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Browsing by Subject "Saliva"

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    A Proteomic Approach to Elucidate the Changes in Saliva and Serum Proteins of Pigs with Septic and Non-Septic Inflammation
    (MDPI, 2022-06-16) López Martínez, María José; Cerón, José J.; Ortin Bustillo, Alba; Escribano, Damián; Kuleš, Josipa; Beletić, Anđelo; Rubić, Ivana; González Sánchez, Juan Carlos; Mrljak, Vladimir; Martínez Subiela, Silvia; Muñoz Prieto, Alberto; Medicina y Cirugía Animal
    Sepsis is a systemic inflammatory response triggered by an infectious agent and is recognized by the World Health Organization as a global concern, since it is one of the major causes of severe illness in humans and animals. The study of the changes that can occur in saliva and serum in sepsis can contribute to a better understanding of the pathophysiological mechanisms involved in the process and also to discover potential biomarkers that can help in its diagnosis and monitoring. The objective of this study was to characterize the changes that occur in the salivary and serum proteome of pigs with experimentally-induced sepsis. The study included five pigs with sepsis induced by LPS administration and five pigs with non-septic inflammation induced by turpentine for comparative purposes. In saliva, there were eighteen salivary proteins differentially expressed in the sepsis condition and nine in non-septic inflammation. Among these, significant increments in aldolase A and serpin B12 only occurred in the sepsis model. Changes in aldolase A were validated in a larger population of pigs with sepsis due to Streptococcus suis infection. In serum, there were 30 proteins differentially expressed in sepsis group and 26 proteins in the non-septic group, and most of the proteins that changed in both groups were related to non-specific inflammation. In the saliva of the septic animals there were some specific pathways activated, such as the organonitrogen compound metabolic process and lipid transport, whereas, in the serum, one of the main activated pathways was the regulation of protein secretion. Overall, saliva and serum showed different proteome variations in response to septic inflammation and could provide complementary information about the pathophysiological mechanisms occurring in this condition. Additionally, salivary aldolase A could be a potential biomarker of sepsis in pigs that should be confirmed in a larger population
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    Application of a score for evaluation of pain, distress and discomfort in pigs with lameness and prolapses: correlation with saliva biomarkers and severity of the disease
    (2019-10) Contreras Aguilar, María Dolores; Escribano Tortosa, Damián; Martínez Miró, Silvia; López Arjona, Marina; Peres Rubio, Camila; Martínez Subiela, Silvia; Cerón Madrigal, José Joaquín; Tecles Vicente, Fernando; Medicina y Cirugía Animal
    A score system was used to evaluate pain, distress and discomfort in healthy pigs and pigs with two different diseases: lameness and rectal prolapse. In addition, correlations between the results of this score and a panel of salivary biomarkers and severity of disease were studied. This panel included biomarkers of stress (cortisol, salivary alpha-amylase (sAA), total esterase activity (TEA), butyrylcholinesterase (BChE) and lipase (Lip)), im- munity (adenosine deaminase isozymes 1 (ADA1) and 2 (ADA2)) and oxidative status (uric acid (UA), Trolox equivalent antioxidant capacity (TEAC), cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of saliva (FRAS), advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2)). Based on their score, diseased animals were subdivided in those without and with evident pain. Lame pigs and prolapsed pigs with pain showed higher salivary levels of cortisol, sAA, TEA, BChE, ADA1 and ADA2 compared with the healthy pigs. In addition, the prolapsed pigs with pain showed higher levels of FRAS, AOPP and H2O2 compared with the healthy animals. Salivary cortisol, TEA, BChE, ADA isozymes 1 and 2, FRAS and AOPP correlated with the pain score. This five-point pain score system can be easily applied to lame and prolapsed pigs, and salivary bio- markers could be used as an additional tool for pain assessment in those pigs.
