bluuebunny/crossref_metadata_2025 huggingface dataset:
P6300Endothelial cell activation by pro-inflammatory cytokines exerts novel paracrine effects on co-cultured cardiomyocytes
Authors: Fourre J , Bardi I , Maughan R T , Terracciano C M , Mason J C
Published: July 2020

Background

Anti-inflammatory therapies have failed to meet expectations in recent clinical trials for heart failure, despite the number of studies demonstrating pro-fibrotic, arrhythmogenic and hypertrophic effects of inflammation. To enlighten this, we sought to examine the contribution of non-myocytes in the cardiac inflammatory response. Endothelial cells (EC) can regulate cardiomyocyte (CM) function with multiple soluble factors. While many studies have shown the effects of pro-inflammatory cytokines on EC or CM separately, it is still unclear how the activation of EC can affect CM function.

Purpose

We studied the effect of pro-inflammatory pre-conditioning of EC on CM in indirect co-culture systems and in an ex vivo model of cardiac tissue. We hypothesised that pro-inflammatory activation of EC would alter the contractility of co-cultured CM.

Methods

Human cardiac microvascular EC were first pre-conditioned for 24h with Cytomix (1 ng/ml TNF-α, 1 ng/ml IL-1β, 25 ng/ml IL-6 Rα/IL-6 chimera) and co-cultured in a transwell system (pore size: 0.4μm) with adult rat ventricular CM. Co-culture supernatants were screened using a Cytokine Profiler Array. In vitro analysis of calcium handling in CM utilised the optical mapping technique and Fluo-4. Contractility of cardiac tissue was measured ex vivo using myocardial slices of 300 μm, prepared from left ventricles of adult rat and cultured for 24h with field stimulation and a fixed stretch producing a sarcomeric length of 2.2 μm.

Results

Treatment of EC by Cytomix prior to co-cultures induced a release of CC and CXC chemokines, G-/GM-CSF, ST2 and the adhesion molecule ICAM-1. Using published RNAseq datasets we noticed that adult CM do not constitutively express the receptors for the chemokines identified with the Profiler Array. However, duration of calcium transients in CM was significantly reduced from 454 to 322 ms in co-cultures with EC pre-conditioned by Cytomix, compared to untreated EC. Amplitude and time to peak were unchanged. In contrast, myocardial slices treated with Cytomix demonstrated a significant increase in contractility compared to control (from 3.3 to 5.3 mN/mm2) but no significant change in the duration of contraction (from 467 to 434 ms) or the rate of relaxation.

Conclusions

Pre-conditioned EC exert paracrine effects on the calcium handling of isolated CM, suggesting pro-inflammatory activation of proximal EC can affect CM function. In myocardial slices, pro-inflammatory stimulation provoked an inotropic response. The divergence of effect with findings in isolated cells may reflect differences in experimental design and multicellularity. Future work will aim to characterise the soluble mediators involved, and selectively target EC in slices to contextualise the effects described in co-cultures. Ultimately, this may inform the development of novel anti-inflammatory strategies for clinical use.

Acknowledgement/Funding

British Heart Foundation
Linking Antarctic krill larval supply and recruitment along the Antarctic Peninsula
Authors: Conroy John A , Reiss Christian S , Gleiber Miram R , Steinberg Deborah K
Published: July 2020

Synopsis

Antarctic krill (Euphausia superba) larval production and overwinter survival drive recruitment variability, which in turn determines abundance trends. The Antarctic Peninsula has been described as a recruitment hot spot and as a potentially important source region for larval and juvenile krill dispersal. However, there has been no analysis to spatially resolve regional-scale krill population dynamics across life stages. We assessed spatiotemporal patterns in krill demography using two decades of austral summer data collected along the North and West Antarctic Peninsula since 1993. We identified persistent spatial segregation in the summer distribution of euphausiid larvae (E. superba plus other species), which were concentrated in oceanic waters along the continental slope, and E. superba recruits, which were concentrated in shelf and coastal waters. Mature females of E. superba were more abundant over the continental shelf than the slope or coast. Euphausiid larval abundance was relatively localized and weakly correlated between the North and West Antarctic Peninsula, while E. superba recruitment was generally synchronized throughout the entire region. Euphausiid larval abundance along the West Antarctic Peninsula slope explained E. superba recruitment in shelf and coastal waters the next year. Given the localized nature of krill productivity, it is critical to evaluate the connectivity between upstream and downstream areas of the Antarctic Peninsula and beyond. Krill fishery catch distributions and population projections in the context of a changing climate should account for ontogenetic habitat partitioning, regional population connectivity, and highly variable recruitment.
Sovereign Wives? An Emotional Politics of Precarity and Resistance in the UK's Military Wives Choir
Authors: Cree Alice
Published: July 2020

This article provides a feminist analysis of the politics of vulnerability and resistance at work in the UK's Military Wives Choir. Military spouses represent vulnerable and “militarized subjects,” providing countless forms of unpaid labor in service of the military that range from the material labor of childcare to representational work in popular culture and everyday life. And yet as scholars of critical military studies and international politics, we so often fall short in our exploration of how military spouses engage with the militaristic processes in which they are embroiled. Indeed, work on the critical and resistant capacity of military wives as political agents is in particularly short supply. This article will use the example of the Military Wives Choir to argue that rather than seeing military wives simply as vulnerable militarized subjects without the capacity for resistance, it is in and through this vulnerability that the possibility of resistance can appear. As such, this work speaks to broader questions regarding the sites in which militarization occurs; even the most militarized of spaces and bodies do not necessarily only provide the preconditions for the emergence of military power. Rather, there is always the possibility for something more.
Circulating Tumor DNA in Cancer Management: A Value Proposition
Authors: Lam Wai Kei Jacky
Published: July 2020

Background

Analysis of circulating tumor DNA (ctDNA) allows the noninvasive molecular profiling of tumor, and such analysis has gained popularity for the detection of mutations with therapeutic implications. A value-based assessment would be useful for an objective evaluation of the benefits of ctDNA testing.

