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BACKGROUND: : Airborne allergens vary from one climatic region to another. Therefore, it is important to analyze the environment of the region to select the most prevalent allergens for the diagnosis and treatment of allergic patients. OBJECTIVE: : To evaluate the prevalence of positive skin tests to pollen and fungal allergens collected from local indigenous plants or isolated molds, as well as other outdoor and indoor allergens in allergic patients in 6 different geographical areas in the Kingdom of Saudi Arabia (KSA), the United Arab Emirates, and Sudan. MATERIAL AND METHODS: : Four hundred ninety-two consecutive patients evaluated at different Allergy Clinics (276 women and 256 men; mean age, 30 years) participated in this study. The selection of indigenous allergens was based on research findings in different areas from Riyadh and adjoining areas. Indigenous raw material for pollen grains was collected from the desert near the capital city of Riyadh, KSA. The following plants were included: Chenopodium murale, Salsola imbricata, Rumex vesicarius, Ricinus communis, Artiplex nummularia, Amaranthus viridis, Artemisia monosperma, Plantago boissieri, and Prosopis juliflora. Indigenous molds were isolated from air sampling in Riyadh and grown to obtain the raw material. These included the following: Ulocladium spp., Penicillium spp., Aspergillus fumigatus, Cladosporium spp., and Alternaria spp. The raw material was processed under Good Manufacturing Practices for skin testing. Other commercially available outdoor (grass and tree pollens) and indoor (mites, cockroach, and cat dander) allergens were also tested. RESULTS: : The highest sensitization to indigenous pollens was detected to C. murale (32%) in Khartoum (Sudan) and S. imbricata (30%) and P. juliflora (24%) in the Riyadh region. The highest sensitization to molds was detected in Khartoum, especially to Cladosporium spp. (42%), Aspergillus (40%), and Alternaria spp. (38%). Sensitization to mites was also very prevalent in Khartoum (72%), as well as in Abu Dhabi (United Arab Emirates) (46%) and Jeddah (KSA) (30%). CONCLUSIONS: : The allergenicity of several indigenous pollens and molds derived from autochthonous sources was demonstrated. Prevalence studies in different regions of KSA and neighbor countries indicate different sensitization rates to these and other outdoor and indoor allergens.
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BACKGROUND: Date fruit and pollen antigens share a number of cross-reactive epitopes. Date pollen has been shown to cross-react with antigens from Artemisia, cultivated rye (Secale cereale), Timothy grass (Phleum pratense), Sydney golden wattle (Acacia longifolia) and Bermuda grass (Cynodon dactylon) pollen. The present study was carried out to examine any cross-reactivities between date palm polypeptides and antigens of some common foods and vegetables that have been implicated in the oral allergy syndrome (OAS). Because most of such cross-reactivities in other allergens are attributable to the presence of carbohydrate chains and profilin, their role was also investigated. METHODS: Fresh extracts of 20 common fruits and vegetables were prepared. Putative date profilins were isolated by affinity chromatography using a poly L-proline column. Date fruit extracts were digested by various endoglycosidases and the immunoglobulin (Ig)E binding of the postdigest products was assessed in immunoblots. Rabbit antisera to whole date fruit extracts, Timothy grass profilin and putative date profilins, as well as human sera from date sensitive individuals were used in immunoblotting, ELISA and in inhibition experiments. RESULTS: IgG, ELISA and immunoblot results with the different rabbit antisera and date-sensitive atopic sera showed several antigenic cross-reactivities and similar cross-reactivities were seen with birch, date and timothy grass profilins. IgE, ELISA and immunoblot experiments with pooled date sensitive human sera showed a range of cross-reactivities with some food extracts. A number of the IgE cross-reactivities could be inhibited after preabsorption of pooled sera with date extracts. Sixty-six percent of individual date hypersensitive human sera bound IgE in putative date fruit profilin and their pooled sera bound IgE in birch pollen profilin. IgE-binding of the endoglycosidase digested date fruit extracts to atopic serum pool was restricted to only a very low molecular weight band of 6.5-8 kDa. CONCLUSION: These results indicate that date palm polypeptides share cross-reactive IgG and IgE epitopes with a number of foods implicated in the oral allergy syndrome, bind to birch and Timothy grass profilins and bind IgE through glycosyl residues. The clinical relevance of these cross-reactivities needs to be further elucidated.
