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Introduction: Long COVID, or post-acute sequelae of SARS-CoV-2 infection (PASC), manifests as persistent and often debilitating symptoms enduring well beyond the initial COVID-19 infection. This disease is especially worrying in children since it can seriously alter their development. Presently, a specific diagnostic test or definitive biomarker set for confirming long COVID is lacking, relying instead on the protracted presence of symptoms post-acute infection. Methods: We measured the levels of 13 biomarkers in 105 saliva samples (49 from children with long COVID and 56 controls), and the Pearson correlation coefficient was used to analyse the correlations between the levels of the different salivary biomarkers. Multivariate logistic regression analyses were performed to determine which of the 13 analysed salivary biomarkers were useful to discriminate between children with long COVID and controls, as well as between children with mild and severe long COVID symptoms. Results: Pediatric long COVID exhibited increased oxidant biomarkers and decreased antioxidant, immune response, and stress-related biomarkers. Correlation analyses unveiled distinct patterns between biomarkers in long COVID and controls. Notably, a multivariate logistic regression pinpointed TOS, ADA2, total proteins, and AOPP as pivotal variables, culminating in a remarkably accurate predictive model distinguishing long COVID from controls. Furthermore, total proteins and ADA1 were instrumental in discerning between mild and severe long COVID symptoms. Discussion: This research sheds light on the potential clinical utility of salivary biomarkers in diagnosing and categorizing the severity of pediatric long COVID. It also lays the groundwork for future investigations aimed at unravelling the prognostic value of these biomarkers in predicting the trajectory of long COVID in affected individuals.
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Biomarcadores , COVID-19 , SARS-CoV-2 , Saliva , Índice de Severidad de la Enfermedad , Humanos , COVID-19/diagnóstico , Saliva/química , Saliva/virología , Biomarcadores/análisis , Niño , Femenino , Masculino , SARS-CoV-2/aislamiento & purificación , Preescolar , AdolescenteRESUMEN
The aim of this study was to evaluate the changes in the serum and salivary inflammatory markers induced by Diabetes mellitus (DM) in dogs and to assess the possible confounding effect of gingivitis. A panel of 13 cytokines was measured in the serum and saliva of dogs diagnosed with DM and compared with healthy dogs without gingivitis (control group 1; CG1) and dogs with gingivitis but otherwise healthy (control group 2; CG2). The results of the present study showed statistically significantly higher levels of IL-8, KC-like and MCP1 in the serum of dogs with DM compared to CG1 dogs. In the case of saliva, the DM group presented statistically higher GM-CSF, IL6, IL15, and MCP1 levels compared to CG1, and lower KC-like chemokine compared to CG2. Finally, gingivitis produced changes in saliva, with salivary levels of GM-CSF, IL-6, IL-7, IL-15, IP-10, KC-like, IL-10, IL-18, MCP1, TNFα being statistically significantly higher in the saliva of CG2 dogs compared to CG1. The results of the present study indicate that dogs with DM have altered cytokine levels in serum and saliva compared to healthy dogs. In addition, this study highlights the importance of taking oral health into account when determining cytokines in dogs, as gingivitis can significantly alter their concentrations. .