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    Automated Spectrophotometric Assays for the Measurement of Ammonia and Bicarbonate in Saliva of Horses: Analytical Validation and Changes in Equine Gastric Ulcer Syndrome (EGUS)
    (MDPI, 2024-02-28) Muñoz-Prieto, Alberto; Llamas-Amor, Eva; Contreras-Aguilar, María Dolores; Ayala de la Peña, Ignacio; Martín Cuervo, María; Cerón, José Joaquín; Hansen, Sanni; Medicina y Cirugía Animal
    Ammonia (NH3) and bicarbonate (HCO3) have been related to gastric ulcers in humans. Ammonia is considered a possible cause of gastric ulcers, whereas bicarbonate has a protective function. The presence of ulcers in the stomach of horses is defined as Equine Gastric Ulcer Syndrome (EGUS), which is a frequent disease in this species, and it has been associated with changes in saliva composition, such as in analytes related to inflammation, immune system and oxidative stress. The objectives of this study were (1) to perform an analytical validation of two automated spectrophotometric assays, one for ammonia and one for bicarbonate, in the horses’ saliva and (2) to evaluate their possible variations with EGUS. Analytical validation of the automated assays for ammonia and bicarbonate in the saliva of horses showed that both assays were precise and accurate. In addition, significantly higher values of ammonia and lower values of bicarbonate were found in the saliva of horses with EGUS compared to healthy horses. It can be concluded that ammonia and bicarbonate can be measured in the saliva of horses and that ammonia increases and bicarbonate decreases in this sample type could be related to the presence of EGUS in this species.
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    Biomarkers of oxidative stress in saliva of sheep: Analytical performance and changes after an experimentally induced stress
    (Elsevier, 2019-04) Peres Rubio, Camila; Contreras-Aguilar, Maria Dolores; Quiles, A.; López-Arjona, Marina; Cerón, J.J.; Martínez-Subiela, Silvia; Hevia, M.L.; Escribano Tortosa, Damián; Tecles, Fernando; Medicina y Cirugía Animal
    Oxidative stress can affect animal's health and the quality of its final products. The oxidative status can be evaluated by the measurement of both oxidant and antioxidant biomarkers. The use of saliva as a sample is preferable to blood, as individuals with limited training can collect it easily and non-invasively with minimal stress to the animal. The aim of this study was to perform an analytical validation of automated assays of the ferric reducing ability of plasma (FRAP), the cupric reducing antioxidant capacity (CUPRAC), the Trolox equivalent antioxidant capacity (TEAC) and uric acid as antioxidant biomarkers and of the advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2) as oxidant biomarkers in saliva samples of sheep, and to evaluate their possible changes after stress induced by shearing. All assays produced acceptable results in the analytical validation, from which it can be concluded that oxidative stress biomarkers such as FRAP, CUPRAC, TEAC, uric acid and AOPP and H2O2 can be measured in sheep saliva. In addition, acute stress due to shearing could produce an oxidative stress response in sheep and subsequently increase antioxidants in order to protect cells from damage.
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    Changes in Calprotectin (S100A8-A9) and Aldolase in the Saliva of Horses with Equine Gastric Ulcer Syndrome
    (MDPI, ) Muñoz-Prieto, Alberto; Contreras-Aguilar, María Dolores; Cerón, José Joaquín; Ayala de la Peña, Ignacio; Martín-Cuervo, María; David Eckersall, Peter; Holm Henriksen, Ida-Marie; Tecles, Fernando; Hansen, Sanni; Medicina y Cirugía Animal
    Equine gastric ulcer syndrome (EGUS) is a highly prevalent disease that affects horses worldwide. Within EGUS, two different forms have been described: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). The associated clinical signs cause detrimental activity performance, reducing the quality of life of animals. Saliva can contain biomarkers for EGUS that could be potentially used as a complementary tool for diagnosis. The objective of this work was to evaluate the measurements of calprotectin (CALP) and aldolase in the saliva of horses as potential biomarkers of EGUS. For this purpose, automated assays for the quantification of these two proteins were analytically validated and applied for detecting EGUS in a total of 131 horses divided into 5 groups: healthy horses, ESGD, EGGD, combined ESGD and EGGD, and horses with other intestinal pathologies. The assays showed good precision and accuracy in analytical validation, and they were able to discriminate between horses with EGUS and healthy horses, especially in the case of CALP, although they did not show significant differences between horses with EGUS and horses with other diseases. In conclusion, salivary CALP and aldolase can be determined in the saliva of horses and further studies are warranted to elucidate the potential of these analytes as biomarkers in EGUS.
