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BACKGROUND: F-actin is involved in the progression of ischemic stroke and is associated with the disruption of the blood-brain barrier. In this article, we evaluated serum F-actin as a biomarker in stroke severity and early neurological deterioration (END) in acute ischemic stroke. METHODS: In this study, serum F-actin was measured in consecutively collected 140 AIS patients and 144 healthy controls matched in gender and age by ELISA. Early neurological deterioration (END) was defined as the deterioration of neurological dysfunction within 72 hours of admission, with an increase of ≥ 4 points in the NIHSS score. Severe stroke was defined as a NIHSS score>8 at admission. RESULTS: The serum F-actin level in AIS was significantly higher than healthy controls (p = 0.041). In large-artery atherosclerosis stroke and cardioembolic stroke, serum F-actin were significantly higher than that in small artery occlusion stroke (padjust = 0.019, padjust < 0.001, respectively).F-actin level above the critical value (>1.37 µg/L) was significantly associated with severe stroke (OR, 3.015; 95 %CI, 1.014-8.963; p = 0.047) . In addition, elevated level of F-actin was significantly associated with END (OR, 1.323; 95 % CI, 1.001-1.747, p = 0.049). When the level of F-actin was above the critical value (>2.17 µg/L), its association with END remained significant (OR, 6.303; 95 %CI, 2.160-18.394; p < 0.001) . CONCLUSION: F-actin is an important blood biomarker in the early stage of AIS, and high levels of F-actin are valuable in determining the severity of stroke and predicting early neurological deterioration.
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Actinas , Biomarcadores , Accidente Cerebrovascular Isquémico , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Actinas/sangre , Biomarcadores/sangre , Estudios de Casos y Controles , Evaluación de la Discapacidad , Progresión de la Enfermedad , Accidente Cerebrovascular Isquémico/sangre , Accidente Cerebrovascular Isquémico/diagnóstico , Valor Predictivo de las Pruebas , Pronóstico , Factores de Riesgo , Índice de Severidad de la Enfermedad , Factores de Tiempo , Regulación hacia ArribaRESUMEN
INTRODUCTION: A major complication of sepsis is the development of acute kidney injury (AKI). Recently, it was shown that intracellular actin released from damaged tissues appears in the urine of patients with multiple organ dysfunction syndrome. Our aims were to measure urinary actin (u-actin) concentrations of septic and control patients and to test if u-actin levels could predict AKI and mortality. METHODS: Blood and urine samples were collected from septic and sepsis-related AKI patients at three time points (T1-3): T1: within 24 hours after admission; T2: second day morning; T3: third day morning of follow-up. Patients with malignancies needing palliative care, end-stage renal disease or kidney transplantation were excluded. Serum and u-actin levels were determined by quantitative Western blot. Patients were categorized by the Sepsis-3 and KDIGO AKI classifications. RESULTS: In our study, 17 septic, 43 sepsis-induced AKI and 24 control patients were enrolled. U-actin levels were higher in septic patients compared with controls during follow-up (p<0.001). At T1, the septic and sepsis-related AKI groups also showed differences (p<0.001), yet this increase was not statistically significant at T2 and T3. We also detected significantly elevated u-actin concentrations in AKI-2 and AKI-3 septic patients compared with AKI-1 septic patients (p<0.05) at T1 and T3, along with a significant increase in AKI-2 septic patients compared with AKI-1 septic patients at T2 (p<0.01). This tendency remained the same when referring u-actin to urine creatinine. Parameters of first-day septic patient samples could discriminate AKI from non-AKI state (AUC ROC, p<0.001): u-actin: 0.876; se-creatinine: 0.875. Derived cut-off value for u-actin was 2.63 µg/L (sensitivity: 86.0%, specificity: 82.4%). CONCLUSION: U-actin may be a complementary diagnostic biomarker to se-creatinine in sepsis-related AKI while higher u-actin levels also seem to reflect the severity of AKI. Further investigations may elucidate the importance of u-actin release in sepsis-related AKI.
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Actinas/orina , Lesión Renal Aguda/diagnóstico , Biomarcadores/orina , Sepsis/patología , Actinas/sangre , Lesión Renal Aguda/etiología , Lesión Renal Aguda/mortalidad , Anciano , Área Bajo la Curva , Estudios de Casos y Controles , Creatinina/sangre , Creatinina/orina , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Curva ROC , Sepsis/complicaciones , Sepsis/diagnóstico , Sepsis/mortalidad , Índice de Severidad de la Enfermedad , Análisis de SupervivenciaRESUMEN
Background/aim: This study aimed to investigate the prevalence of sicca symptoms and secondary Sjögren's syndrome (SjS) in patients with systemic sclerosis (SSc). Also this study aimed to evaluate the expression of α-smooth muscle actin (αSMA) in minor salivary gland (MSG) specimens, a possible marker of fibrosis responsible for myofibroblastic transformation. Materials and methods: Patients with SSc who were followed in Rheumatology outpatient clinic at a university hospital evaluated. The questionnaire of sicca symptoms and classification of SjS were evaluated according to the AmericanEuropean Consensus Group (AECG) criteria. Histopathologic evaluations were done in MSG specimens investigating the presence of focal lymphocytic sialadenitis and glandular fibrosis, also assessing the expression of αSMA. Results: This cross-sectional study included 102 patients with SSc [91 females (89%), mean age 52.5 ± 12 years]. In this cohort 76 (75%) patients had sicca symptoms and 36 (35.3%) patients fulfilled the AECG criteria for SjS; all with limited form. Having SjS found to be associated with older age and the presence of positive anti-SS-A antibodies. On histopathologic examinations, glandular fibrosis was observed in 67 (80%) and lymphocytic sialadenitis was detected in 38 (45%) patients; but only 7 samples were positive for αSMA. Conclusion: This study suggested sicca symptoms were found to be very common among patients with SSc. Also secondary SjS was detected in nearly one-third of patients with SSc; especially in limited subtype. Anti SS-A positivity and older age were detected as predictors for SjS. Histopathologic evaluations showed significant glandular fibrosis but rare α-SMA staining in patients with SSc.