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Proteínas Contráctiles , Reacciones Cruzadas/inmunología , Hipersensibilidad a los Alimentos/etiología , Frutas/efectos adversos , Frutas/inmunología , Péptidos/efectos adversos , Péptidos/inmunología , Alérgenos/administración & dosificación , Alérgenos/efectos adversos , Alérgenos/inmunología , Animales , Relación Dosis-Respuesta Inmunológica , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Epítopos/inmunología , Galectina 3/sangre , Galectina 3/efectos de los fármacos , Galectina 3/inmunología , Glicosilación/efectos de los fármacos , Humanos , Immunoblotting , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Proteínas de Microfilamentos/efectos adversos , Proteínas de Microfilamentos/inmunología , Proteínas de Microfilamentos/aislamiento & purificación , Peso Molecular , Péptidos/administración & dosificación , Polen/efectos adversos , Polen/inmunología , Profilinas , Prolina/efectos adversos , Prolina/inmunología , Prolina/aislamiento & purificación , Conejos , SíndromeRESUMEN
BACKGROUND: Date fruits are allergenic and standardized extracts are required for diagnosis and therapy of this allergy. Since there are several cultivars of dates, this study was carried out to assess the allergenicity of different cultivars in order to select suitable source material for standardization. METHODS: The protein profiles of 18 of the most commonly sold varieties were compared by SDS-PAGE and their relative allergenicity assessed by SPT and IgE-based ELISA and immunoblotting. Thirty-two date fruit-sensitive patients were skin tested with a pooled extract from all the cultivars. Six of the patients with high SPT results (> or =3+) who volunteered were further tested with the 18 cultivars and their sera used in ELISA and immunoblotting. RESULTS: Six of the cultivars gave high SPT-positive reactions in > or =4 of patients. Five of these high SPT-reactive cultivars gave high IgE ELISA scores (> or =0.58) but individual cultivars varied in their number of IgE immunoblot bands. Cultivar-specific IgE-binding patterns indicated that only certain cultivars bound IgE at molecular weights of < or =14.3 and 27-33 kDa whilst all cultivars bound to a 54-58 kDa doublet. Cultivars that bind to the < or =14.3 and 27-33 kDa bands appeared to form the majority of the high SPT-reactive cultivars. When individual sera of 24 of the 32 SPT-positive patients were used in IgE immunoblots with the pooled cultivar extract, all sera bound IgE at < or =14.3 and 27-33 kDa and about 60% of sera bound to a 54-58 kDa doublet bands. CONCLUSIONS: These results indicate that allergenicity of date fruits is a cultivar-specific phenomenon. Sixty to 100% of sera from date fruit-allergic patients bind IgE to three major allergens of < or =14.3, 27-33 and 54-58 kDa. Five of the cultivars that evoke high SPT reactions, high IgE ELISA scores and bind IgE to the major allergens, can be selected for the preparation of 'in-house' allergen extracts and for allergen standardization.
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Alérgenos/análisis , Epítopos/inmunología , Hipersensibilidad a los Alimentos/etiología , Frutas/efectos adversos , Inmunoglobulina E/inmunología , Adulto , Alérgenos/química , Alérgenos/inmunología , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Femenino , Hipersensibilidad a los Alimentos/sangre , Hipersensibilidad a los Alimentos/epidemiología , Frutas/inmunología , Humanos , Immunoblotting , Inmunoglobulina E/sangre , Masculino , Peso Molecular , Extractos Vegetales/análisis , Extractos Vegetales/inmunología , Arabia Saudita/epidemiología , Pruebas CutáneasRESUMEN
To determine the endothelin-1 (ET-1) concentrations in synovial fluid and serum of rheumatoid arthritis (RA) patients, this study was designed to examine if serum ET-1 concentration of control subjects has any correlation either with the ET-1 concentration of synovial fluid or ET-1 concentration of serum from RA patients. Twenty-eight patients were studied of whom eight males and twenty females with confirmed rheumatoid arthritis. Twenty-eight healthy volunteers were also included as controls. The immunoreactive concentration of ET-1 was measured using commercially available radioimmunoassay (RIA) kits (Peninsula Laboratories, Belmont CA) specific for ET-1. All the samples were performed in duplicate and after plotting % B/Bo for each standard directly on Y axis and endothelin concentrations on the X axis, the "best fit" curve was drawn and the amount of ET-1 was calculated. Mean ET-1 level in synovial fluid was 15.53 +/- 2.82 pg/me. In serum samples from RA patients, the mean ET-1 level was detected as 16.42 +/- 3.07 pg/ml (n = 28). Sera from twenty-eight healthy volunteers were analyzed as controls and mean ET-1 concentration was 8.68 +/- 1.96 pg/ml. A significant difference (P < 0.001) was found between ET-1 level of sera from RA patients and ET-1 levels from control sera. Highly significant difference (P < 0.001) was also detected between synovial fluid ET-1 and control ET-1 levels. However, no significant difference was found between ET-1 levels of synovial fluid and serum ET-1 levels of RA patients. Results of this study confirmed the presence of elevated levels of ET-1 concentration in synovial fluid and serum samples of patients with RA. The clinical significance and physiological role of endothelin in synovial fluids and sera of patients suffering from a variety of pathophysiological conditions of arthritis deserves further studies.
