11 research outputs found
A fast and accurate method of detecting Aleutian mink disease virus in blood and tissues of chronically infected mink
No full textThe objective of this study was to assess sensitivity of the Omni Klentaq-LA DNA polymerase for detecting Aleutian mink disease virus (AMDV) in mink blood and tissues by PCR without DNA extraction. The presence of AMDV DNA was directly tested by Klentaq in plasma, serum, whole blood and spleen homogenates of 188 mink 4 and 16 months after inoculation with the virus. Samples from bone marrow, small intestine, liver, lungs, kidneys and lymph nodes of 20 of the same mink were also tested by Klentaq. DNA was extracted from paired samples of plasma and the aforesaid tissues by a commercial nucleic acid extraction kit (Dynabeads Silane) and tested by PCR. Compared with the extracted DNA, Klentaq detected a significantly greater number of samples in the whole blood, serum, plasma, spleen and small intestine. It was concluded that Klentaq is a preferred system for directly detecting AMDV DNA in mink blood and tissues. The lower success rate of extracted DNA compared with Klentaq could be the result of DNA losses during the extraction process. This is an important factor in chronically infected mink which have a low AMDV copy number in the bloodstream. Direct AMDV detection also reduces the cost of PCR amplification and lowers the risk of sample contamination.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author
Accuracy of enzyme-linked immunosorbent assays for quantification of antibodies against Aleutian mink disease virus
No full textAbstractThere is a growing interest among mink ranchers to select their stock for tolerance to the Aleutian mink disease virus (AMDV). Enzyme-linked immunosorbent assays (ELISA) are used to identify mink which have low anti-AMDV antibody titres and are expected to tolerate the AMDV infection. The objective of this study was to calculate the accuracy of three ELISA systems which were performed on blood or serum of AMDV-inoculated American mink (Neovison vison) at five laboratories in Canada, USA, Finland, the Netherlands and Denmark. The accuracy was determined by comparing the ELISA results with antibody titres measured by the counter-immunoelectrophoresis (CIEP) using 10 two-fold serial dilutions of the plasma. Antibody titres of 880 black mink which were inoculated with a spleen homogenate from a naturally infected mink were measured between 16 and 176 weeks post-inoculation. Each ELISA result from every laboratory covered a wide range of antibody titres and the Spearman’s rank correlation coefficients between CIEP and ELISA results from different laboratories varied between 0.41 and 0.83, indicating a low to moderate accuracy of ELISA systems for ranking mink by antibody titre. The recombinant VP2-based ELISA used in the Netherlands and Finland ranked the mink by antibody titres more accurately than did the AMDV-G-based ELISA platforms developed in Denmark and the USA, suggesting that the source of antigen was one of the factors affecting the accuracy of ELISA results. It was concluded that the ELISA systems, particularly those based on AMDV-G antigen, require further refinement to improve their accuracy for ranking mink by antibody titre
Trustworthy provenance framework for document workflow provenance
No full textDocument workflows, which plays a major role in enterprise business process automation, are dynamic and prone to be changed over time. Assuring the provenance of these workflows is important when comes to quality, long term preservation, forensics and regulatory compliance. This research introduces a Provenance Framework for collecting workflow provenance data, storing them in documents as metadata securely and querying stored provenance data. The author uses the concept of making data objects independent and consider provenance data as intrinsic property of the data object, to support long term preservation of documents with provenance and to maintain the link between the data object and its provenance in cloud over time. Provenance data modeling and representation is done according to the W3C PROV Model. XMP framework is used to store and query provenance data as metadata in documents. Document signatures and metadata encryption is used to ensure security of provenance data
Antihypertensive effects of apple peel extract on spontaneously hypertensive rats
No full textBackground and Objectives:
Apple peel is a rich source of biological active phytochemicals such as flavonoids. The present study investigated the antihypertensive effect of flavonoid-rich Apple Peel Extract (APE) on Spontaneously Hypertensive Rats (SHR). Methodology: Three groups of animals: Control, captopril (20 mg kg–1 of body weight/day) and APE (25 mg kg–1 of body weight/day) were fed standard rat chow and their corresponding treatment in sugar-free gelatin, daily for a period of eight weeks. Blood Pressure (BP) was monitored weekly using the tail cuff method. Blood and tissue samples were collected after the eighth week. Results: As expected, treatment with captopril consistently reduced BP (p<0.05). APE treatment reduced both systolic and diastolic BP by 15 and 11 mg Hg, respectively, after 5 weeks of treatment, However, statistical significance was only achieved in systolic BP after eight weeks when compared with control (p<0.05). There were no significant differences in serum and lung ACE activity at week eight. Treatment with APE increased liver superoxide dismutase (SOD) activity by 78% and total reduced Glutathione (GSH) concentrations by 42% when compared to control (p<0.05) but had no effect on the activity of glutathione reductase or peroxidase.
