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Combining microdialysis, NanoLC-MS, and MALDI-TOF/TOF to detect neuropeptides secreted in the crab, Cancer borealis.

ABSTRACT: Microdialysis is a useful technique for sampling neuropeptides in vivo, and decapod crustaceans are important model organisms for studying how these peptides regulate physiological processes. However, to date, no microdialysis procedure has been reported for sampling neuropeptides from crustaceans. Here we report the first application of microdialysis to sample neuropeptides from the hemolymph of the crab, Cancer borealis. Microdialysis probes were implanted into the pericardial region of live crabs, and the resulting dialysates were desalted, concentrated, and analyzed by LC-ESI-QTOF and MALDI-TOF/TOF mass spectrometry. Analysis of in vitro microdialysates of hemolymph revealed more neuropeptides and fewer protein fragments than hemolymph prepared by typical analysis methods. Mass spectra of in vivo dialysates displayed neuropeptides from 10 peptide families, including the RFamide, allatostatin, and orcokinin families. In addition, GAHKNYLRFa, SDRNFLRFa, and TNRNFLRFa were sequenced from hemolymph dialysates. The detection of these neuropeptides in the hemolymph suggests that they are functioning as hormones as well as neuromodulators. In vivo microdialysis offers the capability to further study these and other neuropeptides in crustacean hemolymph, complementing current tissue-based studies and extending our knowledge of hormonal regulation of physiological states.

SUBMITTER: Behrens HL

PROVIDER: S-EPMC2602843 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature

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Combining microdialysis, NanoLC-MS, and MALDI-TOF/TOF to detect neuropeptides secreted in the crab, Cancer borealis.

Behrens Heidi L HL Chen Ruibing R Li Lingjun L

Analytical chemistry 20080814 18

Microdialysis is a useful technique for sampling neuropeptides in vivo, and decapod crustaceans are important model organisms for studying how these peptides regulate physiological processes. However, to date, no microdialysis procedure has been reported for sampling neuropeptides from crustaceans. Here we report the first application of microdialysis to sample neuropeptides from the hemolymph of the crab, Cancer borealis. Microdialysis probes were implanted into the pericardial region of live c ...[more]

PMID: 18700782

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Project description:SARS-CoV-2 has caused a large outbreak since its emergence in December 2019. COVID-19 diagnosis became a priority so as to isolate and treat infected individuals in order to break the contamination chain. Currently, the reference test for COVID-19 diagnosis is the molecular detection (RT-qPCR) of the virus from nasopharyngeal swab (NPS) samples. Although this sensitive and specific test remains the gold standard, it has several limitations, such as the invasive collection method, the relative high cost and the duration of the test. Moreover, the material shortage to perform tests due to the discrepancy between the high demand for tests and the production capacities puts additional constraints on RT-qPCR. Here, we propose a PCR-free method for diagnosing SARS-CoV-2 based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling and machine learning (ML) models from salivary samples. Kinetic saliva samples were collected at enrollment and ten and thirty days later (D0, D10 and D30), to assess the classification performance of the ML models compared to the molecular tests performed on NPS specimens. Spectra were generated using an optimized protocol of saliva collection and successive quality control steps were developed to ensure the reliability of spectra. A total of 360 averaged spectra were included in the study. At D0, the comparison of MS spectra from SARS-CoV-2 positive patients (n = 105) with healthy healthcare controls (n = 51) revealed nine peaks that significantly distinguished the two groups. Among the five ML models tested, support vector machine with linear kernel (SVM-LK) provided the best performance on the training dataset (accuracy = 85.2%, sensitivity = 85.1%, specificity = 85.3%, F1-Score = 85.1%). The application of the SVM-LK model on independent datasets confirmed its performances with 88.9% and 80.8% of correct classification for samples collected at D0 and D30, respectively. Conversely, at D10, the proportion of correct classification had fallen to 64.3%. The analysis of saliva samples by MALDI-TOF MS and ML appears as an interesting supplementary tool for COVID-19 diagnosis, despite the mitigated results obtained for convalescent patients (D10).

Dataset Information

Combining microdialysis, NanoLC-MS, and MALDI-TOF/TOF to detect neuropeptides secreted in the crab, Cancer borealis.

Publications

Combining microdialysis, NanoLC-MS, and MALDI-TOF/TOF to detect neuropeptides secreted in the crab, Cancer borealis.

Similar Datasets

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