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Prohormone-derived neuropeptides act as cell-cell signaling molecules to mediate a wide variety of biological processes in the animal brain. Mass spectrometry-based peptidomic experiments are valuable approaches to gain insight into the dynamics of individual peptides under different physiological conditions or experimental treatments. However, the use of anesthetics during animal procedures may confound experimental peptide measurements, especially in the brain, where anesthetics act. Here, we investigated the effects of the commonly used anesthetics isoflurane and sodium pentobarbital on the peptide profile in the rodent hypothalamus and cerebral cortex, as assessed by label-free quantitative peptidomics. Our results showed that neither anesthetic dramatically alters peptide levels, although extended isoflurane exposure did cause changes in a small number of prohormone-derived peptides in the cerebral cortex. Overall, our results demonstrate that acute anesthetic administration can be utilized in peptidomic experiments of the hypothalamus and cerebral cortex without greatly affecting the measured peptide profiles.
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Anestésicos , Isoflurano , Ratas , Animales , Anestésicos/farmacología , Anestésicos/análisis , Péptidos/química , Hipotálamo/química , Corteza CerebralRESUMEN
During the winter, hibernating mammals undergo extreme changes in physiology, which allow them to survive several months without access to food. These animals enter a state of torpor, which is characterized by decreased metabolism, near-freezing body temperatures, and a dramatically reduced heart rate. The neurochemical basis of this regulation is largely unknown. Based on prior evidence suggesting that the peptide-rich hypothalamus plays critical roles in hibernation, we hypothesized that changes in specific cell-cell signaling peptides (neuropeptides and peptide hormones) underlie physiological changes during torpor/arousal cycles. To test this hypothesis, we used a mass spectrometry-based peptidomics approach to examine seasonal changes of endogenous peptides that occur in the hypothalamus and pituitary of a model hibernating mammal, the thirteen-lined ground squirrel (Ictidomys tridecemlineatus). In the pituitary, we observed changes in several distinct peptide hormones as animals prepared for torpor in October, exited torpor in March, and progressed from spring (March) to fall (August). In the hypothalamus, we observed an overall increase in neuropeptides in October (pre-torpor), a decrease as the animal entered torpor, and an increase in a subset of neuropeptides during normothermic interbout arousals. Notable changes were observed for feeding regulatory peptides, opioid peptides, and several peptides without well-established functions. Overall, our study provides critical insight into changes in endogenous peptides in the hypothalamus and pituitary during mammalian hibernation that were not available from transcriptomic measurements. Understanding the molecular basis of the hibernation phenotype may pave the way for future efforts to employ hibernation-like strategies for organ preservation, combating obesity, and treatment of stroke.
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Hibernación , Neuropéptidos , Hormonas Peptídicas , Animales , Estaciones del Año , Hibernación/fisiología , Transducción de Señal , Hipotálamo , MamíferosRESUMEN
Introduction: Early detection of Pneumocystis jirovecii as an opportunistic pathogen that may endanger predisposed persons, including COVID-19 patients, may help to choose the optimal management. Methods: In this study, 585, including 530 COVID-19 patients, with clinical and radiological evidence of respiratory diseases, were investigated for P. jirovecii screening. Clinical specimens were examined by direct microscopy and PCR, and randomly selected positive PCR products were confirmed through DNA sequence analysis. Results: Thirty-one (5.3%) samples were positive in P. jirovecii-specific nested-PCR, while by direct microscopic tests, Pneumocystis was observed in 22 (3.76%) samples. Males (61.7%) and patients over 50 years old (75.6%) were more commonly affected than others, and malaise and fatigue (84%), and wheezing (75%) were the most common symptoms, followed by fever (40.48%) and dyspnea (39.51%). Among the Pneumocystis-positive patients, three cases had coinfection with Aspergillus fumigatus, A. flavus, and A. niger (each n = 1), as documented by direct microscopy, culture, and species identification by PCR-sequencing. Conclusion: Pneumocystis pneumonia is still a diagnostic challenge; therefore, additional large-scale studies are needed to clarify the epidemiology of the disease in immunocompromised or COVID-19 patients.