Content

The value proposition approach was used to evaluate the benefits of implementing ctDNA testing to inform treatment decisions of targeted therapy. The ctDNA testing was shown to complement tumor biopsy testing for the detection of mutations that are predictive of treatment response. It might be particularly useful for tracking resistance mechanisms among patients who experience disease progression despite treatment.

Patients, clinicians, and laboratory medicine specialists would benefit from the implementation of appropriate ctDNA testing in routine clinical care.
Determination of Thiabendazole in Fruit Juices by a New Monoclonal Enzyme Immunoassay
Authors: Abad Antonio , Manclús Juan J , Moreno María J , Montoya Angel
Published: July 2020

A competitive, indirect enzyme-linked immunosorbent assay (ELISA) for thiabendazole has been developed and applied to the analysis of fruit juices spiked with this fungicide. The immunoassay is based on a new monoclonal antibody derived from a hapten functionalized at the nitrogen atom in the 1-position of the thiabendazole structure. To our knowledge, such a structure has not been previously used to obtain antibodies to thiabendazole. The l50 value and the detection limit of the ELISA for standards were 0.2 and 0.05 ng/mL, respectively. Fruit juices were analyzed by diluting samples in assay buffer, without extraction or cleanup. Samples were not even centrifuged or filtered to remove fruit pulp. Under these conditions, the immunoassay was able to accurately determine thiabendazole down to 1 ng/mL in orange and grapefruit juices, down to 5 ng/mL in banana juice, and down to 20 ng/mL in apple and pear juices. Sensitivity differences of the ELISA were caused by the minimum dilution required by each juice to minimize matrix effects: 1/10 for orange and grapefruit juices, 1/50 for banana juice, and 1/100 for apple and pear juices. In an attempt to further increase the sensitivity of the immunoassay for matrixes showing the strongest interferences, apple and pear juices spiked with thiabendazole at low levels (1–20 ng/mL) were extracted with ethyl acetate before analysis. This simple procedure entailed a significant reduction of matrix effects, which in fact allowed us to determine accurately as low as 5 ng/mL thiabendazole in apple and pear juices. Irrespective of whether samples were analyzed by the direct dilution method or after extraction, the simplicity, sensitivity, and sample throughput of this monoclonal immunoassay makes it a very convenient method for the routine monitoring of thiabendazole residues in fruit juices.
DNA Methods: Critical Review of Innovative Approaches
Authors: Kok Esther J , Aarts Henk J M , Hoef A M Angeline van , Kuiper Harry A
Published: July 2020

The presence of ingredients derived from genetically modified organisms (GMOs) in food products in the market place is subject to a number of European regulations that stipulate which product consisting of or containing GMO-derived ingredients should be labeled as such. In order to maintain these labeling requirements, a variety of different GMO detection methods have been developed to screen for either the presence of DNA or protein derived from (approved) GM varieties. Recent incidents where unapproved GM varieties entered the European market show that more powerful GMO detection and identification methods will be needed to maintain European labeling requirements in an adequate, efficient, and cost-effective way. This report discusses the current state-of-the-art as well as future developments in GMO detection.
Use of Π-Acceptors for Spectrophotometric Determination of Dicyclomine Hydrochloride
Authors: Bebawy Lories I , Issa Yosry M , Moneim Kamal M Abdel
Published: July 2020

Simple, rapid, accurate, and sensitive spectrophotometric methods are described for the determination of dicyclomine hydrochloride. The methods are based on the reaction of this drug as an n-electron donor with 2,3-dichloro-5,6-dicyano-p-benzoqunione (DDQ), p-chloranilic acid (p-CA), and chloranil (CL) as Π-acceptors to give highly colored complex species. The colored products are measured spectrophotometrically at 456, 530, and 650 nm for DDQ, p-CA, and CL, respectively. Optimization of the different experimental conditions were studied. Beer's law was obeyed in concentration ranges of 20–100, 50–250, and 80–600 μg/mL for DDQ, p-CA, and CL, respectively. Colored complexes are produced in organic solvents and are stable for at least 1 h. The methods were applied to Spasmorest™ antispasmotic tablets and ampoules with good accuracy and precision.
Simultaneous Determination of Different Antibiotic Residues in Bovine and in Porcine Kidneys by Solid-Phase Fluorescence Immunoassay
Authors: Okerman Lieve , Wasch Katia De , Hoof Jan Van , Smedts Walter
Published: July 2020

