RESUMEN
The use of pesticides in agricultural production originates residues in the environment where they are applied. Pesticide aerial application is a frequent source of exposure to pesticides by persons dedicated to agricultural practices and those living in neighboring communities of sprayed fields. The aim of the study was to assess the genotoxic effects of pesticides in workers occupationally exposed to these chemicals during their aerial application to agricultural fields of Sinaloa, Mexico. The study involved 30 pilots of airplanes used to apply pesticides via aerial application and 30 unexposed controls. Damage was evaluated through the micronucleus assay and by other nuclear abnormalities in epithelial cells of oral mucosa. The highest frequency ratios (FR) equal to 269.5 corresponded to binucleated cells followed by 54.2, corresponding to cells with pyknotic nuclei, 45.2 of cells with chromatin condensation, 3.7 of cells with broken-egg, 3.6 of cells with micronucleus, and 2.0 of karyolytic cells. Age, worked time, smoking, and alcohol consumption did not have significant influence on nuclear abnormalities in the pilots studied. Pesticide exposure was the main factor for nuclear abnormality results and DNA damage. Marked genotoxic damage was developed even in younger pilots with 2 years of short working period, caused by their daily occupational exposure to pesticides.
Asunto(s)
Daño del ADN , Exposición Profesional , Plaguicidas/toxicidad , Pilotos , Adulto , Agricultura , Consumo de Bebidas Alcohólicas , Núcleo Celular , Estudios de Cohortes , Humanos , Masculino , México , Pruebas de Micronúcleos , Persona de Mediana Edad , Mucosa Bucal/efectos de los fármacos , Exposición Profesional/análisis , Fumar , Adulto JovenRESUMEN
OBJECTIVE: To determine whether a low glycemic index diet is better than a normal glycemic index diet in producing ovulatory cycles in women with polycystic ovary syndrome (PCOS) and anovulation. MATERIALS AND METHODS: A randomized controlled clinical trial involving 37 women with PCOS and anovulation. The authors randomly assigned low glycemic index diets (n = 19) and normal glycemic index (n = 18) diets, and analyzed the number of ovulatory cycles for three months. RESULTS: In patients who consumed a low glycemic index diet, 24.6% (14/57) of the cycles were ovulatory. In those who consumed a normal glycemic index diet, only 7.4% (4/54) of the cycles were ovulatory (p = 0.014). CONCLUSIONS: The difference observed in the number of ovulatory cycles could be related to a decrease in the serum levels of circulating androgens, secondary to an improvement in insulin resistance.
Asunto(s)
Anovulación/dietoterapia , Dieta , Síndrome del Ovario Poliquístico/dietoterapia , Adulto , Andrógenos/sangre , Anovulación/etiología , Femenino , Índice Glucémico , Carga Glucémica , Humanos , Resistencia a la Insulina , Síndrome del Ovario Poliquístico/complicaciones , Adulto JovenRESUMEN
The peritrophic matrix is a chitin-protein structure that envelops the food bolus in the midgut of the majority of insects, but is absent in some groups which have, instead, an unusual extra-cellular lipoprotein membrane named the perimicrovillar membrane. The presence of the perimicrovillar membrane (PMM) allows these insects to exploit restricted ecological niches during all life stages. It is found only in some members of the superorder Paraneoptera and many of these species are of medical and economic importance. In this review we present an overview of the midgut and the digestive system of insects with an emphasis on the order Paraneoptera and differences found across phylogenetic groups. We discuss the importance of the PMM in Hemiptera and the apparent conservation of this structure among hemipteran groups, suggesting that the basic mechanism of PMM production is the same for different hemipteran species. We propose that the PMM is intimately involved in the interaction with parasites and as such should be a target for biological and chemical control of hemipteran insects of economic and medical importance.