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Actinas , Glándulas Salivales Menores , Esclerodermia Sistémica , Sialadenitis , Síndrome de Sjögren , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Actinas/sangre , Biopsia , Estudios Transversales , Prevalencia , Glándulas Salivales Menores/patología , Esclerodermia Sistémica/complicaciones , Esclerodermia Sistémica/epidemiología , Sialadenitis/patología , Síndrome de Sjögren/complicaciones , Síndrome de Sjögren/epidemiologíaRESUMEN
Remote ischemic conditioning (RIC), transient restriction and recirculation of blood flow to a limb after traumatic brain injury (TBI), can modify levels of pathology-associated circulating protein. This study sought to identify TBI-induced molecular alterations in plasma and whether RIC would modulate protein and metabolite levels at 24 h after diffuse TBI. Adult male C57BL/6 mice received diffuse TBI by midline fluid percussion or were sham-injured. Mice were assigned to treatment groups 1 h after recovery of righting reflex: sham, TBI, sham RIC, TBI RIC. Nine plasma metabolites were significantly lower post-TBI (six amino acids, two acylcarnitines, one carnosine). RIC intervention returned metabolites to sham levels. Using proteomics analysis, twenty-four putative protein markers for TBI and RIC were identified. After application of Benjamini-Hochberg correction, actin, alpha 1, skeletal muscle (ACTA1) was found to be significantly increased in TBI compared to both sham groups and TBI RIC. Thus, identified metabolites and proteins provide potential biomarkers for TBI and therapeutic RIC in order to monitor disease progression and therapeutic efficacy.
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Actinas/sangre , Lesiones Traumáticas del Encéfalo , Precondicionamiento Isquémico , Proteómica , Animales , Biomarcadores/sangre , Lesiones Traumáticas del Encéfalo/sangre , Lesiones Traumáticas del Encéfalo/terapia , Modelos Animales de Enfermedad , Masculino , RatonesRESUMEN
BACKGROUND: While tissue injury provokes fibrinolysis shutdown in trauma, the mechanism remains elusive. Cellular death causes release of structural proteins, including actin and myosin, which may interact with clot formation and structure. We hypothesized that tissue injury is associated with high circulating actin and that actin produces a hypercoagulable profile with decreased fibrinolysis in vitro. METHODS: Blood was collected from trauma activation patients at a single Level I trauma center for thrombelastography and proteomics. Proteomic analyses were performed through targeted liquid chromatography coupled with mass spectrometry using isotope-labeled standards for quantification of actin and its endogenous inhibitor gelsolin. Based on the results, we added physiologic concentrations of cytoskeletal G-actin to whole blood from healthy volunteers and analyzed changes in thrombelastography, as well as to plasma and examined clot architecture using confocal microscopy of fluorescently labeled fibrinogen. RESULTS: Overall, 108 trauma patients were included: majority (71%) men, median age of 32.7 years, 66% blunt mechanism, median New Injury Severity Score (NISS) of 41. Compared with patients without severe tissue injury (NISS < 15, n = 10), patients with severe tissue injury (NISS > 15, n = 98) had higher levels of circulating actin (0.0428 vs. 0.0301, p = 0.02). Further, there was a trend toward lower gelsolin levels in patients with fibrinolysis shutdown (0.1844 vs. 0.2052, p = 0.17) and tissue plasminogen activator resistance (0.1676 vs. 0.2188, p = 0.06).Ten healthy volunteers were included in the in vitro experiments (50% male; median age, 31.3 years). Actin significantly increased angle (40.0° to 52.9°, p = 0.002) and decreased fibrinolysis (percent clot lysis 30 minutes after reaching maximum amplitude, 4.0% to 1.6%; p = 0.002), provoking fibrinolytic shutdown in three patients. The addition of actin to control plasma decreased fiber resolvability of fibrin clots, monitored by microscopy, and decreased plasmin-mediated fibrinolysis. CONCLUSION: Actin increases clot propagation and provokes fibrinolysis shutdown in vitro, through a mechanism of plasmin inhibition. High circulating levels of actin are present in trauma patients with severe tissue injury, suggesting actin contributes to fibrinolysis shutdown in the setting of tissue injury.
