scholarly journals Development and Validation of a Fecal Extraction Procedure for the Assessment of Multiple Fecal Biomarkers of Intestinal Inflammation (P13-025-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Erin Lewis ◽  
Weimin Guo ◽  
Lijun Li ◽  
Dayong Wu ◽  
Gerald Combs ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Agricultural Research ◽  
Sandwich Elisa ◽  
Extraction Procedure ◽  
Predictive Biomarkers ◽  
Healthy Adults ◽  
Tween 20 ◽  
Fecal Samples ◽  
Single Extraction ◽  
Fecal Biomarkers

Abstract Objectives Fecal biomarkers have emerged as an important tool to assess intestinal inflammation and permeability. Commonly measured biomarkers include calprotectin (CP), myeloperoxidase (MPO), alpha-1-antitrypsin (AAT) and neopterin (NEO). We sought to develop a simple, fast and cost-effective single extraction procedure for use in determining all four biomarkers of interest. The applicability and sensitivity of this procedure for use in healthy adults was examined. Methods Sample extraction buffers and methods including sample weight, dilution, homogenization and centrifugation were all considered in the development of a single extraction procedure. An extraction buffer that included phosphate-buffered saline, phenylmethylsulfonyl fluoride, bovine serum albumin and Tween 20 was used to extract fecal samples. To assess the applicability and sensitivity of the single extraction procedure, concentrations of CP, MPO, AAT and NEO were measured using commercially available sandwich ELISA kits, according to manufacturer's instructions. Results CP, MPO and AAT concentrations were measured in fecal samples of healthy adults (aged 50–80 years, n = 85) and found to be comparable to findings of previously published studies in healthy populations. Mean concentrations of CP and AAT were 3.6 ± 3.8 µg/g of wet weight (range 0.14–18.0 µg/g) and 2.3 ± 0.73 µg/g (range 0.76–5.2 µg/g), respectively. Mean fecal MPO concentrations were 135 ± 24 ng/g (range 3–1290 ng/g). NEO concentrations were examined in a subset of healthy adults (n = 10), with mean concentrations of 18 ± 1 ng/g (range 17–20 ng/g). Conclusions We demonstrated the efficacy of a single extraction procedure used to assess multiple fecal biomarkers of intestinal inflammation. This simple, fast and inexpensive extraction method will facilitate the determination of multiple fecal biomarkers which is critical in validating their use as clinical or predictive biomarkers of intestinal inflammation. Funding Sources Supported by the U.S. Department of Agriculture – Agricultural Research Service (ARS), under Agreement No. 58–1950-4–003, the Bill & Melinda Gates Foundation and Canadian Institutes of Health Research Postdoctoral Fellowship.

Evaluation of Human Fecal Calprotectin Detection Kits and Their Potential use as Non-Invasive Intestinal Biomarker of Infectious Enterocolitis in Pigs

2021 ◽  
Author(s):  
Jéssica Barbosa ◽  
Lucas Rodrigues ◽  
Daniel Columbus ◽  
Juan Aguirre ◽  
John Harding ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Point Of Care ◽  
Sandwich Elisa ◽  
Fecal Calprotectin ◽  
Field Application ◽  
Dipstick Test ◽  
Infectious Colitis ◽  
Fecal Samples ◽  
Non Invasive ◽  
Infectious Enterocolitis

Abstract Background: Fecal calprotectin is largely applied as a non-invasive intestinal inflammation biomarker in human medicine. Previous studies in pigs investigated the levels of fecal calprotectin in healthy animals only. Thus, there is a knowledge gap regarding its application during infectious diarrhea. This study investigated the usefulness of fecal calprotectin as a biomarker of intestinal inflammation in Brachyspira hyodysenteriae and Salmonella Typhimurium infected pigs. Results: Fecal samples from pigs with colitis (n=18) were collected from animals experimentally inoculated with B. hyodysenteriae G44 or from sham-inoculated controls. Fecal samples from pigs with enteritis (n=14) were collected from animals inoculated with Salmonella enterica serovar Typhimurium or from sham-inoculated controls. For both groups, fecal samples were scored as: 0 = normal; 1 = soft, wet cement; 2 = watery feces; 3 = mucoid diarrhea; and 4 = bloody diarrhea. Fecal calprotectin levels were assayed using a sandwich ELISA, a turbidimetric immunoassay and a point-of-care dipstick test. Fecal calprotectin levels were greater in colitis samples scoring 4 versus ≤ 4 using ELISA, and in feces scoring 3 and 4 versus ≤ 1 using immunoturbidimetry (P < 0.05). No differences were found in calprotectin concentration among fecal scores for enteritis samples, regardless of the assay used. All samples were found below detection limits using the dipstick method.Conclusions: Fecal calprotectin is a potential non-invasive biomarker of infectious colitis, but it is not suitable for detection of enteritis. While practical, the use of commercially available human presents sensitivity limitations. Further studies are needed to validate the field application of calprotectin as a marker.