Asunto(s)
Insectos Vectores/anatomía & histología , Insectos Vectores/fisiología , Reduviidae/anatomía & histología , Reduviidae/fisiología , Animales , Evolución Biológica , Enfermedad de Chagas/transmisión , Tracto Gastrointestinal/anatomía & histología , Tracto Gastrointestinal/fisiología , Hemípteros/anatomía & histología , Hemípteros/fisiología , Microvellosidades/fisiología , Microvellosidades/ultraestructuraRESUMEN
The antiproliferative and cytotoxic activity of glucolaxogenin and its ability to induce apoptosis and autophagy in cervical cancer cells are reported. We ascertained that glucolaxogenin exerts an inhibitory effect on the proliferation of HeLa, CaSki and ViBo cells in a dose-dependent manner. Analysis of DNA distribution in the cell-cycle phase of tumor cells treated with glucolaxogenin suggests that the anti-proliferative activity of this steroid is not always dependent on the cell cycle. Cytotoxic activity was evaluated by detection of the lactate dehydrogenase enzyme in supernatants from tumor cell cultures treated with the steroid. Glucolaxogenin exhibited null cytotoxic activity. With respect to the apoptotic activity, the generation of apoptotic bodies, the presence of active caspase-3 and annexin-V, as well as the DNA fragmentation observed in all tumor lines after treatment with glucolaxogenin suggests that this compound does indeed induce cell death by apoptosis. Also, a significantly increased presence of the LC3-II, LC3 and Lamp-1 proteins was evidenced with the ultrastructural existence of autophagic vacuoles in cells treated with this steroidal glycoside, indicating that glucolaxogenin also induces autophagic cell death. It is important to note that this compound showed no cytotoxic effect and did not affect the proliferative capacity of mononuclear cells obtained from normal human peripheral blood activated by phytohaemagglutinin. Thus, glucolaxogenin is a compound with anti-proliferative properties that induces programmed cell death in cancer cell lines, though it is selective with respect to normal lymphocytic cells. These findings indicate that this glycoside could have a selective action on tumor cells and, therefore, be worthy of consideration as a therapeutic candidate with anti-tumor potential.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Neoplasias del Cuello Uterino/tratamiento farmacológico , Anexina A5/metabolismo , Caspasa 3/metabolismo , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Fragmentación del ADN/efectos de los fármacos , Femenino , Glicósidos/metabolismo , Células HeLa , Humanos , L-Lactato Deshidrogenasa/metabolismo , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Fitohemaglutininas/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Neoplasias del Cuello Uterino/patologíaRESUMEN
Mycobacterium tuberculosis, the primary causative agent of tuberculosis, infects macrophages and transforms the hostile intracellular environment into a permissive niche. M. tuberculosis infects macrophages using a variety of microbial ligand/cell receptor systems. In this study, binding assays with biotin-labelled mycobacterial cell wall proteins revealed five Concanavalin A-reactive proteins that bind macrophages. Among these proteins, we identified PstS-1, a 38-kDa M. tuberculosis mannosylated glycolipoprotein, and characterized it as an adhesin. Inhibition assays with mannan and immunoprecipitation demonstrated that PstS-1 binds the mannose receptor. We purified PstS-1 to 95.9% purity using ion exchange chromatography. The presence of mannose in purified PstS-1 was demonstrated by Concanavalin A interaction, which was abolished in the presence of sodium m-periodate and α-D-mannosidase. Gas chromatography revealed that purified PstS-1 contained 1% of carbohydrates by weight, which was mainly mannose. Finally, we used fluorescent microbeads coated with purified PstS-1 in phagocytosis assays and discovered that microbead uptake was inhibited by the pre-incubation of cells with GlcNAc, mannan and α-methyl mannoside. The interaction of PstS-1 coated beads with the mannose receptor was confirmed by confocal colocalization studies that showed high Pearson and Manders's colocalization coefficients. Our findings contribute to a better understanding of the strategies M. tuberculosis uses to infect host cells, the critical first step in the pathogenesis of tuberculosis.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/inmunología , Adhesinas Bacterianas/inmunología , Proteínas Bacterianas/inmunología , Lectinas Tipo C/inmunología , Macrófagos/inmunología , Lectinas de Unión a Manosa/inmunología , Mycobacterium tuberculosis/inmunología , Receptores de Superficie Celular/inmunología , Acetilglucosamina/farmacología , Aciltransferasas/inmunología , Animales , Antígenos Bacterianos/inmunología , Adhesión Bacteriana/inmunología , Línea Celular Tumoral , Pared Celular/inmunología , Concanavalina A/química , Inmunoprecipitación , Mananos/farmacología , Manosa/metabolismo , Receptor de Manosa , Proteínas de la Membrana/inmunología , Metilmanósidos/farmacología , Ratones , Mycobacterium tuberculosis/patogenicidad , Ácido Peryódico/metabolismo , Fagocitosis/inmunología , Unión Proteica , Tuberculosis Pulmonar/patología , alfa-Manosidasa/metabolismoRESUMEN
Lectins participate in the immune mechanisms of crustaceans. They have been considered as humoral receptors for pathogen-associated molecular patterns; however, some reports suggest that lectins could regulate crustacean cellular functions. In the present study, we purified and characterized a serum lectin (CqL) from the hemolymph of Cherax quadricarinatus by affinity chromatography and determined its participation in the regulation of hemocytes' oxidative burst. CqL is a 290-kDa lectin in native form, constituted by 108, 80, and 29-kDa subunits. It is mainly composed of glycine, alanine, and a minor proportion of methionine and histidine. It showed no carbohydrates in its structure. CqL is composed of several isoforms, as determined by 2D-electrophoresis, and shows no homology with any crustacean protein as determined by Lc/Ms mass spectrometry. CqL agglutinated mainly rat and rabbit erythrocytes and showed a broad specificity for monosaccharides such as galactose, glucose, and sialic acid, as well as for glycoproteins, such as porcine stomach and bovine submaxillary mucin and fetuin. It is a Mn(2+)-dependent lectin. CqL recognized 8% of crayfish granular hemocytes and increased 4.2-fold the production of hemocytes' superoxide anion in vitro assays when compared with non-treated hemocytes. This effect showed the same specificity for carbohydrates as hemagglutination; moreover, superoxide dismutase and diphenyleneiodonium chloride were effective inhibitors of CqL oxidative-activation. The CqL homoreceptor is a 120-kDa glycoprotein identified in the hemocytes lysate. Our results suggest that CqL participates actively in the regulation of the generation of superoxide anions in hemocytes using NADPH-dependent mechanisms.