Parallux®, a solid-phase fluorescence immunoassay (SPFIA) developed for antibiotic residue detection in milk, was used for analysis of bovine and porcine kidney tissue. Four tetracyclines, 2 broad-spectrum cephalosporins, 3 beta-lactam antibiotics, and cephapirin were detected in one run after minimal sample preparation. This commercially available test system is designed as cartridges, each with a combination of 1–4 tests. One cartridge can be used to detect 4 analytes in the same sample, or 1 or 2 analytes in different samples. The cartridge with the combination tetracyclines–ceftiofur–penicillin–cephapirin was selected because tetracyclines, beta-lactam antibiotics as well as cephalosporins, are registered for oral or parenteral use in bovines and pigs in Europe. The test is qualitative and is recommended only for screening. Tetracycline, oxytetracycline, chlortetracycline, and doxycycline were easily detected at 300 ppb with the tetracyclines channel; ceftiofur at 1000 ppb and cefquinome at 200 ppb with the ceftiofur channel; penicillin G, ampicillin, and amoxicillin at 50 ppb with the penicillin channel; and cephapirin at 100 ppb with the cephapirin channel. These levels are equal to or lower than the corresponding maximal residue limits in kidney tissue. Cephalexin was not detected. The SPFIA test can be used as an alternative to classical inhibition tests and for post-screening inhibitor- positive kidneys, because it detects 3 specific groups of antibiotics, which enables selection of specific confirmatory methods for identification and quantification.
Liquid Chromatographic Determination of α-Solasonine in Frozen Green Peas as an Indicator of the Presence of Nightshade Berries
Authors: Cavlovic Peter , Mankotia Mohan , Pantazopoulos Peter , Scott Peter M
Published: July 2020

Nightshade berries containing glycoalkaloids can be a contaminant in green peas. Methodology was developed to detect this contamination. The glycoalkaloid α-solasonine was extracted from frozen green peas with 1% (v/v) acetic acid, cleaned up on a C18 cartridge, and determined by liquid chromatography with UV detection at 200 nm. Method performance characteristics for the determination of α-solasonine include linearity from 140 to 1500 ng injected (r = 0.9996–0.9999); recovery ranging from 68 to 79%; limit of quantitation (LOQ) = 4.5 ppm (280 ng standard), and limit of detection = 0.64 ppm (40 ng standard). At the LOQ, the expanded uncertainty at 95% confidence was 0.38 × the reported value. The method was applied to the detection of α-solasonine in frozen green peas in a 2-year study of 60 samples of frozen green peas from Ontario, Canada. None of the samples contained α-solasonine. No unripe berries of Solanum nigrum were detected visually in the samples.
Liquid Chromatographic Determination of Diminazene Diaceturate (Berenil™) in Raw Bovine Milk
Authors: Roybal José E , Pfenning Allen P , Storey Joseph M , Gonzales Steve A , Turnipseed Sherri B
Published: July 2020

A simple liquid chromatographic (LC) method is presented for the determination of diminazene (DZ) in raw bovine milk. DZ is extracted from raw milk by chilled aqueous centrifugation and is isolated from milk components on a cyano solid-phase extraction column. DZ is eluted by using a methanol-ion pairing reagent. A Phenomenex LUNA CN column and an acetonitrile-buffered mobile phase with a counter ion are used for gradient LC. The LC effluent is monitored at a detection wavelength of 372 nm by using a deuterium lamp. Under the parameters described, the retention time of DZ is 8–10 min with a peak area response of 6.5 mAU/ng. The method demonstrated excellent precision over all levels tested (25–400 ppb) with an overall average recovery of 90.4 ± 14.5%. The method is applicable to the monitoring of milk for DZ residues at the 25 ppb level with a limit of quantitation of 10 ppb.
Liquid Chromatography/Tandem Mass Spectrometry Analysis of Chloramphenicol in Cooked Crab Meat
Authors: Rupp Heidi S , Stuart James S , Hurlbut Jeffrey A
Published: July 2020

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method is described for the extraction, cleanup, determination, and confirmation of chloramphenicol (CAP) in cooked crab meat. The method involves pulverization of cooked crab meat with dry ice; extraction of the CAP into ethyl acetate (EtOAc); evaporation (by N2) of the EtOAc; addition of methanol, aqueous NaCl, and heptane; extraction of the lipids into the heptane, followed by extraction of the aqueous phase with EtOAc; evaporation (by N2) of the EtOAc; dissolution into methanol–water; filtration; and separation/detection/confirmation using LC/MS/MS. Crab meat was fortified at 0.25, 0.50, and 1.0 ng/g (ppb) chloramphenicol. Average absolute recoveries were 67, 84, and 86%, respectively, with relative standard deviation values all less than 1%. Four daughter ions (m/z 152, 176, 194, and 257) were monitored off the m/z 321 precursor ion. Determination was based on a standard curve using the peak areas of the m/z 152 daughter ion (the base peak) for standard solutions equivalent to 0.10, 0.20, 0.50, and 1.0 ppb in tissue (made with control crab extract). A set of 6 matrix controls (unfortified crab meat) was also analyzed, in which no chloramphenicol was detected. For identification purposes, the ion ratios (of each daughter ion versus the base daughter ion) of the fortified crab versus those of the chloramphenicol standards agreed within 10% (relative) at fortified chloramphenicol concentrations of 0.25–1.0 ppb.
Enumeration Procedure for Monitoring Test Microbe Populations on Inoculated Carriers in AOAC Use-Dilution Methods
Authors: Tomasino Stephen F , Fiumara Rebecca M , Cottrill Michele P
Published: July 2020