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BACKGROUND: Hydatid cyst is the larval stage of the tape worm Echinococcus granulosus which is located in human and livestock viscera. There are some scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. In this study, the effects of a fraction of hydatid cyst fluid on colon cancer tumor in BALB/c mice were investigated. MATERIALS AND METHODS: In this experimental work six groups of mice were challenged with mouse colon cancer cells. 5 days later when the sign of tumor growth in mice was seen, group 1-4 were injected with hydatid cyst fluid, the 78 kDa fraction, live protoscolices and BCG respectively. Group five was injected with alum alone and the sixth group left intact without any injection. The size of the tumor was measured and compared in all groups. Then blood samples of mice were evaluated for serum cytokine levels. RESULT: In mice injected with hydatid cyst antigens especially a fraction of hydatid cyst fluid, tumor size was smaller than the that of control groups and the difference of tumor size in cases and control groups was statistically significant. CONCLUSION: The results of this study showed that injection of mice with a fraction of hydatid cyst fluid significantly inhibits the growth of mouse colon cancer and this inhibition may be related to effect of immune response to these antigens.
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Neoplasias del Colon , Equinococosis , Echinococcus granulosus , Echinococcus , Animales , Humanos , Ratones , Equinococosis/terapia , Neoplasias del Colon/terapia , InmunoterapiaRESUMEN
Cell-cell signaling peptides (e.g., peptide hormones, neuropeptides) are among the largest class of cellular transmitters and regulate a variety of physiological processes. To identify and quantify the relative abundances of cell-cell signaling peptides in different physiological states, liquid chromatography-mass spectrometry-based peptidomics workflows are commonly utilized on freshly dissected tissues. In such animal experiments, the administration of general anesthetics is an important step for many research projects. However, acute anesthetic administration may rapidly change the measured abundance of transmitter molecules and metabolites, especially in the brain and endocrine system, which would confound experimental results. The aim of this study was to evaluate the effect of short-term (<5 min) anesthetic administration on the measured abundance of cell-cell signaling peptides, as evaluated by a typical peptidomics workflow. To accomplish this goal, we compared endogenous peptide abundances in the rat pituitary following administration of 5% isoflurane, 200 mg/kg sodium pentobarbital, or no anesthetic administration. Label-free peptidomics analysis demonstrated that acute use of isoflurane changed the levels of a small number of peptides, primarily degradation products of the hormone somatotropin, but did not influence the levels of most other peptide hormones. Acute use of sodium pentobarbital had negligible impact on the relative abundance of all measured peptides. Overall, our results suggest that anesthetics used in pituitary peptidomics studies do not dramatically confound observed results.
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Anestésicos Generales , Isoflurano , Neuropéptidos , Hormonas Peptídicas , Animales , Hormona del Crecimiento , Neuropéptidos/análisis , Neuropéptidos/farmacología , Pentobarbital , Ratas , SodioRESUMEN
Background: Early and cost-effective diagnosis and monitoring of the infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are critically important to anticipate and control the disease. We aimed to set up a SYBR Green-based one-step real-time polymerase chain reaction (PCR) as a lower-cost alternative method to detect the virus. Materials and Methods: An in-house SYBR Green-based PCR assay targeting the envelope (E) and RNA-dependent RNA polymerase (RdRp) genes, was set up to diagnose the infection, and was compared with the reference probe-based PCR method. Results: When the commercial probe-based assay was considered as the reference method, SYBR Green-based PCR had a slightly lower sensitivity (81.98% and 86.25% for E and RdRp targets, respectively) and a good specificity (100% and 94.44% for E and RdRp targets, respectively). For both gene targets, three different melting temperature (Tm) patterns were found in the PCRs of the nasopharyngeal/oropharyngeal swab samples, but no size polymorphism was seen in agarose gel electrophoresis. Conclusion: Further studies to improvement of the assay are needed to make it an inexpensive and reliable tool for the diagnosis of COVID-19.