Asunto(s)
Astacoidea/química , Astacoidea/inmunología , Hemocitos/inmunología , Lectinas/análisis , Pruebas de Aglutinación , Aminoácidos/análisis , Animales , Western Blotting , Cromatografía de Afinidad , Electroforesis en Gel Bidimensional , Hemocitos/metabolismo , Hemolinfa/metabolismo , Inmunohistoquímica , Lectinas/sangre , Lectinas/inmunología , Espectrometría de Masas , Fagocitosis/inmunología , Especies Reactivas de Oxígeno/metabolismo , Estallido Respiratorio/inmunología , Estadísticas no ParamétricasRESUMEN
Objetivo: Determinar las posibles interacciones farmacológicas de las hojas de Maytenus macrocarpa, con fármacos estimulantes e inhibitorios de la motilidad intestinal. Métodos: Se utilizaron 110 ratones albinos machos, con pesos medios de 25 g, se empleó el método de Arbos y col, se administró carbón activado al 5
vía oral, dosis de 0.1ml/10g, como marcador intestinal. Los grupos experimentales fueron: control (agua destilada 0,3ml), hojas de chuchuhuasi 1 (500mg/kg), hojas de chuchuhuasi 2 (3000mg/kg), atropina (1,5mg/kg), loperamida (5mg/kg), neostigmina (0,4mg/kg), metoclopramida (10mg/kg), hojas de chuchuhuasi 1 con metoclopramida, hojas de chuchuhuasi 1 con loperamida, hojas de chuchuhuasi 2 con metoclopramida y hojas de chuchuhuasi 2 con loperamida. Para la validación estadística se usó la prueba de Wilconxon, ANOVA y Tukey. Resultados: El porcentaje de recorrido intestinal de carbón activado fue de 27,04, 34,15, 31,66, 25,57, 15,89, 43,30, 33,99, 32,40, 27,90, 49,34 y 25,36 respectivamente, el test de ANOVA de dos colas revelo una p=0,0007. El test de Tukey indico p<0.05 versus el control para neostigmina, loperamida y la interacción chuchuhuasi 3000 mg/kg con metoclopramida, en este último, el test de Wilconxon presento un valor p<0,05. Conclusiones: Se observó interacciones farmacológicas de antagonismo sobre la motilidad intestinal, entre chuchuhuasi y Loperamida y sinergismo entre chuchuhuasi y metoclopramida.
Objectives: To determine the possible pharmacological interactions from the leaves of Maytenus macrocarpa with inhibitory and stimulating bowel motility drugs. Methods: We used 110 male albino mice with average weight of 25g, Arbos and others method was applied. Activated charcoal was administered at 5
at dose of 0.1ml/10g, as an intestinal marker. The experimental groups included 0.1 ml/10 g of distilled water, leave extract of M. macrocarpa 1 (500mg/kg), leave extract of M. macrocarpa 2 (3000 mg/kg), 1,5mg/kg of atropine, 5mg/kg of loperamide, 0.4mg/kg of neostigmine, 10mg/kg of metoclopramide, leave extract of M. macrocarpa 1 with metoclopramide, leave extract of M. macrocarpa 1 with loperamide, leave extract of M. macrocarpa 2 with metoclopramide and leave extract of M. macrocarpa 2 with loperamide. The statistical validation was based on Wilconxon, ANOVA and Tukey test. Results: The intestinal charcoal run percentage was 27.04, 34.15, 31.66, 25.57, 15.89, 43.30, 33.99, 32.40, 27.9, 49.34 and 25.36 respectively. The ANOVA test result in p= 0.0007. The Tukey test indicated p <0.05 versus the control group for neostigmine, loperamide, and the interaction between leave extract of M. macrocarpa 2 with metoclopramide, for the last the Wilcoxon test result in p <0.05. Conclusions: It was observed antagonism drug interactions on gastrointestinal motility between leaves extract of M. macrocarpa with loperamide and synergism interactions with metoclopramide.
