Browsing by Subject "Proteomics"

Now showing 1 - 17 of 17
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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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    Alterations in haemolymph proteome of Mytilus galloprovincialis mussel after an induced injury
    (Elsevier, 2018-01-31) Franco-Martínez, Lorena; Martínez-Subiela, Silvia; Escribano, Damián; Schlosser, Sarah; Nöbauer, Katharina; Razzazi-Fazeli, Ebrahím; Romero, Diego; Cerón, José Joaquín Cerón; Tvarijonaviciute, Asta; Ciencias Sociosanitarias
    A proteomic and biochemical approach was performed to assess the effects of an induced muscle injury on the haemolymph of bivalve molluscs. For this purpose, Mytilus galloprovincialis were exposed to puncture of adductor muscle for three consecutive days, and their haemolymph proteome was then compared to healthy animals using 2-dimensional electrophoresis (2-DE) to identify proteins that differed significantly in abundance. Those proteins were then subjected to tandem mass spectrometry and 6 proteins, namely myosin, tropomyosin, CuZn super-oxide dismutase (SOD), triosephosphate isomerase, EP protein and small heat shock protein were identified. SOD and tropomyosin changes were verified by spectrophotometric measurements and western blotting, respectively. As some of the proteins identified are related to muscular damage and oxidative stress, other biomarkers associated with these processes that can be evaluated by automatic biochemical assays were measured including troponin, creatine kinase (CK), and aspartate aminotransferase (AST) for muscle damage, and SOD, trolox equivalent antioxidant capacity (TEAC) and esterase activity (EA) for oxidative stress. Significantly higher concentrations of troponin, CK, AST, and TEAC were observed in mussels after puncture, being also possible biomarkers of non-specific induced damage.
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    Boar semen proteomics and sperm preservation
    (Elsevier, 2019-06-02) Parrilla, Inmaculada; Pérez Patiño, Cristina; Li, J.; Barranco Cascales, Isabel; Padilla, L.; Rodriguez-Martínez, Heriberto; Martínez, E. A.; Roca J.; Medicina y Cirugía Animal; Facultad de Veterinaria
    Recently numerous proteomic approaches have been undertaken to identify sperm and seminal plasma (SP) proteins that can be used as potential biomarkers for sperm function, including fertilization ability. This review aims firstly to briefly introduce the proteomic technologies and workflows that can be successfully applied for sperm and SP proteomic analysis. Secondly, we summarize the current knowledge about boar SP and the sperm proteome, focusing mainly on its relevance to sperm preservation procedures (liquid storage or cryopreservation) and their outcomes in terms of sperm function and fertility.
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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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    Colostrum Features of Active and Recovered COVID-19 Patients Revealed Using Next-Generation Proteomics Technique, SWATH-MS
    (2023) Hernández-Caravaca, Ivan; González-Brusi, Leopoldo; Romero de Ávila, Mª José; De Paco Matallana, Catalina; Castaño-Molina, Mª Angeles; Díaz-Meca, Lucía; Sánchez-Romero, Javier; Martínez-Alarcón, Laura; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Moros Nicolás, Carla; Pelegrín Vivancos, Pablo; Biología Celular e Histología
    Colostrum performs nutritional, anti-inflammatory and anti-infective functions and promotes immune system formation and organ development. The new coronavirus, SARS-CoV-2, has generated concerns about viral transmission through human milk, with a lack of evidence about human milk’s protective effects against the infection. This study aimed at analyzing presence of the virus and at identifying the protein expression profile of human colostrum in active and COVID-19- recovered patients. Colostrum samples were collected from women with COVID-19 (n = 3), women recently recovered from the infection (n = 4), and non-infected women (n = 5). The samples were analyzed by means of RT-qPCR to determine presence of the virus and using SWATH-MS for proteomic analysis. Proteomic results were then analyzed using bioinformatic methods. The viral tests were negative for SARS-CoV-2 in the colostrum from COVID-19 patients. The proteomic analysis identified 301 common proteins in all samples analyzed. Nineteen proteins were upregulated and 7 were downregulated in the COVID-19 group versus the control samples, whereas 18 were upregulated and 7 were downregulated when comparing the COVID-19 group to the recovered group. Eleven proteins were biomarkers of active COVID-19 infection. Ten were upregulated: ACTN1, CD36, FAM3B, GPRC5B, IGHA2, IGK, PLTP, RAC1, SDCBP and SERPINF1, and one was downregulated: PSAP. These proteins are mainly related to immunity, inflammatory response and protein transport. In conclusion, the results of this study suggest that colostrum is not a vehicle for mother-to-child SARS-CoV-2 transmission. Moreover, the colostrum’s proteome of active and recuperated patients indicate that it could provide immune benefits to infants.