The AOAC Use-Dilution methods do not provide procedures to enumerate the test microbe on stainless steel carriers (penicylinders) or guidance on the expected target populations of the test microbe (i.e., a performance standard). This report describes the procedures used by the U.S. Environmental Protection Agency to enumerate the test microbe (carrier counts) associated with conducting the Use-Dilution method with Staphylococcus aureus (Method 955.15) and Pseudomonas aeruginosa (Method 964.02) and the examination of historical data. The carrier count procedure involves the random selection of carriers, shearing bacterial cells from the carrier surface through sonication, and plating of serially diluted inoculum on trypticase soy agar. For each Use-Dilution test conducted, the official AOAC method was strictly followed for carrier preparation, culture initiation, test culture preparation, and carrier inoculation steps. Carrier count data from 78 Use-Dilution tests conducted over a 6-year period were compiled and analyzed. A mean carrier count of 6.6 logs (approximately 4.0 × 106colony-forming units/carrier) was calculated for both S. aureus and P. aeruginosa. Of the mean values, 95% fell within ±2 repeatability standard deviations. The enumeration procedure and target carrier counts are desirable for standardizing the Use-Dilution methods, increasing their reproducibility, and ensuring the quality of the data.
Interlaboratory Validation of the Vidas SET2 Kit for Detection of Staphylococcal Enterotoxins in Milk Products
Authors: Hennekinne Jacques-Antoine , Ostyn Annick , Guillier Florence , Gohier Martine , Messio Sabine , Dragacci Sylviane , Krys Sophie , Lombard Bertrand , Bauer M C  C ,   Collaborators:, Alves P , Bauer M C , Blanco-Vidal C , Desfargues C , De Reu K , Eckner K , Economides C , Ferrini A M , Gaillard N , Genot P , Gornard S , Guiliani L , Hummerjohann J , In’t Velt P , Jacobs-Reistma W , Ladstatter J , Montourcy G , Pires-Gomes C , Pittet J L , Rola J , Rubert E , Srete-Lancz Z
Published: July 2020

An earlier intralaboratory validation study based on the EN ISO 16140 Standard conducted by the Community Reference Laboratory for coagulase-positive staphylococci including Staphylocococcus aureus showed that, after an extraction step using dialysis concentration, the Vidas SET2 detection kit could be used to screen staphylococcal enterotoxins in milk and milk products. In order to fully validate Vidas SET2, an interlaboratory study was organized. Six freeze-dried samples and 3 ready-to-use concentrated extracts were analyzed by 21 laboratories according to the method, including a detection with Vidas SET2. Results did not show false-positive or -negative results. Accordance and concordance parameters were equal to 100%, corresponding to a concordance odds ratio of 1. This interlaboratory study confirmed the satisfactory outcome of the preliminary tests and of the intralaboratory study performed previously. The Vidas SET2 detection kit can be used as a method for the detection of staphylococcal enterotoxins in milk and milk products as well as the Transia Plate SET detection kit in the European screening method for official control purposes, after an extraction step followed by dialysis concentration.
Chemical Fingerprinting of Hoodia Species and Related Genera: Chemical Analysis of Oxypregnane Glycosides Using High-Performance Liquid Chromatography with UV Detection in Hoodia gordonii
Authors: Avula Bharathi , Wang Yan-Hong , Pawar Rahul S , Shukla Yatin J , Khan Ikhlas A
Published: July 2020

Hoodia gordonii, family Asclepiadaceae, is a succulent plant and is traditionally used in southern Africa for its appetite-suppressant properties. A high-performance liquid chromatographic (HPLC) method with UV detection for analysis of 11 oxypregnane glycosides from H. gordonii has been developed. The simultaneous analysis of 11 oxypregnane glycosides was achieved with a Phenomenex (Torrance, CA) reversed-phase C18 column using gradient mobile phase of water and acetonitrile, both containing 0.025 trifluoroacetic acid. The developed method was applied to the identification of oxypregnane glycosides in 3 different species of Hoodia and 23 related genera. The HPLC profiles of various plant samples were compared for the presence of oxypregnane glycosides.
Validation of a Biacore Method for Screening Eight Sulfonamides in Milk and Porcine Muscle Tissues According to European Decision 2002/657/EC
Authors: Gaudin Valérie , Hédou Céline , Sanders Pascal
Published: July 2020

Sulfonamides are commonly used for prophylactic or therapeutic purposes in veterinary medicine. A maximum residue limit (MRL) for sulfonamides has been set at 100 g/kg in milk and muscle. A multisulfonamide antibody was used for the development of 2 different Biacore protocols, one for the screening of milk samples, the other for muscle samples. Two different Biacore systems were used: Biacore X system (milk protocol), which is considered a research and development apparatus, and Biacore 3000 system (muscle protocol), which is a completely automated system used for high-throughput screening. This report describes the validation of semiquantitative immunological methods according to the European Decision 2002/657/EC concerning the performance of analytical methods. The different performance characteristics (detection capability CC, specificity/selectivity, precision, stability, and applicability) were determined in relation to the European Union MRL of 100 g/kg for sulfonamides. The applicability of the method to porcine, bovine, and poultry muscle was studied. The detection capabilities CC were calculated to be 40 g/L in milk and 60 g/kg in porcine, bovine, and poultry muscles. Eight different sulfonamides, of which 3 (sulfamethazine, sulfamerazine, and sulfadiazine) are authorized in France, were detected simultaneously, at or below the MRL level, with both Biacore systems.
Stability-Indicating Polarographic Determination of Acyclovir Through Chelation with Nickel(II)
Authors: Sheribah Zeinab A , El-Brashy Amina M , El-Gamal Rania M
Published: July 2020

