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Piglets Are Vulnerable to Intestinal Inflammation and Infection Because of Expression of Low‐Affinity Jejunal Alkaline Phosphatase Activities

2017· article· en· W2938800593 on OpenAlexaffabout
N. Burello, Anna‐Kate Shoveller

Bibliographic record

VenueThe FASEB Journal · 2017
Typearticle
Languageen
FieldMedicine
TopicAlkaline Phosphatase Research Studies
Canadian institutionsUniversity of Guelph
Fundersnot available
KeywordsAlkaline phosphataseInflammationGastroenterologyJejunumInternal medicineChemistryMicrobiologyMedicineBiochemistryBiologyEnzyme

Abstract

fetched live from OpenAlex

Intestinal alkaline phosphatase (IAP) plays a pivotal role in protecting gut health through catalyzing the hydrolytic dephosphorylation of the lipid A moiety of endotoxin lipopolysaccharides (LPS) and other emblematic members of pathogen‐associated‐molecular patterns (PAMPs) such as ATP, ultimately detoxifying endotoxins and preventing gut dysbiosis and disorders. Characterization of IAP hydrolytic activities, including enzyme affinity ( K m ) and the maximum enzyme activity ( V max ), has been reported in pigs and other mammalian species by using the chromogenic synthetic substrate P ‐nitrophenyl phosphate (pNPP) rather than using relevant physiological substrates such as ATP and LPS under the physiological pH conditions. This study was conducted to determine kinetics of young porcine jejunal alkaline phosphatase in hydrolyzing the dephosphorylation of ATP, the lipid A moiety of LPS and pNPP at pH 7.4 and 37 °C. Jejunal samples were collected from four 10‐day old Yorkshire pigs and were pulverized to be homogenous under liquid N 2 . The LPS kinetic experiments were conducted with 11 gradient concentrations of LPS, isolated from E. coli 0111:B4, ranging 0 – 5 mg/mL in incubation media. The ATP kinetic experiments were conducted with 11 gradient concentrations of ATP, ranging 0 – 5 mM in incubation media. Free phosphorus (Pi) release was measured by using the PiColorLock detection reagent (Innova Biosciences). Contributions of the non‐specific background Pi associated with ATP and LPS in the time course and the kinetic experiments were corrected. The pNPP kinetic experiments were conducted with 16 gradient concentrations of pNPP, ranging 0 – 6 mM in incubation media. All enzymatic incubations were conducted with 19 – 27 μg of the homogenized porcine jejunal protein. The kinetics (parameter estimates ± SE, P < 0.05) of young porcine jejunal alkaline phosphatase activities were obtained, including V max values of 2.5 ± 0.4 ug/mg protein·min (n = 11), 1.1 ± 0.2 ug/mg protein·min (n = 11), and 0.12 ± 0. 01 umol/mg protein·min (n = 16); and K m values of 1.255 ± 0.498 mM (n = 11), 2.388 ± 0.794 mg/mL (n = 11), and 0.145 ± 0.036 mM (n = 16), for the hydrolysis of ATP, LPS and pNPP, respectively. Our results reveal the young porcine IAP K m value 41 times higher in comparison with reported K m value of human IAP in dephosphorylating ATP, 3 times higher when compared with reported human IAP dephosphorylating LPS, and 5 times higher when compared with reported human IAP hydrolyzing pNPP. Results of this study show that young porcine jejunal IAP activity affinity is several fold lower in detoxifying endotoxin LPS the other emblematic PAMPs member ATP in comparison with humans, suggesting that young pigs are susceptible to gut inflammation and infection. Support or Funding Information NSERC Discovery Program of Canada and Metagen Enzyme Corporation

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.001
metaresearch head score (Gemma)0.003
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.019
Threshold uncertainty score0.585

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0010.003
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0010.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.001
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.036
GPT teacher head0.329
Teacher spread0.293 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one teacher head, not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

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Citations1
Published2017
Admission routes2
Has abstractyes

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