RESUMO
Introducción: Las membranopatías son anemias hemolíticas hereditarias debidas a anomalías cualitativas o deficiencias cuantitativas de las proteínas del citoesqueleto del glóbulo rojo. Objetivo: Actualizar el diagnóstico de las membranopatías con la inclusión de las últimas recomendaciones del comité de grupos de expertos a nivel nacional e internacional. Métodos: Se realizó una revisión de la literatura en inglés y español, a través del sitio web PubMed y el motor de búsqueda Google académico, de artículos publicados en los últimos cinco años. Análisis y síntesis de la información: Las enfermedades de mayor interés clínico son: la esferocitosis, la eliptocitosis y la estomatocitosis hereditaria. Estas en general se heredan con carácter autosómico dominante pero existen formas que se transmiten con carácter recesivo, sin descartar posible mutación de novo. Para su diagnóstico se utilizan pruebas que incluyen el estudio de la morfología de los glóbulos rojos, la fragilidad osmótica, la lisis de glicerol acidificado, la criohemólisis hipertónica, la prueba de unión a la eosina-5'-maleimida por citometría de flujo, la electroforesis en gel de poliacrilamida con dodecilsulfato sódico y la ectacitometría. Conclusiones: Las membranopatías pueden sospecharse de manera preliminar teniendo en cuenta algunas alteraciones de la morfología eritrocitaria, aunque el diagnóstico se basa en estudios familiares y otros de carácter confirmatorio de la enfermedad, como los estudios moleculares. Los profesionales de la salud que atienden a pacientes jóvenes con anemia deben considerar la posibilidad de una anemia hemolítica por trastornos de la membrana eritrocitaria(AU)
ABSTRACT Introduction: Membranopathies are inherited hemolytic anemias due to qualitative abnormalities or quantitative deficiencies of red blood cell cytoskeletal proteins. Objective: to update the diagnosis of membranopathies with the inclusion of the latest recommendations from the committee of expert groups at the national and international level. Methods: A review of the literature in English and Spanish was carried out, through the PubMed website and the academic search engine Google, in articles published in the last five years. Analysis and synthesis of information: The diseases of greatest clinical interest are: spherocytocis, elliptocytosis and hereditary stomatocytosis. These are generally inherited with an autosomal dominant character but there are forms that are transmitted recessively, without ruling out a possible de novo mutation. For its diagnosis, tests are used that include the study of red blood cell morphology, osmotic fragility, acidified glycerol lysis, hypertonic cryohemolysis, eosin-5'-maleimide binding test by flow cytometry, sodium dodecyl sulfate polyacrylamide gel electrophoresis and ectacytometry. Conclusions: Membranopathies can be preliminarily suspected taking into account some alterations in erythrocyte morphology, although the diagnosis is based on family studies and others confirming the disease, such as molecular studies. Healthcare professionals caring for young patients with anemia should consider the possibility of hemolytic anemia due to red cell membrane disorders(AU)
Assuntos
Humanos , Masculino , Feminino , Fragilidade Osmótica , Pessoal de Saúde , Atenção à Saúde , Eletroforese em Gel de Poliacrilamida , Anemia Hemolítica , Citometria de FluxoRESUMO
Objetivo: Analisar qualitativamente o teste de fragilidade osmótica (F.O.) para amostras a fresco ou após incubação a 37°C. Métodos: Foram processadas 20 amostras de sangue periférico, coletadas em duplicata com 5mL em cada tubo com heparina, de pacientes com solicitação de F.O. como exame de rotina para processamento a fresco e após incubação por 24 horas em banho-maria a 37°C, em 13 tubos com concentrações variáveis de 0,1% a 0,9% de NaCl. Resultados: Foram analisadas 20 amostras de pacientes em sua maioria do gênero feminino 17/20 (85%), com idades entre 3 meses a 75 anos, para realização do teste de F.O. A análise qualitativa dos resultados mostrou que 9/20 (45%) amostras tiveram resultado concordante entre os testes de F.O. para amostras a fresco e após incubação a 37°C. Dos resultados discordantes, 8/11 (72,7%) resultados mostram fragilidade dos eritrócitos à hemólise nas amostras a fresco e curva normal (sem hemólise) após incubação da amostra a 37°C. Outros 3/11 (22,3%) resultados apresentaram curva normal (sem hemólise) no teste com amostra à fresco e resistência à hemólise no teste com a amostra após incubação a 37°C. Com o teste de Extato de Fisher não mostrou diferença estatística (p=0,5743) para as amostras processadas a fresco ou após incubação a 37°C. Conclusão: O teste de F.O. se mostrou mais eficiente quando a amostra testada foi analisada após incubação por 24 horas a 37°C em banho-maria, contudo não houve diferença estatística para resultados processados a fresco ou após incubação a 37°C.
