mg/kg, p.o.) reduced hyperalgesia, as well as spontaneous and palpation-induced nociception. However, pure CARV (50 mg/kg) did not cause significant changes in nociceptive responses. Together, these results produced evidence that the encapsulation of carvacrol in β-cyclodextrin can be useful for the development of new options for pain management.

  20. Seven consecutive successful clinical islet isolations with pancreatic ductal injection.

    PubMed

    Matsumoto, Shinichi; Noguichi, Hirofumi; Shimoda, Masayuki; Ikemoto, Tetsuya; Naziruddin, Bashoo; Jackson, Andrew; Tamura, Yoshiko; Olson, Greg; Fujita, Yasutaka; Chujo, Daisuke; Takita, Morihito; Kobayashi, Naoya; Onaca, Nicholas; Levy, Marlon

    2010-01-01

    Inconsistent islet isolation is one of the issues of clinical islet transplantation. In the current study, we applied ductal injection to improve the consistency of islet isolation. Seven islet isolations were performed with the ductal injection of ET-Kyoto solution (DI group) and eight islet isolations were performed without the ductal injection (standard group) using brain-dead donor pancreata. Isolated islets were evaluated based on the Edmonton protocol for transplantation. The DI group had significantly higher islet yields (588,566 +/- 64,319 vs. 354,836 +/- 89,649 IE, p < 0.01) and viability (97.3 +/- 1.2% vs. 92.6 +/- 1.2%, p < 0.02) compared with the standard group. All seven isolated islet preparations in the DI group (100%), versus only three out of eight isolated islet preparations (38%) in the standard group met transplantation criteria. The islets from the DI group were transplanted into three type 1 diabetic patients and all three patients became insulin independent. Ductal injection significantly improved quantity and quality of isolated islets and resulted in high success rate of clinical islet transplantation. This simple modification will reduce the risk of failure of clinical islet isolation.

  1. A lower content of de-methylesterified homogalacturonan improves enzymatic cell separation and isolation of mesophyll protoplasts in Arabidopsis.

    PubMed

    Lionetti, Vincenzo; Cervone, Felice; De Lorenzo, Giulia

    2015-04-01

    Cell adhesion occurs primarily at the level of middle lamella which is mainly composed by pectin polysaccharides. These can be degraded by cell wall degrading enzymes (CWDEs) during developmental processes to allow a controlled separation of plant cells. Extensive cell wall degradation by CWDEs with consequent cell separation is performed when protoplasts are isolated from plant tissues by using mixtures of CWDEs. We have evaluated whether modification of pectin affects cell separation and protoplast isolation. Arabidopsis plants overexpressing the pectin methylesterase inhibitors AtPMEI-1 or AtPMEI-2, and Arabidopsis pme3 plants, mutated in the gene encoding pectin methylesterase 3, showed an increased efficiency of isolation of viable mesophyll protoplasts as compared with Wild Type Columbia-0 plants. The release of protoplasts was correlated with the reduced level of long stretches of de-methylesterified homogalacturonan (HGA) present in these plants. Response to elicitation, cell wall regeneration and efficiency of transfection in protoplasts from transgenic plants was comparable to those of wild type protoplasts.

  2. National Sample Assessment Protocols

    ERIC Educational Resources Information Center

    Ministerial Council on Education, Employment, Training and Youth Affairs (NJ1), 2012

    2012-01-01

    These protocols represent a working guide for planning and implementing national sample assessments in connection with the national Key Performance Measures (KPMs). The protocols are intended for agencies involved in planning or conducting national sample assessments and personnel responsible for administering associated tenders or contracts,…

  3. Pre-consultation educational group intervention to improve shared decision-making in postmastectomy breast reconstruction: study protocol for a pilot randomized controlled trial