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Diabetes Mellitus , Enfermedades de los Perros , Gingivitis , Perros , Animales , Factor Estimulante de Colonias de Granulocitos y Macrófagos , Saliva , Citocinas , Gingivitis/veterinaria , Diabetes Mellitus/veterinariaRESUMEN
BACKGROUND AND OBJECTIVES: Oral fluid (OF) is an easy-to-collect, inexpensive, fast and non-invasive sample to characterize health and welfare status of the pig. However, further standardisation of the collection methods is needed in order to use it regularly in veterinary practice. Cotton ropes are routinely used to collect OF for pathogen detection but they may not be optimal for biomarker analysis due to sample contamination. This study compared two methods (cotton ropes and sponges) to collect porcine OF for biomarker analysis. A panel of 11 biomarkers of stress, inflammation, sepsis, immunity, redox status and general homeostasis was studied. MATERIALS AND METHODS: Eighteen farrow-to-finish pig farms were included in the study. In each farm, three (for sponges) or four pens of pigs (for ropes) were sampled at four age categories: the week after weaning (5 weeks), before (11-12 weeks) and after (12-13 weeks) moving to finisher facility and the week before slaughter (22-25 weeks). In total, 288 OF samples were collected with cotton ropes and 216 with sponges and analysed for the biomarkers: cortisol, alpha-amylase, oxytocin (stress), haptoglobin (inflammation), procalcitonin (sepsis), adenosine deaminase, immunoglobulin G (immune system), ferric reducing antioxidant power (redox status), and creatine kinase, lactate dehydrogenase and total protein (general homeostasis). Samples were also scored visually for dirtiness using a score from 1 (clean) to 5 (very dirty). RESULTS: Rope-collected OF had higher levels of dirtiness (3.7 ± 0.04) compared to sponge-collected OF (2.7 ± 0.15) and had higher values than sponges for cortisol, procalcitonin, oxytocin, haptoglobin, total protein, lactate dehydrogenase and ferric reducing antioxidant power. All biomarkers decreased in value with age. Immunoglobulin G did not perform well for any of the two collection methods. DISCUSSION AND CONCLUSION: The results showed a clear effect of age on the biomarkers in OF collected with both, sponges or ropes. Sponges provided a cleaner sample than cotton ropes for biomarker analysis. Both methods are easy to apply under the commercial conditions in pig farms although sponges may take more time in early weaner stages. From a practical point of view, sampling with sponges achieved the best combination of reduced sampling time and low contamination.
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Canine obesity is the most common nutritional disorder and is associated with decreased quality of life and longevity as well as comorbidities including cardiorespiratory, endocrine, oncologic, or orthopaedic disorders. Ferritin is a major acute-phase protein in dogs, increasing during inflammation; however, it could also be affected by other conditions, including trauma, iron metabolism dysregulations, neoplasia, or hypoxia. Higher ferritin levels have been reported in obese humans, but ferritin has not been explored in canine obesity. To evaluate the possible changes in serum ferritin in canine obesity, ferritin levels from lean/normal weight (CG, n = 55) and overweight/obese dogs (OG, n = 37) were measured, together with complete hemogram and biochemical analyses. Statistically significant higher ferritin levels (1.2-fold) were found in OG (median, (interquartile range), 204 (166-227.5) µg/L) in comparison to CG animals (172 (137-210) µg/L)), with median levels of ferritin in OG dogs above the reference range for healthy animals in our laboratory (60-190 µg/L). In addition, statistically significant higher mean corpuscular volume (MCV), mean cell haemoglobin concentration (MCHC), total proteins, globulins, haptoglobin, total ferric fixation capacity (TIBC), alkaline phosphatase (ALP), butyrylcholinesterase (BChE), triglycerides, and calcium were observed in OG in comparison to CG. The higher levels in ferritin, together with higher TBIC, haematocrit, and MCV, could indicate tissue hypoxia in obese dogs.
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Acute phase proteins have been used as tools for the diagnosis, monitoring, and prognosis of several diseases in domestic animals. However, the dynamics of these proteins in infection by Trypanosoma cruzi, the causative agent of Chagas disease in dogs, is still unknown. The aim of this study was to determine concentrations of acute phase proteins (C-reactive protein, haptoglobin, ferritin and paraoxonase-1) in dogs in a coastal town of Ecuador, with natural Trypanosoma cruzi infection with or without seroreactivity of Ehrlichia canis, Ehrlichia ewingii, Anaplasma phagocytophilum, Anaplasma platys, Borrelia burgdorferi and Dirofilaria immitis. For the detection of Trypanosoma cruzi serum antibodies, two different antigen-based enzyme-linked immunosorbent assay tests were implemented. For the detection of seroreactivity of Ehrlichia canis, Ehrlichia ewingii, Anaplasma phagocytophilum, Anaplasma platys, Borrelia burgdorferi and Dirofilaria immitis, an IDEXX SNAP® 4Dx® test was used. To determine the concentration of C-reactive protein and ferritin, an immunoturbidimetric assay was used; haptoglobin concentration was measured using a commercial colorimetric method validated in dogs; a spectrophotometric method was used to determine the serum concentration of paraoxonase-1. Results showed a reduction in the serum levels of paraoxonase-1 in Trypanosoma cruzi-seroreactive dogs, either with or without seroreactivity to other vector-borne diseases. A serum ferritin increment was observed in Trypanosoma cruzi-seroreactive dogs with seroreactivity to any other vector-borne diseases. Our findings suggest that paraoxonase-1 levels are reduced in Trypanosoma cruzi-seroreactive dogs without evident clinical signs of Chagas disease, despite their seroreactivity to the other vector-borne diseases studied. These findings could indicate an oxidative stress response in Trypanosoma cruzi-seroreactive dogs with no evident signs of inflammation.