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    Cryopreservation differentially alters the proteome of epididymal and ejaculated pig spermatozoa
    (2019-04-11) Perez-Patiño, Cristina; Barranco, Cascales; Li, Junwei; Padilla, Lorena; Martínez, Emilio A; Rodriguez-Martinez, Heriberto; Roca, Jordi; Parrilla, Inmaculada; Medicina y Cirugía Animal
    Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein-protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.
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    High-resolution proteomics and metabolomics in thyroid cancer: Deciphering novel biomarkers
    (Taylor & Francis, 2017-10-30) Navas-Carrillo, Diana; Rodríguez, José Manuel; Montoro-García, Silvia; Orenes-Piñero, Esteban; Bioquímica y Biología Molecular A
    Thyroid cancer (TC) is the most common endocrine malignancy and its incidence has been increasing sharply since the mid-1990s, being the fastest-increasing cancers in both men and women. Increased medical surveillance, the effect of environmental factors and more sensitive diagnostic tests, such as ultrasound and confirmation via fine-needle aspiration biopsy, are thought to account for this increased incidence. There are several histological types of thyroid cancer, including papillary thyroid carcinoma, follicular thyroid carcinoma, medullary thyroid carcinoma, and anaplastic thyroid carcinoma. Determining the type of thyroid cancer is crucial for the assessment of prognosis and treatment selection. Unfortunately, approximately 20–30% of patients undergoing fine-needle aspiration biopsy have inconclusive or indeterminate results, leading to unnecessary surgical intervention in 80% of patients with benign nodules. To resolve this diagnosis dilemma, new biomarkers of thyroid cancer are needed. Proteomic approaches offer an unbiased platform for the comprehensive analysis of the whole proteome in a certain physiological time. Although mRNA expression is widely considered to be indicative of protein expression, protein levels are the result of protein synthesis and degradation, and RNA levels are not informative of protein degradation. Clinically, there is increasing evidence for the role of proteomic and metabolomic technologies in biomarker discovery, providing novel information on the molecular events associated with TC, and potentially lead to the identification of novel drug targets. In this review, we will thoroughly describe the importance of novel proteomic and metabolomic approaches to identify new biomarkers associated with TC.
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    Identification of changes in serum analytes and possible metabolic pathways associated with canine obesity-related metabolic dysfunction
    (Elsevier, 2019-02) Tvarijonaviciute, Asta; Barić Rafaj, R.; Horvatic, A.; Muñoz Prieto, A.; Guillermin, N.; Tumpa, A.; Cerón, J. J.; Martínez Subiela, S.; Mrljak, V.; Medicina y Cirugía Animal
    The main objective of this study was to identify analytes that could change and that could help to clarify the metabolic and physiopathological changes related to canine obesity-related metabolic dysfunction (ORMD). For this, serum from 35 overweight/obese dogs, with and without ORMD, was submitted to a comprehensive panel of biochemistry analysis, a gel-free tandem mass tag isobaric label-based proteomic analysis, and, finally, selected proteins were used as a starting point for creating a protein interaction network. Dogs with ORMD showed significantly higher serum concentrations of alanine aminotransferase (ALT), alkaline phosphatase (ALP), Ca, total proteins, albumin, total cholesterol, triglycerides, glucose, and butyrylcholinesterase (BChE) activity in comparison with dogs without ORMD. Proteomic analysis revealed that 23 proteins related to lipid metabolism, the complement factor system, cellular adhesion and functionality, inflammation, and coagulation were altered in dogs with ORMD. Finally, the obtained protein interaction network highlighted that the central term of this network was the negative regulation of the immune response. These data suggest that canine ORMD is associated with changes in analytes that reflect altered lipid metabolism, and liver and immune function impairment and suggests the potential for a prothrombotic state and lung function alterations.
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    Laser capture microdissection: Big data from small samples
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2015) Datta, Soma; Malhotra, Lavina; Dickerson, Ryan; Chaffee, Scott; Sen, Chandan K.; Roy, Sashwati
    Any tissue is made up of a heterogeneous mix of spatially distributed cell types. In response to any (patho) physiological cue, responses of each cell type in any given tissue may be unique and cannot be homogenized across cell-types and spatial co-ordinates. For example, in response to myocardial infarction, on one hand myocytes and fibroblasts of the heart tissue respond differently. On the other hand, myocytes in the infarct core respond differently compared to those in the peri-infarct zone. Therefore, isolation of pure targeted cells is an important and essential step for the molecular analysis of cells involved in the progression of disease. Laser capture microdissection (LCM) is powerful to obtain a pure targeted cell subgroup, or even a single cell, quickly and precisely under the microscope, successfully tackling the problem of tissue heterogeneity in molecular analysis. This review presents an overview of LCM technology, the principles, advantages and limitations and its down-stream applications in the fields of proteomics, genomics and transcriptomics. With powerful technologies and appropriate applications, this technique provides unprecedented insights into cell biology from cells grown in their natural tissue habitat as opposed to those cultured in artificial petri dish conditions.