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    Changes in Oxidative Status Biomarkers in Saliva and Serum inthe Equine Gastric Ulcer Syndrome and Colic of IntestinalAetiology: A Pilot Study
    (MDPI, 2022-03-07) Contreras-Aguilar, María Dolores; Peres Rubio, Camila; González-Arostegui, Luis Guillermo; Martín-Cuervo, María; Cerón, Jose J.; Ayala de la Peña, Ignacio; Holm Henriksen, Ida-Marie; Jacobsen, Stine; Hansen, Sanni; Medicina y Cirugía Animal
    Changes in the oxidative status of the blood of horses suffering from gastric ulcers and colicof intestinal aetiology (CIE) have been reported. However, saliva can also be a source of biomarkersof oxidative status. Therefore, this study aims to validate automated assays for the measurement ofoxidative status biomarkers (ferric reducing ability of saliva/serum—FRAS/FRAP, cupric reducingantioxidant capacity—CUPRAC, the Trolox equivalent antioxidant capacity—TEAC, uric acid, andadvanced oxidation protein products—AOPP) in the saliva and serum of horses, to assess theirchanges in the different ulcer gastric diseases (squamous—ESGD and glandular—EGGD) and CIE,and to evaluate their relationship with serum amyloid A (SAA), adenosine deaminase (ADA), andthe systemic inflammatory response syndrome (SIRS) status. The assays showed a low imprecisionand good linearity with enough sensitivity in both fluids. In EGGD, higher levels of FRAS, uric acid,and AOPP in saliva were observed compared to the healthy group, correlating with the salivaryADA levels. Horses with CIE showed increases in uric acid concentrations in serum associated with heir SIRS status and outcome of the disease. In conclusion, analytes related to the oxidative statuscan be measured in the saliva and serum from horses by automated assays, and some of them canpotentially be assessed as biomarkers in horses with gastric ulcers and CIE
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    Changes in Proteins in Saliva and Serum in Equine Gastric Ulcer Syndrome Using a Proteomic Approach
    (MDPI, 2022-05-02) Muñoz-Prieto, Alberto; Contreras-Aguilar, Maria Dolores; Cerón, Jose Joaquín; Ayala de la Peña, Ignacio; Martin-Cuervo, María; González-Sánchez, Juan Carlos; Jacobsen, Stine; Kuleš, Josipa; Beletić, Anđelo; Rubić, Ivana; Mrljak, Vladimir; Tecles, Fernando; Hansen, Sanni; Medicina y Cirugía Animal
    Changes in the salivary proteome in 12 horses with the two diseases included in equine gastric ulcer syndrome (EGUS), equine glandular gastric disease (EGGD) (n = 6) and equine squamous gastric disease (ESGD) (n = 6), were evaluated using a high-resolution LC-MS/MS analysis of TMT-labelled peptides and compared to 10 healthy control horses. Serum was also analysed for comparative purposes. The comparison between the horses with EGGD and controls showed significant changes in 10 salivary proteins, whereas 36 salivary proteins were differently abundant between ESGD and control groups. The most upregulated proteins in the case of EGGD were relatedto immune activation whereas, in horses with ESGD, the most significantly changed proteins were associated with squamous cell regulation and growth. Compared to serum, saliva showed a higher number of proteins with significant changes and a different pattern of changes. The proteins identified in our study, in addition to providing new information about the pathophysiological mechanisms in these diseases, could have the potential to be novel biomarkers for the diagnosis or monitoring of EGGD and ESGD.