Asunto(s)
Humanos , Interacciones Farmacológicas , Loperamida , Maytenus , Metoclopramida , Motilidad Gastrointestinal , Plantas Medicinales , Antagonismo de Drogas , Sinergismo FarmacológicoRESUMEN
When Avibacterium paragallinarum reference strain 0083 (serovar A) was grown in an iron-restricted culture medium, the expression of the 60, 68 and 93 kDa outer membrane proteins increased as compared with normal media. Sera of chickens experimentally infected with Av. paragallinarum recognized these iron-restriction induced proteins, suggesting their expression in vivo. The three outer membrane proteins were identified as transferrin receptor and iron transport proteins by mass spectroscopy and a search in sequence databases. As these proteins have been reported to be regulated by the Fur protein in many bacteria, we investigated, through molecular methods, the presence of the fur gene in Av. paragallinarum. A candidate fur gene of Av. paragallinarum was amplified by polymerase chain reaction using complementary primers to conserved regions of fur gene sequences from members of the Pasteurellaceae family. The nucleotide sequence of the cloned gene, from ATG to TAA stop codon, was 453 base pairs in length and the deduced amino acid sequence showed 94% identity with Fur sequences of Actinobacillus pleuropneumoniae and Haemophilus ducreyi. The Av. paragallinarum deduced Fur protein (17.8 kDa) amino acid sequence contains the N-terminal helix-turn-helix DNA-binding domain and the two iron-binding sites in the C-terminal end, typical of other described Fur proteins. The study of iron-restriction-induced proteins and the mechanism regulating their expression could lead to an understanding of the responses of Av. paragallinarum to survive in an iron-restricted environment on host mucosal surfaces.
Asunto(s)
Proteínas Bacterianas/genética , Pasteurellaceae/genética , Receptores de Transferrina/metabolismo , Proteínas Represoras/genética , Secuencia de Aminoácidos , Proteínas Bacterianas/metabolismo , Secuencia de Bases , Western Blotting , Clonación Molecular , Biología Computacional , Cartilla de ADN/genética , Electroforesis en Gel de Poliacrilamida , Hierro/metabolismo , Espectrometría de Masas , Datos de Secuencia Molecular , Receptores de Transferrina/genética , Proteínas Represoras/metabolismo , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
One of the functions of the immune system is to recognize and destroy abnormal or infected cells to maintain homeostasis. This is accomplished by cytotoxic lymphocytes. Cytotoxicity is a highly organized multifactor process. Here, we reviewed the apoptosis pathways induced by the two main cytotoxic lymphocyte subsets, natural killer (NK) cells and CD8+ T cells. In base to recent experimental evidence, we reviewed NK receptors involved in recognition of target-cell, as well as lytic molecules such as perforin, granzymes-A and -B, and granulysin. In addition, we reviewed the Fas-FasL intercellular linkage mediated pathway, and briefly the cross-linking of tumor necrosis factor (TNF) and TNF receptor pathway. We discussed three models of possible molecular interaction between lytic molecules from effector cytotoxic cells and target-cell membrane to induction of apoptosis.
Asunto(s)
Apoptosis/inmunología , Citotoxicidad Inmunológica , Células Asesinas Naturales/inmunología , Modelos Biológicos , Linfocitos T Citotóxicos/inmunología , Animales , Antígenos de Diferenciación de Linfocitos T/metabolismo , Exocitosis/inmunología , Proteína Ligando Fas/metabolismo , Proteína de Dominio de Muerte Asociada a Fas/metabolismo , Granzimas/metabolismo , Humanos , Perforina/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Galectin-3 has been implicated in tumor progression. We demonstrated immunohistochemically that galectin-3 was negative in normal breast tissue, but it was highly increased in breast cancer and in metastatic tissues to brain. Similarly, histochemistry with mucin-specific lectins showed increased recognition in breast tumor and metastasis with Machaerocereus eruca agglutinin (Fualpha 1,2 (GalNAcalpha 1,3) Galss1,4 in complex mucin) but not for Amaranthus leucocarpus (Galss1,3-GalNAc-alpha 1,0-Ser/Thr) and Arachis hypogaea lectins (Galss1,3GalNAc/Galss1,4GlcNAc). Mucin-type glycans and galectin-3 colocalized in breast cancer and metastasis, but not in normal tissue, suggesting upregulated biosynthesis of complex O-glycosidically linked glycans and galectin-3 favor breast cancer progression and brain metastasis.