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    Proteome changes induced by a short, non-cytotoxic exposure to the mycoestrogen zearalenone in the pig intestine
    (Elsevier, 2020-07-30) Soler, Laura; Stella, Alexandre; Pallarés, Francisco José; Lahjouji, Tarek; Burlet Schiltz, Odile; Oswald, Isabelle P.; Seva Alcaraz, Juan; Anatomía y Anatomía Patológica Comparadas
    Intestinal epithelial homeostasis is regulated by a complex network of signaling pathways. Among them is estrogen signaling, important for the proliferation and differentiation of epithelial cells, immune signaling and metabolism. The mycotoxin zearalenone (ZEN) is an estrogen disruptor naturally found in food and feed. The exposure of the intestine to ZEN has toxic effects including alteration of the immune status and is possibly implicated in carcinogenesis, but the molecular mechanisms linked with these effects are not clear. Our objective was to explore the proteome changes induced by a short, non-cytotoxic exposure to ZEN in the intestine using pig jejunal explants. Our results indicated that ZEN promotes little proteome changes, but significantly related with an induction of ERα signaling and a consequent disruption of highly interrelated signaling cascades, such as NF-κB, ERK1/2, CDX2 and HIF1α. The toxicity of ZEN leads also to an altered immune status characterized by the activation of the chemokine CXCR4/SDF-1 axis and an accumulation of MHC-I proteins. Our results connect the estrogen disrupting activity of ZEN with its intestinal toxic effect, associating the exposure to ZEN with cell-signaling disorders similar to those involved in the onset and progression of diseases such as cancer and chronic inflammatory disorders.
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    Proteomic analysis in usual and nonspecific interstitial pneumonia
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Ohara, Ichiyo; Aida, Shinsuke; Shimazaki, Hideyuki; Kobayashi, Hideo; Tsuda, Hitoshi; Toda, Tosifusa; Nakanishi, Kuniaki; Tamai, Seiichi
    Differentiating nonspecific interstitial pneumonia (NSIP) from usual interstitial pneumonia (UIP) is important for the determination of both treatment and prognosis. Using two-dimensional fluorescence difference gel electrophoresis (2D-DIGE), we examined 8 UIPs, 8 NSIPs, and 30 normal lung tissues. Comparisons with control in 2D-DIGE showed that (a) in UIP, nine protein spots were significantly upregulated and seven were significantly downregulated, (b) in NSIP, four protein spots were significantly upregulated and nine were significantly downregulated. The detected proteins were analyzed by MALDI-TOF mass spectrometry, allowing qualitative differences in vimentin subtypes to be characterized. One vimentin subtype was upregulated in UIP, while another one was downregulated in NSIP (vs. control). These different characteristics were partially supported by the results of Western blot analysis. Our immunohistochemistry revealed vimentin expression within fibroblasts (a) in fibroblastic foci in UIP and (b) in fibrotic alveolar walls in NSIP. Differences in vimentin subtypes may provide useful biomarkers for separating NSIP from UIP, alongside differences in histological characteristics.
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    Proteomic analysis of the lung in rats with hypobaric hypoxia-induced pulmonary hypertension
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2013) Ohata, Yuichiro; Ogata, Sho; Nakanishi, Kuniaki; Kanazawa, Fumiko; Uenoyama, Maki; Hiroi, Sadayuki; Tominaga, Susumu; Toda, Tosifusa; Kawai, Toshiaki
    Experimental pulmonary hypertension that develops in hypobaric hypoxia is characterized by structural remodeling of the lung. Proteomics - which may be the most powerful way to uncover unknown remodeling proteins involved in enhancing cardiovascular performance - was used to study 150 male Wistar rats housed for up to 21 days in a chamber at the equivalent of 5500 m altitude level. After 14 days’ exposure to hypobaric hypoxia, pulmonary arterial pressure (PAP) was significantly increased. In lung tissue, about 140 matching protein spots were found among 8 groups (divided according to their hypobaric period) by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) (pH4.5-pH6.5, 30 kDa-100 kDa). In hypobaric rats, three spots were increased two-fold or more (vs. control rats) in two-dimensional differential in-gel electrophoresis (2D-DIGE). The increased proteins were identified, by matrix-assisted laser desorption ionization time of flight (MALDI-TOF), as one isoform of heat shock protein 70 (HSP70) and two isoforms of protein disulfide isomerase associated 3. This result was confirmed by Western blotting analysis of 2D-PAGE. Conceivably, HSP70 and PDIA3 may play roles in modulating the lung structural remodeling that occurs due to pulmonary hypertension in hypobaric hypoxia.