    PubMed Central

    2013-01-01

    Background The Pre-Consultation Educational Group Intervention pilot study seeks to assess the feasibility and inform the optimal design for a definitive randomized controlled trial that aims to improve the quality of decision-making in postmastectomy breast reconstruction patients. Methods/design This is a mixed-methods pilot feasibility randomized controlled trial that will follow a single-center, 1:1 allocation, two-arm parallel group superiority design. Setting: The University Health Network, a tertiary care cancer center in Toronto, Canada. Participants: Adult women referred to one of three plastic and reconstructive surgeons for delayed breast reconstruction or prophylactic mastectomy with immediate breast reconstruction. Intervention: We designed a multi-disciplinary educational group workshop that incorporates the key components of shared decision-making, decision-support, and psychosocial support for cancer survivors prior to the initial surgical consult. The intervention consists of didactic lectures by a plastic surgeon and nurse specialist on breast reconstruction choices, pre- and postoperative care; a value-clarification exercise led by a social worker; and discussions with a breast reconstruction patient. Control: Usual care includes access to an informational booklet, website, and patient volunteer if desired. Outcomes: Expected pilot outcomes include feasibility, recruitment, and retention targets. Acceptability of intervention and full trial outcomes will be established through qualitative interviews. Trial outcomes will include decision-quality measures, patient-reported outcomes, and service outcomes, and the treatment effect estimate and variability will be used to inform the sample size calculation for a full trial. Discussion Our pilot study seeks to identify the (1) feasibility, acceptability, and design of a definitive RCT and (2) the optimal content and delivery of our proposed educational group intervention. Thirty patients have been

  4. Effect of stacking interactions on the thermodynamics and kinetics of lumiflavin: a study with improved density functionals and density functional tight-binding protocol.

    PubMed

    Bresnahan, Caitlin G; Reinhardt, Clorice R; Bartholow, Thomas G; Rumpel, John P; North, Michael; Bhattacharyya, Sudeep

    2015-01-01

    The π-π stacking interaction between lumiflavin and a number of π-electron-rich molecules has been studied by density functional theory using several new-generation density functionals. Six known lumiflavin-aromatic adducts were used and the models were evaluated by comparing the geometry and energetics with experimental results. The study found that dispersion-corrected and hybrid functionals with larger (>50%) Hartree-Fock exchanges produced superior results in modeling thermodynamic characteristics of these complexes. The functional producing the best energetics for these model systems was used to study the stacking interactions of lumiflavin with biologically relevant aromatic groups. Additionally, the reduction of flavin-in the presence of both a hydride donor and a nondonor π-electronic system was also studied. Weak interactions were observed in the stacked lumiflavin complexes of benzene, phenol, and indole, mimicking phenyl alanine, tryptophan, and tyrosine side chains, respectively, of an enzyme. The stacked complex of naphthalene and flavin showed little change in flavin's redox potential indicating insignificant effect on the thermodynamics of the hydride transfer reaction. In contrast, the hydride transfer reaction with the hydride donor N-methyl nicotinamide tells a different story, as the transition state was found to be strongly impacted by the stacking interactions. A comparison of performance between the density functional theory (DFT) and the computationally less expensive dispersion-corrected self-consistent density functional tight-binding (SCC-DFTB-D) theory revealed that the latter produces consistent energetics for this hydride transfer reaction and additional DFT-computed perturbative corrections could significantly improve these results.

  5. Improving adolescent mental health and resilience through a resilience-based intervention in schools: study protocol for a randomised controlled trial

    PubMed Central

    2014-01-01

    Background Research investigating the effectiveness of universal interventions to reduce the risk of mental health problems remains limited. Schools are a promising setting within which adolescents can receive interventions aimed at promoting their mental health. The aim of this study is to assess the effectiveness of a resilience-based prevention-focused intervention in reducing the risk of mental health problems among adolescents attending secondary school in socio-economically disadvantaged areas. Methods/design A cluster randomised control trial will be conducted, with schools as the unit of randomisation. Initially, 32 secondary schools will be randomly allocated to a control or intervention group (12 control and 20 intervention). An intervention focused on improving student internal and external resilience factors will be implemented in intervention schools. A survey of students in Grade 7 in both intervention and control schools will be conducted (baseline) and repeated three years later when the students are in Grade 10. The Strengths and Difficulties Questionnaire will be used to measure the risk of mental health problems. At follow-up, the risk of mental health problems will be compared between Grade 10 students in intervention and control schools to determine intervention effectiveness. Discussion The study presents an opportunity to determine the effectiveness of a comprehensive resilience-based intervention in reducing the risk of mental health problems in adolescents attending secondary schools. The outcomes of the trial are of importance to youth, schools, mental health clinicians and policymakers. Trial registration Australian New Zealand Clinical Trials Registry, ACTRN12611000606987, registered 14 June 2011. PMID:25037455

  6. A cost-effectiveness analysis of provider and community interventions to improve the treatment of uncomplicated malaria in Nigeria: study protocol for a randomized controlled trial