Asunto(s)
Neoplasias Encefálicas/química , Neoplasias de la Mama/química , Galectina 3/análisis , Mucinas/análisis , Arachis , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/secundario , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Femenino , Glicoproteínas , Glicosilación , Histocitoquímica/métodos , Humanos , Inmunohistoquímica , Mucinas/metabolismo , Neuraminidasa/metabolismo , Lectinas de Plantas , Manejo de Especímenes/métodos , Tripsina/metabolismo , Regulación hacia ArribaRESUMEN
Latex from Hevea brasiliensis contains several allergenic proteins that are involved in type I allergy. One of them is Hev b 2, which is a beta-1,3-glucanase enzyme that exists in different isoforms with variable glycosylation content. Two glucanase isoforms were isolated from trees of the GV-42 clone by gel filtration, affinity and ion-exchange chromatography. Isoform I had a carbohydrate content of about 20%, with N-linked N-acetyl-glucosamine, N-acetyl-galactosamine, fucose and galactose residues as the main sugars, while isoform II showed 6% carbohydrate content consisting of N-acetyl-glucosamine, fucose, mannose and xylose. Both isoforms were crystallized by the hanging-drop vapour-diffusion method. Isoform I crystals were grown using 0.2 M trisodium citrate dihydrate, 0.1 M Na HEPES pH 7.5 and 20%(v/v) 2-propanol, but these crystals were not appropriate for data collection. Isoform II crystals were obtained under two conditions and X-ray diffraction data were collected from both. In the first condition (0.2 M trisodium citrate, 0.1 M sodium cacodylate pH 6.5, 30% 2-propanol), crystals belonging to the tetragonal space group P4(1) with unit-cell parameters a = b = 150.17, c = 77.41 A were obtained. In the second condition [0.2 M ammonium acetate, 0.1 M trisodium citrate dihydrate pH 5.6, 30%(w/v) polyethylene glycol 4000] the isoform II crystals belonged to the monoclinic space group P2(1), with unit-cell parameters a = 85.08, b = 89.67, c = 101.80 A, beta = 113.6 degrees. Preliminary analysis suggests that there are four molecules of isoform II in both asymmetric units.
Asunto(s)
Alérgenos/química , Proteínas de Plantas/química , Alérgenos/genética , Alérgenos/aislamiento & purificación , Secuencia de Aminoácidos , Antígenos de Plantas , Glicoproteínas/química , Glicoproteínas/aislamiento & purificación , Hevea , Datos de Secuencia Molecular , Monosacáridos/análisis , Fragmentos de Péptidos/química , Proteínas de Plantas/genética , Proteínas de Plantas/aislamiento & purificación , Polimorfismo Genético , Isoformas de Proteínas/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Difracción de Rayos XRESUMEN
We investigated the proportion, phenotype, and cytotoxicity of CD8+CD57+ and CD57- T cells in peripheral blood from 20 tuberculosis (TB)-patients and 20 healthy tuberculin skin test-positive donors. Our results showed an increase in CD8+CD57+ T cells from TB-patients as compared with those from age-matched healthy donors (p<0.0001). CD8+CD57+ T cells from TB-patients expressed CD69, perforin, granzyme-A, and a CD28-CD62L-CD161- phenotype without recognition for the alpha-galactosylceramide-CD1d complex. This cell subset also expressed TNF-alpha and IFN-gamma, under phorbol-myristate-acetate/ionomycin stimulation. Interestingly, the cytotoxicity against autologous monocytes was higher in CD57- cells from TB-patients and donors than their CD57+ counterparts, in the presence of Mycobacterium tuberculosis H37Rv culture filtrate. However, only CD8+CD57+ T cells from TB-patients exhibited spontaneous cytotoxicity against monocytes in the absence of antigen. Our results suggest that CD8+CD57+ T cells are a subset of effector cells that could be helpful to evaluate the cell-mediated immune response to M. tuberculosis.