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Actinas/sangre , Fibrinólisis , Heridas y Lesiones/sangre , Adulto , Cromatografía Liquida , Femenino , Gelsolina/sangre , Humanos , Técnicas In Vitro , Puntaje de Gravedad del Traumatismo , Masculino , Espectrometría de Masas , Microscopía Confocal , Proteómica , Tromboelastografía , Centros TraumatológicosRESUMEN
BACKGROUND: The effect of bowel inflammation and cancer on the expression of the most prevalent internal controls: ACTB, GAPDH and B2M in whole blood is unknown, although at least GAPDH occurred to be tightly regulated and suspected of supporting cancer growth, challenging its suitability as a reference. OBJECTIVES: To evaluate the effect of colorectal cancer (CRC) and active inflammatory bowel disease (IBD) on the stability of ACTB, B2M, GAPDH, HPRT1, SDHA, and TBP leukocyte expression. MATERIAL AND METHODS: Gene expression in controls and CRC and IBD patients (n = 21/18/25) was evaluated in real-time quantitative polymerase chain reaction (RT-qPCR) using NormFinder, geNorm, BestKeeper, and comparative ΔCt method, and validated by comparison with absolute quantification of interleukin 1ß (IL-1ß) and CCL4. RESULTS: HPRT1, SDHA and TBP were superior normalizers in CRC and IBD. The highest expression variability was noted in active IBD. B2M was significantly lower in CRC but higher in IBD. GAPDH was higher in CRC and IBD. ACTB and GAPDH corresponded with CRC advancement (ρ = 0.52 and ρ = 0.53) and with clinical activity in Crohn's disease (ρ = 0.44 and ρ = 0.57) and ulcerative colitis (GAPDH: ρ = 0.72). ACTB, B2M and GAPDH correlated with circulating inflammatory/angiogenic indices, differently in IBD and CRC. CONCLUSIONS: Leukocyte GAPDH, ACTB, and B2M expression is affected by bowel inflammation and cancer, rendering them unsuitable as a reference in CRC and IBD.
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Actinas/sangre , Biomarcadores de Tumor/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Perfilación de la Expresión Génica/métodos , Gliceraldehído-3-Fosfato Deshidrogenasa (Fosforilante)/sangre , Enfermedades Inflamatorias del Intestino/genética , Enfermedades Inflamatorias del Intestino/patología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Microglobulina beta-2/sangre , Actinas/metabolismo , Inductores de la Angiogénesis/sangre , Quimiocina CCL4 , Neoplasias Colorrectales/metabolismo , Expresión Génica , Regulación Neoplásica de la Expresión Génica , Gliceraldehído-3-Fosfato Deshidrogenasa (Fosforilante)/metabolismo , Humanos , Enfermedades Inflamatorias del Intestino/metabolismo , Interleucina-1beta , Fragmentos de Péptidos , Estándares de Referencia , Microglobulina beta-2/metabolismoRESUMEN
Objective: We previously demonstrated that plasma levels of F-actin and Thymosin Beta 4 differs among patients with septic shock, non-infectious systemic inflammatory syndrome and healthy controls and may serve as biomarkers for the diagnosis of sepsis. The current study aims to determine if these proteins are associated with or predictive of illness severity in patients at risk for sepsis in the Emergency Department (ED).Methods: Prospective, biomarker study enrolling patients (>18 years) who met the Shock Precautions on Triage Sepsis rule placing them at-risk for sepsis.Results: In this study of 203 ED patients, F-actin plasma levels had a linear trend of increase when the quick Sequential Organ Failure Assessment (qSOFA) score increased. F-actin was also increased in patients who were admitted to the Intensive Care Unit (ICU) from the ED, and in those with positive urine cultures. Thymosin Beta 4 was not associated with or predictive of any significant outcome measures.Conclusion: Increased levels of plasma F-actin measured in the ED were associated with incremental illness severity as measured by the qSOFA score and need for ICU admission. F-actin may have utility in risk stratification of undifferentiated patients in the ED presenting with signs and symptoms of sepsis.
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Actinas/sangre , Inflamación/sangre , Sepsis/sangre , Choque Séptico/sangre , Timosina/sangre , Adulto , Anciano , Infecciones Bacterianas/sangre , Infecciones Bacterianas/mortalidad , Infecciones Bacterianas/patología , Biomarcadores/sangre , Servicio de Urgencia en Hospital , Femenino , Hospitalización , Humanos , Inflamación/microbiología , Inflamación/patología , Unidades de Cuidados Intensivos , Masculino , Persona de Mediana Edad , Enfermedades no Transmisibles/epidemiología , Puntuaciones en la Disfunción de Órganos , Pronóstico , Factores de Riesgo , Sepsis/microbiología , Sepsis/patología , Choque Séptico/microbiología , Choque Séptico/patologíaRESUMEN
BACKGROUND: Endoscopic tonsillectomy is associated with postoperative pain. Postoperative pain management remains to be improved in children. We aimed to investigate oxycodone preemptive analgesia in children undergoing endoscopic plasma total adenotonsillectomy. METHODS: 166 children with adenotonsillar hypertrophy were recruited at Wuhan Children's Hospital between 08/2016 and 03/2017. They were randomly assigned to receive SPOA (postoperative sufentanil), SPEA+SPOA (preemptive sufentanil and postoperative sufentanil), and OPEA+SPOA (preemptive oxycodone and postoperative sufentanil). The primary endpoint was serum c-fos levels. The secondary endpoints were the response entropy (RE) value, Pediatric Anesthesia Emergence Delirium (PAED) score, FLACC score, and adverse events. RESULTS: c-fos mRNA levels were increased significantly after surgery in the SPOA and SPEA+SPOA groups (Pâ<â.05). Postoperatively, c-fos mRNA levels were higher in the SPOA group compared with the OPEA+SPOA group (Pâ=â.044). The RE values increased in all groups after surgery (Pâ<â.05). At extubation, RE values were higher in the SPOA group compared with the SPEA+SPOA and OPEA+SPOA groups (Pâ<â.05). The PAED scores were higher in the SPOA group compared with the OPEA+SPOA group (Pâ=â.045). In the SPOA group, the FLACC scores were decreased at 24âh after surgery vs 4âhours (Pâ=â.044). Prediction probability (Pk) values indicated that RE and c-fos mRNA levels were quantitative predictors for early postoperative stress reaction after surgery. CONCLUSIONS: The subanalgesic dose of oxycodone (0.1âmg/kg) as preemptive analgesia could improve pain after endoscopic plasma total adenotonsillectomy in children.
