Autonomous Microfluidic Sample Preparation System for Protein Profile-Based Detection of Aerosolized Bacterial Cells and Spores

2007 ◽  
Vol 79 (15) ◽  
pp. 5763-5770 ◽  
Author(s):  
Jeanne C. Stachowiak ◽  
Erin E. Shugard ◽  
Bruce P. Mosier ◽  
Ronald F. Renzi ◽  
Pamela F. Caton ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Protein Profile ◽  
Bacterial Cells

Automated Sample Preparation System for Rapid Biological Threat Detection

2005 ◽  
Author(s):  
Jeanne C. Stachowiak ◽  
Erin E. Shugard ◽  
Pamela Caton ◽  
Bruce P. Mosier ◽  
Ron Renzi ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Bacterial Species ◽  
Fused Silica ◽  
Fluorescent Labeling ◽  
Control Flow ◽  
Size Exclusion ◽  
Bacterial Cells ◽  
Fused Silica Capillaries ◽  
Exclusion Chromatography ◽  
Automated Sample Preparation

Rapid, automated sample preparation of bacterial cells and spores is required for threat analysis by remotely deployed chemical and biological warning systems. Sandia is designing, building, and testing an automated front-end sample preparation system based on miniature and microfluidic components, with the goal of concentrating bacterial species collected from the air, harvesting and solubilizing proteins from them, and delivering them to Sandia’s MicroChemLab capillary gel electrophoresis system1,2 for analysis (Fig. 1). Miniature, motorized valves and pumps control flow between system components connected by fused silica capillaries (Fig. 4). Sample processing modules include concentration by dielectrophoresis in an array of insulating posts or by mechanical filtration; heat-activated chemical lysis; mechanical filtration; removal of chemical lysis agents by size exclusion chromatography (SEC); and in-capillary fluorescent labeling.

scholarly journals Ketamine used in the therapy of depressive disorders impacts protein profile, proliferation rate and phagocytosis resistance of enterococci

2021 ◽  
Author(s):  
Tomasz Artur Jarzembowski ◽  
Agnieszka Daca ◽  
Wiesław J. Cubała ◽  
Marek Bronk ◽  
Łukasz Naumiuk
Keyword(s):  
Depressive Disorders ◽  
Early Stage ◽  
Protein Profile ◽  
Proliferation Rate ◽  
Bacterial Suspension ◽  
Bacterial Cells ◽  
High Concentrations ◽  
Etiological Agents ◽  
Different Response

Abstract Background: Ketamine is known to cause rapid anti-depressive effect. Additionally, it has been also proved that at high concentrations ketamine inhibits bacterial growth. It is also widely known that even sub-inhibitory concentration of chemicals, as concentration of ketamine used in therapy of depression, may change bacterial properties, including their virulence. The knowledge about possible influence of ketamine on bacterial commensals seems to be essential, as the mechanism of ketamine’s action in depression is believed to result also from its’ anti-inflammatory activity. In the current study we aimed to evaluate the in vitro influence of ketamine on proliferation rate, phagocytosis resistance and toxicity of enterococci. Results: The studied enterococcal strains were isolated as etiological agents of infection and collected in the Department of Medical Microbiology, Medical University of Gdansk. To measure metabolic activity of Enterococcus faecalis 10µM of CFDA-SE was added to bacterial suspension. The number of bacterial cells and fluorescence of particles were determined using FACSVerse flow cytometer. Additionally, for the determination of phagocytosis resistance, THP-1 human monocytes cell line from ATCC was used. Suspension of monocytes which engulfed bacteria was then stained with propidium iodide to determine cells’ membrane permeability and to evaluate cytotoxicity of enterococci. The result of the study proved diverse influence of therapeutic concentration of ketamine on Enterococci. In 23.1% of strains both proliferation rate and metabolism activity were inhibited. This group of strains was more susceptible to phagocytosis and had lower cytotoxicity than in culture without ketamine. Different response of isolates to ketamine was also visible in changes of protein profile determined by MALDI TOF. Conclusions: The analysis of bacteria at early stage of growth curve demonstrate the bacterial diversity in response to ketamine and let us set the hypothesis that microbiome susceptibility to ketamine may be one of the elements which should be taken into consideration when planning the successful pharmacotherapy of depression.

scholarly journals THE INFLUENCE OF ORGANIC SOLVENT PROTEIN PRECIPITATION ON SDS PAGE PROTEIN PROFILE IN SERUM

2010 ◽  
Vol 7 (3) ◽  
pp. 337-341
Author(s):  
Tri Joko Raharjo ◽  
Rusmiati Suprihatin ◽  
Deni Pranowo
Keyword(s):  
Sample Preparation ◽  
Organic Solvent ◽  
Serum Protein ◽  
Relative Concentration ◽  
Protein Profile ◽  
Protein Precipitation ◽  
Sds Page ◽  
Minor Protein

A study on the influence of organic solvent protein precipitation to the profile of the serum protein has been accomplished. The expected conditions were precipitation of abundant proteins present in serum result in increasing relative concentration of minor protein which can be useful for sample preparation for biomarker studies. The serum were precipitated with various diluted (

scholarly journals Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Hao-Tsai Cheng ◽  
Sen-Yung Hsieh ◽  
Chang-Mu Sung ◽  
Betty Chien-Jung Pai ◽  
Nai-Jen Liu ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Protein Extraction ◽  
Protein Profile ◽  
Two Dimensional Gel Electrophoresis ◽  
Protein Preparation ◽  
Preparation Methods ◽  
Human Bile ◽  
Acetone Precipitation ◽  
Exogenous Compounds ◽  
Dimensional Electrophoresis