    PubMed Central

    2012-01-01

    Background There is mounting evidence of poor adherence by health service personnel to clinical guidelines for malaria following a symptomatic diagnosis. In response to this, the World Health Organization (WHO) recommends that in all settings clinical suspicion of malaria should be confirmed by parasitological diagnosis using microscopy or Rapid Diagnostic Test (RDT). The Government of Nigeria plans to introduce RDTs in public health facilities over the coming year. In this context, we will evaluate the effectiveness and cost-effectiveness of two interventions designed to support the roll-out of RDTs and improve the rational use of ACTs. It is feared that without supporting interventions, non-adherence will remain a serious impediment to implementing malaria treatment guidelines. Methods/design A three-arm stratified cluster randomized trial is used to compare the effectiveness and cost-effectiveness of: (1) provider malaria training intervention versus expected standard practice in malaria diagnosis and treatment; (2) provider malaria training intervention plus school-based intervention versus expected standard practice; and (3) the combined provider plus school-based intervention versus provider intervention alone. RDTs will be introduced in all arms of the trial. The primary outcome is the proportion of patients attending facilities that report a fever or suspected malaria and receive treatment according to malaria guidelines. This will be measured by surveying patients (or caregivers) as they exit primary health centers, pharmacies, and patent medicine dealers. Cost-effectiveness will be presented in terms of the primary outcome and a range of secondary outcomes, including changes in provider and community knowledge. Costs will be estimated from both a societal and provider perspective using standard economic evaluation methodologies. Trial registration Clinicaltrials.gov NCT01350752 PMID:22682276

  7. Montreal protocol: Business opportunites

    SciTech Connect

    1998-12-31

    The Montreal Protocol on Substances that Deplete the Ozone Layer was signed by 24 countries in 1987, establishing measures for controlling the production and consumption of ozone-depleting substances. This publication begins with some background information on ozone depletion and the history of the Protocol. It then describes aspects of the Protocol`s Multilateral Fund, created to assist developing countries to meet Protocol deadlines: Its administration, structure, and how projects are initiated. Names, addresses, and phone/fax numbers of Fund contacts are provided. Canadian projects under the Fund are then reviewed and opportunities for Canadian environmental companies are noted. Finally, information sheets are presented which summarize Fund-related Canadian bilateral projects undertaken to date.

  8. Reliable broadcast protocols

    NASA Technical Reports Server (NTRS)

    Joseph, T. A.; Birman, Kenneth P.

    1989-01-01

    A number of broadcast protocols that are reliable subject to a variety of ordering and delivery guarantees are considered. Developing applications that are distributed over a number of sites and/or must tolerate the failures of some of them becomes a considerably simpler task when such protocols are available for communication. Without such protocols the kinds of distributed applications that can reasonably be built will have a very limited scope. As the trend towards distribution and decentralization continues, it will not be surprising if reliable broadcast protocols have the same role in distributed operating systems of the future that message passing mechanisms have in the operating systems of today. On the other hand, the problems of engineering such a system remain large. For example, deciding which protocol is the most appropriate to use in a certain situation or how to balance the latency-communication-storage costs is not an easy question.

  9. Lay health supporters aided by a mobile phone messaging system to improve care of villagers with schizophrenia in Liuyang, China: protocol for a randomised control trial

    PubMed Central

    Gong, Wenjie; Caine, Eric D; Xiao, Shuiyuan; Hughes, James P; Ng, Marie; Simoni, Jane; He, Hua; Smith, Kirk L; Brown, Henry Shelton; Gloyd, Stephen

    2016-01-01

    Introduction Schizophrenia is a severe, chronic and disabling mental illness. Non-adherence to medication and relapse may lead to poorer patient function. This randomised controlled study, under the acronym LEAN (Lay health supporter, e-platform, award, and iNtegration), is designed to improve medication adherence and high relapse among people with schizophrenia in resource poor settings. Methods/analysis The community-based LEAN has four parts: (1) Lay health supporters (LHSs), mostly family members who will help supervise patient medication, monitor relapse and side effects, and facilitate access to care, (2) an E-platform to support two-way mobile text and voice messaging to remind patients to take medication; and alert LHSs when patients are non-adherent, (3) an Award system to motivate patients and strengthen LHS support, and (4) iNtegration of the efforts of patients and LHSs with those of village doctors, township mental health administrators and psychiatrists via the e-platform. A random sample of 258 villagers with schizophrenia will be drawn from the schizophrenic ‘686’ Program registry for the 9 Xiang dialect towns of the Liuyang municipality in China. The sample will be further randomised into a control group and a treatment group of equal sizes, and each group will be followed for 6 months after launch of the intervention. The primary outcome will be medication adherence as measured by pill counts and supplemented by pharmacy records. Other outcomes include symptoms and level of function. Outcomes will be assessed primarily when patients present for medication refill visits scheduled every 2 months over the 6-month follow-up period. Data from the study will be analysed using analysis of covariance for the programme effect and an intent-to-treat approach. Ethics and dissemination University of Washington: 49464 G; Central South University: CTXY-150002-6. Results will be published in peer-reviewed journals with deidentified data made available on