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Adenoidectomía , Analgésicos Opioides/uso terapéutico , Oxicodona/uso terapéutico , Dolor Postoperatorio/prevención & control , Tonsilectomía , Actinas/sangre , Adenoidectomía/efectos adversos , Adenoidectomía/métodos , Niño , Endoscopía/efectos adversos , Endoscopía/métodos , Femenino , Humanos , Masculino , Proteínas Proto-Oncogénicas c-fos/sangre , Reacción en Cadena en Tiempo Real de la Polimerasa , Sufentanilo/uso terapéutico , Tonsilectomía/efectos adversos , Tonsilectomía/métodosRESUMEN
Bench press (i.e. arm-based) and half-squat (i.e. leg-based) are exercises commonly used to increase and evaluate muscular strength. In addition to differences in the location of the muscles that participate in each exercise, the total muscle mass required for the latter is larger than that involved in the former. The aim of this study is to analyze the effects of a maximal incremental strength test when performed by bench press and by half-squat on myocellular damage, oxidative damage and the inflammatory cytokine response. Ten male athletes were subjected to half-squat and bench press incremental strength tests. Blood samples were collected at rest, 15-minutes and 24â¯h post-test. Hydroperoxide and malondialdehyde concentrations were determined as lipid peroxidation markers. Lactate dehydrogenase (LDH) and creatine kinase isoenzyme MB (CK-MB) activities were determined as markers of muscle damage. α-Actin concentration was determined as a marker of sarcomeric damage. Serum interleukin (IL) 6, IL10, and tumor necrosis factor alpha (TNFα) were determined to assess the inflammatory response. LDH and CK-MB values were greater at 15â¯min and 24â¯h post bench press exercise (pâ¯<â¯0.05). No differences were found in lipid peroxidation or α-actin. Interestingly, IL10 values were greater in response to the press bench at 24â¯h post-test (pâ¯<â¯0.05). Our results suggest that, at equivalent workloads, an arm-based exercise induced higher anti-inflammatory effects and more severe muscle damage compared with a leg-based exercise.
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Mediadores de Inflamación/sangre , Interleucina-10/sangre , Músculo Esquelético/fisiología , Entrenamiento de Fuerza/métodos , Actinas/sangre , Brazo/fisiología , Forma MB de la Creatina-Quinasa/sangre , Humanos , Peróxido de Hidrógeno/sangre , Interleucina-6/sangre , L-Lactato Deshidrogenasa/sangre , Pierna/fisiología , Masculino , Malondialdehído/sangre , Contracción Muscular/fisiología , Fuerza Muscular/fisiología , Factor de Necrosis Tumoral alfa/sangre , Adulto JovenRESUMEN
BACKGROUND: Circulating cell-free DNA (cfDNA) is not found in healthy subjects, but is readily detected after thermal injury and may contribute to the risk of multiple organ failure. The hypothesis was that a postburn reduction in DNase protein/enzyme activity could contribute to the increase in cfDNA following thermal injury. METHODS: Patients with severe burns covering at least 15 per cent of total body surface area were recruited to a prospective cohort study within 24 h of injury. Blood samples were collected from the day of injury for 12 months. RESULTS: Analysis of blood samples from 64 patients revealed a significant reduction in DNase activity on days 1-28 after injury, compared with healthy controls. DNase protein levels were not affected, suggesting the presence of an enzyme inhibitor. Further analysis revealed that actin (an inhibitor of DNase) was present in serum samples from patients but not those from controls, and concentrations of the actin scavenging proteins gelsolin and vitamin D-binding protein were significantly reduced after burn injury. In a pilot study of ten military patients with polytrauma, administration of blood products resulted in an increase in DNase activity and gelsolin levels. CONCLUSION: The results of this study suggest a novel biological mechanism for the accumulation of cfDNA following thermal injury by which high levels of actin released by damaged tissue cause a reduction in DNase activity. Restoration of the actin scavenging system could therefore restore DNase activity, and reduce the risk of cfDNA-induced host tissue damage and thrombosis.