Conclusion:
Long term intake of APE reduces high blood pressure in SHR possibly through endogenous antioxidant pathways. This preclinical trial suggests that APE as a dietary supplement could be effective in managing early stages of hypertension.
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Monitoring emerging pathogens using negative nucleic acid test results from endemic pathogens in pig populations: Application to porcine enteric coronaviruses
No full textThis study evaluated the use of endemic enteric coronaviruses polymerase chain reaction (PCR)-negative testing results as an alternative approach to detect the emergence of animal health threats with similar clinical diseases presentation. This retrospective study, conducted in the United States, used PCR-negative testing results from porcine samples tested at six veterinary diagnostic laboratories. As a proof of concept, the database was first searched for transmissible gastroenteritis virus (TGEV) negative submissions between January 1st, 2010, through April 29th, 2013, when the first porcine epidemic diarrhea virus (PEDV) case was diagnosed. Secondly, TGEV- and PEDV-negative submissions were used to detect the porcine delta coronavirus (PDCoV) emergence in 2014. Lastly, encountered best detection algorithms were implemented to prospectively monitor the 2023 enteric coronavirus-negative submissions. Time series (weekly TGEV-negative counts) and Seasonal Autoregressive-Integrated Moving-Average (SARIMA) were used to control for outliers, trends, and seasonality. The SARIMA’s fitted and residuals were then subjected to anomaly detection algorithms (EARS, EWMA, CUSUM, Farrington) to identify alarms, defined as weeks of higher TGEV-negativity than what was predicted by models preceding the PEDV emergence. The best-performing detection algorithms had the lowest false alarms (number of alarms detected during the baseline) and highest time to detect (number of weeks between the first alarm and PEDV emergence). The best-performing detection algorithms were CUSUM, EWMA, and Farrington flexible using SARIMA fitted values, having a lower false alarm rate and identified alarms 4 to 17 weeks before PEDV and PDCoV emergences. No alarms were identified in the 2023 enteric negative testing results. The negative-based monitoring system functioned in the case of PEDV propagating epidemic and in the presence of a concurrent propagating epidemic with the PDCoV emergence. It demonstrated its applicability as an additional tool for diagnostic data monitoring of emergent pathogens having similar clinical disease as the monitored endemic pathogens.This article is published as Serafini Poeta Silva, Ana Paula, Guilherme Arruda Cezar, Edison Sousa Magalhães, Kinath Rupasinghe, Srijita Chandra, Gustavo S. Silva, Marcelo Almeida et al. "Monitoring emerging pathogens using negative nucleic acid test results from endemic pathogens in pig populations: Application to porcine enteric coronaviruses." PloS one 19, no. 7 (2024): e0306532. doi: https://doi.org/10.1371/journal.pone.0306532. © 2024 Serafini Poeta Silva et al. This is an open access article distributed under the terms of the (http://creativecommons.org/licenses/by/4.0/). which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Allyl isothiocyanate regulates lysine acetylation and methylation marks in an experimental model of malignant melanoma
No full textObjective(s): Isothiocyanates (ITCs) are biologically active plant secondary metabolites capable of mediating various biological effects including modulation of the epigenome. Our aim was to characterize the effect of allyl isothiocyanate (AITC) on lysine acetylation and methylation marks as a potential epigenetic-induced anti-melanoma strategy. Methods: Our malignant melanoma model consisted of (1) human (A375) and murine (B16-F10) malignant melanoma as well as of human; (2) brain (VMM1) and lymph node (Hs 294T) metastatic melanoma; (3) non-melanoma epidermoid carcinoma (A431) and (4) immortalized keratinocyte (HaCaT) cells subjected to AITC. Cell viability, histone deacetylases (HDACs) and acetyltransferases (HATs) activities were evaluated by the Alamar blue, Epigenase HDAC Activity/Inhibition and EpiQuik HAT Activity/Inhibition assay kits, respectively, while their expression levels together with those of lysine acetylation and methylation marks by western immunoblotting. Finally, apoptotic gene expression was assessed by an RT-PCR-based gene expression profiling methodology. Results: AITC reduces cell viability, decreases HDACs and HATs activities and causes changes in protein expression levels of various HDACs, HATs, and histone methyl transferases (HMTs) all of which have a profound effect on specific lysine acetylation and methylation marks. Moreover, AITC regulates the expression of a number of genes participating in various apoptotic cascades thus indicating its involvement in apoptotic induction. Conclusions: AITC exerts a potent epigenetic effect suggesting its potential involvement as a promising epigenetic-induced bioactive for the treatment of malignant melanoma. © 2019, The Author(s)