  10. SMS reminders to improve the tuberculosis cure rate in developing countries (TB-SMS Cameroon): a protocol of a randomised control study

    PubMed Central

    2014-01-01

    . This study explores the potential of mHealth to support DOTS strategy. It will gather new evidence on the effectiveness of mHealth-based interventions and SMS reminders in the improvement of treatment adherence and the cure rate of tuberculosis patients, especially in a low-resource country such as Cameroon. Trial registration The trial is registered on the Pan-African Clinical Trials Registry (http://www.pactr.org) under unique identification number: PACTR201307000583416. PMID:24460827

  11. ‘Placement budgets’ for supported employment – improving competitive employment for people with mental illness: study protocol of a multicentre randomized controlled trial

    PubMed Central

    2012-01-01

    Background Vocational integration of people with mental illness is poor despite their willingness to work. The ‘Individual Placement and Support’ (IPS) model which emphasises rapid and direct job placement and continuing support to patient and employer has proven to be the most effective vocational intervention programme. Various studies have shown that every second patient with severe mental illness was able to find competitive employment within 18 months. However, the goal of taking up employment within two months was rarely achieved. Thus, we aim to test whether the new concept of limited placement budgets increases the effectiveness of IPS. Methods/Design Six job coaches in six out-patients psychiatric clinics in the Canton of Zurich support unemployed patients of their clinic who seek competitive employment. Between June 2010 and May 2011 patients (N=100) are randomly assigned to three different placement budgets of 25h, 40h, or 55h working hours of job coaches. Support lasts two years for those who find a job. The intervention ends for those who fail to find competitive employment when the respective placement budgets run out. The primary outcome measure is the time between study inclusion and first competitive employment that lasted three months or longer. Over a period of three years interviews are carried out every six months to measure changes in motivation, stigmatization, social network and social support, quality of life, job satisfaction, financial situation, and health conditions. Cognitive and social-cognitive tests are conducted at baseline to control for confounding variables. Discussion This study will show whether the effectiveness of IPS can be increased by the new concept of limited placement budgets. It will also be examined whether competitive employment leads in the long term to an improvement of mental illness, to a transfer of the psychiatric support system to private and vocational networks, to an increase in financial independence

  12. Improving the care for people with acute low-back pain by allied health professionals (the ALIGN trial): A cluster randomised trial protocol

    PubMed Central

    2010-01-01

    Background Variability between clinical practice guideline recommendations and actual clinical practice exists in many areas of health care. A 2004 systematic review examining the effectiveness of guideline implementation interventions concluded there was a lack of evidence to support decisions about effective interventions to promote the uptake of guidelines. Further, the review recommended the use of theory in the development of implementation interventions. A clinical practice guideline for the management of acute low-back pain has been developed in Australia (2003). Acute low-back pain is a common condition, has a high burden, and there is some indication of an evidence-practice gap in the allied health setting. This provides an opportunity to develop and test a theory-based implementation intervention which, if effective, may provide benefits for patients with this condition. Aims This study aims to estimate the effectiveness of a theory-based intervention to increase allied health practitioners' (physiotherapists and chiropractors in Victoria, Australia) compliance with a clinical practice guideline for acute non-specific low back pain (LBP), compared with providing practitioners with a printed copy of the guideline. Specifically, our primary objectives are to establish if the intervention is effective in reducing the percentage of acute non-specific LBP patients who are either referred for or receive an x-ray, and improving mean level of disability for patients three months post-onset of acute LBP. Methods The design of the study is a cluster randomised trial. Restricted randomisation was used to randomise 210 practices (clusters) to an intervention or control group. Practitioners in the control group received a printed copy of the guideline. Practitioners in the intervention group received a theory-based intervention developed to address prospectively identified barriers to practitioner compliance with the guideline. The intervention primarily consisted of