ANTECEDENTES: El ADN libre de las células circulantes (circulating cell-free DNA, cfDNA) no se encuentra en sujetos sanos, pero se detecta fácilmente después de una lesión térmica y puede contribuir al riesgo de fallo multiorgánico. La hipótesis fue que una disminución en la actividad de la proteína/enzima ADNasa tras la lesión térmica podría contribuir a la elevación del cfDNA que ocurre tras la misma. MÉTODOS: Los pacientes con quemaduras graves con una extensión ≥ 15% del área de superficie corporal total (total body surface area, TBSA) se incluyeron en un estudio prospectivo de cohortes durante las primeras 24 horas posteriores a la lesión. Se recogieron muestras de sangre desde el día de la lesión hasta los 12 meses posteriores a la misma. RESULTADOS: El análisis de muestras de sangre de 64 pacientes reveló una reducción significativa de la actividad de la ADNasa en los días 1 a 28 después de la lesión, en comparación con los controles sanos. Los niveles de proteína ADNasa no se vieron afectados, lo que sugiere la presencia de un inhibidor enzimático. Un análisis adicional reveló que la actina (un inhibidor de la ADNasa) estaba presente en las muestras de suero de los pacientes, pero no en los controles, y las concentraciones de la gelsolina, proteína que causa la disociación de la actina, y la proteína de unión a la vitamina D se redujeron significativamente después de la lesión térmica. En un estudio piloto de 10 pacientes con politrauma por lesiones militares, la administración de hemoderivados produjo un aumento en la actividad de la ADNasa y de los niveles de gelsolina. CONCLUSIÓN: Este estudio sugiere un nuevo mecanismo biológico para la acumulación de cfDNA después de una lesión térmica, por el cual los altos niveles de actina liberada por el tejido dañado causarían una reducción en la actividad de la ADNasa. La restauración del sistema eliminador de actina podría, por lo tanto, restaurar la actividad de la ADNasa y reducir el riesgo de daño tisular y trombosis en el huésped inducido por el cfDNA.
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Actinas/metabolismo , Quemaduras/metabolismo , Desoxirribonucleasas/metabolismo , Actinas/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Quemaduras/sangre , Quemaduras/enzimología , Estudios de Casos y Controles , Ácidos Nucleicos Libres de Células/sangre , Ácidos Nucleicos Libres de Células/metabolismo , Desoxirribonucleasas/sangre , Femenino , Fluorometría/métodos , Gelsolina/sangre , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Proteína de Unión a Vitamina D/sangre , Adulto JovenRESUMEN
BACKGROUND: Platelet-rich plasma (PRP) is frequently used in the treatment of acute and chronic wounds. One of the major problems concerning the use of PRP is the absence of a well-characterized and standardized product, which leads to a high variety in study outcomes. Therefore, more studies on the composition and standardization of PRP in wound healing are needed. STUDY DESIGN AND METHODS: Platelet concentrates derived from healthy blood donors were made in plasma (PC-plasma) or platelet additive solution (PC-PAS). The effects of PC-plasma, PC-PAS, and plasma were then tested on proliferation, differentiation, and migration of fibroblasts, as well as sprouting of endothelial cells in fibrin gels and chemotaxis of white blood cells (WBCs). RESULTS: PC-plasma stimulates the migration and proliferation of human dermal fibroblasts more than plasma or platelets alone. Furthermore, platelet factors decrease the expression of α-smooth muscle actin in dermal fibroblast cultures. PC-plasma also stimulates sprouting of endothelial cells. Finally, PC-plasma also acts as a strong chemoattractant for WBCs. CONCLUSIONS: Allogeneic PC-plasma has beneficial effects on various aspects of wound healing in vitro and is superior to plasma or platelets alone. PC-plasma is an attractive candidate for further in vivo evaluation.
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Plaquetas/fisiología , Quimiotaxis de Leucocito , Fibroblastos/fisiología , Neovascularización Fisiológica , Plasma/fisiología , Plasma Rico en Plaquetas/fisiología , Cicatrización de Heridas/fisiología , Actinas/sangre , Movimiento Celular , Proliferación Celular , Células Cultivadas , Humanos , Péptidos y Proteínas de Señalización Intercelular/sangreRESUMEN
BACKGROUND AND OBJECTIVE: Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic disease that has a poor 3-year median survival rate with unclear pathophysiology. Radiological features include bibasal, subpleural fibrosis and honeycombing while its pathology is characterized by fibroblastic foci and honeycombing. Proteomic analysis of circulating molecules in plasma may identify factors that characterize IPF and may assist in the diagnosis, prognostication and determination of pathogenic pathways in this condition. METHODS: Two independent quantitative proteomic techniques were used, isobaric tags for relative and absolute quantitation (iTRAQ) and multiple reaction monitoring (MRM), to identify differentially expressed plasma proteins in a group of IPF patients in comparison to healthy controls with normal lung function matched for age and gender. RESULTS: Five proteins were identified to be differentially expressed in IPF compared to healthy controls (upregulation of platelet basic protein and downregulation of actin, cytoplasmic 2, antithrombin-III, extracellular matrix protein-1 and fibronectin). CONCLUSION: This study further validates the combinational use of non-targeted discovery proteomics (iTRAQ) with targeted quantitation by mass spectrometry (MRM) of soluble biomarkers to identify potentially important molecules and pathways for pulmonary diseases such as IPF.