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Intercellular signaling events mediated by neuropeptides and peptide hormones represent important targets for both basic science and drug discovery. For many bioactive peptides, the protein receptors that transmit information across the receiving cell membrane are not known, severely limiting these signaling pathways as potential therapeutic targets. Identifying the receptor(s) for a given peptide of interest is complicated by several factors. Most notably, cell-cell signaling peptides are generated through dynamic biosynthetic pathways, can act on many different families of receptor proteins, and can participate in complex ligand-receptor interactions that extend beyond a simple one-to-one archetype. Here, we discuss recent methodological advances to identify signaling partners for bioactive peptides. Recent efforts have centered on methods to identify candidate receptors via transcript expression, methods to match peptide-receptor pairs through high throughput screening, and methods to capture direct ligand-receptor interactions using chemical probes. Future applications of the receptor identification approaches discussed here, as well as technical advancements to address their limitations, promise to lead to a greater understanding of how cells communicate to deliver complex physiologies. Importantly, such advancements will likely provide novel targets for the treatment of human diseases within the central nervous and endocrine systems.
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Neuropéptidos/metabolismo , Hormonas Peptídicas/metabolismo , Receptores Acoplados a Proteínas G/análisis , Receptores de Neuropéptido/análisis , Animales , Humanos , Ligandos , Aprendizaje Automático , Sondas Moleculares/química , Receptores Acoplados a Proteínas G/química , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Neuropéptido/química , Receptores de Neuropéptido/metabolismoRESUMEN
AIM: Chronic liver injury and cirrhosis leads to liver failure. Hyperammonemia is a deleterious consequence of liver failure. On the other hand, oxidative stress seems to play a pivotal role in the pathogenesis of liver fibrosis as well as in the cytotoxic mechanism of ammonia. There is no promising therapeutic agent against ammonia-induced complications. The present study was conducted to evaluate the role of carnosine (CA) administration on liver pathological changes, elevated plasma ammonia, and its consequent events in cirrhotic rats. METHODS: Bile duct ligated (BDL) rats were used as a model of cirrhosis. CA (250, 500, and 1000mg/kg, daily, i.p) was administered for 28 consecutive days to BDL animals. At the end of treatments, markers of oxidative stress and liver fibrosis was determined in liver and serum biomarkers of liver injury and plasma ammonia was assessed. Moreover, changes in animals' locomotor activity were monitored. RESULTS: Severe bridging fibrosis, inflammation, and necrosis in liver, along with elevated serum biomarkers of liver injury were evident in BDL animals. Furthermore, plasma ammonia was drastically elevated in cirrhotic rats and animals' locomotor activity was suppressed. It was found that CA (250, 500, and 1000mg/kg, daily, i.p) significantly alleviated liver injury and its consequent events in cirrhotic rats. The data suggested that CA is not only a useful and safe agent to preserve liver function, but also prevented hyperammonemia and brain damage as a deleterious consequence of cirrhosis and liver failure.
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Carnosina/uso terapéutico , Hiperamonemia/tratamiento farmacológico , Cirrosis Hepática/tratamiento farmacológico , Animales , Hiperamonemia/complicaciones , Cirrosis Hepática/complicaciones , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Hyperammonemia is associated with chronic and acute liver injury. There is no promising therapeutic agent against ammonia-induced complications. Hence, finding therapeutic molecules with safe profile of administration has clinical value. The present study was conducted to evaluate the role of taurine (TA) administration on plasma and brain ammonia and its consequent events in different models of chronic and acute liver injury and hyperammonemia. Bile duct ligated (BDL) rats were used as a model of chronic liver injury. Thioacetamide and acetaminophen-induced acute liver failure were used as acute liver injury models. A high level of ammonia was detected in blood and brain of experimental groups. An increase in brain ammonia level coincided with a decreased total locomotor activity of animals and significant changes in the biochemistry of blood and also liver tissue. TA administration (500 and 1000 mg/kg, i.p), effectively alleviated liver injury and its consequent events including rise in plasma and brain ammonia and brain edema. The data suggested that TA is not only a useful and safe agent to preserve liver function, but also prevented hyperammonemia as a deleterious consequence of acute and chronic liver injury.
