Physical-chemical analysis of the cerebrospinal fluid of healthy dogs submitted to different storage periods and temperatures

Rosato, Paula Nunes; Gama, Fernanda Gomes Velasque; Santana, Áureo Evangelista

doi:10.1590/s0103-84782006000600021

Cited by 4 publications

(9 citation statements)

References 13 publications

(11 reference statements)

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“…All 49 metabolites that presented significant differences between the time groups are listed in Table S. 23. in Supporting Information.…”

Section: Effect Of Delay Time Between Sample Collection and Freezing ...mentioning

confidence: 99%

“…[13][14][15][16][17] There are a few reports on the stability of single proteins in CSF connected to AD [18][19][20][21] as well as on the effect of storage temperature (4 or 23 °C) on the proteome profile of CSF by analyzing intact proteins and peptides with surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). 20,22,20 Two studies on canine CSF report the effect of storage temperature on protein concentration, glucose level, pH and enzyme activity 23 as well as the effect of storage temperature, with or without addition of stabilizing agent, on the cell count. 24 Furthermore, guidelines have been published concerning preanalytical factors that should be examined (e.g., cytological examination and glucose concentration measurement) leading to recommendations for proteomic CSF analysis.…”

Section: Introductionmentioning

confidence: 99%

“…Two studies on canine CSF report the effect of storage temperature on protein concentration, glucose level, pH and enzyme activity as well as the effect of storage temperature, with or without addition of stabilizing agent, on the cell count . Furthermore, guidelines have been published concerning preanalytical factors that should be examined (e.g., cytological examination and glucose concentration measurement) leading to recommendations for proteomic CSF analysis. – …”

Section: Introductionmentioning

confidence: 99%

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The Effect of Preanalytical Factors on Stability of the Proteome and Selected Metabolites in Cerebrospinal Fluid (CSF)

et al. 2009

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To standardize the use of cerebrospinal fluid (CSF) for biomarker research, a set of stability studies have been performed on porcine samples to investigate the influence of common sample handling procedures on proteins, peptides, metabolites and free amino acids. This study focuses at the effect on proteins and peptides, analyzed by applying label-free quantitation using microfluidics nanoscale liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (chipLC-MS) as well as matrix-assisted laser desorption ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FT-ICR-MS) and Orbitrap LC-MS/MS to trypsin-digested CSF samples. The factors assessed were a 30 or 120 min time delay at room temperature before storage at -80 degrees C after the collection of CSF in order to mimic potential delays in the clinic (delayed storage), storage at 4 degrees C after trypsin digestion to mimic the time that samples remain in the cooled autosampler of the analyzer, and repeated freeze-thaw cycles to mimic storage and handling procedures in the laboratory. The delayed storage factor was also analyzed by gas chromatography mass spectrometry (GC-MS) and liquid chromatography mass spectrometry (LC-MS) for changes of metabolites and free amino acids, respectively. Our results show that repeated freeze/thawing introduced changes in transthyretin peptide levels. The trypsin digested samples left at 4 degrees C in the autosampler showed a time-dependent decrease of peak areas for peptides from prostaglandin D-synthase and serotransferrin. Delayed storage of CSF led to changes in prostaglandin D-synthase derived peptides as well as to increased levels of certain amino acids and metabolites. The changes of metabolites, amino acids and proteins in the delayed storage study appear to be related to remaining white blood cells. Our recommendations are to centrifuge CSF samples immediately after collection to remove white blood cells, aliquot, and then snap-freeze the supernatant in liquid nitrogen for storage at -80 degrees C. Preferably samples should not be left in the autosampler for more than 24 h and freeze/thaw cycles should be avoided if at all possible.

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“…All 49 metabolites that presented significant differences between the time groups are listed in Table S. 23. in Supporting Information.…”

Section: Effect Of Delay Time Between Sample Collection and Freezing ...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The Effect of Preanalytical Factors on Stability of the Proteome and Selected Metabolites in Cerebrospinal Fluid (CSF)

et al. 2009

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“…Cerebrospinal fluid (CSF) is a plasma ultrafiltrate produced by the choroid plexuses and contains few nucleated cells and a low protein concentration (Christman 1992, Di Terlizzi & Platt 2009. Protein concentration and enzyme activity are relatively stable with time (Rosato et al 2006) but differential cell percentages and cell morphology deteriorate in a time-dependent manner in dogs and humans after CSF is collected (Bienzle et al 2000, Fry et al 2006, De Graaf et al 2011. Samples with a protein content of ≥50 mg/dL are less susceptible to cell deterioration (Fry et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Diagnostic utility of canine cerebrospinal fluid preserved by 10% buffered formalin