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BACKGROUND: Streptococcus suis can cause meningitis, polyarthritis and acute death in piglets. However, the risk factors associated with S. suis infection remain incompletely understood. Therefore, a longitudinal study was carried out, in which six batches from two Spanish pig farms with S. suis problems were repeatedly examined to determine possible risk factors. METHODS: A prospective case-control study was conducted, and potential risk factors were evaluated using mixed-effects logistic regression models. The explanatory variables included: (a) concomitant pathogens; (b) biomarkers associated with stress, inflammation and oxidative status; (c) farm environmental factors; and (d) parity and S. suis presence in sows. Three models were built to study the effect of these variables, including two to assess the risk factors involved in the subsequent development of disease. RESULTS: Risk factors for S. suis-associated disease included porcine reproductive and respiratory syndrome virus co-infection at weaning (odds ratio [OR] = 6.69), sow parity (OR = 0.71), haptoglobin level before weaning (OR = 1.01), relative humidity (OR = 1.11) and temperature (OR = 0.13). LIMITATIONS: Laboratory diagnosis was done at the batch level, with individual diagnosis based on clinical signs only. CONCLUSIONS: This study confirms the multifactorial nature of S. suis-associated disease, with both environmental factors and factors related to the host involved in disease development. Controlling these factors may, therefore, help prevent the appearance of disease.
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Infecciones Estreptocócicas , Streptococcus suis , Enfermedades de los Porcinos , Embarazo , Animales , Porcinos , Femenino , Enfermedades de los Porcinos/epidemiología , Granjas , España/epidemiología , Estudios Longitudinales , Estudios de Casos y Controles , Factores de Riesgo , Infecciones Estreptocócicas/epidemiología , Infecciones Estreptocócicas/veterinariaRESUMEN
The effects of filtration (F) and alpha-amylase depletion (AD) were assessed in n = 34 saliva samples. Each saliva sample was split into three aliquots and treated as follows: (1) no treatment; (2) 0.45µm commercial filter; and (3) 0.45µm commercial filter and affinity depletion of alpha-amylase. Then, a panel of biochemical biomarkers consisting of amylase, lipase, alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), creatine kinase (CK), calcium, phosphorus, total protein, albumin, urea, creatinine, cholesterol, triglycerides, and uric acid was measured. Differences between the different aliquots were observed in all measured analytes. The most marked changes were found in triglycerides and lipase data for filtered samples, and in alpha-amylase, uric acid, triglycerides, creatinine, and calcium results in alpha-amylase-depleted aliquots. In conclusion, the salivary filtration and amylase depletion methods employed in this report caused significant changes in saliva composition measurements. Based on these results, it would be recommended to consider the possible effects of these treatments in salivary biomarkers when filtration or amylase depletion is performed.
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Ácido Úrico , alfa-Amilasas , Creatinina , Calcio , Triglicéridos , Biomarcadores , AmilasasRESUMEN
The present study aims to evaluate the possible effects of the presence of pen faeces and feed on the measurement of a panel of biomarkers in porcine oral fluid. For this, clean porcine oral fluid was pooled and incubated with two different concentrations of pen faeces or feed representing a high or low level of contamination with each material. In addition, these pools were aliquoted and subjected to centrifugation, filtration or chemical clarification to evaluate if these techniques could revert the effects of those contaminants in biomarker evaluation. A panel of 21 biomarkers that assessed stress, inflammation, immune system and redox homeostasis among others, were measured for all aliquots. Changes of statistical relevance (p < 0.05) in oral fluid contaminated with pen faeces or feed versus untreated samples were observed for all methods employed with the exception of adenosine deaminase (ADA) and creatine kinase (CK) in oral fluid contaminated with pen faeces or feed. Pen faeces did not affect the measurement of haptoglobin, superoxide dismutase, CK, lactate dehydrogenase (LDH), ADA and cortisol (when the latter is measured by chemiluminescence); while uric acid, LDH, CK, ADA, and hydrogen peroxide methods were not affected by the presence of feed in oral fluid. The effects of centrifugation, filtration or chemical clarification with chitosan in these contaminated samples were modest and for most cases did not caused baseline levels on the measured biomarkers. In conclusion, the presence of pen faeces or feed in porcine oral fluid can interfere with the results obtained when analytes are measured.