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    Proteomic profiling of porcine seminal extracellular vesicles reveals potential in vivo fertility biomarkers
    (Wiley, 2025-07-04) Barranco Cascales, Isabel; Martínez Díaz, Pablo; Parra, Ana; Martínez-Alborcia, María José; Lucas Arjona, Xiomara; Rodríguez-Martínez, Heriberto; Roca, Jordi; Medicina y Cirugía Animal; Facultad de Veterinaria
    Background: Predicting male fertility in farm animals remains a challenge. Seminal plasma (SP) contains a high amount of heterogeneous seminal extracellular vesicles (sEVs), believed involved in reproductive processes and maybe key to understanding male fertility. Aims: To identify the sEV proteins that are differentially expressed between more and less fertile boars and that could be candidates for fertility biomarkers in boars used in artificial insemination (AI) programs. Materials and methods: Small (S) and large (L) sEV subsets from SP samples of AI boars with differences in fertility: high (H) or low (L) farrowing rate (FR) and large (L) or small (S) litter size (LS). The S- and L-sEV subsets were isolated by size exclusion chromatography and characterized according to the Minimal Information for Studies of Extracellular Vesicles (MISEV2023) guidelines. Proteomic analyses (three biological replicates per fertility group and sEV subset) were performed using a Bruker timsTOF fleX™ instrument with data-independent acquisition parallel accumulation-serial fragmentation (diaPASEF) technology. Results: A total of 470 and 726 proteins were quantified in S-sEVs and 1801 and 1834 proteins in L-sEVs from FR and LS boars, respectively. Differentially expressed sEV proteins (log2fold change ≥±1, p ≤ 0.05 and effect size d of Cohen >2.0) were found between the fertility groups: seven in S-sEVs and 52 in L-sEVs between H-FR and L-FR boars, and 47 in S-sEVs and 52 in L-sEVs between L-LS and S-LS boars. Many of these differentially expressed sEV proteins are involved in reproductive processes, particularly in sperm function and sperm-zona pellucida binding, but also in embryo development and implantation. Conclusions: The sEV proteome differs between more and less fertile boars, with many of the differentially expressed proteins known as involved in reproductive processes. This would suggest that sEVs may be involved in male fertility and that some of the differentially expressed sEV proteins could be potential fertility markers for AI boars.
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    Proteomics-Based Identification of Salivary Changes in Patients with Burning Mouth Syndrome
    (MDPI, 2021-05-01) Castillo-Felipe, candela; Franco-Martínez, Lorena; Tvarijonaviciute, Asta; Lopez Jornet, Pia; Lamy, Elsa; Dermatología, Estomatología, Radiología y Medicina Física
    Burning mouth syndrome (BMS) is a chronic oral condition characterized by an intraoral burning sensation, taste alterations, and dry mouth sensations. The disease affects 0.7–15% of the general population, being most common in post-menopausal women. Although BMS is related to anxiety and/or depression and sleep disturbances, its etiology as well as its diagnosis remain unclear. Therefore, the present study aimed to contribute to the knowledge about this syndrome and to look for objective diagnostic tools. Therefore, whole saliva proteomes of patients suffering from BMS were compared to those of healthy persons. The results of this study manifest alterations in salivary proteins related to stress, immune system, and inflammation and, therefore, suggest implication of these pathways in BMS development. Moreover, biomarkers related to stress, immune system, and inflammation, such as salivary amyloid A, immunoglobulins, or leukocyte elastase inhibitors, among others, could contribute to BMC management, although further research is needed to confirm these suppositions.