scholarly journals Experimental infectious challenge in pigs leads to elevated fecal calprotectin levels following colitis, but not enteritis

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Jéssica A. Barbosa ◽  
Lucas A. Rodrigues ◽  
Daniel A. Columbus ◽  
Juan C. P. Aguirre ◽  
John C. S. Harding ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Point Of Care ◽  
Sandwich Elisa ◽  
Fecal Calprotectin ◽  
Field Application ◽  
Dipstick Test ◽  
Fecal Samples ◽  
Brachyspira Hyodysenteriae ◽  
Non Invasive ◽  
Serovar Typhimurium

Abstract Background Fecal calprotectin is largely applied as a non-invasive intestinal inflammation biomarker in human medicine. Previous studies in pigs investigated the levels of fecal calprotectin in healthy animals only. Thus, there is a knowledge gap regarding its application during infectious diarrhea. This study investigated the usefulness of fecal calprotectin as a biomarker of intestinal inflammation in Brachyspira hyodysenteriae and Salmonella Typhimurium infected pigs. Results Fecal samples from pigs with colitis (n = 18) were collected from animals experimentally inoculated with B. hyodysenteriae (n = 8) or from sham-inoculated controls (n = 3). Fecal samples from pigs with enteritis (n = 14) were collected from animals inoculated with Salmonella enterica serovar Typhimurium (n = 8) or from sham-inoculated controls (n = 4). For both groups, fecal samples were scored as: 0 = normal; 1 = soft, wet cement; 2 = watery feces; 3 = mucoid diarrhea; and 4 = bloody diarrhea. Fecal calprotectin levels were assayed using a sandwich ELISA, a turbidimetric immunoassay and a point-of-care dipstick test. Fecal calprotectin levels were greater in colitis samples scoring 4 versus ≤ 4 using ELISA, and in feces scoring 3 and 4 versus ≤ 1 using immunoturbidimetry (P < 0.05). No differences were found in calprotectin concentration among fecal scores for enteritis samples, regardless of the assay used. All samples were found below detection limits using the dipstick method. Conclusions Fecal calprotectin levels are increased following the development of colitis, but do not significantly change due to enteritis. While practical, the use of commercially available human kits present sensitivity limitations. Further studies are needed to validate the field application of calprotectin as a marker of intestinal inflammation.

Quantitation and Confirmation of Chloramphenicol, Florfenicol, and Nitrofuran Metabolites in Honey Using LC-MS/MS

2018 ◽  
Vol 101 (3) ◽  
pp. 897-903 ◽  
Author(s):  
Brian T Veach ◽  
Renea Anglin ◽  
Thilak K Mudalige ◽  
Paula J Barnes
Keyword(s):  
Extraction Procedure ◽  
Extraction Methods ◽  
Liquid Extraction ◽  
Tandem Ms ◽  
Robust Method ◽  
Calibration Standards ◽  
Single Extraction ◽  
Liquid Liquid Extraction ◽  
Lc Tandem Ms ◽  
Nitrofuran Metabolites

Abstract This paper describes a rapid and robust method utilizing a single liquid–liquid extraction for the quantitation and confirmation of chloramphenicol, florfenicol, and nitrofuran metabolites in honey. This methodology combines two previous extraction methods into a single extraction procedure and utilizes matrix-matched calibration standards and stable isotopically labeled standards to improve quantitation. The combined extraction procedure reduces the average extraction time by &gt;50% when compared with previously used procedures. The drug residues were determined using two separate LC-tandem MS conditions. Validation of all the analytes was performed, with average quantitation ranging from 92 to 105% for all analytes and the RSDs for all analytes being ≤12%.

scholarly journals Removal of Direct Yellow 12 from Water Samples by Cloud Point Extraction Using Triton X-100 as Nonionic Surfactant

2011 ◽  
Vol 8 (4) ◽  
pp. 1588-1595 ◽  
Author(s):  
Shahnaz Abedi ◽  
Farzin Nekouei
Keyword(s):  
Nonionic Surfactant ◽  
Cloud Point ◽  
Salt Concentration ◽  
Water Samples ◽  
Cloud Point Extraction ◽  
Extraction Procedure ◽  
Optimum Conditions ◽  
Single Extraction ◽  
Triton X

A surfactant mediated cloud point extraction (CPE) procedure has been developed to remove color from wastewater containing direct yellow 12 (Chrysophenine G), using triton x-100 (TX-100) as nonionic surfactant. The effects of the concentration of the surfactant, pH, temperature and salt concentration on the different concentration of dye have been studied and optimum conditions were obtained for the removal of direct yellow 12 (DY 12). The concentration of DY 12 in the dilute phase was measured using UV-Vis spectrophotometer. It was found that the separation of phases was complete and the recovery of DY 12 was very effective in the presence of NaCl as an electrolyte. The results showed that up to 600 mg L−1of DY 12 can quantitatively be removed (>96%) by Cloud point extraction procedure in a single extraction using optimum conditions.