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Hidrocortisona , Porcinos , Animales , Heces , BiomarcadoresRESUMEN
Saliva from pigs is gaining attention as an easy sample to obtain, being a source of biomarkers that can provide information on animal health and welfare. This study aimed to evaluate the changes that can occur in salivary biomarkers of the redox status of pigs with an experimentally induced sepsis. For that, the cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of saliva (FRAS), Trolox equivalent antioxidant capacity (TEAC), advanced oxidation protein products (AOPP), ferrous oxidation-xylenol orange (FOX), peroxide activity (POX-Act), and reactive oxygen-derived compounds (d-ROMs) were measured in the saliva of pigs with experimentally induced sepsis by endotoxin lipopolysaccharide (LPS), non-septic inflammation induced by turpentine, and in healthy individuals before and after 3 h, 6 h, 24 h, and 48 h. AOPP, POX-Act, and d-ROMs in the sepsis group were higher than in the control from 3 h to 24 h after the inoculation. CUPRAC, FRAS, and TEAC were higher in sepsis than the control group at 24 h. These changes were of higher magnitude than those that occurred in the turpentine group. In conclusion, our findings reveal that sepsis produces changes in salivary biomarkers of redox status, which opens the possibility of using them as potential biomarkers in this species.
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OBJECTIVES: To develop and evaluate a new highly sensitive assay to detect IgG anti-SARS-CoV-2 RBD in saliva samples. METHODS: A two-step sandwich type immunoassay based on the amplified luminescent proximity homogeneous technology was developed and an analytical validation was performed. As a part of this validation, the influence of factors, such as different sampling conditions (stimulated saliva and passive drool) and the correction of values by total protein content, in the ability of saliva to detect increases in antibodies after an immune stimulus and be an alternative to serum, was evaluated. For this purpose, paired samples of saliva and serum at different times after vaccination were used. RESULTS: Saliva concentrations were lower than serum, but both fluids showed similar kinetics, with higher correlations when saliva was obtained by passive flow and the results were not corrected by protein. CONCLUSIONS: The developed method showed a good analytical performance and can properly measure antibody concentrations in saliva of vaccinated individuals. However, saliva could have a lower sensitivity compared to serum at initial stages of the immune response and also when the antibody response decreased after a stimulus.
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COVID-19 , Saliva , Anticuerpos Antivirales , Humanos , Inmunoglobulina G , SARS-CoV-2RESUMEN
A comprehensive panel of 29 salivary analytes was measured in fattening pigs to evaluate its possible changes along their productive cycle. The identification of those changes would allow a better interpretation of the results according to the productive phase of the animal. Saliva samples were obtained from 49 Large-White pigs (24 females, 25 males) in suckling phase, at the beginning and the end of the nursery phase, and at the beginning and the end of the growing phase. Several analytes changed according to the phase of the productive cycle, with most of the analytes showing higher values at lactation and at the beginning of nursery. Additionally, differences were seen due to sex. When possible relations between performance parameters and analytes were evaluated, significant positive but weak relationships were found between weight at birth and salivary γ-glutamyl transferase, and between back-fat thickness and salivary lactate dehydrogenase. In conclusion, differences in the values of salivary analytes can be found in fattening pigs depending on the productive phase and sex of the animals.
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African swine fever (ASF) is a notifiable viral disease of domestic and wild suids. Despite intensive research efforts, the pathogenesis of the disease is still far from being understood. Analysis of biomarkers in different body fluids may supplement traditional pathogenesis studies. As reliable protocols are often established in laboratories with lower biosafety, the reliable inactivation of samples is crucial. The objective of this study was to find a procedure that inactivates the virus while preserving the biomarkers for downstream analyses. To this means, three different inactivation protocols were employed, namely Tergitol-type NP-40 (NP-40), polyoxyethylene-p-t-octylphenol (Triton X-100) and one with 95 °C heating. It could be demonstrated that all samples treated with 0.5% (v/v) concentration of both detergents showed an absence of virus infectivity. The same was true for heated samples. However, heated serum was not suitable for analyses. Next, the impact of treatment on biomarker readouts was assessed. While all protocols had an impact on the detection of biomarkers, correlation was retained. In particular, NP-40 may be the desired detergent for more accurate measurements while achieving efficient virus inactivation. Based on these studies, samples can be reliably inactivated for most biomarker analyses, and thus broader interdisciplinary cooperation is possible.