A simple and sensitive, stability-indicating polarographic method was developed for the determination of acyclovir (ACV) in raw materials and dosage forms. The proposed method relies on the chelation of ACV with nickel(II) in Britton Robinson buffer (pH 5) and measuring the resulting polarographic wave either in the direct current (DCt) or differential pulse (DPP) modes. The polarographic wave has been characterized as being catalytic reduction prewave. Different experimental parameters affecting the formation of the NiACV chelate and its polarographic activity were studied and optimized. The current concentration relationship was found to be linear over the range of 0.88 and 18 g/mL, with minimum detectabilities of 0.10 and 0.19 g/mL using DPP and DCt modes, respectively. The method was used to investigate the kinetics of the acid-induced degradation of the drug. The apparent first-order rate constants and half-life times were calculated.
Determination of Allergenic Egg Proteins in Food by Protein-, Mass Spectrometry-, and DNA-Based Methods
Authors: Lee Ji-Yun , Kim Chang Jong
Published: July 2020

Egg allergy is one of the most common food allergies in both adults and children, and foods including eggs and their byproducts should be declared under food allergen labeling policies in industrial countries. Therefore, to develop and validate a sensitive and specific method to detect hidden egg allergens in foods, we compared immunochemical, DNA-based, and proteomic methods for detecting egg allergens in foods using egg allergen standards such as egg whole protein, egg white protein, egg yolk protein, ovomucoid, ovalbumin, ovotransferrin, lysozyme, and -livetin. Protein-based immunochemical methods, including ELISA as an initial screening quantitative analysis and immunoblotting as a final confirmatory qualitative analysis, were very sensitive and specific in detecting potentially allergenic egg residues in processed foods in trace amounts. In contrast, the proteomics-based, matrix-assisted laser desorption/ionization time-of-flight MS and LC-tandem quadrupole time-of-flight MS methods were not able to detect some egg allergens, such as ovomucoid, because of its nondenaturing property under urea and trypsin. The DNA-based PCR method could not distinguish between egg and chicken meat because it is tissue-nonspecific. In further studies for the feasibility of these immunochemical methods on 100 real raw dietary samples, four food samples without listed egg ingredients produced a positive response by ELISA, but exhibited negative results by immunoblotting.
Analysis of Quinapril by Two Solvent-Saving Methods: Application of Capillary Column High-Performance Liquid Chromatography with Ultraviolet Absorbance Detection and LDI-TOF-MS
Authors: Lu Chi-Yu , Wang Yi-Rou , Chen Su-Hwei , Feng Chia-Hsien
Published: July 2020

A capillary column high-performance liquid chromatography (CapLC) method and a laser desorption ionization-time of flight (LDI-TOF)-MS method are described for the determination of quinapril, an angiotensin-converting enzyme inhibitor. Effective separation was achieved by using a C18 capillary column at a flow rate of 10 L/min. For CapLC, quinapril and 7-hydroxycoumarin (internal standard) were detected at 210 and 320 nm, respectively. Phenformin replaced 7-hydroxycoumarin as the internal standard for the LDI-TOF-MS method successfully developed to detect quinapril. The calibration curves showed good linearity in the range of 1100 g/mL in these two methods. For high throughput purposes, the LDI-TOF-MS method was simpler and faster than the CapLC method. Both green methods were suitably validated and successfully applied to determine quinapril in commercial tablets.
Validation of a Method for the Determination of Chloramphenicol in Poultry and Swine Liver by Ultra-Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry
Authors: Xia Xi , Li Xiaowei , Ding Shuangyang , Shen Jianzhong
Published: July 2020

A sensitive and reliable method has been developed and validated for the determination of chloramphenicol in poultry and swine liver using SPE and ultra-performance liquid chromatography (UPLC)/MS/MS. The liver samples were extracted with ethyl acetate, defatted with n-hexane, and further cleaned up using SPE cartridges with polymeric sorbent. An Acquity BEH C18 column was used for gradient UPLC separation, with water and acetonitrile as the mobile phase. The multiple reaction monitoring mode was used for two precursor-product ion transitions for chloramphenicol and one for the internal standard. The method was validated at 0.1, 0.3, and 1.0 µg/kg. Mean recoveries from fortified samples ranged from 95.5 to 106.7% with an RSD of 12.2%. The method LOD was <0.02 µg/kg.
Development and Validation of a Stability-Indicating High-Performance Liquid Chromatography Method for the Simultaneous Determination of Albuterol, Budesonide, and Ipratropium Bromide in Compounded Nebulizer Solutions
Authors: Blewett Anthony J , Varma Deepti , Gilles Tiffany , Butcher Rashidi , Jacob Jaini , Amazan Jean , Jansen Susan A
Published: July 2020