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Artritis Reumatoide/metabolismo , Endotelina-1/metabolismo , Líquido Sinovial/metabolismo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Endotelina-1/sangre , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
FHIT, a candidate tumour suppressor gene, has recently been identified at chromosomal region 3p14.2, and deletions of the gene have been reported in many types of human cancers. Loss of heterozygosity (LOH) at this region has also been found frequently in follicular thyroid carcinoma (FTC). To investigate the potential role of FHIT in thyroid tumorigenesis, we examined 57 thyroid tumour specimens (eight benign adenomas, 40 papillary, four follicular and five anaplastic carcinomas), and two thyroid carcinoma cell lines (NPA, SW579) for genetic alterations by using reverse transcription-polymerase chain reaction (RT-PCR), PCR product sequencing, single-strand conformation polymorphism (SSCP) and Southern blot analysis. Two cervical carcinoma cell lines (C-33A, HeLa) were included as positive controls. We detected truncated FHIT transcripts in three of eight (38%) benign adenomas, nine of 40 (23%) papillary, and two of five (40%) anaplastic carcinomas, and in three cell lines (SW579, C-33A, HeLa). Most of the truncated transcripts lacked exons 4 or 5 to 7 or 8 of the gene and were presumably non-functional as the translation start site is located in exon 5. SSCP analysis of the coding exons failed to detect any point mutations among the samples without abnormal FHIT transcripts. Southern blot analysis demonstrated either loss or reduced intensity of major Bam HI restriction fragments in the three cell lines found to have abnormal FHIT transcripts, indicating, respectively, either intragenic homozygous or heterozygous deletions of the FHIT gene. Intragenic homozygous deletions were also found in two papillary thyroid carcinoma specimens: one was missing a 13 kb Bam HI fragment which contains exon 4, the other had deletions of 15.5, 13 and 4.2 kb fragments which contain exons 2 and 9, 4, and 5, respectively. The absence of a defective FHIT gene in FTC indicates that an additional tumour suppressor gene may reside in this region and be involved in the development of FTC. Given that defective FHIT genes were found in both benign and malignant thyroid tumours, the inactivation of this putative tumour suppressor gene is likely to be an early event in the pathogenesis of some forms of thyroid neoplasms.
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Ácido Anhídrido Hidrolasas , Genes Supresores de Tumor/genética , Proteínas de Neoplasias , Proteínas/genética , Neoplasias de la Tiroides/genética , Southern Blotting , Humanos , Pérdida de Heterocigocidad , Mutación/genética , Polimorfismo Conformacional Retorcido-Simple , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
Whole Nigella sativa (N. sativa) proteins were purified on a DEAE Sephadex A50 ion exchange column. Complete fractionation was achieved in four peaks. Analysis of the purified peaks was carried out by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Whole N. sativa showed a number of protein bands ranging from 94-10 kDa molecular mass. In mixed lymphocyte cultures (MLC), whole N. sativa and its purified proteins were found stimulatory as well as suppressive and this effect varied from one donor to another. Maximum stimulation (mean + S.E. of % relative index was 63.73 + 20.78) was observed with fractionated N. sativa proteins (P1) (10 microg/ml) in MLC. In MLC, also N. sativa peaks (P1 and P2) were stimulatory at all concentrations (10 microg/ml, 1 microg/ml or 0.1 microg/ml) used. However, a uniformly suppressive effect of N. sativa and its all four peaks at a concentration of 10 microg/ml was noticed when lymphocytes were activated with pokeweed mitogen (PWM). The effect of N. sativa proteins was further evaluated on the production of cytokines which were measured by using specific enzyme-linked immunosorbent assay. Large quantities of IL-1beta were secreted by whole N. sativa in culture medium with non-activated peripheral blood mononuclear cells (PBMC) (450 pg/ml) and with allogeneic cells (410 pg/ml). Fractionated N. sativa was less effective when compared with whole N. sativa proteins. No effect on IL-4 secretion was seen either by using non-activated, PWM-activated or allogeneic-cells. Whole N. sativa suppressed as well as stimulated the production of IL-8 in non-activated and PWM-activated PBMC respectively. All N. sativa peaks with protein concentration of 2 microg/ml were stimulatory for the induction of IL-8 by PWM-activated cells. However, no effect on IL-8 was seen either with whole N. sativa or its peaks when allogeneic PBMC were used. Stimulatory effect of whole N. sativa and fractionated proteins was also noticed on the production of TNF-alpha either using non-activated or mitogen activated cells.