Objective: Qualitatively analyze the osmotic fragility test (O.F.) for samples fresh or after incubation at 37°C. Methods: Twenty peripheral blood samples were processed, collected in duplicate with 5 ml in each tube with heparin, from patients with O.F. request. as a routine examination for fresh processing and after incubation for 24 hours in a water bath at 37°C, in 13 tubes with varying concentrations of 0.1% to 0.9% NaCl. Results: Twenty samples of patients were analyzed, mostly female, 17/20 (85%), aged between 3 months to 75 years, for the O.F. test. The qualitative analysis of the results showed that only 9/20 (45%) samples had a consistent result between the F.O. tests for fresh samples and after incubation at 37°C. From the discordant results, 8/11 (72.7%) results show fragility of erythrocytes to hemolysis in fresh samples and normal curve (without hemolysis) after sample incubation at 37o C. Other 3/11 (22.3%) results showed normal curve (without hemolysis) in the test with fresh sample and resistance to hemolysis in the test with the sample after incubation at 37o C. With the Fisher Extact test showing no statistical difference (p=0.5743) for samples processed fresh or after incubation at 37°C. Conclusion: The O.F. proved to be more efficient when the tested sample was analyzed after incubation for 24h at 37°C in a water bath, however, there was no statistical difference for results processed fresh or after incubation at 37°C.
Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Adulto Jovem , Fragilidade Osmótica , Anemia Hemolítica , HemóliseRESUMO
This study aimed to investigate the relationship between red cell distribution width (RDW) and erythrocyte osmotic stability in non-diabetic and diabetic individuals in both sexes. The study sample (N = 122) was constituted by 53 type 2 diabetics (DM) and 69 non-diabetics (ND), being 21 and 22 men in each group, respectively. The osmotic stability of erythrocytes was obtained by the variation in saline concentration (dX) capable of determining hypoosmotic lysis. Higher RDW values and lower serum iron concentrations were found in the diabetic group when compared to the non-diabetic volunteers. In the group of diabetic women, RDW was positively correlated with the reticulocyte index, and both RDW and dX were negatively correlated with iron, haemoglobin, transferrin saturation index, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. In all the groups studied, RDW was positively correlated with dX, especially in the diabetic group, where the correlation was the strongest. RDW elevation in both women and men with type 2 diabetes mellitus was associated with decreased serum iron indicators. Furthermore, RDW has a similar meaning to dX, as small erythrocytes have less haemoglobin, resulting in both an increase of RDW and dX.
Assuntos
Diabetes Mellitus Tipo 2/sangue , Índices de Eritrócitos , Fragilidade Osmótica , Idoso , Biomarcadores/sangue , Contagem de Células Sanguíneas , Diabetes Mellitus Tipo 2/metabolismo , Eritrócitos/metabolismo , Feminino , Hemoglobinas/metabolismo , Humanos , Ferro/metabolismo , Cinética , Masculino , Pessoa de Meia-IdadeRESUMO
Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous and complex disease, both from a clinical and molecular point of view. The prolonged use of alcohol and tobacco, along with the release of tumor secretions can modulate blood cells, such as erythrocytes. Here, this study was conducted with 24 patients diagnosed with HNSCC and an equal number of healthy individuals are matched by age and gender. The levels of lipid peroxidation were measured using the individual plasma, while for lipid concentrations, identification and quantification Na, K-ATPase activity and osmotic fragility, the red blood cell concentrate were used. The release of TBARS was significantly higher in patients with HNSCC. The lipid profile assays demonstrated a rearrangement of the erythrocyte membrane due to a decrease in total phospholipids and phosphatidylethanolamine followed by an increase in total cholesterol and phosphatidylcholine. Na, K-ATPase activity also increased. Erythrocytes were more fragile in patients with HNSCC than in health individuals. Therefore, the membrane of erythrocytes were rearranged and Na, K-ATPase function altered in the HNSCC patients. Our findings suggests that the alcohol, tobacco and tumor secretion modulate in a specific manner that the erythrocytes membranes of these patients making this system a potential tool for HNSCC biomarker of tumor progression.
Assuntos
Membrana Eritrocítica/química , Membrana Eritrocítica/metabolismo , Neoplasias de Cabeça e Pescoço/metabolismo , Biomarcadores , Estudos de Casos e Controles , Humanos , Peroxidação de Lipídeos , Lipídeos de Membrana/metabolismo , Fragilidade Osmótica , Estresse Oxidativo , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
ABSTRACT Background: The osmotic fragility test (OFT), conventionally used for assisting the diagnosis of many erythrocyte disorders, is a manual and time-consuming analysis not daily performed in many medical laboratories. This study was aimed at defining the stability of whole blood samples used for assessing erythrocyte osmotic resistance. Methods: Twenty-one consecutive routine whole blood samples collected into 5.4 mg K2EDTA were tested immediately after collection (day 0) and at different time intervals afterward (day 1, 2, 3, 4, 7, 10 and 14) after storage at 4 °C. The OFT was performed with the Osmored Monotest (1.3% glycerol; Eurospital, Trieste, Italy). Results at the different time points were compared with those obtained at day 0 and with the reference change value (i.e., 33%). Results: The median value of both hyperosmolar and hyposmolar resistance increased from baseline, reaching statistical significance at day 7 for hyperosmolar resistance and at day 1 for hyposmolar resistance, respectively. The median relative increase of hemolysis percentage values become greater than the reference change value at day 3 for hyposmolar resistance, while this limit was never overcome for hyperosmolar resistance. A significant inverse association was found between the mean increase in hyperosmolar resistance and the baseline value of hyperosmolar resistance (r = −0.92), mean corpuscular volume (MCV; r = −0.46) or mean corpuscular hemoglobin (MCH; r = −0.44), as well as between the mean increase in hyposmolar resistance and the baseline value of hyposmolar resistance (r = −0.86), or patient age (r = −0.56). Conclusions: The sample stability seems critical for the OFT. Whole blood specimens should not be stored refrigerated at 4 °C for >2 days before testing.