Asunto(s)
Antígenos CD57/inmunología , Mycobacterium tuberculosis/inmunología , Linfocitos T Citotóxicos/inmunología , Tuberculosis Pulmonar/inmunología , Tuberculosis Pulmonar/microbiología , Adulto , Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos T/metabolismo , Biomarcadores , Femenino , Granzimas/metabolismo , Humanos , Interferón gamma/metabolismo , Lectinas Tipo C , Masculino , Glicoproteínas de Membrana/metabolismo , Persona de Mediana Edad , Perforina , Fenotipo , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Linfocitos T Citotóxicos/metabolismo , Tuberculosis Pulmonar/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: Airway allergic diseases are regulated by interleukin (IL)-5, which causes infiltration of eosinophils into the bronchial epithelium, and by IL-4 which increases serum immunoglobulin E (IgE) production and promotes CD30 expression on Th cells. CD30 generates a costimulatory signal involved in apoptosis or cell proliferation, depending on the microenvironment. Our aims were: (i) to analyze if CD4+ CD30+ T cells from allergic patients proliferate in response to Dermatophagoides pteronyssinus, and (ii) if upon stimulation this cell population produces IL-4 and IL-5. METHODS: Peripheral blood mononuclear cell (PBMC) from 17 allergic rhinitis and mild allergic asthma patients and 12 healthy nonallergic individuals were stimulated with allergen in the presence or absence of anti-IL-4, anti-IL-5 or anti-IL-4Ralpha monoclonal antibodies (mAbs). TdT-mediated dUTP nick end-labeling (TUNEL) assay, 7-aminoactinomycin-D (7-AAD) intercalation, and flow cytometry were used to determine the CD4+ CD30+ blasts percentage, cell proliferation, apoptosis, and intracellular cytokines after 7 culture days. RESULTS: Cell proliferation induced with allergen showed that 90% of the allergen-stimulated blasts were CD4+, 50% of which were CD30+. Allergen-stimulated PBMC showed a progressive increase (mean: from 7% to 23%) of CD4+ CD30+IFN-gamma+ and CD4+ CD30+IL-4+ blasts which diminished (mean: 6%) after 5 culture days. In contrast, CD4+ CD30+IL-5+ blasts showed a continuous progression (from 12% to 24%) that maintained after 7 culture days. The vast majority of CD4+ CD30+ blasts were negative to 7-AAD or TUNEL. Additionally, a significant decrease (34%) was observed in the number of CD4+ CD30+ blasts when IL-4 was neutralized. CONCLUSIONS: These data suggest that specific allergen stimulation of PBMC isolated from allergic patients generates a nonapoptotic CD4+ CD30+ blast subset that produces IL-5.
Asunto(s)
Alérgenos/farmacología , Antígenos Dermatofagoides/inmunología , Linfocitos T CD4-Positivos/inmunología , Dermatophagoides pteronyssinus , Interleucina-5/biosíntesis , Adolescente , Adulto , Apoptosis/inmunología , Asma/sangre , Estudios de Casos y Controles , Proliferación Celular , Células Cultivadas , Femenino , Citometría de Flujo , Humanos , Antígeno Ki-1/inmunología , Activación de Linfocitos , Masculino , Probabilidad , Rinitis Alérgica Estacional/sangre , Muestreo , Sensibilidad y Especificidad , Estadísticas no ParamétricasRESUMEN
OBJECTIVE: The aim was to explore the role of prolactine (PRL) in the lymphocyte activation process in active and inactive systemic lupus erythematosus (SLE) patients in an in vitro model. METHODS: Peripheral blood mononuclear cells (PBMNC) were isolated from SLE patients and healthy individuals. The mRNA for prolactine and its receptor, obtained by standard techniques with an appropriate primer, were subjected to PCR and visualised. The PBMC were cultured with: a) medium alone as a negative control, b) unspecific mitogen as a positive control (PMA-ionomycin for CD154 or concanavalin A for CD69), c) PRL alone, d) mitogen plus PRL, e) mitogen plus antibody anti-PRL (1:50) and f) mitogen plus an unrelated antibody. Then CD69 and CD154 were determined by flow cytometry analysis. RESULTS: Twelve inactive and 15 active SLE patients were studied. 25% of the active patients displayed hyperprolactinemia. Under basal conditions, CD69 expression was associated with disease activity. In contrast, CD154 did not show this association. The PBMNC activated in vitro were capable of producing and secreting prolactine as measured by mRNA and Nb2 assay. In the same way the mRNA for prolactine receptor was visualized. Cells from SLE patients cultivated with PRL alone did not display increased CD69 or CD154 expression. The addition of PRL to the unspecific stimulated culture did not have an additive effect. In contrast, the addition of antibodies against PRL, in order to block the autocrine prolactine, resulted in a striking reduction in CD69 and CD154 expression. CONCLUSIONS: PRL is produced and secreted by the immune cell and acts just after the first trigger signal of activation in an autocrine way. The expression of CD69 and CD154 molecules depend partially on the prolactine.