  13. Physiotherapy to improve physical activity in community-dwelling older adults with mobility problems (Coach2Move): study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background Older adults can benefit from physical activity in numerous ways. Physical activity is considered to be one of the few ways to influence the level of frailty. Standardized exercise programs do not necessarily lead to more physical activity in daily life, however, and a more personalized approach seems appropriate. The main objective of this study is to investigate whether a focused, problem-oriented coaching intervention (‘Coach2Move’) delivered by a physiotherapist specializing in geriatrics is more effective for improving physical activity, mobility and health status in community-dwelling older adults than usual physiotherapy care. In addition, cost-effectiveness will be determined. Methods/Design The design of this study is a single-blind randomized controlled trial in thirteen physiotherapy practices. Randomization will take place at the individual patient level. The study population consists of older adults, ≥70 years of age, with decreased physical functioning and mobility and/or a physically inactive lifestyle. The intervention group will receive geriatric physiotherapy according to the Coach2Move strategy. The control group will receive the usual physiotherapy care. Measurements will be performed by research assistants not aware of group assignment. The results will be evaluated on the amount of physical activity (LASA Physical Activity Questionnaire), mobility (modified ‘get up and go’ test, walking speed and six-minute walking test), quality of life (SF-36), degree of frailty (Evaluative Frailty Index for Physical Activity), fatigue (NRS-fatigue), perceived effect (Global Perceived Effect and Patient Specific Complaints questionnaire) and health care costs. Discussion Most studies on the effect of exercise or physical activity consist of standardized programs. In this study, a personalized approach is evaluated within a group of frail older adults, many of whom suffer from multiple and complex diseases and problems. A complicating

  14. A stepped wedge, cluster controlled trial of an intervention to improve safety and quality on medical wards: the HEADS-UP study protocol

    PubMed Central

    Pannick, Samuel; Beveridge, Iain; Ashrafian, Hutan; Long, Susannah J; Athanasiou, Thanos; Sevdalis, Nick

    2015-01-01

    Introduction The majority of preventable deaths in healthcare are due to errors on general wards. Staff perceptions of safety correlate with patient survival, but effectively translating ward teams’ concerns into tangibly improved care remains problematic. The Hospital Event Analysis Describing Significant Unanticipated Problems (HEADS-UP) trial evaluates a structured, multidisciplinary team briefing, capturing safety threats and adverse events, with rapid feedback to clinicians and service managers. This is the first study to rigorously assess a simpler intervention for general medical units, alongside an implementation model applicable to routine clinical practice. Methods/analysis 7 wards from 2 hospitals will progressively incorporate the intervention into daily practice over 14 months. Wards will adopt HEADS-UP in a pragmatic sequence, guided by local clinical enthusiasm. Initial implementation will be facilitated by a research lead, but rapidly delegated to clinical teams. The primary outcome is excess length of stay (a surplus stay of 24 h or more, compared to peer institutions’ Healthcare Resource Groups-predicted length of stay). Secondary outcomes are 30-day readmission or excess length of stay; in-hospital death or death/readmission within 30 days; healthcare-acquired infections; processes of escalation of care; use of traditional incident-reporting systems; and patient safety and teamwork climates. HEADS-UP will be analysed as a stepped wedge cluster controlled trial. With 7840 patients, using best and worst case predictions, the study would achieve between 75% and 100% power to detect a 2–14% absolute risk reduction in excess length of stay (two-sided p<0.05). Regression analysis will use generalised linear mixed models or generalised estimating equations, and a time-to-event regression model. A qualitative analysis will evaluate facilitators and barriers to HEADS-UP implementation and impact. Ethics and dissemination Participating

  15. Snakebite management in Iran: Devising a protocol

    PubMed Central

    Monzavi, Seyed Mostafa; Dadpour, Bita; Afshari, Reza

    2014-01-01

    Background: Snakebite in Iran has been a health concern. However, management of snakebite is not standardized and varies from center to center. This study is aimed at devising an evidence-based comprehensive protocol for snakebite management in Iran, to reduce unnecessary variations in practice. Materials and Methods: A narrative search in electronic databases was performed. Fifty peer-reviewed articles, guidelines, and textbooks were reviewed and practical details were extracted. Our currently used protocol in the Mashhad Toxicology Center was supplemented with this information. Consequently an improved wide-range protocol was developed. The protocol was then discussed and amended within a focus group comprised of medical toxicologists and internal medicine specialists. The amended version was finally discussed with expert physicians specialized in different areas of medicine, to be optimized by supplementing other specific considerations. Results: During a one-year process, the protocol was finalized. The final version of the protocol, which was designed in six steps, comprised of three components: A schematic algorithm, a severity grading scale, and instructions for supportive and adjunctive treatments. The algorithm pertains to both Viperidae and Elapidae snakebite envenomations and consists of a planned course of action and dosing of antivenom, based on the severity of the envenomation. Conclusion: Snakebite envenomation is a clinical toxicologic emergency, which needs to be treated in a timely and organized manner. Hence, a multi-aspect protocol was designed to improve the clinical outcomes, reduce unnecessary administration of antivenom, and help physicians make more proper clinical judgments. PMID:24778670