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    Changes in Saliva Analytes Correlate with Horses’ Behavioural Reactions to An Acute Stressor: A Pilot Study
    (2019-11-18) Contreras Aguilar, María Dolores; Henry, Séverine; Coste, Caroline; Tecles Vicente, Fernando; Escribano Tortosa, Damián; Cerón Madrigal, José Joaquín; Hausberger, Martine; Medicina y Cirugía Animal; Université de Rennes, Université de Normandie, CNRS, Laboratoire Ethologie et humaine - UMR CNRS 6552, Station Biologique
    Acute stress induces an array of behavioural reactions in horses that vary between individuals. Attempts to relate behavioural patterns and physiological responses have not always given clear-cut results. Here, we measured the changes in a panel of salivary components: salivary alpha-amylase (sAA), lipase, total esterase (TEA), butyrylcholinesterase (BChE), adenosine deaminase (ADA), and cortisol, and their potential link with horses’ behaviours after acute stress. Saliva samples were collected in nine riding horses subjected to a test consisting of opening an umbrella. Saliva sampling was obtained at a basal time point in the stall (T1), in the test indoor arena (T2), at a time of stress (T3), and 30 min (T4) and 60 min (T5) later. The horses’ behaviour was recorded at T3 for 1 min. sAA, lipase, TEA, and BChE showed significant changes along time, increasing at T3 for BChE, and decreasing at T4 for sAA and BChE. Butyrylcholinesterase appeared to be the most reliable predictor of behavioural responses, as it correlated with the index of emotionality, of laterality, and the occurrence of alarm signals, while sAA decreased when horses expressed quieter behaviours. These first results bring promising lines for novel, more precise physiological markers of acute stress in horses that can bridge the gap between behaviour and physiology.
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    Changes in saliva analytes in equine acute abdominal disease: a sialochemistry approach
    (2019-06-06) Contreras Aguilar, María Dolores; Escribano Tortosa, Damián; Martínez Subiela, Silvia; Martín Cuervo, María; Lamy, Elsa; Tecles Vicente, Fernando; Cerón Madrigal, José Joaquín; Medicina y Cirugía Animal
    Background: The biochemical components of saliva can change in certain pathologies in horses, for example in acute abdominal disease. The aim of this study was (1) to evaluate if a panel of biochemical analytes usually used in serum can be measured in saliva of horses and (2) to study the possible changes of these biochemical analytes in saliva of horses affected by acute abdominal disease. A panel of 23 analytes was analytically validated in saliva of horses and possible changes in these analytes in a pilot study with six healthy horses and six horses with acute abdominal disease were evaluated. The analytes with significant changes were then evaluated in a larger population of 20 healthy and 37 diseased horses. Results: Seven analytes showed significant increases in the pilot study which were confirmed in the larger population. The analytes which showed significant changes, and their median fold increase and significance shown in the larger population were salivary γ-glutamyl transferase (gGT, 2.3 fold, P = 0.001), creatine kinase (CK, 6.2 fold, P < 0.001), urea (2.3 fold, P = 0.001), total bilirubin (2.6 fold, P < 0.001), total proteins (3.2 fold, P < 0.001), phosphorus (P, 4.5 fold, P < 0.001) and alpha-amylase (sAA, 8.5 fold, P < 0.001). Total proteins, P and sAA showed sensitivities higher than 70% at their optimal cut-off points and a specificity of 100% in differentiating between healthy horses and those with acute abdominal disease. Conclusions: A panel of 23 biochemical analytes can be measured in saliva of horses, where gGT, CK, urea, total bilirubin, total protein, P and sAA levels are raised in horses with acute abdominal disease.