Asunto(s)
Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos T/metabolismo , Linfocitos T CD4-Positivos/fisiología , Ligando de CD40/metabolismo , Lupus Eritematoso Sistémico/inmunología , Lupus Eritematoso Sistémico/fisiopatología , Prolactina/genética , Adulto , Comunicación Autocrina/inmunología , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/metabolismo , Células Cultivadas , Humanos , Lectinas Tipo C , Activación de Linfocitos , Persona de Mediana Edad , Prolactina/metabolismo , ARN Mensajero/análisis , Receptores de Prolactina/genéticaRESUMEN
Alzheimer's disease (AD) is a pathological process characterized by neuron degeneration and, as recently suggested, brain plasticity. In this work, we compared the reactive plasticity in AD brains associated to O-glycosydically linked glycans, recognized by lectins from Amaranthus leucocarpus (ALL) and Macrobrachium rosenbergii (MRL), and the tau neuritic degeneration. The neuritic degenerative process was evaluated by the quantification of aggregated neuritic structures. Lesions were determined using antibodies against hyperphosphorylated-tau (AD2), amyloid-beta, and synaptophysin. In these conditions, we classified and quantified three pathological structures associated to the neuritic degenerative process: 1) Amyloid-beta deposits (AbetaDs), 2) Classic neuritic plaques (NPs), and 3) Dystrophic neurites clusters (DNCs) lacking amyloid-beta deposits. Reactive plasticity structures were constituted by meganeuritic clusters (MCs) and peri-neuronal sprouting in neurons of the CA4 region of the hippocampus, immunoreactive to synaptophysin (exclusively in AD brains) and GAP-43. Besides, MCs were associated to sialylated O-glycosydically linked glycans as determined by positive labeling with ALL and MRL. Considering that these lectins are specific for the synaptic sprouting process in AD, our results suggest the co-occurrence of of several areas of reactive plasticity and neuron degeneration in AD.
Asunto(s)
Enfermedad de Alzheimer/patología , Encéfalo/patología , Degeneración Nerviosa/patología , Plasticidad Neuronal , Anciano , Anciano de 80 o más Años , Enfermedad de Alzheimer/metabolismo , Encéfalo/metabolismo , Estudios de Casos y Controles , Femenino , Histocitoquímica , Humanos , Inmunohistoquímica , Lectinas , Masculino , Persona de Mediana Edad , Placa Amiloide/patología , Polisacáridos/metabolismoRESUMEN
The ability of yeast cells of Histoplasma capsulatum to attach and agglutinate human erythrocytes has been described. This is the first report involving these yeasts in the hemagglutination phenomenon. Results revealed that the yeast cells were able to bind to erythrocytes irrespective of blood groups and to agglutinate them when a high density of yeast cells was used. Assays on the inhibition of yeast attachment to erythrocytes were also performed, using sugar-treated yeast cells. Results indicate that galactose (Gal), mainly the beta-anomer, specially inhibited yeast attachment. Disaccharides (Gal-derivatives) and glycosaminoglycans containing Gal residues, mainly chondroitin sulfate C, promote this type of inhibition. In addition, preliminary data of inhibition assays also involved a probable ionic strength driven mechanism mediated by sialic acid and heparan sulfate, suggesting that yeast binding to erythrocytes could be associated with negative charges of both molecules.
Asunto(s)
Eritrocitos/microbiología , Hemaglutinación , Histoplasma/patogenicidad , Antígenos de Grupos Sanguíneos , Adhesión Celular , Sulfatos de Condroitina/metabolismo , Disacáridos/metabolismo , Agregación Eritrocitaria , Galactosa/metabolismo , Glicosaminoglicanos/metabolismo , Heparitina Sulfato/química , Humanos , Ácido N-Acetilneuramínico/químicaRESUMEN
The protozoan parasite Entamoeba histolytica is the etiological agent of human amebiasis. The pathology of the disease starts with the cytolysis of the host target cells by amoebae. It is initiated by the adhesion of trophozoites to the host cells, through surface lectin via specific receptors. These adherence lectins have been demonstrated to be highly conserved, and can be recognised by serum antibodies from patients with invasive amebiasis. Some of these molecules have been used as antigens in serologic studies, which has been very helpful in the diagnosis of invasive intestinal amebiasis. However, false-positive serologic reactivity can occur using E. histolytica extracts and purified antigens. Additional problems are because the extracts display a great enzymatic activity. Several diagnostic methods, using different molecules and techniques, have been described. However, the problem still remains since these tests are not capable of differentiating between amoebic liver abscess (ALA) and intestinal amebiasis.Here, the research has been addressed to the 66-kDa antigen, which is a part of the outer membrane proteins from the E. histolytica strain HM1-IMSS trophozoites. First of all, we characterized the 66-kDa antigen in order to prove the relevance. We found that the 66-kDa antigen is a part of the plasma membranes and is distributed rather homogeneously on the cell surface of trophozoites. Apparently, the 66-kDa antigen is a glycoprotein. Using a monoclonal antibody (MAb), we found 25% of inhibition in the erythrophagocytosis by the trophozoites. Starting form one monoclonal antibody, we prepared an anti-idiotype (anti-Id) antibody reagent, with the purpose of searching for the different expressions of the idiotype between the sera from ALA and the intestinal amebiasis patients. Moreover, we produced the antibody Ab3 that is capable of recognising the 66-kDa antigen; it means that the Ab2 displays the internal image of the antigen. We found that 91.6% of the serum from ALA patients displayed the expression of the Id. In contrast, 15.7% of the E. histolytica asymtomatic cyst carriers displayed the Id expression, 6.6% of the patients with another parasite infection, and 11% of the negative controls (serum from umbilical cords of newborn babies). Our results showed that the expression of the Id could be differentiated among the AHA patients from the other groups with a 91.6% sensibility and 88.3% specificity.