Assuntos
Humanos , Masculino , Feminino , Idoso , Fragilidade Osmótica , Eritrócitos , Fase Pré-AnalíticaRESUMO
Hydroxyurea (HU) is used as a therapy in sickle cell anemia (SCA). Many studies have established that HU improves patient quality of life by reducing symptoms. However, the effect of HU on erythrocytes is not well-described. We evaluated several parameters related to oxidative stress and total lipid content of erythrocytes in patients with SCA. The patient cohort consisted of 7 SCA patients treated with HU, 17 untreated SCA patients, and 15 healthy subjects. Erythrocytes from patients with SCA displayed increased oxidative stress relative to the control group, including higher thiobarbituric acid reactive substances (TBARS), Fe3+ content, and osmotic fragility, and decreased total cholesterol. We observed that treatment of SCA patients with HU increased Fe3+ content and activity of glutathione peroxidase, and decreased glutathione reductase activity, glutathione levels, total cholesterol, and phospholipid content comaperaded to patients untreated with HU. Thus, HU alters biochemical characteristics of erythrocytes; future studies will determine whether they are beneficial or not.
Assuntos
Anemia Falciforme , Eritrócitos Anormais/metabolismo , Hidroxiureia/administração & dosagem , Estresse Oxidativo/efeitos dos fármacos , Adulto , Anemia Falciforme/sangue , Anemia Falciforme/tratamento farmacológico , Colesterol/sangue , Feminino , Humanos , Masculino , Fragilidade Osmótica/efeitos dos fármacos , Fosfolipídeos/sangue , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Plant species have been used for therapeutic purposes since ancient times and are still in use today since these products represent a source of raw material for the production of phytotherapeutic formulations. Screening and investigation of plants with pharmacological potential require the evaluation of characteristics related to their action, efficacy and safety in different steps. Among these steps, pre- clinical trials are used to evaluate the properties of the test product in in vitro experiments, such as cytotoxicity assays. Within this context, this study consists of a bibliometric analysis of some in vitro cytotoxicity and toxicity assays in erythrocytes used during bioprospecting of medicinal plants. The results demonstrated the wide application of erythrocytes to evaluate the biological effects of medicinal plant extracts. The methods were found to be valid and effective for the preliminary investigation of the in vitro cytotoxicity and toxicity of plant products.
El uso de especies vegetales para fines terapeÌuticos es una praÌctica histoÌrica y todaviÌa bastante actual, ya que estos productos pueden representar una fuente de materia prima para la produccioÌn de formulaciones fitoteraÌpicas. En investigacioÌn de plantas con potencial farmacoloÌgico requiere la evaluacioÌn de su accioÌn, eficacia y seguridad, a traveÌs de diferentes etapas. Entre estas, en los ensayos precliÌnicos se evaluÌan las propiedades del producto-prueba en experimentos in vitro, tales como ensayos de citotoxicidad, entre otros. En este aspecto, el presente estudio consiste en un anaÌlisis bibliomeÌtrico acerca de algunas pruebas de citotoxicidad y toxicidad in vitro en eritrocitos realizados en los ensayos de bioprospeccioÌn de plantas medicinales. Los resultados evidencian la amplia utilizacioÌn de eritrocitos para la evaluacioÌn de los efectos bioloÌgicos de extractos de plantas medicinales, apuntaÌndolos como meÌtodos vaÌlidos y eficaces para la investigacioÌn preliminar de la citotoxicidad y toxicidad in vitro de productos vegetales.