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Actinas/sangre , Antitrombina III/análisis , Proteínas de la Matriz Extracelular/sangre , Fibronectinas/sangre , Fibrosis Pulmonar Idiopática , Proteómica/métodos , beta-Tromboglobulina/análisis , Biomarcadores/sangre , Femenino , Regulación de la Expresión Génica , Humanos , Fibrosis Pulmonar Idiopática/diagnóstico , Fibrosis Pulmonar Idiopática/metabolismo , Masculino , Espectrometría de Masas/métodos , Persona de Mediana EdadRESUMEN
Hand, foot and mouth disease (HFMD) is an infectious disease caused by a variety of enterovirus infections, and the most common types of virus infections are the newenterovirus71 (EV71) and coxsackievirus A group 16 (CoxA16). A small fraction of HFMD will cause further severe HFMD. A rapid and accurate diagnosis biomarker of severe HFMD is important for the timely treatment. In the study, we conducted a clinical biomarker discovery study using iTRAQ combined with MS. Serum proteome alterations in severe HFMD group (nâ¯=â¯32) and health control group (nâ¯=â¯32) were analyzed. 47 proteins were upregulated (fold changeâ¯>â¯1.5) between the severe HFMD group and HC group. The identified proteins were classified into different groups according to the molecular function, biology processes, cellular component. During the up-regulated proteins, serum amyloid A (SAA) and human ß-actin (ACTB), were confirmed in the serum of the severe HFMD and HC by ELISA assay. SAA and ACTB levels were significantly higher in the sever HFMD patients (Pâ¯<â¯.01), consistent with iTRAQ-LC-MS/MS analysis. In summary, Our results showed that SAA and human ß-actin (ACTB) may be served as a potential biomarker of the clinical diagnosis of severe HFMD.
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Actinas/sangre , Enterovirus Humano A , Enfermedad de Boca, Mano y Pie/sangre , Proteína Amiloide A Sérica/metabolismo , Índice de Severidad de la Enfermedad , Biomarcadores/sangre , Preescolar , Femenino , Enfermedad de Boca, Mano y Pie/patología , Enfermedad de Boca, Mano y Pie/virología , Humanos , Masculino , ProteómicaRESUMEN
BACKGROUND The aim of this study was to determine whether senescence in renal glomeruli is involved in lupus nephritis (LN); the expression of senescence-associated ß-galactosidase (SA-ß-Gal) and its association with glomerular lesions were investigated in a mouse model of LN. MATERIAL AND METHODS Eighteen MRL/lpr mice with severe proteinuria were randomly divided into 2 equal groups and intraperitoneally injected with dexamethasone (DEX) or saline; 4 age-matched mice with mild proteinuria served as controls. Serum creatinine and urinary protein levels were analyzed, and kidney histological changes were observed by periodic acid-Schiff and Sirius Red staining. SA-ß-Gal was detected via histochemistry. Glomerular expression of collagen IV, α-SMA, and nephrin was analyzed by immunohistochemistry, and glomerular complement C3 deposition was tested by immunofluorescence. The relationships between SA-ß-Gal expression and renal function or glomerular lesion markers were determined by Spearman's correlation analysis. RESULTS Mice with severe proteinuria exhibited glomerular segmental sclerosis and endothelial cell proliferation. DEX administration suppressed these lesions but had no significant effect on 24-hour urinary protein levels. The elevated glomerular expression of SA-ß-Gal in proteinuric mice was attenuated by DEX treatment. In addition, DEX treatment markedly downregulated glomerular C3 deposition and collagen IV and α-SMA expression, while significantly increasing nephrin expression. Furthermore, SA-ß-Gal expression was positively correlated with urinary protein levels and expression of α-SMA. CONCLUSIONS Accelerated senescence of glomerular cells may contribute to glomerular injury in LN.
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Glomérulos Renales/patología , Nefritis Lúpica/patología , Actinas/sangre , Animales , Senescencia Celular/fisiología , Colágeno Tipo IV/sangre , Creatinina/sangre , Dexametasona/farmacología , Modelos Animales de Enfermedad , Femenino , Inmunohistoquímica , Glomérulos Renales/efectos de los fármacos , Glomérulos Renales/metabolismo , Nefritis Lúpica/sangre , Nefritis Lúpica/inducido químicamente , Nefritis Lúpica/metabolismo , Proteínas de la Membrana/sangre , Ratones , Ratones Endogámicos MRL lpr , Proteinuria/patología , beta-Galactosidasa/metabolismoRESUMEN
PURPOSE: Aging changes the balance of sex hormones and causes endothelial dysfunction in the penis, both of which are important determinants of erectile dysfunction (ED). The purpose of this study was to evaluate whether exercise training could protect against erectile dysfunction by increasing serum testosterone and penile eNOS levels in aging rats. METHODS: A total of 14 young (2-month-old) and 14 middle-aged (18-month-old) Sprague Dawley rats were randomly assigned to either untrained control (young control, [YC], middle-aged control, [MC]) or endurance exercise-trained (young exercise, [YE], middle-aged exercise, [ME]) groups with seven rats per group. The exercise groups trained with treadmill running for 6â¯weeks. Body composition parameters (body weight, heart mass, liver mass, and testicular mass), serum sex hormone levels (testosterone, luteinizing hormone, follicle-stimulating hormone, and prolactin), endothelial function-related parameters in the penis (endothelial nitric oxide synthase [eNOS], CD31, alpha smooth muscle actin [α-SMA]), and maximal intracavernous pressure measure (ICP) and total ICP were analyzed in middle-aged rats. RESULTS: The middle-aged groups showed increased body weight, as compared with the young groups, but exercise training attenuated the aging-induced increase in body weight. The middle-aged groups had lower testicular mass compared with the young groups, but exercise training attenuated aging-induced decreases in testicular mass. Exercise training increased serum testosterone levels in both the young and middle-aged groups. However, there were no changes in the levels of luteinizing hormone, follicle-stimulating hormone, and prolactin among the groups. MC group showed decreased protein levels of p-eNOS, as compared with the YC group. However, exercise training protected against aging-induced decrease in eNOS and p-eNOS protein levels in the penis. Interestingly, exercise training also increased protein levels of α-SMA and maximal ICP in the middle-aged group. CONCLUSIONS: Exercise training has beneficial effects on erectile function in aged rats through increased testosterone production from the testis and strengthening of the cavernous endothelium with activation of eNOS. Therefore, exercise training may be a therapeutic modality for improving erectile dysfunction associated with aging.