Asunto(s)
Anticuerpos Antiidiotipos/inmunología , Antígenos de Protozoos/inmunología , Entamoeba histolytica/inmunología , Entamoeba histolytica/aislamiento & purificación , Entamebiasis/diagnóstico , Animales , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Antígenos de Protozoos/análisis , Entamebiasis/inmunología , Humanos , Inmunoensayo , Ratones , Ratones Endogámicos BALB C , Sensibilidad y EspecificidadRESUMEN
We purified and characterized a lectin from the corn coleoptyle (Zea mays). The lectin (CCL) was purified by affinity chromatography on a Lactosyl-Sepharose 4B column. It is a glycoprotein of 88.7 kDa, composed mainly by glutamic, aspartic, glycine, and Ser residues; in a minor proportion, it contained methionine and cysteine residues. Carbohydrates that constituted 12% of the total weight comprised galactose, mannose, and N-acetyl-D-glucosamine. The lectin contained the blocked amino-terminus. Analysis of the lectin, determined from peptides obtained after trypsin digestion by MALDI-TOF (matrix-assisted laser desorption ionization-time of flight), indicated that CCL has 18% homology with a putative calcium-dependent Ser/Thr protein kinase, from Arabidopsis thaliana, and 39% homology with a NADPH-dependent reductase from Z. mays. The lectin showed hemagglutinating activity toward several erythrocytes, including human A, B, and O. Hapten inhibition assays indicated that the lectin interacts specifically with the OH on C4 from galactose residues. OH- on C1 plays a relevant role in the interaction with CCL, since beta-galactose residues are better recognized than those from the anomeric alpha-galactose. Lack of lectin activity was observed in corn extracts; the highest specific activity was obtained from coleoptyle obtained at the 7th day after seeding.
Asunto(s)
Lectinas/aislamiento & purificación , Zea mays/química , Secuencia de Aminoácidos , Aminoácidos/análisis , Cromatografía de Afinidad , Cotiledón/química , Lectinas/química , Datos de Secuencia Molecular , Fitohemaglutininas/aislamiento & purificación , Lectinas de Plantas , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Factores de Tiempo , TripsinaRESUMEN
The atopic diseases are mediated by type I hypersensitivity. These disorders are multifactorial, although, the inherency is probably the most important cause. The disregulation of the Th1/Th2 immune response is one of the predominant factors on the generation and maintenance on the atopic process. We analyze the mechanisms of the regulation of the response Th1/Th2, the recent theories of the disregulation on this response as an etiology, the soluble factors as cytokines, chemokines and their receptors, surface cell markers and transcriptional factors recently mentioned to be important on this disregulation mechanism. Otherwise, we propose some general considerations and perspectives.
Asunto(s)
Hipersensibilidad Inmediata/inmunología , Membrana Celular/inmunología , Quimiocinas/inmunología , Citocinas/inmunología , Humanos , Células TH1/inmunología , Células Th2/inmunologíaRESUMEN
In this work we identified in adult and juvenile freshwater prawn, Macrobrachium rosenbergii, three major type of circulating hemocytes: fusiform; rounded; and large ovoid hemocytes. Rounded and large hemocytes represent the first defense line, since this type of cells exerts phagocytic activity as well as lectin synthesis. Considering that glycosylation plays important roles in cell communication and as a target for pathogenic microorganisms, in this report was also described the main glycosidic modifications that occur in the large and rounded hemocytes from the freshwater prawn during maturation as determined with lectins. Neu5Acalpha2,6Gal, was identified homogeneously distributed in the membrane in 90% of hemocytes from juvenile organisms. Maturation of the freshwater prawn induced a decrease or complete loss of Neu5Acalpha2,6Gal residues that were replaced with Neu5Acalpha2,3 molecules in practically all hemocytes from adult organisms. This change was paralleled by a diminution in 9-O-acetyl-neuraminic acid (Neu5,9Ac(2)) expression. T and Tn antigens (Galbetal,3 GalNAcalpha1-0-Ser/Thr or GalNAcalpha1-0-Ser/Thr, respectively), as well as N-glycosidically linked glycans, seem to be highly conserved throughout maturation. Our results show that sialylation of freshwater prawn hemocytes is modulated throughout the maturation process.