Assuntos
Bioensaio/métodos , Extratos Vegetais/toxicidade , Eritrócitos/efeitos dos fármacos , Antioxidantes/toxicidade , Fragilidade Osmótica , Estresse Oxidativo , Eritrócitos/citologia , Bioprospecção , Hemólise/efeitos dos fármacosRESUMO
BACKGROUND: The cytometric flow osmotic fragility test (FC-OFT) was recently introduced. However, the test is still under development and some variables have not yet been fully tested. METHODS: The osmotic fragility of hereditary spherocytosis (HS) cases and healthy controls were evaluated by FC-OFT using a series of tubes containing decreasing concentrations of NaCl. The analyses were executed in fresh and incubated (37°C for 24 h) blood samples anticoagulated with EDTA and heparin. The percentages of residual red blood cells were used to plot the osmotic fragility curves. The OF curves of each tested condition were compared using the median corpuscular fragility (MCF). ROC curve analyses identified the most accurate NaCl concentrations for differentiation between HS cases and healthy controls. RESULTS: FC-OFT curves assumed a sigmoidal dose-response shape and the MCF of cases and controls were different in all instances. MCF comparisons revealed that incubation and anticoagulant have major and minor effects on the FC-OFT, respectively. One hundred percent of sensitivity and specificity was obtained from 5.5 to 6.0 g/L of NaCl in EDTA-treated fresh blood, from 6.0 to 8.0 g/L of NaCl in EDTA-treated incubated blood, and in none of the tested NaCl concentration in heparinized blood. CONCLUSIONS: EDTA is the anticoagulant of choice for the assay. Incubation at 37°C for 24 h increased its diagnostic capability. The most reliable NaCl concentration for the discrimination of HS case from controls was 6.0 g/L of NaCL in fresh EDTA-treated blood, and was 7.5 g/L of NaCl in incubated EDTA-treated blood. © 2018 International Clinical Cytometry Society.
Assuntos
Anticoagulantes/farmacologia , Eritrócitos/efeitos dos fármacos , Fragilidade Osmótica/efeitos dos fármacos , Cloreto de Sódio/análise , Esferocitose Hereditária/tratamento farmacológico , Adulto , Estudos de Casos e Controles , Feminino , Citometria de Fluxo , Humanos , Masculino , Curva ROC , Esferocitose Hereditária/diagnósticoAssuntos
Clorpirifos/toxicidade , Eritrócitos/efeitos dos fármacos , Sangue Fetal/efeitos dos fármacos , Inseticidas/toxicidade , Neonicotinoides/toxicidade , Aldeídos/toxicidade , Catalase/metabolismo , Eritrócitos/enzimologia , Eritrócitos/metabolismo , Sangue Fetal/enzimologia , Sangue Fetal/metabolismo , Humanos , Fragilidade Osmótica/efeitos dos fármacos , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismoRESUMO
ResumenLas alteraciones del sodio son un factor primordial o agravante que podrían condicionar o causar la muerte tanto en personas conocidas sanas como que en aquellas que presentan algún tipo de patología ya sea aguda o crónica. Dichas alteraciones en muchas ocasiones podrían pasar desapercibidas, dado su grandísima complejidad de detección, de tratamiento y en ocasiones de manejo clínico. Siendo estos factores que vuelven a dichas alteraciones un eje de la medicina clínica olvidada en muchas ocasiones por parte de los peritos encargados de realizar autopsias tanto clínicas como aquellas en las que priva algún interés de índole médico legal.
AbstractSodium alterations are a primary or aggravating factor that could cause death in both well-known and wellknown people, as well as those that present some type of pathology, either acute or chronic. These alterations in many occasions could go unnoticed, given their great complexity of detection, treatment and sometimes clinical management. These factors make these alterations an axis of clinical medicine often forgotten by experts in charge of performing clinical autopsies as well as those in which it deprives some interest of a legal medical nature
Assuntos
Humanos , Fragilidade Osmótica , Sódio , Causas de Morte , Mielinólise Central da Ponte , Patologia Legal , Medicina Legal , Hipernatremia , HiponatremiaRESUMO
This study aimed to evaluate the influence of acute and chronic exercise on erythrocyte membrane stability and various blood indices in a population consisting of five national-level male swimmers, over 18 weeks of training. The evaluations were made at the beginning and end of the 1st, 7th, 13th and 18th weeks, when volume and training intensity have changed. The effects manifested at the beginning of those weeks were considered due to chronic adaptations, while the effects observed at the end of the weeks were considered due to acute manifestations of the exercise load of that week. Acute changes resulting from the exercise comprised increases in creatine kinase activity (CK) and leukocyte count (Leu), and decrease in hematocrit (Ht) and mean corpuscular volume (MCV), at the end of the first week; increase in the activities of CK and lactate dehydrogenase (LDH), in the uric acid (UA) concentration and Leu count, at the end of the seventh week; increases in CK and LDH activities and in the mean corpuscular hemoglobin concentration (MCHC), at the end of the 13th week; and decrease in the value of the osmotic stability index 1/H50 and increases in the CK activity and platelets (Plt) count, at the end of the 18th week. Chronic changes due to training comprised increase in the values of 1/H50, CK, LDH, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), serum iron (Fe), MCV and Plt. Although acute training has resulted in decrease in the osmotic stability of erythrocytes, possibly associated with exacerbation of the oxidative processes during intense exercise, chronic training over 18 weeks resulted in increased osmotic stability of erythrocytes, possibly by modulation in the membrane cholesterol content by low and high density lipoproteins.