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    Changes in saliva proteins in two conditions of compromised welfare in pigs: T an experimental induced stress by nose snaring and lameness
    (Elsevier, 2019-06-19) Escribano Tortosa, Damián; Horvatić, Anita,; Contreras Aguilar, María Dolores; Guillemin, Nicolas,; Cerón Madrigal, José Joaquín; Tecles Vicente, Fernando; Martínez Miró, Silvia; Eckersall, Peter David; Manteca, Xavier; Mrljak, Vladimir; Producción Animal
    The aim of this study was to identify biological pathways and proteins differentially expressed in saliva of pigs in two conditions of compromised welfare: an acute stress consisting of restraint with a nose snare and in pigs with lameness which is a highly frequent problem in the swine industry. For this purpose, high-resolution quantitative proteomics based on Tandem Mass Tags labelling was used. Four proteins showed significant differences in the conditions of compromised welfare, namely cornulin, the heat shock protein 27 and lactate dehydrogenase (LDH), that showed significant increases, whereas immunoglobulin J chain showed a significant decrease. LDH, which was the protein that showed the highest differences, was selected for validation and clinical evaluation as a diagnostic biomarker. Significant changes in this protein were observed between pigs restrained with a nose snare and pigs with lameness compared with healthy pigs when measured with available commercial assays in a larger population of pigs. In conclusion, this study reports that in situations of compromised welfare on farm, such as acute stress and lameness in pigs, there are changes in proteins and metabolic pathways in saliva, and describes a series of proteins that could potentially be used as biomarkers for both short term acute stress and longer term chronic stress of lameness. These biomarkers would have the advantage of being measured in saliva by a noninvasive and not stressful collection sampling procedure.
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    Changes in salivary proteins can reflect beneficial physiological effects of ejaculation in the dog
    (2021-01-28) Muñoz-Prieto, Alberto; Escribano, Damián; Horvatić, Anita; Contreras-Aguilar, María Dolores; Bernal, Luis; Rubić, Ivana; Cerón, Jose Joaquín; Dabrowski, Roman; Mrljak, Vladimir; Medicina y Cirugía Animal
    The objective of this study was to study the changes in salivary proteins that occur in the dog after the ejaculation process. Saliva samples from eight dogs before and after induced ejaculation were analyzed by proteomic using Tandem Mass Tag (TMT) labeling and LC-MS/MS analysis. A total of 33 salivary proteins showed significant changes after the ejaculation process. The up-regulated proteins that showed changes of higher magnitude were mucin-7 (MUC-7), peroxiredoxin-4 (PRDX4) and galectin-3 (LEGALS3) whereas proteins such as alpha-1-acid glycoprotein (A1G1) and alpha-1B-glycoprotein (A1BG) were the most down-regulated. MUC-7 and PRDX4 expression in saliva after ejaculation could be associated with the protective “environment” created by the organism to exert pr 3o-fertility activities and antioxidants benefits in spermatozoa. Also LEGALS3 increment could be associated with an improvement of wellbeing and could contribute to a positive global effect in the body. Down-regulations of A1G1 and A1GB proteins found in saliva after ejaculation could be associated with a reduction in systemic inflammation. Overall it can be concluded that, changes in proteins in saliva that are produced after ejaculation can reflect a state of increase immune defenses, improvement of antioxidant status and low inflammation.
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    Changes of the salivary and serum proteome in canine hypothyroidism
    (Elsevier, 2024-01) González Arostegui, L. G.; Muñoz Prieto, Alberto; Rubio, C. P.; Cerón, J. J.; Bernal, L.; Rubić, I.; Mrljak, V.; González Sánchez, J. C.; Tvarijonaviciute, Asta; Medicina y Cirugía Animal
    In this study, changes in salivary and serum proteome of dogs with hypothyroidism were studied using tandem mass tags (TMT) labelling and liquid chromatography-mass spectrometry (LC-MS/MS). Saliva and serum proteome from 10 dogs with hypothyroidism were compared with 10 healthy dogs. In saliva, a total of seven proteins showed significant changes between the two groups, being six downregulated and one upregulated, meanwhile, in serum, a total of six proteins showed significant changes, being five downregulated and one upregulated. The altered proteins reflected metabolic and immunologic changes, as well as, skin and coagulation alterations, and these proteins were not affected by gender. One of the proteins that were downregulated in saliva, lactate dehydrognease (LDH), was measured by a spectrophotometric assay in saliva samples from 42 dogs with hypothyroidism, 42 dogs with non-thyroid diseases and 46 healthy dogs. The activity of LDH was lower in the saliva of hypothyroid dogs when compared to non-thyroid diseased dogs and healthy controls. This study indicates that canine hypothyroidism can produce changes in the proteome of saliva and serum. These two sample types showed different variations in their proteins reflecting physiopathological changes that occur in this disease, mainly related to the immune system, metabolism, skin and coagulation. In addition, some of the proteins identified in this study, specially LDH in saliva, should be further explored as potential biomarkers of canine hypothyroidism.