  16. Protocol to Exploit Waiting Resources for UASNs.

    PubMed

    Hung, Li-Ling; Luo, Yung-Jeng

    2016-01-01

    The transmission speed of acoustic waves in water is much slower than that of radio waves in terrestrial wireless sensor networks. Thus, the propagation delay in underwater acoustic sensor networks (UASN) is much greater. Longer propagation delay leads to complicated communication and collision problems. To solve collision problems, some studies have proposed waiting mechanisms; however, long waiting mechanisms result in low bandwidth utilization. To improve throughput, this study proposes a slotted medium access control protocol to enhance bandwidth utilization in UASNs. The proposed mechanism increases communication by exploiting temporal and spatial resources that are typically idle in order to protect communication against interference. By reducing wait time, network performance and energy consumption can be improved. A performance evaluation demonstrates that when the data packets are large or sensor deployment is dense, the energy consumption of proposed protocol is less than that of existing protocols as well as the throughput is higher than that of existing protocols.

  17. Protocol to Exploit Waiting Resources for UASNs.

    PubMed

    Hung, Li-Ling; Luo, Yung-Jeng

    2016-01-01

    The transmission speed of acoustic waves in water is much slower than that of radio waves in terrestrial wireless sensor networks. Thus, the propagation delay in underwater acoustic sensor networks (UASN) is much greater. Longer propagation delay leads to complicated communication and collision problems. To solve collision problems, some studies have proposed waiting mechanisms; however, long waiting mechanisms result in low bandwidth utilization. To improve throughput, this study proposes a slotted medium access control protocol to enhance bandwidth utilization in UASNs. The proposed mechanism increases communication by exploiting temporal and spatial resources that are typically idle in order to protect communication against interference. By reducing wait time, network performance and energy consumption can be improved. A performance evaluation demonstrates that when the data packets are large or sensor deployment is dense, the energy consumption of proposed protocol is less than that of existing protocols as well as the throughput is higher than that of existing protocols. PMID:27005624

  18. Quantum key distribution protocol using random bases

    NASA Astrophysics Data System (ADS)

    Meslouhi, A.; Amellal, H.; Hassouni, Y.; El Baz, M.; El Allati, A.

    2016-04-01

    In order to enhance the quantum key distribution (QKD) security, a new protocol, “QKDPRB” based on random bases is proposed. It consists of using standard encoding bases moving circularly with a variable rotational angle α which depends on angular velocity ω(t); thus, the traditional bases turn into relative ones. To prove the security and the efficiency of the protocol, we present a universal demonstration which proves a high level security of the proposed protocol, even in the presence of the intercept and resend attack. Finally, the QKDPRB may improve the security of QKD.

  19. Isolation of ribosomes and polysomes.

    PubMed

    Rivera, Maria C; Maguire, Bruce; Lake, James A

    2015-03-01

    Here we describe a preparative differential centrifugation protocol for the isolation of ribosomes from a crude cell homogenate. The subcellular fraction obtained is enriched in ribosome monomers and polysomes. The protocol has been optimized for the homogenization and collection of the ribosomal fraction from prokaryotic cells, mammalian and plant tissues, reticulocytes, and chloroplasts. The quality of the ribosomal preparation is enhanced by the removal of the remaining cellular components and adsorbed proteins by pelleting through a sucrose cushion with a high concentration of monovalent salts, NH4Cl or KCl. The different components of the ribosomal fraction isolated using this protocol can be further purified by sucrose gradient centrifugation.

  20. Reliable multicast protocol specifications protocol operations

    NASA Technical Reports Server (NTRS)

    Callahan, John R.; Montgomery, Todd; Whetten, Brian

    1995-01-01

    This appendix contains the complete state tables for Reliable Multicast Protocol (RMP) Normal Operation, Multi-RPC Extensions, Membership Change Extensions, and Reformation Extensions. First the event types are presented. Afterwards, each RMP operation state, normal and extended, is presented individually and its events shown. Events in the RMP specification are one of several things: (1) arriving packets, (2) expired alarms, (3) user events, (4) exceptional conditions.

  1. An improved method of DNA isolation suitable for PCR-based detection of begomoviruses from jute and other mucilaginous plants.