Assuntos
Membrana Eritrocítica/metabolismo , Natação/fisiologia , Atletas , Creatina Quinase/sangue , Índices de Eritrócitos/fisiologia , Hematócrito , Humanos , L-Lactato Desidrogenase/sangue , Contagem de Leucócitos , Masculino , Fragilidade Osmótica/fisiologia , Contagem de Plaquetas , Ácido Úrico/sangue , Adulto JovemRESUMO
Methylglyoxal (MG) is a reactive dicarbonyl metabolite originated mainly from glucose degradation pathway that plays an important role in the pathogenesis of diabetes mellitus (DM). Reactions of MG with biological macromolecules (proteins, DNA and lipids) can induce cytotoxicity and apoptosis. Here, human erythrocytes, leukocytes and platelets were acutely exposed to MG at concentration ranging from 0.025 to 10 mM. Afterwards, hemolysis and osmotic fragility in erythrocytes, DNA damage and cell viability in leukocytes, and the activity of purinergic ecto-nucleotidases in platelets were evaluated. The levels of glycated products from leukocytes and free amino groups from erythrocytes and platelets were also measured. MG caused fragility of membrane, hemolysis and depletion of amino groups in erythrocytes. DNA damage, loss of cell viability and increased levels of glycated products were observed in leukocytes. In platelets, MG inhibited the activity of enzymes NTPDase, 5'-nucleotidase and adenosine deaminase (ADA) without affecting the levels of free amino groups. Our findings provide insights for understanding the mechanisms involved in MG acute toxicity towards distinct blood cells.
Assuntos
Plaquetas/efeitos dos fármacos , Dano ao DNA , Eritrócitos/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Aldeído Pirúvico/toxicidade , 5'-Nucleotidase/metabolismo , Adenosina Desaminase/metabolismo , Adulto , Plaquetas/enzimologia , Plaquetas/patologia , Sobrevivência Celular/efeitos dos fármacos , Ensaio Cometa , Relação Dose-Resposta a Droga , Eritrócitos/enzimologia , Eritrócitos/patologia , Feminino , Hemólise/efeitos dos fármacos , Humanos , Leucócitos/enzimologia , Leucócitos/patologia , Masculino , Fragilidade Osmótica/efeitos dos fármacosRESUMO
Eugenia uniflora is used in the Brazilian folk medicine to treat intestinal disorders and hypertension. However, scanty information exist on its potential toxicity to human, and little is known on its antioxidant activity in biological system. Hence, we investigated for the first time the potential toxic effects of ethanolic extract (EtOH) of E. uniflora (EEEU) in human leukocytes and erythrocytes, as well as its influence on membrane erythrocytes osmotic fragility. In addition, EEEU was chemically characterized and its antioxidant capacity was evaluated. We found that EEEU (1-480µg/mL) caused neither cytotoxicity nor DNA damage evaluated by Trypan blue and Comet assay, respectively. EEEU (1-480µg/mL) did not have any effect on membrane erythrocytes fragility. In addition, EEEU inhibited Fe2+-induced lipid peroxidation in rat brain and liver homogenates, and scavenged the DPPH radical. EEEU presented some polyphenolic compounds with high content such as quercetin, quercitrin, isoquercitrin, luteolin and ellagic acid, which may be at least in part responsible for its beneficial effects. Our results suggest that consumption of EEEU at relatively higher concentrations may not result in toxicity. However, further in vitro and in vivo studies should be conducted to ascertain its safety.
Assuntos
Antioxidantes/farmacologia , Eritrócitos/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Solventes/química , Antioxidantes/isolamento & purificação , Antioxidantes/toxicidade , Compostos de Bifenilo/química , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Ensaio Cometa , Relação Dose-Resposta a Droga , Eritrócitos/metabolismo , Etanol/química , Eugenia/química , Humanos , Leucócitos/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fragilidade Osmótica/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fitoterapia , Picratos/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/toxicidade , Folhas de Planta , Plantas Medicinais , Polifenóis/isolamento & purificação , Polifenóis/farmacologia , Medição de Risco , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Aquaporin-1 (AQP1) is the membrane water channel responsible for changes in erythrocyte volume in response to the tonicity of the medium. As the aberrant distribution of proteins in hereditary spherocytosis (HS) generates deficiencies of proteins other than those codified by the mutated gene, we postulated that AQP1 expression might be impaired in spherocytes. AQP1 expression was evaluated through flow cytometry in 5 normal controls, 1 autoimmune hemolytic anemia, 10 HS (2 mild, 3 moderate, 2 severe, and 3 splenectomized), and 3 silent carriers. The effect of AQP1 inhibitors was evaluated through water flow-based tests: osmotic fragility and hypertonic cryohemolysis. Serum osmolality was measured in 20 normal controls and 13 HS. The effect of erythropoietin (Epo) on AQP1 expression was determined in cultures of erythroleukemia UT-7 cells, dependent on Epo to survive. Independent of erythrocyte size, HS patients showed a lower content of AQP1 in erythrocyte membranes which correlated with the severity of the disease. Accordingly, red blood cells from HS subjects were less sensitive to cryohemolysis than normal erythrocytes after inhibition of the AQP1 water channel. A lower serum osmolality in HS with respect to normal controls suggests alterations during reticulocyte remodeling. The decreased AQP1 expression could contribute to explain variable degrees of anemia in hereditary spherocytosis. The finding of AQP1 expression induced by Epo in a model of erythroid cells may be interpreted as a mechanism to restore the balance of red cell water fluxes.