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Adyuvantes Inmunológicos/farmacología , Proteínas de Plantas/farmacología , Adyuvantes Inmunológicos/aislamiento & purificación , Cromatografía por Intercambio Iónico/métodos , Electroforesis en Gel de Poliacrilamida , Humanos , Interleucinas/biosíntesis , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Activación de Linfocitos/efectos de los fármacos , Prueba de Cultivo Mixto de Linfocitos , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Lectinas de Plantas , Proteínas de Plantas/aislamiento & purificación , Plantas Medicinales/química , Mitógenos de Phytolacca americana/farmacología , Semillas/química , Dodecil Sulfato de Sodio , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
Tumour cell invasion and metastasis is a multistep process that involves the degradation of extracellular matrix proteins by matrix metalloproteinases (MMPs). Tissue inhibitors of metalloproteinases (TIMPs) act as negative regulators of MMPs and thus prevent tumour cell invasion and metastasis by preserving extracellular matrix (ECM) integrity. In the present study we examined the expression of one member of TIMPs, TIMP-1, in 39 thyroid tumour specimens and two thyroid carcinoma cell lines (NPA and SW579). We also investigated the effect of high TIMP-1 expression on the invasive potential of NPA cells. Northern blot analysis showed that TIMP-1 mRNA levels correlated directly with tumour aggressiveness: the highest number of TIMP-1 transcripts was found in stages III and IV vs benign goitre (P < 0.0001). However, TIMP-1 expression was not increased in NPA and SW579 cells, both of which are derived from poorly differentiated thyroid tumours. Immunohistochemical study showed strong TIMP-1 staining in the stroma cells of advanced stages of carcinomas. Overexpression of TIMP-1 by gene transfer resulted in a significant suppression of the malignant phenotype of NPA cells as judged by an in vitro tumour invasion assay. These results suggest that high levels of TIMP-1 transcripts in advanced stages of thyroid carcinoma likely come from stroma rather than thyroid cancer cells, and TIMP-1 may function as a thyroid tumour invasion/metastasis suppressor.
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Carcinoma Papilar/metabolismo , Proteínas de Neoplasias/metabolismo , ARN Mensajero/metabolismo , Neoplasias de la Tiroides/metabolismo , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Adulto , Anciano , Carcinoma/metabolismo , Carcinoma/patología , Carcinoma Papilar/patología , Femenino , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Vectores Genéticos/administración & dosificación , Bocio Nodular/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Proteínas de Neoplasias/genética , Estadificación de Neoplasias , Neoplasias de la Tiroides/patología , Inhibidor Tisular de Metaloproteinasa-1/genética , TransfecciónRESUMEN
BACKGROUND: Date-palm (Phoenix dactylifera L.) fruits are eaten daily by most inhabitants of the Middle East and the neighboring countries. Recent reports have indicated that dates are allergenic. This study aimed to investigate the antigenic and allergenic potential of date fruits. METHODS: Date-fruit extracts from eight cultivars were evaluated in skin prick tests (SPT) in an atopic population, used to produce antisera, analyzed by SDS-PAGE, and fractionated by gel-filtration chromatography. Sera from SPT-positive individuals were evaluated by ELISA and RAST, and in anti-igE immunoblot experiments. RESULTS: About 13% of patients were SPT-positive for at least two extracts. SDS-PAGE of whole extracts revealed 15-18 protein bands of 6.5->100 kDa, and Sephacryl S-200 fractions gave distinct peptide bands. RAST and anti-IgE ELISA gave a range of positive results, which could be abrogated when sera were preabsorbed with fruit extracts. IgE immunoblots of different extracts with pooled positive sera revealed different anti-IgE-binding immunoprints. All the positive sera from fruit-allergic and pollen-allergic individuals bound strongly to two anti-IgE reactive bands of 6.5 to 12-14 kDa and 28-33 kDa, respectively, and about 50% of sera bound to a 54-58-kDa band. CONCLUSIONS: These results strongly indicate that 1) date-palm fruit is a potent allergen 2) sera from fruit-allergic as well as pollen-allergic patients recognize common fruit-specific epitopes 3) there is heterogeneity in patient responses to the different extracts.