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    Circadian pattern of acute phase proteins in the saliva of growing pigs
    (Elsevier, 2013-05) Gutiérrez Montes, Ana María; Escribano Tortosa, Damián; Fuentes Rubio, María; Cerón Madrigal, José Joaquín; Producción Animal
    The circadian rhythm of the acute phase proteins (APPs) haptoglobin (Hp) and C-reactive protein (CRP) was assessed in saliva samples from 18- and 21-week old pigs. Saliva was collected at 07.00, 11.00, 15.00 and 19.00 h on two consecutive days and the Hp and CRP concentrations were quantified using two species-specific, time-resolved immunofluorometric assays. Salivary Hp levels were significantly higher (P < 0.001) in the morning compared to late afternoon (0.68 and 0.37 lg/mL, respectively) although the magnitude of the difference was much lower than is produced by inflammatory conditions. No significant differences were observed in CRP concentrations. Although the concentration of both APPs was higher in the 21- compared to the 18-week old pigs (P < 0.0001), no differences were observed in the circadian rhythm of these APPs when the two age groups were com- pared. Animal gender did not influence the circadian pattern of either APP, although the mean salivary CRP levels were higher in females (P < 0.05).
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    Correlated color temperature and light intensity: complementary features in non-visual light field
    (Public Library of Science (PLoS), 2021-07-12) Arguelles Prieto, Raquel; Madrid, Juan Antonio; Rol de Lama, María de los Ángeles; Bonmatí Carrión, María de los Ángeles; Fisiología
    An appropriate exposure to the light-dark cycle, with high irradiances during the day and darkness during the night is essential to keep our physiology on time. However, considering the increasing exposure to artificial light at night and its potential harmful effects on health (i.e. chronodisruption and associated health conditions), it is essential to understand the non-visual effects of light in humans. Melatonin suppression is considered the gold standard for nocturnal light effects, and the activation of intrinsically photosensitive retinal ganglion cells (ipRGCs) through the assessment of pupillary light reflex (PLR) has been recently gaining attention. Also, some theoretical models for melatonin suppression and retinal photoreceptors activation have been proposed. Our aim in this study was to determine the influence of correlated color temperature (CCT) on melatonin suppression and PLR, considering two commercial light sources, as well as to explore the possible correlation between both processes. Also, the contribution of irradiance (associated to CCT) was explored through mathematical modelling on a wider range of light sources. For that, melatonin suppression and PLR were experimentally assessed on 16 healthy and young volunteers under two light conditions (warmer, CCT 3000 K; and cooler, CCT 5700 K, at ~5·1018 photons/cm2/sec). Our experimental results yielded greater post-stimulus constriction under the cooler (5700 K, 13.3 ± 1.9%) than under the warmer light (3000 K, 8.7 ± 1.2%) (p < 0.01), although no significant differences were found between both conditions in terms of melatonin suppression. Interestingly, we failed to demonstrate correlation between PLR and melatonin suppression. Although methodological limitations cannot be discarded, this could be due to the existence of different subpopulations of Type 1 ipRGCs differentially contributing to PLR and melatonin suppression, which opens the way for further research on ipRGCs projection in humans. The application of theoretical modelling suggested that CCT should not be considered separately from irradiance when designing nocturnal/diurnal illumination systems. Further experimental studies on wider ranges of CCTs and light intensities are needed to confirm these conclusions.