Asunto(s)
Disfunción Eréctil/prevención & control , Óxido Nítrico Sintasa de Tipo III/metabolismo , Erección Peniana/fisiología , Condicionamiento Físico Animal , Testosterona/sangre , Actinas/sangre , Envejecimiento/fisiología , Animales , Masculino , Pene/inervación , Pene/fisiología , Estimulación Física , Presión , Distribución Aleatoria , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To investigate the possible role of signal transducer and activator of transcription 5 (STAT5) in the pathogenesis of liver fibrosis in Egyptian patients with chronic hepatitis C (CHC) virus infection and its relation to hepatic stellate cells (HSC). SUBJECTS AND METHODS: Sixty-five patients (46 males and 19 females) were divided into 4 groups based on the severity of fibrosis as detected by Fibroscan as follows: F1, n = 15; F2, n = 21; F3, n = 13; and F4, n = 16. Twenty age- and gender-matched healthy persons volunteered as controls. The serum levels of STAT5, TGF-ß1, α-smooth muscle actin (α-SMA), fasting blood sugar, and fasting insulin, as well as homeostasis model assessment of insulin resistance (HOMA-IR), were determined and compared for all groups. The usefulness of the studied serum biomarkers for predicting liver fibrosis was evaluated using a receiver operating characteristic curve. RESULTS: Serum levels of STAT5 were significantly lower in patients compared to controls (9.69 ± 5.62 vs. 14.73 ± 6.52, p ≤ 0.001); on the contrary, TGF-ß1, α-SMA, and HOMA-IR were significantly higher in patients compared to controls (mean: 1,796.04 vs. 1,636.94; 14.94 vs. 8.1; and 7.91 vs. 4.18; p ≤ 0.01 and 0.001, respectively). TGF-ß1 and α-SMA showed a progressive increase with advancing severity of hepatic fibrosis (mean TGF-ß1: 2,058.4 in F1-F2 and 1,583.8 in F3-F4, p ≤ 0.04; mean α-SMA: 13.59 in F1-F2 and 16.62 in F3-F4, p ≤ 0.05). STAT5 had a significant negative correlation with TGF-ß1 (p ≤ 0.001), while no correlation was detected with α-SMA (p ≤ 0.8). CONCLUSIONS: STAT5 may play a significant role in hepatic fibrogenesis through the induction of TGF-ß1 but not through the activation of hepatic stellate cells.
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Actinas/sangre , Biomarcadores/sangre , Cirrosis Hepática/sangre , Factor de Transcripción STAT5/sangre , Factor de Crecimiento Transformador beta1/sangre , Adulto , Anciano , Estudios de Casos y Controles , Egipto , Femenino , Células Estrelladas Hepáticas , Hepatitis C Crónica/complicaciones , Humanos , Cirrosis Hepática/etiología , Masculino , Persona de Mediana Edad , Curva ROC , Adulto JovenRESUMEN
OBJECTIVE: The is to report immunohistochemical observations of aortic α-smooth muscle actin (SMA) expressions in patients with aortic aneurysm, acute aortic dissection, and coronary artery disease and to discuss phenotypic switching of smooth muscle cells (SMCs) of these lesions. METHODS: Forty-nine consecutive patients scheduled for surgical treatment for acute type A aortic dissection (20 patients), aortic aneurysm (9 patients), and coronary artery disease (20 patients) were included. Surgical specimens of the aorta were obtained and prepared for hematoxylin and eosin and immunohistochemical stainings. RESULTS: A comparison of aortic structural changes between the three groups showed that patients with coronary artery disease had the least severe aorta degeneration with the most intense α-SMA positivity. Aortic structural impairment was the most severe in patients with aortic dissection, whereas α-SMA positivity was more intense in patients with aortic dissection than in those with aortic aneurysm. CONCLUSION: Disparities in α-SMA expressions in the aortic tissues of the three groups represent the extent of SMC degenerations or a phenotypic switching between contractile and synthetic SMCs. The results imply severe SMC degenerations in patients with aortic aneurysm, which may be beneficial because of the production of extracellular matrix necessary for healing of the vascular wall, but severe disruptions in elastic fibers in patients with aortic dissection. Patients with coronary artery disease show slight SMC degeneration and phenotypic switching among the three groups. The possible apoptotic and genetic mechanisms of aortic structural impairments warrant further elaborations.