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Alérgenos/efectos adversos , Antígenos/efectos adversos , Hipersensibilidad a los Alimentos , Frutas/efectos adversos , Alérgenos/química , Alérgenos/inmunología , Antígenos/química , Antígenos/inmunología , Western Blotting , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Frutas/química , Frutas/inmunología , Humanos , Inmunoglobulina E/sangre , Extractos Vegetales/efectos adversos , Extractos Vegetales/química , Extractos Vegetales/inmunología , Prueba de Radioalergoadsorción , Pruebas CutáneasRESUMEN
BACKGROUND: Allergenicity to Prosopis juliflora pollen antigen has been reported from only a few countries, including the US, South Africa, India and Kuwait. In some parts of Saudi Arabia, species of Prosopis have been introduced by the millions as roadside ornamentation. There appear to be four flowering seasons during which pollen grains float in all directions. However, the role of Prosopis pollen as the sensitizing and/or triggering agent of allergic asthma and/or rhinitis in the Kingdom has never been evaluated. PATIENTS AND METHODS: A total of 473 allergic patients suffering from bronchial asthma in four different geographical regions (Abha, Qassim, Hofuf and Gizan), and attending allergy clinics and chest disease centers of university and Ministry of Health hospitals in the region were tested for immediate hypersensitivity reaction to Prosopis juliflora allergens. Airborne pollen grains at one center were also studied for one full year, using volumetric sampling techniques. RESULTS: A total of 76.1% patients in Qassim, 37.5% in Gizan, 29% in Abha and 11% in Hofuf reacted positively to Prosopis antigen. Multiple sensitivities to other pollen antigens were detected in all patients. The level of airborne Prosopis pollen detected in Gizan exceeded 90 grains m -3 of air. CONCLUSION: In view of the documented evidence of Prosopis-involved allergenicity, the role of Prosopis pollen as a sensitizing factor in Saudi Arabia has been confirmed. However, the cause of elicitation of symptoms in many multiple sensitive patients, together with the question of cross-reactivities, needs thorough and detailed investigation. In vitro confirmation of all positive results is also required to incriminate Prosopis as one of the major allergens in parts of Saudi Arabia.
RESUMEN
The cytokine IL-12 is proposed to play a bridging role between innate and adaptive immunity. Here we demonstrate that IL-12 binds specifically to human neutrophils. This binding leads to a transient increase in 1) intracellular free calcium due to its release from membrane-enclosed stores and its influx from extracellular medium, 2) actin polymerization, and 3) tyrosine phosphorylation. IL-12 treatment also leads to a concentration-dependent increase in reactive oxygen metabolite production. The effect of IL-12 is blocked by neutralizing Abs to IL-12. Inhibition of either calcium transient or tyrosine phosphorylation causes inhibition of reactive oxygen metabolite production. However, inhibition of actin polymerization enhances IL-12-induced oxidase activation. Our data suggest 1) a direct role for IL-12 in the activation of human neutrophils, and 2) a calcium-dependent signaling pathway for IL-12.
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Calcio/metabolismo , Interleucina-12/fisiología , Neutrófilos/metabolismo , Transducción de Señal/inmunología , Tirosina/metabolismo , Actinas/metabolismo , Calcio/fisiología , Humanos , Interleucina-12/metabolismo , Activación Neutrófila/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Fosforilación/efectos de los fármacos , Polímeros/metabolismo , Unión Proteica/inmunología , Transducción de Señal/efectos de los fármacos , Tirosina/fisiologíaRESUMEN
Tumor necrosis factor-alpha (TNF-alpha) is associated with several acute and chronic inflammatory conditions. New therapies directed at inhibiting TNF-alpha will be important in treating pathological processes mediated by TNF-alpha. In this study, we studied and compared the effect of the carbocyclic nucleoside analogue (9-[(1R, 3R)-trans-cyclopentan-3-ol] adenine) with pentoxifylline on modulating TNF-alpha production. The carbocyclic nucleoside analogue inhibited TNF-alpha production in a dose-dependent manner (1 microM-1 mM) by stimulated peripheral blood mononuclear cells and cell lines of both monocyte (THP-1) and T-lymphocyte phenotypes (CEM x 174). The drug potently inhibited TNF production in cells stimulated by endotoxin, the superantigen (staphylococci enterotoxin A), the mitogen (phytohemagglutinin), and the protein kinase C activator (phorbol myristate acetate) with ED50 ranging from 5 to 30 microM. At moderate concentrations, the carbocyclic nucleoside analogue inhibited superantigen (ED50 = 300 microM) and alloantigen (mixed lymphocyte reaction) T cell proliferative responses (ED50 = 150 microM). The involvement of protein kinase C and prostaglandin E2 (PGE2), mediators that regulate TNF-alpha production, was also investigated. Unlike PTX, the nucleoside analogue did not upregulate PGE2 production. The inhibition of TNF-alpha production appeared to be mediated at least partly by PKC, since the nucleoside analogue caused suppression of PKC activity in stimulated cells. The results show that the carbocyclic nucleoside analogue is a TNF-alpha inhibitor that may be appropriate in the therapy of TNF-alpha-associated complications. The suppressive properties of the carbocyclic nucleoside analogue on antigen and alloantigen (mixed lymphocyte reaction) responses may be appropriate in disease conditions in which inhibiting both TNF-alpha and T-cell reactivity is desirable.