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    Data for compounds included in the article: "A sensitive immunoassay for the quantitation of Pig-MAP in pig saliva samples"
    (2024-10-05) Piñeiro, M; Matas-Quintanilla, M; Miralles, A; Gutiérrez, AM; Medicina y Cirugía Animal
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    Detection and measurement of alpha-amylase in canine saliva and changes after an experimentally induced sympathetic activation
    (2017-08-22) Contreras Aguilar, María Dolores; Tecles Vicente, Fernando; Martínez Subiela, Silvia; Escribano Tortosa, Damián; Bernal Gambín, Luis Jesús; Cerón Madrigal, José Joaquín; Medicina y Cirugía Animal; Department of Food and Animal Science, School of Veterinary Medicine, Universitat Autònoma de Barcelona
    Background: Salivary alpha-amylase (sAA) is considered a biomarker of sympathetic activation in humans, but there is controversy regarding the existence of sAA in dogs. The hypothesis of this study was that sAA exists in dogs and it could change in situations of sympathetic stimulation. Therefore, the aims of this study were: 1) to demonstrate the presence of alpha-amylase in saliva of dogs by Western-Blot, 2) to validate an spectrophotometric method for the measurement of sAA activity and 3) to evaluate the possible changes in sAA activity after the induction of an ejaculation in dogs which is known to produce a sympathetic activation. Results: Western-Blot demonstrated a band in dog saliva specimens between 60 kDa and 50 kDa, similar to purified sAA. The spectrophotometric assay validated showed an adequate inter- and intra-assay precision, and a high correlation coefficient (r = 0.999) in the linearity under dilution study. sAA median activity significantly increased just after ejaculation compared with just before the ejaculation (2.06-fold, P =0.005). Conclusions: This study demonstrated the existence of alpha-amylase in saliva of dogs and that this enzyme can be measured by a spectrophotometric assay. In addition, results showed that sAA increase after a sympathetic activation and could be potentially used as non-invasive biomarker of sympathetic activity in this species.
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    Development of a time-resolved fluorometry based immunoassay for the determination of canine haptoglobin in various body fluids
    (INRA / EDP Sciences, 2005) Parra Muñoz, María Dolores; Väisänen, Ville; Cerón Madrigal, José Joaquín; Ciencias Sociosanitarias
    A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of haptoglobin (Hp) in canine serum. Haptoglobin was purified from canine acute phase serum by ammonium sulphate precipitation followed by gel filtration. This isolated dog Hp was used as the standard to calibrate the assay. Intra- and inter-assay coefficients of variation of the assay were, respectively, 5.7% and 16.6% at 0.51 mg/mL, 2.4% and 10.6% at 2.1 mg/mL and 10.5% and 11.9% at 32.5 mg/mL. The dilution of serum samples with high Hp concentrations resulted in linear regression equations with R2 of 0.99 and 0.97. A high correlation was found in serum Hp measurements by TR-IFMA and a commercial assay based on peroxidase activity of haemoglobin bound to haptoglobin (R2 = 0.96). The limit of detection for the TR-IFMA method was 0.002 µg/mL. The addition of fresh haemolysate to serum samples did not affect the haptoglobin concentration (P = 0.694). Statistical differences (P < 0.003) were found between healthy dogs and dogs with different pathological processes. In whole blood, Hp concentrations were much lower than in serum but closely related (R2 = 0.84) whereas saliva Hp concentrations were poorly related with serum concentrations (R2 =0.53). However, the concentration of Hp in saliva was significantly (P < 0.039) higher in dogs with pathological processes compared to healthy dogs. The assay sensitivity was adequate to also be applied to whole blood and saliva specimens.
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    Effect of oral antiseptics in reducing SARS-CoV-2 infectivity: Evidence from a randomized double- blind clinical trial
    (Taylor and Francis Group, 2022-07-27) Sánchez Barrueco, Álvaro; Mateos-Moreno, Maria Victoria; Martínez- Beneyto, Yolanda; García-Vázquez, Elisa; Campos González, Alfonso; Zapardiel Ferrero, Javier; Bogoya Castaño, Abel; Alcalá Rueda, Ignacio; Villacampa Aubá, José Miguel; Cenjor Español, Carlos; Moreno-Parrado, Laura; Ausina- Márquez, Verónica; García-Esteban, Sandra; Artacho, Alejandro; López- Labrador, F. Xavier; Mira, Alex; Ferrer, María D.; Medicina
    Background: In vitro studies have shown that several oral antiseptics have virucidal activity against SARS-CoV-2. Thus, mouthwashes have been proposed as an easy to implement strategy to reduce viral transmission. However, there are no data measuring SARS-CoV-2 viability after mouthwashes in vivo. Methods: In this randomized double-blind, five-parallel-group, placebo-controlled clinical trial, SARS-CoV-2 salivary viral load (by quantitative PCR) and its infectious capacity (incubating saliva in cell cultures) have been evaluated before and after four different antiseptic mouthwashes and placebo in 54 COVID-19 patients. Results: Contrary to in vitro evidence, salivary viral load was not affected by any of the four tested mouthwashes. Viral culture indicated that cetylpyridinium chloride (CPC) significantly reduced viral infectivity, but only at one-hour post-mouthwash. Conclusion: These results indicate that some of the mouthwashes currently used to reduce viral infectivity are not efficient in vivo and, furthermore, that this effect is not immediate, generating a false sense of security.