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The use of mannequins to practice different clinical procedures in undergraduate students complies with the 3R principle (Replacement, Reduction, Refinement) without affecting professional competencies. However, commercial solutions are often expensive and therefore, not available for many schools. The main aim of this study was to describe the development and validation of an economical Do-It-Yourself mannequin for jugular blood collection from dogs. The use of the mannequin was evaluated by (1) assessment of the opinion of students and experts and (2) by conducting a pilot study where salivary biomarkers of stress were determined in students and dogs. The costs of the materials needed for the mannequin confection were less than 60 Euros and it was easily built in less than 1 h. The mannequin was very well accepted and scored by both, students and experts, being mostly liked a lot and considered it to be very useful for the practices (Scored 8, 9 or 10/10). Students that could first practice with the mannequin reported a self-perceived higher level of confidence and had lower levels of alpha-amylase in saliva after the procedure. Overall, the mannequin enables the initiation of blood sampling skills in agreement with 3R principles and is easy to perform, economically affordable and sustainable. This model could be adapted to other vein simulations and animal species, and has the potential to help students deal with stressful situations such as taking blood samples from a live animal.
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Competencia Clínica , Maniquíes , Animales , Perros , Humanos , Proyectos Piloto , EstudiantesRESUMEN
A career in journalism can be very stressful, as journalists frequently have to deal with uncontrolled situations such as conducting live interviews. Therefore, training is essential during their career, both for the development of communication skills and for the improvement of the real and effective capacity to perform the tasks of their professional activity. The aim of this study was to assess the levels of stress in students before and after a practical training in a professional television set using subjective (State-Trait Anxiety Inventory (STAI) and Likert scale) and objective (salivary cortisol and alpha-amylase) methods. The results indicate that a live interview produces stress in the students as revealed by increased concentrations of cortisol and alpha amylase in saliva. Furthermore, students with lower initial concentrations of these biomarkers obtained better grades in evaluation, suggesting that greater control of anticipatory stress could be associated with a better activity performance.
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Ansiedad , Estrés Psicológico , Biomarcadores , Humanos , Hidrocortisona , Saliva , EstudiantesRESUMEN
Classically, the need for highly sophisticated instruments with important economic costs has been a major limiting factor for clinical pathology laboratories, especially in developing countries. With the aim of making clinical pathology more accessible, a wide variety of free or economical technologies have been developed worldwide in the last few years. 3D printing and Arduino approaches can provide up to 94% economical savings in hardware and instrumentation in comparison to commercial alternatives. The vast selection of point-of-care-tests (POCT) currently available also limits the need for specific instruments or personnel, as they can be used almost anywhere and by anyone. Lastly, there are dozens of free and libre digital tools available in health informatics. This review provides an overview of the state-of-the-art on cost-effective alternatives with applications in routine clinical pathology laboratories. In this context, a variety of technologies including 3D printing and Arduino, lateral flow assays, plasmonic biosensors, and microfluidics, as well as laboratory information systems, are discussed. This review aims to serve as an introduction to different technologies that can make clinical pathology more accessible and, therefore, contribute to achieve universal health coverage.
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Patología Clínica , Análisis Costo-Beneficio , Laboratorios , Microfluídica , Pruebas en el Punto de AtenciónRESUMEN
Burning mouth syndrome (BMS) is a chronic oral condition characterized by an intraoral burning sensation, taste alterations, and dry mouth sensations. Although a number of factors have been closely related to the appearance of the symptoms, including anxiety, depression, and sleep disturbances, the etiology of BMS remains unclear. Furthermore, currently no objective diagnostic tools exist, making its diagnosis challenging. Therefore, to contribute to the knowledge about BMS etiology and look for objective tools for its diagnosis, the present study was conducted. Thus, the aim of this study was to analyze the proteomic profile of the resting whole saliva of patients with BMS and age and sex-matched controls using two-dimensional electrophoresis. The results showed evidence of changes in saliva at the level of proteins related to important pathways such as stress (sAA), immune system (Ig), and inflammation (leukocyte elastase inhibitor). While some of our findings have been previously described others, such as the deregulation of the coiled-coin domain containing protein 25 in BMS, are presented here for the first time to our knowledge. Thus, saliva provides us with relevant information about BMS pathophysiology and could be considered a suitable biofluid for its study and/or diagnosis.