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Adenina/análogos & derivados , Inmunosupresores/farmacología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Adenina/farmacología , Adenina/toxicidad , Dinoprostona/fisiología , Humanos , Interleucina-8/metabolismo , Isoantígenos/farmacología , Leucocitos Mononucleares/efectos de los fármacos , Prueba de Cultivo Mixto de Linfocitos , Pentoxifilina/farmacología , Inhibidores de Fosfodiesterasa/farmacología , Proteína Quinasa C/fisiología , Superantígenos/farmacología , Linfocitos T/química , Linfocitos T/inmunología , Linfocitos T/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
PROBLEM: Cytokines have been shown to be present in human follicular fluid and have regulatory functions on follicular maturation. The presence of leukemia inhibitory factor (LIF) and interleukin (IL)-12 in human follicular fluid obtained at different stages of maturation was investigated. METHOD OF STUDY: Follicular fluids and granulosa cells were obtained from preovulatory and immature follicles. Follicular fluids from both groups were assayed for IL-12 and LIF by enzyme-linked immunosorbent assay. Granulosa cells from preovulatory and immature follicles were treated with human chorionic gonadotropin (hCG) in vitro and subsequent LIF and IL-12 production were measured. RESULTS: The average concentration of LIF was significantly higher in preovulatory follicles (7.6 +/- 1.3 pg/ml, n = 24) than in immature follicles (2.0 +/- 1.3 pg/ml, n = 6). The concentration of IL-12 was significantly higher in follicular fluid obtained from immature follicles (10.9 +/- 5.0 pg/ml) than in preovulatory follicles (1.3 +/- 0.4 pg/ml). hCG only stimulated LIF production from mature granulosa cells; it had no effect on IL-12 production. CONCLUSIONS: IL-12 and LIF are present in follicular fluid and their levels are regulated differently during follicular maturation. hCG stimulates LIF production from granulosa cells in vitro.
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Líquido Folicular/metabolismo , Inhibidores de Crecimiento/biosíntesis , Interleucina-2/biosíntesis , Interleucina-6 , Linfocinas/biosíntesis , Células Cultivadas , Gonadotropina Coriónica/farmacología , Femenino , Fertilización In Vitro , Células de la Granulosa/metabolismo , Inhibidores de Crecimiento/análisis , Humanos , Infertilidad Femenina/terapia , Factor Inhibidor de Leucemia , Linfocinas/análisis , Folículo Ovárico/metabolismo , EmbarazoRESUMEN
Cyclin D1 plays a key role in the regulation of the G1/S transition through the cell cycle. Deregulation of cyclin D1, most often leading to overexpression of the gene, has been reported in many tumor types. It has been suggested that cyclin D1 overexpression could be an alternative mechanism for pRb inactivation. We have previously found Rb gene mutations in 55% of malignant thyroid tumors. In the present study, we examined the cyclin D1 gene expression and amplification in 24 tumor samples (two of them are benign goiters) randomly selected from the same series of thyroid tumors, to see whether cyclin D1 overexpression is present in those specimens without Rb gene mutations. We found a four- to fivefold increase in cyclin D1 expression in 7 of 22 thyroid carcinomas as compared with that in benign nodular goiters. Six of them were found in carcinomas without Rb gene mutations. Among the remaining 15 thyroid carcinoma samples, 11 were found previously to have Rb gene mutations. The association between increased cyclin D1 expression and absence of Rb mutation is statistically significant (p < 0.05). We found no evidence of the cyclin D1 gene amplification or rearrangement to account for such an increase in cyclin D1 expression. We conclude that cyclin D1 overexpression may be relevant to thyroid carcinogenesis. Two mechanisms may be involved in the inactivation of pRb: one is through Rb gene mutations, and the other is by cyclin D1 overexpression.