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    Saliva changes in composition associated to COVID-19: a preliminary study
    (Nature Research, 2022-06-27) Muñoz Prieto, Alberto; Rubić, Ivana; González Sánchez, Juan Carlos; Kuleš, Josipa; Martínez Subiela, Silvia; Cerón, José Joaquín; Bernal, Enrique; Torres Cantero, Alberto; Vicente Romero, María Rosario; Mrljak, Vladimir; Tvarijonaviciute, Asta; Medicina y Cirugía Animal
    The coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV 2), is usually associated with a wide variety of clinical presentations from asymptomatic to severe cases. The use of saliva as a diagnostic and monitoring fluid has gained importance since it can be used to investigate the immune response and to direct quantification of antibodies against COVID-19. Additionally, the use of proteomics in saliva has allowed to increase our understanding of the underlying pathophysiology of diseases, bringing new perspectives on diagnostics, monitoring, and treatment. In this work, we compared the salivary proteome of 10 patients with COVID-19, (five patients with mild and five patients with severe COVID-19) and ten control healthy patients. Through the application of proteomics, we have identified 30 proteins whose abundance levels differed between the COVID-19 groups and the control group. Two of these proteins (TGM3 and carbonic anhydrase-CA6) were validated by the measurement of gGT and TEA respectively, in 98 additional saliva samples separated into two groups: (1) COVID-19 group, integrated by 66 patients who tested positive for COVID-19 (2) control group, composed of 32 healthy individuals who did not show any sign of disease for at least four weeks and were negative for COVID-19 in RT-PCR. In the proteomic study there were observed upregulations in CAZA1, ACTN4, and ANXA4, which are proteins related to the protective response against the virus disturbance, and the upregulation of TGM3, that is correlated to the oxidative damage in pulmonary tissue. We also showed the downregulation in cystatins and CA6 that can be involved in the sensory response to stimulus and possibly related to the presence of anosmia and dysgeusia during the COVID-19. Additionally, the presence of FGB in patients with severe COVID-19 but not in mild COVID-19 patients could indicate a higher viral aggregation and activation in these cases. In conclusion, the salivary proteome in patients with COVID-19 showed changes in proteins related to the protective response to viral infection, and the altered sensory taste perception that occur during the disease. Moreover, gGT and TEA could be potential biomarkers of respiratory complications that can occurs during COVID 19 although further larger studies should be made to corroborate this.
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    Sperm proteome after interaction with reproductive fluids in porcine: from the ejaculation to the fertilization site
    (MDPI, 2020-08-22) Luongo, Chiara; Gónzalez-Brusi, Leopoldo; García-Vázquez, Francisco Alberto; Cots Rodríguez, Paula; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Biología Celular e Histología
    Ejaculated sperm are exposed to different environments before encountering the oocyte. However, how the sperm proteome changes during this transit remains unsolved. This study aimed to identify proteomic changes in boar sperm after incubation with male (seminal plasma, SP) and/or female (uterine fluid, UF; and oviductal fluid, OF) reproductive fluids. The following experimental groups were analyzed: (1) SP: sperm + 20% SP; 2) UF: sperm + 20% UF; 3) OF: sperm + 20% OF; 4) SP + UF: sperm + 20% SP + 20% UF; and (5) SP+OF: sperm + 20% SP + 20% OF. The proteome analysis, performed by HPLC-MS/MS, allowed the identification of 265 proteins. A total of 69 proteins were detected in the UF, SP, and SP + UF groups, and 102 proteins in the OF, SP, and SP + OF groups. Our results showed a higher number of proteins when sperm were incubated with only one fluid than when they were co-incubated with two fluids. Additionally, the number of sperm-interacting proteins from the UF group was lower than the OF group. In conclusion, the interaction of sperm with reproductive fluids alters its proteome. The description of sperm-interacting proteins in porcine species after co-incubation with male and/or female reproductive fluids may be useful to understand sperm transport, selection, capacitation, or fertilization phenomena.
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    The hepatic stellate cell in the post-genomic era
    (Murcia : F. Hernández, 2002) Okuyama, H.; Shimahara, Y.; Kawada, N.
    The draft human genome sequence was published on February 15, 2001, which will provide a huge amount of information on human genetics, human disease, and human cell biology. Now, medical scientists and cell biologists are turning their attention to illustrating gene expression pattern using gene microarray and to identifying the functions and the expression patterns of proteins encoded by the genes. Hepatic stellate cell is one of the sinusoidconstituent cells that play multiple roles in the liver p a t h o p h y s i o l o g y. Transformation of stellate cells from the vitamin A-storing phenotype to the “myofibroblastic” one closely correlates to hepatic fibrosis during chronic liver trauma. Analyses of the molecular mechanisms of stellate cell activation have made a great progress, in particular, in the field of intracellular signal transduction of transforming growth f a c t o r-ß and platelet-derived growth factor, integrin signaling related to cell-adhesion, and cell motilityassociated Rho and focal-adhesion kinase. Accumulation of the information on the stellate cell activation would shed light on the establishment of a novel therapeutic strategy against fibrosis of human liver disease.