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OBJECTIVE: To compare the changes in anti-malondialdehyde-modified low-density lipoprotein (MDA-LDL) IgG levels among patients undergoing off-pump and on-pump coronary artery bypass grafting (CABG) or valvuloplasty. SUBJECTS AND METHODS: A total of 38, 39 and 34 patients who underwent off-pump CABG, on-pump CABG and valvuloplasty, respectively, were enrolled in this study. Serum anti-MDA-LDL IgG values were measured 24 h before and after the operative procedures and at discharge. Echocardiography was also done before surgery and before discharge. The results were compared with values from 50 healthy controls. RESULTS: In all patients, a reduction in antibody titers was observed post-operatively. However, the decrease was significant only in the off-pump CABG - before surgery: 42.33 (25.83-58.51), after surgery: 30.86 (16.36-51.33) and at discharge: 10.96 (6.82-23.57; p = 0.027). There was a significant positive association between anti-MDA-LDL IgG levels and ejection fraction (r = 0.248, p = 0.036) and a negative association with E/E', a marker of pulmonary capillary wedge pressure, in the coronary patients (r = -0.345, p = 0.012), but no significant associations were found in patients with valvular heart disease. CONCLUSIONS: Serum anti-MDA-LDL IgG levels were associated with cardiac function indices in coronary patients undergoing CABG.
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Autoanticuerpos/inmunología , Procedimientos Quirúrgicos Cardíacos/métodos , Enfermedad de la Arteria Coronaria/cirugía , Enfermedades de las Válvulas Cardíacas/cirugía , Inmunoglobulina G/inmunología , Lipoproteínas LDL/inmunología , Malondialdehído/análogos & derivados , Adulto , Anciano , Biomarcadores , Gasto Cardíaco , Anuloplastia de la Válvula Cardíaca/métodos , Puente de Arteria Coronaria/métodos , Ecocardiografía , Femenino , Humanos , Masculino , Malondialdehído/inmunología , Persona de Mediana EdadRESUMEN
BACKGROUND AND AIM OF THE STUDY: Cardiopulmonary bypass (CPB) is used during on-pump coronary artery bypass grafting (CABG) and heart valvular replacement surgery, and is associated with the induction of oxidative stress. The aim of the study was to assess the association between indices of cardiac function and prooxidant-antioxidant balance (PAB) values in patients undergoing valve replacement surgery and on-or off-pump CABG. METHODS: Data were obtained from 44, 33, and 41 patients undergoing off-pump CABG, on-pump CABG, and valve replacement surgery, respectively. The PAB values were measured 24 h before and after the operative procedure, and at the time of discharge. Echocardiography was performed before surgery and before discharge. RESULTS: The changes in E/E', end-diastolic volume, end-systolic volume, left ventricular diastolic and systolic diameter were significantly related to baseline PAB values. In the valve replacement group, neither baseline nor changes in PAB values were associated with echocardiographic measurements. Also, neither off-pump nor on-pump CABG were significantly different in inducing oxidative stress (p = 0.596). When PAB values were measured in CABG patients, there was a significant difference in values between the three time points (p = 0.013). In the valve replacement group, PAB values were not significantly different between the preoperative and postoperative samples. CONCLUSION: The inverse association between the level of oxidative stress and cardiac function measurement may indicate that high levels of oxidative stress may be a predictor of the deterioration of cardiac function in CABG patients. However, in valvular heart disease patients the serum PAB value was not associated with changes in cardiac function. Levels of oxidative stress, as assessed by the PAB assay, were not significantly different for patients undergoing surgery with or without CPB.