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Ciclina D1/genética , Expresión Génica , Genes de Retinoblastoma/genética , Mutación , Neoplasias de la Tiroides/genética , Adenocarcinoma Folicular/genética , Adulto , Anciano , Northern Blotting , Southern Blotting , Carcinoma/genética , Carcinoma Papilar/genética , Femenino , Bocio Nodular/genética , Humanos , Masculino , Persona de Mediana EdadRESUMEN
We examined the specificity of the recently reported alterations in circulating lymphocytes in heatstroke by determining lymphocyte subsets in 14 consecutive heatstroke patients before and after cooling and in 7 heat-stressed controls using single- or two-color immunofluorescence flow cytometry. The relationship with catecholamine levels was also studied. In heatstroke, percentages of T (CD3(+)/CD19(-)), T-helper (CD4(+)/CD8(-)), T-inactive [CD3(+)/human leukocyte antigen-DR-], CD11a+, CD11c+, and CD44(+) lymphocytes were significantly decreased, whereas percentages of T-suppressor-cytotoxic (CD8(+)/CD4(-)), natural killer (NK; CD3(-)/CD16(+) or CD56(+)), CD3(+)/CD16(+) or CD56(+), and CD54(+) lymphocytes were significantly increased, compared with 11 normal controls. The changes in the absolute numbers of lymphocyte subsets were in the same direction and were significant for T-helper, T-suppressor-cytotoxic, NK, CD3(+)/CD16(+) or CD56(+), and CD11c+ lymphocytes. Milder but significant changes in percentages of T-helper, T-suppressor-cytotoxic, CD11c+, and CD44(+) lymphocytes were seen in heat stress. Cooling was associated with partial or complete normalization, further derangement (CD11a+, CD11c+), or overcorrection (NK, T-suppressor-cytotoxic, CD11b+) of abnormal percentages of lymphocyte subsets. Norepinephrine levels were significantly elevated in heatstroke (4.7-fold) and heat stress (3.2-fold), but did not significantly correlate with lymphocyte subsets. We conclude that heatstroke is associated with significant changes in percentages and in absolute numbers of a wide range of circulating lymphocyte subsets that are not related to elevated catecholamine levels or totally normalized by cooling. Similar, albeit milder, changes are seen in heat stress, suggesting that the two syndromes represent a continuum.
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Moléculas de Adhesión Celular/metabolismo , Trastornos de Estrés por Calor/fisiopatología , Golpe de Calor/fisiopatología , Subgrupos Linfocitarios/fisiología , Antígenos CD/inmunología , Catecolaminas/metabolismo , Femenino , Humanos , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Norepinefrina/metabolismo , Arabia Saudita , Linfocitos T/fisiología , TemperaturaRESUMEN
Aeroallergens and antigens in sandstorm dust, extracts of which were skin prick test (SPT) positive in allergic patients, were detected by rocket immunoelectrophoresis and ELISA. Fungi and bacteria isolated by agar settle plates and soil dilution and soil washing methods were enumerated and identified. Cat dander, Acacia, Alternaria, Aspergillus, Chenopodium, Cladosporium, Bermuda grass, Pithecellobium, Prosopis, Rumex, cultivated rye, and Washingtonia palm allergens were detected by both methods. Viable microbes including 1892 +/- 325 colony-forming units (cfu) of bacteria, and 869 +/- 75 cfu of fungi were isolated per gram of dust by the soil dilution method. Randomly selected microbial colonies on streaking and subculture were found to consist of between two and seven mixed colonies. Fungi including Alternaria, Aspergillus, Botrytis, Cladosporium, Mortierella, Mucor, Mycelia sterilia, Penicillium, Pythium, Ulocladium, Verticillium, and some yeasts were isolated. Actinomyces, Bacillus, Pseudomonas, and mostly coagulase-negative Staphylococcus species were identified, but the bulk of unidentified bacterial isolates were mainly mixed colonies of rods, cocci, coccobacilli, and some filamentous types. Six-hour agar settle-plate counts during sandstorms were 100 and 40% higher for bacteria and fungi, respectively, than without sandstorms. The most abundant aeroallergens were those of Acacia, Alternaria, Aspergillus, Bermuda grass, Cladosporium, cultivated rye, Prosopis, and cat dander. Pithecellobium dulce, Rumex crispus, and Washingtonia palm allergens were detectable for the first time in Riyadh. IgE reactivities of the dust in man were demonstrated by ELISA using sera from atopic, exposed, and normal subjects. These results indicate that sandstorm dust is a prolific source of potential triggers of allergic and nonallergic respiratory ailments, and the methods mentioned here should be routinely used for quick sampling of the environment.