In recent years, there has been a large increase in the use of pharmaceutical compounding to prepare medications that are not commercially available. The treatment of asthma typically includes the use of albuterol (ALB), ipratropium bromide (IPB), and/or budesonide (BUD) nebulizer solutions. There is currently no commercially available nebulizer solution containing all three of these compounds, and patients must rely on often-unregulated compounding. There is a distinct need for methodologies that can be used to analyze compounded formulations to ensure patient safety. We report an HPLC-UV method to separate and quantitate ALB, IPB, and BUD in nebulizer solutions. The method used a gradient elution to achieve separation via an RP C18 column. The method was validated, showed good selectivity, and was linear over several orders of magnitude. The method was applied to the analysis of nebulizer solutions and determination of their storage stability. Significant ALB-dependent degradation occurred within 5 h in solutions formulated with the free base of ALB, while those containing the sulfate salt of ALB produced no degradation. Alkali solutions can cause base-catalyzed hydrolysis of IPB and degradation of BUD. Compounded formulations containing ALB need to include an acid to control pH and prevent degradation.
A Non-targeted Approach to Chemical Discrimination Between Green Tea Dietary Supplements and Green Tea Leaves by HPLC/MS
Authors: Sun Jianghao , Chen Pei , Lin Long-Ze , Harnly James M
Published: July 2020

Green tea-based dietary supplements (GTDSs) have gained popularity in the U.S. market in recent years. This study evaluated the phytochemical composition difference of GTDS in comparison with green tea leaves using an HPLC/MS fingerprinting technique coupled with chemometric analysis. Five components that are most responsible for class separation among samples were identified as (-) epicatechin gallate, strictinin, trigalloylglucose, quercetin-3-O-glucosylrhamnosylglucoside, and kaempferol-3-O-galactosyl-rhamnosylglucoside, according to the accurate mass measurements and MS/MS data. The similarity coefficients between the GTDSs in solid form with green tea were 0.55 to 0.91, while for the GTDSs in liquid form they were 0.12 to 0.89, which suggested that chemical composition variance across the GTDSs was significant. Flavonol aglycone concentrations were higher in GTDSs than in tea leaves, indicating the degradation of flavonol glycosides or the oxidation of catechin during the manufacturing and storage processes. In some GTDS samples, compounds were identified that were on the label. The results demonstrate the urgency of QC for GTDS products.
159 Relationships among circulating prolactin concentrations, hair coat score, and weights of grazing cows
Authors: Davis Erin , Cauble Reagan , Powell Jeremy G , Lester Toby D , Gragg Walker , Kutz Bryan , Rosenkrans Charles
Published: July 2020

Prolactin (PRL) is a pleiotropic protein hormone synthesized and released from the anterior pituitary. Concentrations of PRL can serve as a biomarker of cattle exposure to ergot alkaloids. Our objective was to determine relationships among prolactin concentrations, hair coat score, and body weights of non-lactating crossbred beef cows (n = 71). Cows were weighed, hair coat scored (1 = 100% shed, and 5 = 0% shed), and blood samples collected in May, June, and July. Cows were grazing mixed grass pastures that included native endophyte-infected tall fescue. Serum PRL concentrations were determined by validated RIA. Based on May samples, cows were categorized as high (n = 11; 159 ± 29 ng/mL PRL), medium (n = 48; 51 ± 4 ng/mL PRL), or low (n = 12; 21 ± 4 ng/ mL PRL). Data were analyzed with Pearson correlations, and repeated measures ANOVA with month and prolactin category (PRLCAT) as main effects. Concentrations of PRL were correlated (r > 0.53; P < 0.0001) over the three months, and May PRL concentrations were correlated (r > 0.29; P < 0.02) with cow body weights in May, June, and July. Cows in the low PRLCAT had lower (P < 0.01) concentrations of PRL in all three months. Hair coat score decreased (P < 0.0001) each month (4.3, 2.7, and 1.3; respectively, May, June, July), and was higher (P < 0.05) for cows with low PRLCAT (3.3 vs. 2.7 and 2.4 for low, medium, and high, respectively). Cow body weight increased (P < 0.0001) from May to July. Cows in the low PRLCAT had lower (P < 0.05) body weights than medium and high PRLCAT cows (484 vs. 555, and 575 kg; respectively). Concentrations of PRL in May could be useful in identifying cattle with slick hair coats and heavier body weights.
161 Impact of insoluble dietary fiber on the efficacy of the phytase enzyme in nursery pigs
Authors: Acosta Jesus A , Patience John F
Published: July 2020

The objective of this experiment was to determine the effect of insoluble fiber on the efficacy of phytase in nursery pig diets. A total of 480 weaned pigs (BW 5.48 ± 0.14 kg; L337 X Camborough, PIC, Hendersonville, TN) were blocked by BW and randomly allotted to 48 pens. A common nutrient-adequate diet was fed from d 0–9. From d9-14, 2 phosphorus [P] deficient basal diets were fed: a low fiber [LF] corn-soybean meal diet (neutral detergent fiber [NDF]=7.5%; standardized total tract digestible [STTD] P = 0.16%) or a higher fiber [HF] corn-soybean meal plus 20% corn bran diet (NDF=15.2%; STTD P = 0.14%). From d 15–35, pigs received 8 dietary treatments (6 pens per treatment): 4 increasing levels of added phytase (0, 109, 218, and 327 phytase units [FTU]/kg; Quantum-Blue-5G, AB-Vista, Wiltshire, UK) applied to either the LF or HF basal diets. Titanium dioxide served as a digestibility marker. Fecal samples were collected on d 32–34. On d 35, one pig per pen was euthanized and fibulas harvested for bone ash assay. Data were analyzed using PROC MIXED of SAS. There were no interactions between fiber and phytase (P > 0.100). Adding phytase increased ADG (0.268, 0.282, 0.291, 0.306 kg/d; P < 0.001) and G:F (0.52, 0.55, 0.56, 0.58; P < 0.001) for the 4 increasing phytase levels, respectively. Fiber decreased ADG (0.282 vs 0.291 kg/d; P = 0.033) and tended to decrease G:F (0.54 vs 0.56; P = 0.097). There were no effects of phytase or fiber on ADFI (P > 0.100). Phytase increased bone ash (P = 0.005), but there was no effect of fiber (P = 0.949). Phytase did not affect the ATTD of DM, NDF, or ADF (P > 0.100) while fiber decreased the ATTD of DM, NDF and ADF (P < 0.001). In conclusion, increasing insoluble fiber did not affect the ability of phytase to improve growth performance and bone mineralization when pigs were fed a P deficient diet.
The Case Against the Doctors: Gender, Authority, and Critical Science Writing in the 1960s
Authors: O’Donnell Kelly
Published: July 2020