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OBJECTIVES: To evaluate four sample treatments in a safe and straightforward procedure to detect SARS-CoV-2 in saliva. METHODS: Four sample treatments were evaluated in a 3-step procedure to detect SARS-CoV-2 in saliva: 1) heating at 95 °C for 5 min for sample inactivation; 2) sample treatment; 3) analysis by reverse-transcription loop-mediated isothermal amplification (LAMP). Saliva samples used were from infected individuals or were spiked with known quantities of viral particles. RESULTS: Three treatments had a limit of detection (LOD) of 500.000 viral particles per ml of saliva and could be used to detect individuals with potential to transmit the disease. The treatment of phosphate buffer, dithiothreitol, ethylenediaminetetraacetic acid and proteinase K, with an additional 95 °C heating step, yielded a lower LOD of 95; its sensitivity ranged from 100% in patients with nasopharyngeal swab reverse-transcriptase polymerase chain reaction cycle threshold values <20 to 47.8% for values >30. CONCLUSIONS: This report highlights the importance of an adequate sample treatment for saliva to detect SARS-CoV-2 and describes a flexible procedure that can be adapted to point-of-care. Although its sensitivity when LAMP is used is lower than reverse-transcriptase polymerase chain reaction, this procedure can contribute to COVID-19 control by detecting individuals able to transmit the disease.
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COVID-19 , SARS-CoV-2 , Humanos , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , ARN Viral/genética , Saliva , Sensibilidad y EspecificidadRESUMEN
OBJECTIVES: The aim of the present study was to validate a commercially available automated assay for the measurement of total adenosine deaminase (tADA) and its isoenzymes (ADA1 and ADA2) in saliva in a fast and accurate way, and evaluate the possible changes of these analytes in individuals with SARS-CoV-2 infection. METHODS: The validation, in addition to the evaluation of precision and accuracy, included the analysis of the effects of the main procedures that are currently being used for SARS-CoV-2 inactivation in saliva and a pilot study to evaluate the possible changes in salivary tADA and isoenzymes in individuals infected with SARS-CoV-2. RESULTS: The automated assay proved to be accurate and precise, with intra- and inter-assay coefficients of variation below 8.2%, linearity under dilution linear regression with R2 close to 1, and recovery percentage between 80 and 120% in all cases. This assay was affected when the sample is treated with heat or SDS for virus inactivation but tolerated Triton X-100 and NP-40. Individuals with SARS-CoV-2 infection (n=71) and who recovered from infection (n=11) had higher mean values of activity of tADA and its isoenzymes than healthy individuals (n=35). CONCLUSIONS: tADA and its isoenzymes ADA1 and ADA2 can be measured accurately and precisely in saliva samples in a rapid, economical, and reproducible way and can be analyzed after chemical inactivation with Triton X-100 and NP-40. Besides, the changes observed in tADA and isoenzymes in individuals with COVID-19 open the possibility of their potential use as non-invasive biomarkers in this disease.
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Adenosina Desaminasa/metabolismo , Bioensayo/métodos , Biomarcadores/metabolismo , COVID-19/diagnóstico , SARS-CoV-2/enzimología , Saliva/enzimología , Adulto , COVID-19/virología , Estudios de Casos y Controles , Femenino , Humanos , Isoenzimas , Masculino , Persona de Mediana Edad , Proyectos Piloto , Adulto JovenRESUMEN
Here, we report on the role of haptoglobin (Hp), whose expression depends on the synthesis of interleukin 6 (IL-6), related to the pathogenesis of multiple sclerosis (MS), as a possible marker of muscle improvement achieved after treatment with the polyphenol epigallocatechin gallate (EGCG) and an increase in the ketone body beta-hydroxybutyrate (BHB) in the blood. After 4 months of intervention with 27 MS patients, we observed that Hp does not significantly increase, alongside a significant decrease in IL-6 and a significant increase in muscle percentage. At the same time, Hp synthesis is considerably and positively correlated with IL-6 both before and after treatment; while this correlation occurs significantly reversed with muscle percentage before treatment, no correlation is evident after the intervention. These results seem to indicate that Hp could be a marker of muscle status and could be a diagnosis tool after therapeutic intervention in MS patients.