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    Effect of repeated administration of lipopolysaccharide on inflammatory and stress markers in saliva of growing pigs
    (Elsevier, 2014-04-16) Campos, Paulo H.R.F.; Gutiérrez Montes, Ana María; Le Floc’h, Nathalie; Cerón Madrigal, José Joaquín; Merlot, Elodie; Escribano Tortosa, Damián; Producción Animal
    Although saliva could be considered to be an ideal biological sample for evaluation of biomarkers relat- ing to stress and inflammatory responses in pigs, little is known about how these might be influenced by the presence of endotoxaemia. In the present study, the response to repeated administrations of li- popolysaccharide (LPS) was investigated, using a panel of salivary stress markers such as chromogranin A (CgA) and cortisol, as well as inflammatory/immune markers such as haptoglobin (Hp), C-reactive protein (CRP) and immunoglobulin A (IgA). Sixteen growing pigs were adapted to experimental conditions for 3 weeks, after which, 10 of the pigs were selected to receive three doses of LPS at 48 h intervals. Saliva samples were taken from all pigs prior to any LPS administration (baseline) and at time points corresponding to 3 h after each injection of LPS (T1, T2 and T3). Results showed that repeated administration of LPS induced significant elevation of salivary markers of hypothalamic-pituitary-adrenal (cortisol) and immune (Hp, CRP and IgA) activity compared to base- line levels (P < 0.05). However, rectal temperature, CRP and cortisol data suggested that the amplitude of the inflammatory response decreased with successive LPS administrations. Thus, measurement of sal- ivary biomarkers could be a practical tool for evaluating the inflammatory response to endotoxaemia in pigs. In the case of chronic inflammatory states, salivary Hp and IgA might be more sensitive markers than CRP or cortisol.
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    Effect of thermal and chemical treatments used for SARS-COV-2 inactivation in the measurement of saliva analytes.
    (2022-06-08) Lamey, Elsa; Rubio, Camila P; Carreira, Laura; Capela e Silva, Fernando; Martinez Subiela, Silvia; Tecles, Fernando; Lopez Jornet, Pia; Ceron, Jose; Tvarijonaviciute, Asta; Dermatología, Estomatología, Radiología y Medicina Física
    The present study aims to assess the effects of thermal and chemical inactivating procedures, that can be used for SARS-CoV-2 inactivation, on different salivary analytes. SDS–Polyacrylamide Gel Electrophoresis (SDS-PAGE) protein profile and a panel of 25 specific biomarkers of oxidative status, stress, metabolism and tissue damage were evaluated in samples subjected to different treatments: thermal (65 °C or 92 °C) and chemical with detergents [sodium dodecyl sulphate (SDS), Triton X-100 or NP-40]. Salivary SDS-PAGE profile was most affected by heating at 92 °C, with three and two protein bands decreasing and increasing their expression levels, respectively. This treatment also affected the results of several enzymes, with some of them being also affected by heating at 65 °C and incubation with SDS. The use of Triton X-100 or NP-40 resulted in increased values of cortisol, triglycerides and glucose, not affecting the other tested biomarkers. The present results will help researchers and clinicians to select the best protocols to work in safe conditions with saliva, taking into account the target analyte planned to be measured.
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