Aims. Bile is an important body fluid which assists in the digestion of fat and excretion of endogenous and exogenous compounds. In the present study, an improved sample preparation for human bile was established.Methods and Material. The method involved acetone precipitation followed by protein extraction using commercially available 2D Clean-Up kit. The effectiveness was evaluated by 2-dimensional electrophoresis (2DE) profiling quality, including number of protein spots and spot distribution.Results. The total protein of bile fluid in benign biliary disorders was 0.797 ± 0.465 μg/μL. The sample preparation method using acetone precipitation first followed by 2D Clean-Up kit protein extraction resulted in better quality of 2DE gel images in terms of resolution as compared with other sample preparation methods. Using this protocol, we obtained approximately 558 protein spots on the gel images and with better protein spots presentation of haptoglobin, serum albumin, serotransferrin, and transthyretin.Conclusions. Protein samples of bile prepared using acetone precipitation followed by 2D Clean-Up kit exhibited high protein resolution and significant protein profile. This optimized protein preparation protocol can effectively concentrate bile proteins, remove abundant proteins and debris, and yield clear presentation of nonabundant proteins and its isoforms on 2-dimensional electrophoresis gel images.

Microbially Induced Flotation of Galena and Quartz from Pyrite

2009 ◽  
Vol 71-73 ◽  
pp. 509-512
Author(s):  
H. Sarvamangala ◽  
K.A. Natarajan
Keyword(s):  
Bacillus Subtilis ◽  
Protein Profile ◽  
Selective Separation ◽  
Extracellular Protein ◽  
Bacterial Cells ◽  
Presence And Absence

Selective separation of pyrite from galena and quartz was achieved through microbiologically induced flotation in presence of Bacillus subtilis. B. subtilis functions as a depressant for pyrite while it promotes the flotation of galena and quartz. Bacterial extracellular protein (EP) was isolated and the protein profile of bacterial cells grown in presence and absence of minerals established.

scholarly journals Effects of Long Term Exposure to Aspirin on Growth, Functionality and Protein Profile of Lactobacillus rhamnosus (LGG) (ATCC 53103)

2016 ◽  
Vol 5 (4) ◽  
pp. 46 ◽  
Author(s):  
Temitayo Obanla ◽  
Sarah Adjei-Fremah ◽  
Rabin Gyawali ◽  
Tahl Zimmerman ◽  
Mulumebet Worku ◽  
...  
Keyword(s):  
Protein Profile ◽  
Agar Plate ◽  
Lactobacillus Rhamnosus ◽  
Exposure Period ◽  
The Elderly ◽  
Bacterial Cells ◽  
Bacterial Populations ◽  
Gastrointestinal Health ◽  
Mrs Broth

<p>In the present study, we determined the effects of long-term exposure to aspirin on the growth and functionality of <em>Lactobacillus rhamnosus</em>. One isolated colony of <em>L. rhamnosus</em> was propagated three times in deMan, Rogosa and Sharpe (MRS) broth and incubated at 37°C. The active strain was then harvested and washed with 0.1% peptone water. Cells were transferred into 9 mL MRS broth containing approximately 6 mg/mL of aspirin, vigorously mixed and incubated for 4 h. The cells were further harvested, transferred into MRS broth and incubated at 37°C. The exposure protocol was repeated for five sequential transfers a week, and the exposed strain cell was surface plated onto MRS agar containing equal amount of aspirin. One isolated colony of aspirin exposed <em>L. rhamnosus</em> from an aspirin containing agar plate was further activated in MRS broth. This procedure was repeated sequentially for 12 consecutive weeks. Bacterial populations, <em>beta</em>-galactosidase activity (<em>beta</em>-gal), and protein expression were determined. Bacterial cells unexposed to aspirin were used as the control. Our results showed that <em>L. rhamnosu</em>s could survive after long term exposure to a sub-inhibitory concentration of aspirin. The average <em>beta</em>-gal activity of <em>L. rhamnosus</em> in unexposed cells was 153 ± 2.5 Gal U.; however, <em>beta</em>-gal activity was completely inhibited in exposed cells throughout the exposure period. There was approximately 54% more protein expressed in the long term aspirin-exposed strain compared to the short term exposed (4 h) strain as determined by BCA assay. In addition, the SDS-page gel indicated the synthesis of more protein in the exposed <em>L. rhamnosus</em>. The onset of common or chronic illnesses, especially among the elderly population, is often treated with common medications. However, regular the intake of medical drugs such as aspirin could affect beneficial gut microflora. Therefore, foods containing probiotics such as yogurt and other functional foods are important for the maintenance of optimal gastrointestinal health.</p><p><em><br /></em></p>

Pollutant Identification by Selected Area Electron Diffraction

1974 ◽  
Vol 32 ◽  
pp. 532-533
Author(s):  
R. E. Ferrell ◽  
G. G. Paulson ◽  
C. W. Walker
Keyword(s):  
Thermal Treatment ◽  
Electron Diffraction ◽  
Sample Preparation ◽  
Crystal Structures ◽  
Water Samples ◽  
Environmental Samples ◽  
Short Review ◽  
Selected Area Electron Diffraction ◽  
Multiple Component

Selected area electron diffraction (SAD) has been used successfully to determine crystal structures, identify traces of minerals in rocks, and characterize the phases formed during thermal treatment of micron-sized particles. There is an increased interest in the method because it has the potential capability of identifying micron-sized pollutants in air and water samples. This paper is a short review of the theory behind SAD and a discussion of the sample preparation employed for the analysis of multiple component environmental samples.

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