In the 1960s, widespread popular-cultural deference to the authority of science and medicine in the United States began to wane as a generation of journalists and activists reevaluated and criticized researchers and physicians. This article uses the career of feminist journalist Barbara Seaman to show the role that the emerging genre of critical science writing played in this broader cultural shift. First writing from her position as a mother, then as the wife of a physician, and finally as a credentialed science writer, Seaman advanced through distinct categories of journalistic authority throughout the 1960s. An investigation of Seaman’s early years in the profession also vividly demonstrates the roles that gender and professional expertise played in both constricting and permitting new forms of critique during this era.
Fluorescence characterization and microbial degradation of dissolved organic matter leached from salt marsh plants in the Yellow River Delta
Authors: Zhang Tao , Feng Zhihua , Luo Chunle , Sun Yixin , Li Jinzhen , Xu Juntian , Wang Xuchen
Published: July 2020

Aims

Salt marsh vegetation is an important contributor of dissolved organic matter (DOM) to coastal waters. The dynamics of DOM leaching from different marsh plants, however, have not been well studied or compared.

Methods

In this study, we conducted laboratory experiments to investigate the processes of DOM leaching from three common marsh plants (Phragmites australis, Suaeda salsa and Aeluropus littoralis) collected from the Yellow River Delta (YRD) salt marsh in October 2016. The YRD is one of the largest and most well-protected coastal ecosystems on the east coast of China.

Important Findings

We found that the plant leaves released DOM at much higher concentrations than the plant roots or stems, as measured by the dissolved organic carbon (DOC) and dissolved nitrogen (DN). On average, 15% of the biomass C and 30% of the biomass N were released from the plant leaves as DOC and DN during the 27-day incubation period. The DOM released from the plants was very labile, and 92.4%–98.1% of the DOC and 88.0%–94.6% of the DN released from the plants were consumed by bacteria during the 27-day incubation period. The fluorescence characteristics of the plant-released DOM indicated that chromophoric dissolved organic matter was a major fraction of the DOM and that protein-like components were the primary organic fractions released from the plants. Bacterial degradation altered both the fluorescence properties and the chemical composition of the DOM. The results of the laboratory experiments were well supported by the field investigation, which indicated that a large amount of DOM was outwelled from the YRD salt marshes in late autumn. Our study suggests that the DOM released from the biomass of salt marsh plants provides an important source of both DOC and DN for marsh and coastal waters. The highly labile DOC and DN provide essential food sources to support microbial communities in the YRD salt marsh and adjacent coastal waters.
COVID-19 Case and Contact Investigation in an Office Workspace
Authors: Hall Matthew T , Bui Han Q , Rowe John , Do Tai A
Published: July 2020

This investigation report describes a case of COVID-19 in a combined military and civilian office workspace and the contact investigation and mitigation efforts that followed. This office space included an embedded public health officer who was able to conduct the contact investigation and advise on the outbreak response. Over a 3-day period, the index case unintentionally exposed 150 coworkers to SARS-CoV-2 through participation in carpools, conferences, and small meetings. Of these exposures 37 were considered medium risk at the time and 113 were considered low risk. A total of 5 contacts reported COVID-like-symptoms at the time of the investigation and another 5 developed symptoms during the 14-day quarantine period and all were directed to self-isolate. None of the contacts required hospitalization and all the symptomatic contacts tested negative for SARS-CoV-2. With the advice and aid of the embedded public health officer, the office authorized telework, conducted thorough cleaning of spaces, distributed informative messaging, conducted virtual question-and-answer forums, and evaluated outbreak policies. This report demonstrates that the close integration of public health and office management can lead to rapid identification of those at risk of infection and implementation of mitigation and control efforts to stop the spread of disease.
High contrast at small separation – II. Impact on the dark hole of a realistic optical set-up with two deformable mirrors
Authors: Beaulieu M , Martinez P , Abe L , Gouvret C , Baudoz P , Galicher R
Published: July 2020