    PubMed

    Ghosh, Raju; Paul, Sujay; Ghosh, Subrata Kumar; Roy, Anirban

    2009-07-01

    A relatively quick and inexpensive modified cetyl trimethylammonium bromide method for extraction of DNA from leaf materials containing large quantities of mucilage is described. The modification including use of more volume of extraction buffer and dissolving crude nucleic acid pellet in 1 M NaCl, reduced markedly the viscosity of the mucilage and thus in the final purification step yielded a larger quantity of mucilage-free DNA suitable for subsequent PCR-based detection of begomoviruses. The method was standardized with jute samples with yellow mosaic disease and validated with different other mucilaginous-hosts with low titre of begomoviruses. DNA isolated using this method showed consistency in yield and compatibility with PCR for detection of begomoviruses from different mucilaginous plant species. The method was compared for efficacy with other reported methods and it was found to be superior over the existing methods described for isolation of DNA from mucilaginous hosts. Thus the method described could be used on a wider scale for reliable and consistent detection of begomoviruses from mucilaginous hosts for characterization and variability study.

  2. Odor-enriched environment rescues long-term social memory, but does not improve olfaction in social isolated adult mice.

    PubMed

    Gusmão, Isabela D; Monteiro, Brisa M M; Cornélio, Guilherme O S; Fonseca, Cristina S; Moraes, Márcio F D; Pereira, Grace S

    2012-03-17

    Prolonged permanence of animals under social isolation (SI) arouses a variety of psychological symptoms like aggression, stress, anxiety and depression. However, short-term SI is commonly used to evaluate social memory. Interestingly, the social memory cannot be accessed with delays higher than 30min in SI mice. Our hypothesis is that SI with intermediate duration, like one week (1w), impairs the long-term storage of new social information (S-LTM), without affecting anxiety or other types of memories, because the SI compromises the olfactory function of the animal. Our results demonstrated that SI impaired S-LTM, without affecting other kinds of memory or anxiety. In addition, the SI increased the latency in the buried-food finding task, but did not affect the habituation or the discrimination of odors. Next, we postulated that if continuous input to the olfactory system is fundamental for the maintenance of the olfactory function and social memory persistence, isolated mice under odor-enriched environment (OEE) should behave like group-housed (GH) animals. In fact, the OEE prevented the S-LTM deficit imposed by the SI. However, OEE did not restore the SI mice olfaction to the GH mice level. Our results suggest that SI modulates olfaction and social memory persistence, probably, by independent mechanisms. We also showed for the first time that OEE rescued S-LTM in SI mice through a mechanism not necessarily involved with olfaction.

  3. An improved method of DNA isolation suitable for PCR-based detection of begomoviruses from jute and other mucilaginous plants.

    PubMed

    Ghosh, Raju; Paul, Sujay; Ghosh, Subrata Kumar; Roy, Anirban

    2009-07-01

    A relatively quick and inexpensive modified cetyl trimethylammonium bromide method for extraction of DNA from leaf materials containing large quantities of mucilage is described. The modification including use of more volume of extraction buffer and dissolving crude nucleic acid pellet in 1 M NaCl, reduced markedly the viscosity of the mucilage and thus in the final purification step yielded a larger quantity of mucilage-free DNA suitable for subsequent PCR-based detection of begomoviruses. The method was standardized with jute samples with yellow mosaic disease and validated with different other mucilaginous-hosts with low titre of begomoviruses. DNA isolated using this method showed consistency in yield and compatibility with PCR for detection of begomoviruses from different mucilaginous plant species. The method was compared for efficacy with other reported methods and it was found to be superior over the existing methods described for isolation of DNA from mucilaginous hosts. Thus the method described could be used on a wider scale for reliable and consistent detection of begomoviruses from mucilaginous hosts for characterization and variability study. PMID:19442842

  4. Improved isolation and purification of functional human Fas receptor extracellular domain using baculovirus-silkworm expression system.

    PubMed

    Muraki, Michiro; Honda, Shinya

    2011-11-01

    To achieve an efficient isolation of human Fas receptor extracellular domain (hFasRECD), a fusion protein of hFasRECD with human IgG1 heavy chain Fc domain containing thrombin cleavage sequence at the junction site was overexpressed using baculovirus-silkworm larvae expression system. The hFasRECD part was separated from the fusion protein by the effective cleavage of the recognition site with bovine thrombin. Protein G column treatment of the reaction mixture and the subsequent cation-exchange chromatography provided purified hFasRECD with a final yield of 13.5mg from 25.0 ml silkworm hemolymph. The functional activity of the product was examined by size-exclusion chromatography analysis. The isolated hFasRECD less strongly interacted with human Fas ligand extracellular domain (hFasLECD) than the Fc domain-bridged counterpart, showing the contribution of antibody-like avidity in the latter case. The purified glycosylated hFasRECD presented several discrete bands in the disulphide-bridge non-reducing SDS-PAGE analysis, and virtually all of the components were considered to participate in the binding to hFasLECD. The attached glycans were susceptible to PNGase F digestion, but mostly resistant to Endo Hf digestion under denaturing conditions. One of the components exhibited a higher susceptibility to PNGase F digestion under non-denaturing conditions.