Assuntos
Aquaporina 1/biossíntese , Eritrócitos/metabolismo , Regulação da Expressão Gênica , Esferocitose Hereditária/sangue , Adolescente , Adulto , Anemia Hemolítica Autoimune/sangue , Anemia Hemolítica Autoimune/genética , Aquaporina 1/sangue , Aquaporina 1/genética , Transporte Biológico , Água Corporal , Linhagem Celular , Criança , Pré-Escolar , Membrana Eritrocítica/metabolismo , Eritrócitos/patologia , Eritropoetina/farmacologia , Hemólise , Heterozigoto , Humanos , Leucemia Linfocítica Crônica de Células B/sangue , Pessoa de Meia-Idade , Concentração Osmolar , Fragilidade Osmótica , Esferocitose Hereditária/genética , Esferocitose Hereditária/cirurgia , EsplenectomiaRESUMO
Phytochemicals such as plant essential oils (EOs) have been reported to favour various activities in the innate immune system of fish. Thus, the aim of this study was to verify the in vitro effect of three different plant EOs (Ocimum americanum, Cymbopogon flexuosus and Melaleuca alternifolia) on non-specific immune parameters and erythrocyte osmotic fragility of red drum, Sciaenops ocellatus. Concentrations of each plant EO evaluated in preparations of head-kidney macrophages, blood leucocytes and blood plasma were as follows: 0.0 (control), 1.0, 2.0, 4.0, 8.0, and 16.0 µg/ml. Red drum head-kidney macrophages significantly increased extracellular superoxide anion production when exposed (20 h) to O. americanum EO (1.0-8.0 µg/ml) and C. flexuosus EO (2.0 and 4.0 µg/ml). The respiratory burst of blood leucocytes (NBT test) significantly increased in all concentrations when compared to the respective control group, for all EOs. At the highest concentration (16.0 µg/ml), C. flexuosus EO significantly inhibited the haemolytic activity of complement system in red drum blood after 1 h exposure. None of the tested concentrations significantly altered plasma lysozyme activity or erythrocyte osmotic fragility after exposing (1 h) red drum whole blood to each EO. This study demonstrated that these plant EOs are capable of triggering superoxide anion production in red drum leucocytes (head-kidney macrophages and/or blood leucocytes). In vivo studies are warranted to address their potential as immunostimulants in the diet of red drum and other aquacultured species.
Assuntos
Ração Animal/análise , Dieta/veterinária , Peixes/imunologia , Óleos Voláteis/farmacologia , Óleos de Plantas/farmacologia , Animais , Cymbopogon/química , Eritrócitos/fisiologia , Rim Cefálico/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/fisiologia , Melaleuca/química , Ocimum/química , Óleos Voláteis/administração & dosagem , Fragilidade Osmótica/efeitos dos fármacos , Óleos de Plantas/administração & dosagemRESUMO
OBJECTIVE: To evaluate the osmotic fragility of red blood cells and the level of lipid peroxidation, the Ca(2+)-ATPase activity of red cell ghosts and placental homogenates from salt-loaded pregnant rats. METHODS: Salt-loaded pregnant rats received 1.8% NaCl solution ad libitum as a beverage for seven days, starting on 15th day of pregnancy. Then, it was evaluated the level of lipid peroxidation and the Ca(2+)-ATPase activity of placental homogenates and red blood cell ghosts from control and experimental rats. Furthermore, the osmotic fragility of the red blood cells was evaluated by measuring the lysis of these cells when incubated with a NaCl solution with different osmolarities. RESULTS: It was found that placental homogenates and red blood cell ghosts from experimental pregnant rats showed an increased level of lipid peroxidation and a lowered Ca(2+)-ATPase activity, as compared to control pregnant rats. They also presented an increased osmotic fragility of their red blood cells. CONCLUSIONS: Salt-loaded pregnant rats showed, similar to preeclamptic women, an increased level of lipid peroxidation and a lowered Ca(2+)-ATPase activity in placental and red blood cells membranes, as well as an increased osmotic fragility of the red blood cells.