Sancho

Adamantos

Holmes

2021

J of Small Animal Practice

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To examine the diagnostic utility of canine cerebrospinal fluid samples collected into tubes containing ethylenediaminetetraacetic acid (EDTA) with and without the addition of 10% buffered formalin analysed within 6 to 20 hours.Materials and MethOds: Inclusion criteria were dogs presenting to a referral hospital with neurological signs and having cerebrospinal fluid analysis performed. Samples were submitted to an external laboratory in tubes containing ethylenediaminetetraacetic acid as paired-samples; one with the addition of one drop of 10% buffered formalin and the other without formalin. Cytology report, total nucleated cell counts, and protein concentration were reviewed. Three different categories of cell preservation were defined: diagnostic, non-diagnostic and unclassified. Each sample was included in one of these categories depending on cytological features, and the diagnostic quality between samples was compared. Samples were further divided in two groups depending on protein concentration using 50 mg/dL as cut-off value and the diagnostic quality between samples was compared. results: 254 samples from 127 dogs were included. 47% of samples without formalin were non-diagnostic, 46% diagnostic and 6% unclassified. In the formalin group, 2% samples were non-diagnostic, 92% diagnostic and 6% unclassified. Samples with formalin preservation were statistically more likely to be diagnostic than samples without formalin preservation. In both protein groups (≥50 and <50 mg/ dL) formalin samples were statistically more likely to be diagnostic as well.clinical significance: The addition of one drop of 10% formalin is a simple, widely available method which can help to improve the accuracy of cytological assessment in canine cerebrospinal fluid by preserving cellular morphology when analysis is performed within 6 to 20 hours.

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“…Cerebrospinal fluid (CSF), formed by plasma ultrafiltration and active transport mechanism of substances across the blood–brain barrier, circulates in the subarachnoid space of the entire central nervous system (CNS) [ 1 – 5 ]. Besides its protective function toward the CNS, CSF serves to transport neurotransmitters, maintain ionic balance, and excrete cerebral metabolites [ 3 – 6 ].…”

Section: Introductionmentioning

confidence: 99%

Analysis of cerebrospinal fluid from cattle with central nervous system disorders after storage for 24 hours with autologous serum

et al. 2015

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BackgroundWe compared the changes in cell morphology, total and differential cell counts between cerebrospinal fluid (CSF) samples analyzed within an hour of collection (fresh sample) and after the addition of autologous serum and storage for 24 h (stored sample) in 27 cattle with central nervous system disorders.ResultsThere was a positive linear correlation between total and differential cell counts in the fresh and the stored samples. Cell morphology was preserved in all stored samples, except for increased vacuolization of mononuclear cells and cleaved nuclei of some small mononuclear cells. In the stored CSF samples, the total nucleated cell count and monocyte percentage were decreased (P = 0.01; P = 0.03), while the lymphocyte percentage was increased (P = 0.04). Mononuclear pleocytosis diagnosed in 20 fresh samples was cytologically confirmed in 12 of the 20 stored samples. In the remaining eight stored samples, the number of total nucleated cells was within the normal range. Neutrophilic pleocytosis was confirmed in all seven stored samples.The overall agreement rate between cytologic interpretation of the fresh and the stored CSF samples was 70 % (100 % for neutrophilic pleocytosis and 60 % for mononuclear pleocytosis).ConclusionsAdding 11 % of autologous serum to CSF samples might allow delayed analysis with a good agreement rate for CSF cytological interpretation. Caution is nonetheless warranted, as animal age, anamnesis, and neurological presentation need to be considered when interpreting stored CSF without pleocytosis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-015-0502-x) contains supplementary material, which is available to authorized users.

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Physical-chemical analysis of the cerebrospinal fluid of healthy dogs submitted to different storage periods and temperatures

Abstract: Disorders involving the nervous system have a great importance in veterinary medicine because they have a high incidence and few auxiliary tools for their diagnostic, prognostic and evaluation of the employed therapy. Today

Cited by 4 publications

References 13 publications

The Effect of Preanalytical Factors on Stability of the Proteome and Selected Metabolites in Cerebrospinal Fluid (CSF)

The Effect of Preanalytical Factors on Stability of the Proteome and Selected Metabolites in Cerebrospinal Fluid (CSF)

Diagnostic utility of canine cerebrospinal fluid preserved by 10% buffered formalin

Analysis of cerebrospinal fluid from cattle with central nervous system disorders after storage for 24 hours with autologous serum

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