Atividade antioxidante in vitro e in vivo de diferentes cultivares de maçã (Malus domestica Borkh) do estado de Santa Catarina
No full textTese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias, Programa de Pós-Graduação em Ciência dos Alimentos, Florianópolis, 2010.A maçã (Malus domestica Borkh) muito consumida em diversas regiões do mundo é considerada uma fruta rica em compostos fenólicos, os quais variam conforme a cultivar e a parte da fruta analisada. Este estudo objetivou analisar as características físico-químicas de diferentes cultivares de maçãs colhidas no sul do Brasil, nos anos de 2008 e 2009; comparar o conteúdo de compostos fenólicos e a atividade antioxidante na polpa, fruta inteira e casca destas cultivares; e investigar o efeito do consumo agudo de duas cultivares de maçã sobre a atividade antioxidante e a oxidação lipídica em humanos. Entre as duas safras, foram estudadas, no total, 22 cultivares de maçãs colhidas nas Estações Experimentais da Empresa de Pesquisa Agropecuária e Extensão Rural de Santa Catarina (EPAGRI/SC). O conteúdo de matéria seca, sólidos solúveis totais, pH, açúcares totais, acidez titulável, fenólicos totais, flavanóis totais, antocianinas monoméricas totais e atividade antioxidante foram determinados nas amostras. Foram observadas diferenças estatisticamente significativas entre as cultivares, em todos os parâmetros de composição química analisados. As concentrações de fenólicos totais, flavanóis totais, antocianinas monoméricas totais e atividade antioxidante medida pelos métodos ABTS, DPPH e FRAP também diferiram significativamente entre as cultivares e foram maiores na casca, seguidas da fruta inteira e polpa. Independente do ano de colheita, a cultivar Catarina apresentou significativamente as maiores concentrações de fenólicos totais, flavanóis totais e maior atividade antioxidante, enquanto que a cultivar Golden Delicious, em 2008, apresentou as menores concentrações, e em 2009, esteve entre as cultivares com menores conteúdos destes parâmetros. Foi observada uma relação positiva entre o conteúdo de fenólicos totais e a atividade antioxidante dos extratos; entre flavanóis totais e atividade antioxidante; e entre fenólicos totais e flavanóis totais na polpa, fruta inteira e na casca. Nove mulheres saudáveis, não fumantes, participaram de um estudo de intervenção controlado para verificar o efeito do consumo agudo de 300 mL de suco de maçã da cultivar Golden Delicious, suco de maçã da cultivar Catarina ou Água, sobre o estado antioxidante e a oxidação lipídica sérica. Uma hora após o consumo de ambos os sucos, houve significativo aumento na capacidade antioxidante, no ácido ascórbico e ácido úrico séricos, e diminuição da oxidação lipídica comparado com o grupo controle que consumiu apenas água. Nenhuma diferença significativa foi observada entre o efeito do consumo agudo do suco da Golden Delicious e Catarina; contudo, as modificações observadas após o consumo dos sucos foram estatisticamente diferentes das observadas após o consumo de água. As concentrações séricas de fenólicos totais não foram afetadas em nenhum dos tratamentos. Nenhum efeito significante sobre a inibição da oxidação lipídica ex vivo foi observado. Após a intervenção com suco de maçã Golden Delicious e Catarina, o aumento da atividade antioxidante e à redução da oxidação lipídica sérica foram relacionados ao aumento da concentração sérica de ácido úrico. Nenhuma relação foi observada entre as alterações da concentração de ácido ascórbico e atividade antioxidante e oxidação lipídica sérica. Os resultados deste estudo indicam que o genótipo de maçãs substancialmente influencia a composição química da fruta, especialmente quanto ao conteúdo de compostos fenólicos, entretanto o efeito do consumo agudo de maçãs sobre o estado antioxidante e oxidação lipídica em humanos independe da genética desta fruta
LINE PARAMETERS FOR THE OXYGEN A BAND
No full textAuthor Institution: Department of Physics, College of William and Mary, Williamsburg, VA; Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA; Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD; California Institute of Technology, Pasadena, CASimulation of the oxygen A band to a level that is sufficient for accurate studies of the Earth's atmosphere is complex in that not only are Doppler and Lorentz broadening important, but also Dicke narrowing, pressure shifts, line mixing and speed dependence. In addition all of these parameters except the speed dependence require temperature dependence parameters as well. To measure all of the required line parameters with the multispectrum nonlinear least squares fitting technique, spectra were acquired by the Bruker IFS125-HR Fourier Transform Spectrometer at the Jet Propulsion Laboratory in combination with various multpass cells, a cavity ring down spectrometer at NIST and a photoacoustic spectrometer at the California Institute of Technology. The combination of the data from these three very different types of spectrometers in a single simultaneous fit of the entire band enables the measurement of all of these quantities. The results to this point will be summarized.