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Antioxidantes , Puente Cardiopulmonar/efectos adversos , Puente de Arteria Coronaria , Implantación de Prótesis de Válvulas Cardíacas , Oxidantes/metabolismo , Estrés Oxidativo , Anciano , Antioxidantes/análisis , Antioxidantes/metabolismo , Bencidinas , Enfermedades Cardiovasculares/metabolismo , Enfermedades Cardiovasculares/cirugía , Compuestos Cromogénicos , Puente de Arteria Coronaria/efectos adversos , Puente de Arteria Coronaria/métodos , Femenino , Pruebas de Función Cardíaca/métodos , Implantación de Prótesis de Válvulas Cardíacas/efectos adversos , Implantación de Prótesis de Válvulas Cardíacas/métodos , Humanos , Masculino , Persona de Mediana Edad , Cuidados Posoperatorios , Cuidados Preoperatorios , Estadística como Asunto , Resultado del TratamientoRESUMEN
BACKGROUND: It has been reported that obesity is associated with higher levels of oxidative stress. We aimed to evaluate the hypothesis that pro-oxidant antioxidant balance (PAB) values could be affected by adiposity and to assess the association between PAB levels and indices of obesity. METHODS: Subjects (n = 733) were recruited and tlen were divided into 3 groups of normal-weight (BMI < 25, n = 207), overweight (25 < BMI < 30, n = 375), and obese (BMI > 30, n = 151). PAB values were measured in all participants. RESULTS: There was a significant association between PAB values and weight when the correlation was determined for all subjects (p < 0.05). Obese subject had significantly higher levels of PAB values [40.8 (34.3 - 51.1) HK unit] compared with overweight [37.5 (29.7 - 47.3) HK unit] and normal-weight subjects [37.2 (29.6 - 45.2) HK unit] (p < 0.05 in each case). However, there was no significant difference in PAB values between normal and over-weight subjects (p > 0.05). On performing the analysis of covariance, low-density lipoprotein-cholesterol (LDL-C) was found to have a significant independent association with PAB values (j = 0.046, p = 0.04). CONCLUSIONS: The high levels of PAB values in obese subjects without overt signs/symptoms of cardiovascular disease may be related to a heightened state of oxidative stress associated with obesity.
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Antioxidantes/metabolismo , Peso Corporal , Especies Reactivas de Oxígeno/sangre , Adulto , Estudios de Casos y Controles , Humanos , Persona de Mediana Edad , Obesidad , Valores de ReferenciaRESUMEN
BACKGROUND: Inflammatory states are known to cause an imbalance in the redox status. We aimed to study the possible associations between pro-oxidant-antioxidant balance (PAB) and serum high-sensitive C-reactive protein (hs-CRP) concentrations and traditional cardiovascular disease (CVD) risk factors in an unselected Iranian population and in groups of individuals with specific disease. METHODS: The study was conducted among an unselected population of 758 male subjects. Biochemical markers, including hs-CRP and PAB values, were measured. RESULTS: Serum hs-CRP concentrations were positively associated with serum PAB values (r = 0.260, P ≤ 0.001). Univariate analysis showed that PAB values were statistically higher in individuals who were obese or smokers compared with non-obese and non-smokers individuals (P < 0.01). While serum hs-CRP concentrations were significantly higher in older subjects, subjects with obesity, diabetes mellitus, metabolic syndrome, central obesity, hypertriglyceridaemia, higher low-density lipoprotein cholesterol concentrations, positive history of CVD and lower physical activity than subjects without these risk factors (P < 0.05). Comparing hs-CRP and PAB values in different CVD risk score subgroups showed a significant incremental rise in both parameters as CVD risk score increased (P < 0.05). Using multiple linear regression analysis we found a strong association between PAB values and hs-CRP concentrations. CONCLUSIONS: This study showed that among Iranian subjects, the inflammatory marker, hs-CRP, was strongly and positively associated with a marker of oxidative stress and also with several traditional risk factors of CVD. Moreover, the impact of traditional cardiovascular risk factors on hs-CRP concentrations and PAB values differed.
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Antioxidantes/metabolismo , Proteína C-Reactiva/metabolismo , Enfermedades Cardiovasculares/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Adulto , Anciano , Biomarcadores/metabolismo , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/epidemiología , Humanos , Irán/epidemiología , Masculino , Persona de Mediana Edad , Estrés Oxidativo , Factores de Riesgo , Adulto JovenRESUMEN
The relationship between serum anti-heat shock protein (Hsp)27 antibody and high sensitive C-reactive protein (hs-CRP) levels and indices of cardiac function were investigated in patients undergoing coronary artery bypass grafting (CABG) or heart valve replacement. The changes in anti-Hsp27 antibody titers and hs-CRP levels were compared among patients undergoing off-pump and on-pump CABG or valvular heart replacement. Fifty-three patients underwent off-pump, on-pump CABG, and heart valvular replacement in each group. Serum anti-Hsp27 titers and hs-CRP values were measured 24 h before and after the operation and at discharge. Echocardiography was performed before surgery and before discharge. The results were compared with values from 83 healthy controls. hs-CRP levels increased and anti-Hsp27 antibody decreased following surgery (P < 0.001 and P < 0.05, respectively), although these changes were independent of operative procedure (P = 0.361 and P = 0.120, respectively). Anti-Hsp27 antibody levels were higher at the time of discharge (P = 0.016). Only in coronary patients were anti-Hsp27 antibody levels negatively associated with E/E' (r = -0.268, P = 0.022), a marker of pulmonary capillary wedge pressure. In conclusions, anti-Hsp27 antibody levels are associated with indices of cardiac function in coronary patients. Cardiopulmonary bypass had no significant effect on the induction of changes in anti-Hsp27 levels. Moreover, anti-Hsp27 antibody levels fell in all groups postoperatively; this may be due to the formation of immune complexes of antigen-antibody, and antibody levels were higher at the time of discharge.