  5. Potential anesthesia protocols for space exploration missions.

    PubMed

    Komorowski, Matthieu; Watkins, Sharmila D; Lebuffe, Gilles; Clark, Jonathan B

    2013-03-01

    In spaceflight beyond low Earth's orbit, medical conditions requiring surgery are of a high level of concern because of their potential impact on crew health and mission success. Whereas surgical techniques have been thoroughly studied in spaceflight analogues, the research focusing on anesthesia is limited. To provide safe anesthesia during an exploration mission will be a highly challenging task. The research objective is thus to describe specific anesthesia procedures enabling treatment of pre-identified surgical conditions. Among the medical conditions considered by the NASA Human Research Program Exploration Medical Capability element, those potentially necessitating anesthesia techniques have been identified. The most appropriate procedure for each condition is thoroughly discussed. The substantial cost of training time necessary to implement regional anesthesia is pointed out. Within general anesthetics, ketamine combines the unique advantages of preservation of cardiovascular stability, the protective airway reflexes, and spontaneous ventilation. Ketamine side effects have for decades tempered enthusiasm for its use, but recent developments in mitigation means broadened its indications. The extensive experience gathered in remote environments, with minimal equipment and occasionally by insufficiently trained care providers, confirms its high degree of safety. Two ketamine-based anesthesia protocols are described with their corresponding indications. They have been designed taking into account the physiological changes occurring in microgravity and the specific constraints of exploration missions. This investigation could not only improve surgical care during long-duration spaceflights, but may find a number of terrestrial applications in isolated or austere environments.

  6. Improving mood with psychoanalytic and cognitive therapies (IMPACT): a pragmatic effectiveness superiority trial to investigate whether specialised psychological treatment reduces the risk for relapse in adolescents with moderate to severe unipolar depression: study protocol for a randomised controlled trial

    PubMed Central

    2011-01-01

    Background Up to 70% of adolescents with moderate to severe unipolar major depression respond to psychological treatment plus Fluoxetine (20-50 mg) with symptom reduction and improved social function reported by 24 weeks after beginning treatment. Around 20% of non responders appear treatment resistant and 30% of responders relapse within 2 years. The specific efficacy of different psychological therapies and the moderators and mediators that influence risk for relapse are unclear. The cost-effectiveness and safety of psychological treatments remain poorly evaluated. Methods/Design Improving Mood with Psychoanalytic and Cognitive Therapies, the IMPACT Study, will determine whether Cognitive Behavioural Therapy or Short Term Psychoanalytic Therapy is superior in reducing relapse compared with Specialist Clinical Care. The study is a multicentre pragmatic effectiveness superiority randomised clinical trial: Cognitive Behavioural Therapy consists of 20 sessions over 30 weeks, Short Term Psychoanalytic Psychotherapy 30 sessions over 30 weeks and Specialist Clinical Care 12 sessions over 20 weeks. We will recruit 540 patients with 180 randomised to each arm. Patients will be reassessed at 6, 12, 36, 52 and 86 weeks. Methodological aspects of the study are systematic recruitment, explicit inclusion criteria, reliability checks of assessments with control for rater shift, research assessors independent of treatment team and blind to randomization, analysis by intention to treat, data management using remote data entry, measures of quality assurance, advanced statistical analysis, manualised treatment protocols, checks of adherence and competence of therapists and assessment of cost-effectiveness. We will also determine whether time to recovery and/or relapse are moderated by variations in brain structure and function and selected genetic and hormone biomarkers taken at entry. Discussion The objective of this clinical trial is to determine whether there are specific

  7. Data-centric privacy protocol for intensive care grids.

    PubMed

    Luna, Jesus; Dikaiakos, Marios; Marazakis, Manolis; Kyprianou, Theodoros

    2010-11-01

    Modern e-Health systems require advanced computing and storage capabilities, leading to the adoption of technologies like the grid and giving birth to novel health grid systems. In particular, intensive care medicine uses this paradigm when facing a high flow of data coming from intensive care unit's (ICU) inpatients just like demonstrated by the ICGrid system prototyped by the University of Cyprus. Unfortunately, moving an ICU patient's data from the traditionally isolated hospital's computing facilities to dat