Assuntos
ATPases Transportadoras de Cálcio/metabolismo , Membrana Eritrocítica/enzimologia , Placenta/enzimologia , Pré-Eclâmpsia/enzimologia , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Animais , Modelos Animais de Doenças , Feminino , Peroxidação de Lipídeos , Fragilidade Osmótica , Pré-Eclâmpsia/sangue , Gravidez , Ratos Sprague-Dawley , Cloreto de SódioRESUMO
Raphiodon echinus (R. echinus) is used in Brazilian folk medicine for the treatment of inflammation, coughs, and infectious diseases. However, no information is available on the potential antioxidant, cytotoxicity and genotoxicity of this plant. In this study, the polyphenolic constituents, antioxidant capacity and potential toxic effects of aqueous and ethanolic extracts of R. echinus on human erythrocytes and leukocytes were investigated for the first time. R. echinus extracts showed the presence of Gallic, chlorogenic, caffeic and ellagic acids, rutin, quercitrin and quercetin. Aqueous and ethanolic extracts of R. echinus exhibited antioxidant activity in DPPH radical scavenging with IC50 = 111.9 µg/mL (EtOH extract) and IC50 = 227.9 µg/mL (aqueous extract). The extracts inhibited Fe(2+) (10 µM) induced thiobarbituric acid reactive substances (TBARS) formation in rat brain and liver homogenates. The extracts (30-480 µg/mL) did not induce genotoxicity, cytotoxicity or osmotic fragility in human blood cells. The findings of this present study therefore suggest that the therapeutic effect of R. echinus may be, in part, related to its antioxidant potential. Nevertheless, further in vitro and in vivo studies are required to ascertain the safety margin of its use in folk medicine.
Assuntos
Antioxidantes/farmacologia , DNA/efeitos dos fármacos , Lamiaceae/química , Fragilidade Osmótica/efeitos dos fármacos , Polifenóis/química , Animais , Antioxidantes/química , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , DNA/sangue , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Extratos Vegetais/análise , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , RatosRESUMO
Clinical hematology facilitates the diagnosis of disease and can act as a prognostic indicator of pathological conditions in fish. The aim of the present study was to evaluate hematological parameters of Nile tilapia (Oreochromis niloticus) subjected to different anesthetics and anticoagulants. Thirty apparently healthy fishes (average weight of 473 ± 35. 50 g and mean total length of 29. 33 ± 0. 37 cm), were selected from the local commercial fish farm in the Lages municipality (Santa Catarina, Brazil). The animals were randomly divided into three groups of 10. In two groups, anesthesia was induced with eugenol (70 mg·L- 1) (EG) and Benzocaine hydrochloride (100 mg·L-1) (BG), respectively. Anesthesia was not administered to fish of the third group (CG/control group). Blood samples were obtained by venipuncture of the caudal vessels and placed into microtubes containing sodium heparin or Na2EDTA for further analysis. The results were analyzed by Sigma Stat for Windows, the paired t-test for significant differences between anticoagulants of the same group, and analysis of variance followed by the Tukey test for comparison of means between groups (p ? 0. 05). Most of the observed changes in the erythrogram were significantly higher for the anticoagulant heparin and benzocaine group in comparison to the control group. However, the values obtained for the leukogram were significantly higher for all groups subjected to the Na2EDTA anticoagulant, suggesting that heparin may cause cell clumping. The results suggest that the anesthetics under investigation effectively minimizes the effects of stress caused by handling and invasive procedures, and that the anticoagulant heparin causes less hemolysis in comparison to Na2EDTA for Nile tilapia. Thus, the hematological variations attributed to different anesthetic protocols and/or different anticoagulantsshould be considered for the species Oreochromis niloticus.(AU)
A hematologia clínica é uma ferramenta que auxilia na realização do diagnóstico de doenças e pode atuar como um indicador prognóstico das condições patológicas de peixes. O objetivo deste estudo foi avaliar os valores hematológicos de tilápias do Nilo (Oreochromis niloticus) submetidas a diferentes anestésicos e amostras obtidas com diferentes anticoagulantes. Foram utilizados trinta peixes, com peso médio de 473±35,50g e comprimento total médio de 29,33±0,37cm, adquiridos de piscicultura comercial localizada no município de Lages (SC). Os animais foram divididos aleatoriamente em três grupos (n=10) conforme a forma de indução: grupo eugenol (GE) na concentração de 70mg L-1, grupo cloridrato de benzocaína (GB) na concentração de 100mg L-1 e grupo controle (GC) sem o uso de anestésicos. As amostras de sangue foram obtidas por venopunção dos vasos caudais e acondicionadas em microtubos contendo heparina sódica ou Na2EDTA e posteriormente processadas. Os resultados foram avaliados pelo Sigma Stat, sendo utilizado o teste-t pareado para dados entre diferentes anticoagulantes pertencentes ao mesmo grupo e análise de variância, seguida de teste de Tukey para comparação das médias entre grupos (p?0,05). A maioria das alterações observadas no eritrograma foram superiores com o anticoagulante heparina e no grupo benzocaína comparados com o grupo controle. Porém no leucograma, os valores obtidos foram maiores com o anticoagulante Na2EDTA em todos os grupos, sugerindo que a heparinapode causar aglomeração celular. Baseado nos resultados encontrados conclui-se que os anestésicos testados minimizaram o efeito do estresse causado por manipulações e procedimentos invasivos quandocomparado ao grupo controle e a heparina foi o anticoagulante que causou menor hemólise quando comparado com o Na2EDTA para esta espécie no teste de fragilidade osmótica eritrocitária. (AU)