THE EFFECTIVE HAMILTONIAN FOR THE GROUND STATE OF AND NEW MEASUREMENTS OF THE FINE STRUCTURE SPECTRUM NEAR 1.2 .
No full textAuthor Institution: Department of Physics and Astronomy, Pomona College, Claremont, CA 91711; Chemistry Department Brookhaven National Laboratory, Upton, NY 11973 and Stony Brook University, Stony Brook, NY 11794; Homer L. Dodge Department of Physics and Astronomy, University of Oklahoma, Norman, OK; Gottfreid-Wilhelm-Liebniz-Universitat, Institut fur Physikalische Chemie \& Elektrochemie, D-30167 Hannover, GermanyWe have measured rotational transitions in the ground, , electronic state of naturally occuring isotopomers of PbF in a supersonic free jet Fourier transform microwave spectrometer. The data for is particularly interesting because it is a candidate for a future experimental e-EDM measurement. To fit the data for this species to the measurement precision, the nuclear spin-spin dipolar interaction and a second term that can be equivalently viewed as a centrifugal distortion correction to the familiar Frosch and Foley hyperfine coupling terms, or an dependent correction to the nuclear spin-rotational coupling are required, in addition to the standard terms. To characterize the higher component of the ground state of PbF, we are attempting a direct measurement of transitions between the two components in a slit jet-cooled sample using a frequency comb-referenced extended cavity diode laser. This spectrum was originally detected in a hot source by Fourier transform near-infrared spectroscopy, nderline{\textbf{191}}, 108 1998.} but low transitions were unresolved at that time. Acknowledgments: Work at Brookhaven National Laboratory was carried out under Contract No. DE-AC02-98CH10886 with the U.S. Department of Energy. Work by N. E. Shafer-Ray was performed with support from the National Science Foundatation award NSF-0855431. J.-U. Grabow ackonwledges funding from the Deutsche Forschungsgemeinschaft and the Land Niedersachsen
Measurement of tissue azithromycin levels in self-collected vaginal swabs post treatment using liquid chromatography and tandem mass spectrometry (LC-MS/MS)
No full text© 2017 Vodstrcil et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Background Azithromycin is recommended for the treatment of uncomplicated urogenital chlamydia infection although the standard 1gram dose sometimes fails to eradicate the infection (treatment failure). One hypothesis proposed for treatment failure has been insufficient levels of the antibiotic at the site of infection. We developed an assay using liquid chromatography and tandem mass spectrometry (LC-MS/MS) to measure azithromycin concentration in high-vaginal swabs and monitor how concentration changes over time following routine azithromycin treatment. Methods Azithromycin concentrations were measured in two groups of women either within the first 24h of taking a 1g dose (N = 11) or over 9 days (N = 10). Azithromycin concentrations were normalised to an internal standard (leucine enkephalin), and the bulk lipid species phosphatidylcholine [PC(34:1)], using an Agilent 6490 triple quadrupole instrument in positive ionisation mode. The abundances of azithromycin, PC(34:1), and leu-enkephalin were determined by multiple reaction monitoring and absolute levels of azithromycin estimated using standard curves prepared on vaginal specimens. Results Vaginal azithromycin concentrations of women were rapidly obtained after 5h post-treatment (mean concentration = 1031mcg/mg of lipid, range = 173-2693mcg/mg). In women followed for 9 days, peak concentrations were highest after day 2 (mean concentration = 2206mcg/mg, range = 721-5791mcg/mg), and remained high for at least 9 days with a mean concentration of 384mcg/mg (range = 139-1024mcg/mg) on day 9. Conclusion Our study confirmed that a single 1g dose of azithromycin is rapidly absorbed and remains in the vagina at relatively high levels for at least a week, suggesting that poor antibiotic absorption is unlikely to be an explanation for treatment failure