Asunto(s)
Actinas/metabolismo , Aneurisma de la Aorta Torácica/metabolismo , Disección Aórtica/metabolismo , Enfermedad de la Arteria Coronaria/metabolismo , Actinas/sangre , Disección Aórtica/sangre , Aneurisma de la Aorta Torácica/sangre , Enfermedad de la Arteria Coronaria/sangre , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana EdadRESUMEN
The short F-actins in the red blood cell (RBC) membrane skeleton are coated along their lengths by an equimolar combination of two tropomyosin isoforms, Tpm1.9 and Tpm3.1. We hypothesized that tropomyosin's ability to stabilize F-actin regulates RBC morphology and mechanical properties. To test this, we examined mice with a targeted deletion in alternatively spliced exon 9d of Tpm3 (Tpm3/9d-/- ), which leads to absence of Tpm3.1 in RBCs along with a compensatory increase in Tpm1.9 of sufficient magnitude to maintain normal total tropomyosin content. The isoform switch from Tpm1.9/Tpm3.1 to exclusively Tpm1.9 does not affect membrane skeleton composition but causes RBC F-actins to become hyperstable, based on decreased vulnerability to latrunculin-A-induced depolymerization. Unexpectedly, this isoform switch also leads to decreased association of Band 3 and glycophorin A with the membrane skeleton, suggesting that tropomyosin isoforms regulate the strength of F-actin-to-membrane linkages. Tpm3/9d-/- mice display a mild compensated anemia, in which RBCs have spherocytic morphology with increased osmotic fragility, reduced membrane deformability, and increased membrane stability. We conclude that RBC tropomyosin isoforms directly influence RBC physiology by regulating 1) the stability of the short F-actins in the membrane skeleton and 2) the strength of linkages between the membrane skeleton and transmembrane glycoproteins.
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Actinas/sangre , Eritrocitos/citología , Eritrocitos/metabolismo , Tropomiosina/sangre , Citoesqueleto de Actina/metabolismo , Animales , Masculino , Ratones , Ratones Noqueados , Polimerizacion , Unión Proteica , Isoformas de Proteínas , Tropomiosina/genética , Tropomiosina/metabolismoRESUMEN
BACKGROUND: Nonocclusive mesenteric ischemia can cause intestinal infarction but the diagnosis is challenging. This prospective study evaluated three plasma biomarkers of intestinal infarction after cardiac surgery. MATERIALS AND METHODS: Patients were recruited after cardiac surgery if they required laparotomy (with or without intestinal resection) for suspected nonocclusive mesenteric ischemia. Plasma levels of D-lactate, intestinal fatty acid-binding protein (i-FABP), and smooth muscle actin (SMA) before laparotomy were measured. RESULTS: Twenty patients were recruited (68 ± 9 y, EuroSCORE: 8.7 ± 2.8, mortality 70%). A positive laparotomy (n = 13) was associated with no change in D-lactate (P = 0.95), decreased i-FABP (P = 0.007), and increased SMA (P = 0.01). All patients with high SMA had a positive laparotomy. A subgroup analysis was undertaken in the eight patients who required multiple laparotomies. D-lactate increased between the two laparotomies in nonsurvivors (n = 4). Plasma i-FABP (P = 0.008) and SMA (P = 0.036) significantly decreased after the bowel resection, regardless of survival outcome. CONCLUSIONS: None of the biomarkers were accurate enough to reliably diagnose intestinal infarction. However, all patients with high values of SMA developed intestinal infarction, thus warranting further investigation. An increasing D-lactate after intestinal resection suggests impending death.
Asunto(s)
Actinas/sangre , Procedimientos Quirúrgicos Cardíacos , Proteínas de Unión a Ácidos Grasos/sangre , Infarto/diagnóstico , Ácido Láctico/sangre , Isquemia Mesentérica/diagnóstico , Complicaciones Posoperatorias/diagnóstico , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Femenino , Humanos , Infarto/sangre , Infarto/etiología , Infarto/cirugía , Intestinos/irrigación sanguínea , Laparotomía , Masculino , Isquemia Mesentérica/sangre , Isquemia Mesentérica/etiología , Isquemia Mesentérica/cirugía , Persona de Mediana Edad , Proyectos Piloto , Complicaciones Posoperatorias/sangre , Complicaciones Posoperatorias/cirugía , Estudios Prospectivos , Curva ROCRESUMEN
This review considers the functions of extracellular actin - cell surface bound, associated with extracellular matrix, or freely circulating. The role of this protein in different pathological processes is analyzed: its toxic effects and involvement in autoimmune diseases as an autoantigen. The extracellular actin clearance system and its role in protection against the negative effects of actin are characterized. Levels of free-circulating actin, anti-actin immunoglobulins, and components of the actin clearance system as prognostic biomarkers for different diseases are reviewed. Experimental approaches to protection against excessive amounts of free-circulating F-actin are discussed.