Assuntos
Animais , Ciclídeos/sangue , Anestesia/veterinária , Anticoagulantes , Fragilidade OsmóticaRESUMO
Clinical hematology facilitates the diagnosis of disease and can act as a prognostic indicator of pathological conditions in fish. The aim of the present study was to evaluate hematological parameters of Nile tilapia (Oreochromis niloticus) subjected to different anesthetics and anticoagulants. Thirty apparently healthy fishes (average weight of 473 ± 35. 50 g and mean total length of 29. 33 ± 0. 37 cm), were selected from the local commercial fish farm in the Lages municipality (Santa Catarina, Brazil). The animals were randomly divided into three groups of 10. In two groups, anesthesia was induced with eugenol (70 mg·L- 1) (EG) and Benzocaine hydrochloride (100 mg·L-1) (BG), respectively. Anesthesia was not administered to fish of the third group (CG/control group). Blood samples were obtained by venipuncture of the caudal vessels and placed into microtubes containing sodium heparin or Na2EDTA for further analysis. The results were analyzed by Sigma Stat for Windows, the paired t-test for significant differences between anticoagulants of the same group, and analysis of variance followed by the Tukey test for comparison of means between groups (p ? 0. 05). Most of the observed changes in the erythrogram were significantly higher for the anticoagulant heparin and benzocaine group in comparison to the control group. However, the values obtained for the leukogram were significantly higher for all groups subjected to the Na2EDTA anticoagulant, suggesting that heparin may cause cell clumping. The results suggest that the anesthetics under investigation effectively minimizes the effects of stress caused by handling and invasive procedures, and that the anticoagulant heparin causes less hemolysis in comparison to Na2EDTA for Nile tilapia. Thus, the hematological variations attributed to different anesthetic protocols and/or different anticoagulantsshould be considered for the species Oreochromis niloticus.
A hematologia clínica é uma ferramenta que auxilia na realização do diagnóstico de doenças e pode atuar como um indicador prognóstico das condições patológicas de peixes. O objetivo deste estudo foi avaliar os valores hematológicos de tilápias do Nilo (Oreochromis niloticus) submetidas a diferentes anestésicos e amostras obtidas com diferentes anticoagulantes. Foram utilizados trinta peixes, com peso médio de 473±35,50g e comprimento total médio de 29,33±0,37cm, adquiridos de piscicultura comercial localizada no município de Lages (SC). Os animais foram divididos aleatoriamente em três grupos (n=10) conforme a forma de indução: grupo eugenol (GE) na concentração de 70mg L-1, grupo cloridrato de benzocaína (GB) na concentração de 100mg L-1 e grupo controle (GC) sem o uso de anestésicos. As amostras de sangue foram obtidas por venopunção dos vasos caudais e acondicionadas em microtubos contendo heparina sódica ou Na2EDTA e posteriormente processadas. Os resultados foram avaliados pelo Sigma Stat, sendo utilizado o teste-t pareado para dados entre diferentes anticoagulantes pertencentes ao mesmo grupo e análise de variância, seguida de teste de Tukey para comparação das médias entre grupos (p?0,05). A maioria das alterações observadas no eritrograma foram superiores com o anticoagulante heparina e no grupo benzocaína comparados com o grupo controle. Porém no leucograma, os valores obtidos foram maiores com o anticoagulante Na2EDTA em todos os grupos, sugerindo que a heparinapode causar aglomeração celular. Baseado nos resultados encontrados conclui-se que os anestésicos testados minimizaram o efeito do estresse causado por manipulações e procedimentos invasivos quandocomparado ao grupo controle e a heparina foi o anticoagulante que causou menor hemólise quando comparado com o Na2EDTA para esta espécie no teste de fragilidade osmótica eritrocitária.
Assuntos
Animais , Anestesia/veterinária , Anticoagulantes , Ciclídeos/sangue , Fragilidade OsmóticaRESUMO
This study evaluated the effects of 2 different types of acute aerobic exercise on the osmotic stability of human erythrocyte membrane and on different hematological and biochemical variables that are associated with this membrane property. The study population consisted of 20 healthy and active men. Participants performed single sessions of 2 types of exercise. The first session consisted of 60 min of moderate-intensity continuous exercise (MICE). The second session, executed a week later, consisted of high-intensity interval exercise (HIIE) until exhaustion. The osmotic stability of the erythrocyte membrane was represented by the inverse of the salt concentration (1/H50) at the midpoint of the sigmoidal curve of dependence between the absorbance of hemoglobin and the NaCl concentration. The values of 1/H50 changed from 2.29±0.1 to 2.33±0.09 after MICE and from 2.30±0.08 to 2.23±0.12 after HIIE. During MICE mean corpuscular volume increased, probably due to in vivo lysis of older erythrocytes, with preservation of cells that were larger and more resistant to in vitro lysis. The study showed that a single bout of acute exercise affected erythrocyte stability, which increased after MICE and decreased after HIIE.