Sa2051 Safety and Tolerability of a 2-Month Course of Lactobacillus Reuteri and Effect on Circulating and Fecal Biomarkers in Healthy Adults

2012 ◽  
Vol 142 (5) ◽  
pp. S-390
Author(s):  
Nisha Mangalat ◽  
Yuying Liu ◽  
Nicole Y. Fatheree ◽  
Melissa R. Van Arsdall ◽  
Michael J. Ferris ◽  
...  
Keyword(s):  
Lactobacillus Reuteri ◽  
Healthy Adults ◽  
Fecal Biomarkers

scholarly journals Surrogate Fecal Biomarkers in Inflammatory Bowel Disease: Rivals or Complementary Tools of Fecal Calprotectin?

2017 ◽  
Vol 24 (1) ◽  
pp. 78-92 ◽  
Author(s):  
Mirko Di Ruscio ◽  
Filippo Vernia ◽  
Antonio Ciccone ◽  
Giuseppe Frieri ◽  
Giovanni Latella
Keyword(s):  
Inflammatory Bowel Disease ◽  
Bowel Disease ◽  
Intestinal Inflammation ◽  
Fecal Calprotectin ◽  
Response To Therapy ◽  
Polymorphonuclear Neutrophil ◽  
Cochrane Library ◽  
Clinical Relapse ◽  
Inflammatory Bowel ◽  
Fecal Biomarkers

Abstract Background Current noninvasive methods for assessing intestinal inflammation in inflammatory bowel disease (IBD) remain unsatisfactory. Along with C-reactive protein and erythrocyte sedimentation rate, fecal calprotectin (FC) is the standard test for assessing IBD activity, even though its specificity and accuracy are not optimal and it lacks a validated cutoff. Over the past few decades, several fecal markers released from intestinal inflammatory cells have been investigated in IBD; they are the subject of this systematic review. Methods A systematic electronic search of the English literature up to April 2017 was performed using Medline and the Cochrane Library. Only papers written in English that analyzed fecal biomarkers in IBD were included. In vitro studies, animal studies, studies on blood/serum samples, and studies analyzing FC or fecal lactoferrin alone were excluded. Results Out of 1023 citations, 125 eligible studies were identified. Data were grouped according to each fecal marker including S100A12, high-mobility group box 1, neopterin, polymorphonuclear neutrophil elastase, fecal hemoglobin, alpha1-antitrypsin, human neutrophil peptides, neutrophil gelatinase-associated lipocalin, chitinase 3-like-1, matrix metalloproteinase 9, lysozyme, M2-pyruvate kinase, myeloperoxidase, fecal eosinophil proteins, human beta-defensin-2, and beta-glucuronidase. Some of these markers showed a high sensitivity and specificity and correlated with disease activity, response to therapy, and mucosal healing. Furthermore, they showed a potential utility in the prediction of clinical relapse. Conclusions Several fecal biomarkers have the potential to become useful tools complementing FC in IBD diagnosis and monitoring. However, wide variability in their accuracy in assessment of intestinal inflammation suggests the need for further studies.

scholarly journals Exosomes in Intestinal Inflammation

2021 ◽  
Vol 12 ◽  
Author(s):  
Kanchana K. Ayyar ◽  
Alan C. Moss
Keyword(s):  
Inflammatory Bowel Disease ◽  
Monoclonal Antibodies ◽  
Breast Milk ◽  
Nucleic Acids ◽  
Bowel Disease ◽  
Intestinal Inflammation ◽  
Fecal Samples ◽  
Inflammatory Bowel

Exosomes are 30–150 nm sized vesicles released by a variety of cells, and are found in most physiological compartments (feces, blood, urine, saliva, breast milk). They can contain different cargo, including nucleic acids, proteins and lipids. In Inflammatory Bowel Disease (IBD), a distinct exosome profile can be detected in blood and fecal samples. In addition, circulating exosomes can carry targets on their surface for monoclonal antibodies used as IBD therapy. This review aims to understand the exosome profile in humans and other mammals, the cargo contained in them, the effect of exosomes on the gut, and the application of exosomes in IBD therapy.

scholarly journals Ferrous Sulfate-Iron (60 mg/d) Does Not Increase Risks to Malarial Infectivity, Pathogenic Bacterial Proliferation or Other Adverse Effects in Non-Anemic Healthy Adults