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Anticuerpos/sangre , Proteína C-Reactiva/análisis , Enfermedades Cardiovasculares/metabolismo , Proteínas de Choque Térmico HSP27/inmunología , Adulto , Anciano , Válvula Aórtica/cirugía , Biomarcadores/sangre , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/fisiopatología , Enfermedades Cardiovasculares/cirugía , Puente de Arteria Coronaria Off-Pump , Ecocardiografía , Femenino , Implantación de Prótesis de Válvulas Cardíacas , Humanos , Masculino , Persona de Mediana Edad , Periodo PosoperatorioRESUMEN
OBJECTIVES: We studied the association between the prooxidant-antioxidant balance (PAB), anti-malondialdehyde-modified low-density lipoprotein (oxidized LDL, ox-LDL) IgG antibody and indices of cardiac function (systolic and diastolic function) in patients with coronary artery disease (CAD). METHODS: Fifty-five patients with established CAD were selected, and serum levels of anti-ox-LDL IgG and PAB values were measured and compared with 40 matched healthy controls. Systolic and diastolic functions were determined for all patients. RESULTS: PAB values were significantly higher in patients than in controls (p < 0.001), whilst serum anti-ox-LDL concentrations were not statistically different between the 2 groups (p = 0.821). However, after adjustment for high-density lipoprotein cholesterol, the patients had higher anti-ox-LDL levels (p = 0.04). Total PAB values were inversely associated with ejection fraction (r = -0.326, p = 0.031), but this was not the case for anti-ox-LDL in either group (p > 0.05). CONCLUSION: Serum concentrations of a marker of oxidative stress (PAB values) are inversely associated with cardiac function. PAB is a relatively simple index that could be incorporated into risk assessment in CAD patients. Anti-ox-LDL IgG antibody concentration does not appear to reflect total oxidative stress as assessed by PAB.
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Anticuerpos/metabolismo , Antioxidantes/metabolismo , Enfermedad de la Arteria Coronaria/fisiopatología , Inmunoglobulina G/metabolismo , Lipoproteínas LDL/inmunología , Biomarcadores/metabolismo , Enfermedad de la Arteria Coronaria/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estrés Oxidativo/inmunología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
We studied the immune responses to heat shock protein (Hsp)-27 and pro-oxidant-antioxidant balance (PAB) values in patients with valvular heart disease, but free of angiographically evident coronary artery disease (CAD). Patients who were candidates for valvuloplasty surgery and 30 healthy matched controls were recruited. The anti-Hsp-27 antibody titers were 0.35 ± 0.04 absorbency units (AU) in the valvuloplasty group, being significantly higher than for the controls (0.11 ± 0.02 AU; P < .05). The PAB values were significantly higher in cases (134.67 ± 13.69 Hamidi-Koliakos(HK) unit) when compared with controls (49.78 ± 6.75 HK unit; P < .05). In cases, the ejection fraction was inversely correlated with anti-Hsp-27 antibody (P < .05) but was not significantly related to PAB values (P > .05). Based on the echocardiographic findings, the patients had no evident heart failure, but the high levels of anti-Hsp-27 and PAB values in patients with valvular heart disease may indicate that these variables can be used as markers of heart failure. However, a longitudinal study is required to confirm this hypothesis.