Future large space- or ground-based telescopes will offer the resolution and sensitivity to probe the habitable zone of a large sample of nearby stars for exo-Earth imaging. To this end, such facilities are expected to be equipped with a high-contrast instrument to efficiently suppress the light from an observed star to image these close-in companions. These observatories will include features such as segmented primary mirrors, secondary mirrors, and struts, leading to diffraction effects on the star image that will limit the instrument contrast. To overcome these constraints, a promising method consists in combining coronagraphy and wavefront shaping to reduce starlight at small separations and generate a dark region within the image to enhance the exoplanet signal. We aim to study the limitations of this combination when observing short-orbit planets. Our analysis is focused on SPEED, the Nice test bed with coronagraphy, wavefront shaping with deformable mirrors (DMs), and complex telescope aperture shape to determine the main realistic parameters that limit contrast at small separations. We develop an end-to-end simulator of this bench with Fresnel propagation effects to study the impact of large phase and amplitude errors from the test-bed optical components and defects from the wavefront shaping system on the final image contrast. We numerically show that the DM finite stroke and non-functional actuators, coronagraph manufacturing errors, and near-focal-plane phase errors represent the major limitations for high-contrast imaging of exoplanets at small separations. We also show that a carefully defined optical set-up opens the path to high contrast at small separation.
2147. Human Infections due to Actinotignum Species: A 5-Year Retrospective Review at Mayo Clinic Rochester, Minnesota
Authors: Syed Sadia , Sohail Muhammad R
Published: July 2020

Background

We aim to investigate the incidence, clinical presentation, management, and outcome of infections due to Actinotignum species observed at Mayo Clinic Rochester over the last 5 years.

Methods

We searched the clinical microbiology laboratory database to identify isolates of Actinotignum spp. from all body sites between January 1, 2014 and December 31, 2018.

Results

Fifty-four patients with positive culture with Actinotignum were identified. Mean age was 67 years and 27 (50%) had an underlying urogenital condition. Actinotignum was isolated in 26 urine cultures, 6 blood cultures, 12 abscess fluid cultures, and 10 bone/joint tissue cultures (Table 1). Fifteen (28%) specimens were monomicrobial while 39 (72%) were polymicrobial. Recovery from urine cultures was interpreted as colonization in 11 (20%) cases. Of the 54 patients with positive cultures, 43 patients had Actinotignum-associated clinical infection; 15 (35%) with urinary tract infections (11 with cystitis and 4 with pyelonephritis), 12 (28%) with abscesses (skin, intraabdominal, and surgical site infections), 10 (23%) with bone/joint infection, and 6 (14%) with bacteremia (Table 2). Most frequently isolated species was A. schaalii (n = 40); followed by 2 cases of A. sanguinis. Susceptibility testing (n = 40) showed that all stains were susceptible to penicillin (MIC< = 0.5), 36% were susceptible to clindamycin (MIC < = 2) and 10% susceptible to metronidazole (MIC < = 8). There was no recurrence of Actinotignum-related infections in any of the treated cases. Two patients with bone/joint infection underwent repeat surgical intervention due to worsening infection while on antibiotic treatment prior to resolution of infection. There was 1 death in a patient with bacteremia (polymicrobial) who had presented with a massive stroke (Table 3).

Conclusion

A. schaalii was most commonly associated with urinary tract infections followed by abscesses and bone/joint infections in elderly population. Majority of the infections were polymicrobial. All tested isolated were susceptible to penicillin; however, resistance was frequent for clindamycin and metronidazole. All appropriately treated patients had resolution of infection without recurrence from Actinotignum, except for one patient with bacteremia who died from massive stroke

Disclosures

All authors: No reported disclosures.
641. Evaluation of the FilmArray Pneumonia Panel and Potential Impact of Antimicrobial Use on Patients in a Trauma and Medical Intensive Care Unit
Authors: Krupinski- Shaw Kathy , Lopez Lauren , Orlinski Halle , Ozbolt Patrick , Antonara Stella
Published: July 2020

Background

Organisms causing infections of the lower respiratory tract in hospitalized patients can lead to high morbidity and mortality. Identification of the agents of pneumonia allows implementation of appropriate antimicrobial therapy and fast and accurate results are essential for the application of the correct antimicrobial regimen.

Methods

For 6 months results of quantitative bronchioalveolar lavage (Q-BALs) respiratory cultures, ordered as a standard of care for patients in our intensive care unit, were compared with the results obtained by a new multiplex molecular assay for the detection of lower respiratory tract pathogens, the FilmArray pneumonia panel (PP). The panel offers semi-quantitation of the bacterial targets that were compared with the quantitative results of the Q-BALs. Additionally, a retrospective chart review was performed to examine whether there would be any difference in the timing of appropriate antimicrobial therapy if the results of the panel were to be available for those patients. Appropriate antimicrobial therapy was determined according to the institution protocol for treatment of patients for ventilator-associated pneumonia based on the results of the quantitative cultures

Results

Thirty-six unique patients Q-BALs were run and of those there was 82% agreement on the detected targets between cultures and PP. Six targets were not detected by the panel (yeast, S. maltophilia, Streptococci, Salmonella spp.). M. catarrhalis, S. agalactiae and 3 viral targets were detected only by the panel. There was 100% agreement between the panel detected resistance markers and the culture isolates susceptibilities. Of the 36 patients, 12 were excluded because their medical records were not available for review. Of the 24 reviewed, 8 (33.3%) would have de-escalation in their antibiotics use at least 24h earlier due to the PP result. Eight (33.3%) would have no potential change in therapy and 8 (33.3%) could have inappropriate escalation or continuation due to reporting of potential pathogens by the PP but recorded as normal flora by cultures.

Conclusion

The use of PP would lead to a reduction of unnecessary antimicrobial therapy in 1/3 of the patients examined. However, quantification of organisms otherwise reported as normal flora may lead to unnecessary treatment and requires education of staff to understand the results of the assay.

Disclosures

All authors: No reported disclosures.