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1820-1820
Author(s):  
Erin Lewis ◽  
Athar Chishti ◽  
Christopher Schwake ◽  
John Leong ◽  
Marcia Osburne ◽  
...  
Keyword(s):  
Adverse Events ◽  
Ferrous Sulfate ◽  
Agricultural Research ◽  
Ex Vivo ◽  
Iron Status ◽  
Bacterial Species ◽  
Healthy Adults ◽  
Klebsiella Pneumonia ◽  
Bacterial Proliferation ◽  
Markers Of Inflammation

Abstract Objectives A double-blind, placebo-controlled, randomized clinical trial was conducted to determine the effects of different modalities of supplementation with ferrous sulfate (FeSO4)-iron on the safety profiles of non-anemic, healthy adults. Methods Eligible volunteers (194) were screened for general health; 108 were enrolled and randomized to four treatment groups (n = 27): a placebo (corn starch), daily doses of FeSO4 providing 60 mg Fe/d with or without a 15-component multiple micronutrient powder (UNICEF), or single weekly doses of FeSO4 providing 420 mg Fe. Treatment materials were provided in opaque capsules that were physically similar. The following safety profile indicators were assessed both before and after 28 days of intervention: infectivity of subjects’ RBCs to Plasmodium falciparum determined ex vivo; proliferation potential of 5 species of pathogenic bacteria (Escherichia coli, Acinetobacter baumannii, Klebsiella pneumonia, Staphylococcus aureus, and Salmonella typhimurium) in subjects’ sera determined ex vivo; markers of inflammation (fecal calprotectin; serum myeloperoxidase, alpha-1 antitrypsin and TNFα); parameters of iron status (serum Fe, Hb, transferrin, % transferrin saturation, and ferritin). Adverse events were assessed by the study physician. Results Results showed that no treatment affected: i) 28-day changes in either fasting or post-prandial levels of any parameter of Fe status; ii) 28-day changes in markers of gut or systemic inflammation; iii) RBC susceptibility to P. falciparum; iv) ex vivo proliferation of and bacterial species except A. baumannii, for which the 28-day change in post-prandial peak growth rate index was less than that of the placebo treatment group; or v) subject reported adverse events, which were minimal. Conclusions These results indicate that administration of FeSO4 in these modalities to non-anemic, healthy adults are well tolerated and do not increase their risks to malarial infection and bacterial proliferation. Funding Sources The Bill and Melinda Gates Foundation U.S. Department of Agriculture – Agricultural Research Service (ARS), under Agreement No. #58–1950-4–003.

scholarly journals One-Single Extraction Procedure for the Simultaneous Determination of a Wide Range of Polar and Nonpolar Organic Contaminants in Seawater

2018 ◽  
Vol 5 ◽  
Author(s):  
Vincent Fauvelle ◽  
Javier Castro-Jiménez ◽  
Natascha Schmidt ◽  
Benoit Carlez ◽  
Christos Panagiotopoulos ◽  
...  
Keyword(s):  
Simultaneous Determination ◽  
Organic Contaminants ◽  
Extraction Procedure ◽  
Single Extraction ◽  
Wide Range ◽  
Nonpolar Organic

scholarly journals Resistance to barley yellow dwarf luteovirus in Aegilops species

1994 ◽  
Vol 74 (3) ◽  
pp. 631-634 ◽  
Author(s):  
K. M. Makkouk ◽  
W. Ghulam ◽  
A. Comeau
Keyword(s):  
Triticum Aestivum ◽  
Agricultural Research ◽  
Sandwich Elisa ◽  
Yellow Dwarf ◽  
Aegilops Species ◽  
Elite Group ◽  
Barley Yellow Dwarf ◽  
Tissue Blot Immunoassay ◽  
Das Elisa ◽  
Barley Yellow Dwarf Luteovirus

One thousand and ninety-seven Aegilops accessions were evaluated for their reaction to a PAV serotype of barley yellow dwarf luteovirus (BYDV). The accessions tested belong to the species bicornis, biuncialis, caudata, crassa, columnaris, comosa, cylindrica, kotschyi, longissima, mutica, neglecta (= triaristata 4 ×), ovata, peregrina, searsii, sharonensis, speltoides, tauschii (= squarrosa), triuncialis, umbellulata, uniaristata, vavilovii and ventricosa. The first evaluation of virus levels in the different accessions was conducted at International Center for Agricultural Research in the Dry Areas (ICARDA), Aleppo, Syria, using double antibody sandwich ELISA (DAS-ELISA). Accession reaction ranged from highly resistant to highly susceptible. Thirty-eight Aegilops accessions resistant at ICARDA, were evaluated at Sainte-Foy, Quebec, Canada, by tissue-blot immunoassay. Diversity of response to BYDV infection was again observed in this elite group. Seven accessions belonging to the species biuncialis, caudata, neglecta and triuncialis were highly BYDV resistant at both locations; five of these originated from Bulgaria. Key words: Introgression, interspecific, Triticum aestivum, BYDV, ELISA, immunoassay, tissue blot

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