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Contaminantes Atmosféricos/análisis , Alérgenos/análisis , Bacterias/aislamiento & purificación , Polvo , Hongos/aislamiento & purificación , Hipersensibilidad Respiratoria/etiología , Enfermedades Respiratorias/etiología , Viento , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoelectroforesis , Arabia Saudita , Pruebas CutáneasRESUMEN
The purpose of this study was to investigate prostanoid synthesis in different segments of the umbilicoplacental vascular tree and its relationship to impaired maternal glucose tolerance. Segments from the umbilical artery and vein, allantochorionic artery branches, and the cotyledon artery from 21 women with diabetes or impaired glucose tolerance and 10 healthy women were studied. Production of prostacyclin (PGI2) and thromboxane (TxA2) metabolites was determined. The Mann-Whitney U test, Wilcoxon signed-ranks matched-pairs test, Kruskal-Wallis test, analysis of variance, and simple linear regression analysis were used. A two-tailed P value of < 0.05 was considered statistically significant. From the umbilical artery distal to the cotyledon artery, the PGI2 synthesis decreased and the TxA2 synthesis increased gradually towards the periphery in normal pregnancy. The PGI2/TxA2 ratio was more than 200 times higher in the umbilical artery than in the cotyledon artery. The TxA2 production tended in general to be higher in the diabetic group than in the control group, resulting in significantly lower PGI2/TxA2 ratios in some vessels. The prostanoid production was not significantly correlated to maternal HbA1c or cord C-peptide concentrations. The balance between vascular prostacyclin and thromboxane synthesis in the umbilicoplacental arterial tree changed gradually towards the periphery: the more peripheral, the lower the prostacyclin and the higher the thromboxane production. The physiological role of this phenomenon is unknown, but may be of importance for the equilibration of vascular tone between arteries of different calibers. The altered prostanoid balance found in diabetic pregnancy was not directly attributable to the degree of maternal glycemic control, but may reflect increased free radical activity and peroxide production in diabetes.
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6-Cetoprostaglandina F1 alfa/biosíntesis , Diabetes Mellitus Tipo 2/embriología , Diabetes Gestacional/embriología , Intolerancia a la Glucosa/embriología , Placenta/irrigación sanguínea , Tromboxano A2/biosíntesis , Arterias Umbilicales/metabolismo , Estudios de Cohortes , Técnicas de Cultivo , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Gestacional/metabolismo , Femenino , Intolerancia a la Glucosa/metabolismo , Prueba de Tolerancia a la Glucosa , Humanos , Embarazo , Tercer Trimestre del Embarazo , Valores de Referencia , Arterias Umbilicales/anatomía & histología , Arterias Umbilicales/citologíaRESUMEN
The immune response identified by the induction of Th1/Th2 cells plays a critical role in the pathogenesis of various inflammatory and immune disorders. We have determined that in children with systemic onset juvenile rheumatoid arthritis (JRA), peripheral blood mononuclear cells (PBMC) constitutively and after stimulation with various antigens in vitro induce a higher secretion of interleukin-4 (IL-4) and IL-10 with a characteristic deficiency of IL-2 and interferon-gamma (IFN-gamma). This cytokine pattern is a representative of a mixed Th1/Th2 cell response in JRA. The CD3/CD28 costimulatory molecule was found to be a potent inducer of IL-4 and IL-10 secretion. PBMC-derived augmented IL-10 secretion was inhibited by exogenous Th1 cell type recombinant cytokines (IL-2, IL-12, and IFN-gamma). Although IL-10 inhibits PBMC-induced proinflammatory IL-1alpha and tumor necrosis factor-alpha secretion, it had no major effect on IL-6 production. The finding of a distinctly enhanced mixed Th1/Th2 cell response cytokine (IL-4 and IL-10) pattern in JRA provides a framework for developing strategies for immunologic intervention in this rheumatic disorder in children.
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Artritis Juvenil/inmunología , Citocinas/inmunología , Interleucina-10/inmunología , Células TH1/inmunología , Células Th2/inmunología , Estudios de Casos y Controles , Células Cultivadas , Niño , Preescolar , Citocinas/biosíntesis , Femenino , Humanos , Interleucina-10/biosíntesis , Leucocitos Mononucleares/inmunología , MasculinoRESUMEN
OBJECTIVE: To determine Th1/Th2 cell response cytokine production profile in Behçet's disease (BD). METHODS: Interleukin 2 (IL-2), interferon-gamma (IFN-gamma) and IL-12 (Th1 cell response), and IL-4, IL-10, and IL-13 (Th2 cell response) cytokine production from anti-CD3/anti-CD40 stimulated peripheral blood mononuclear cells (PBMC) were measured by ELISA. RESULTS: PBMC from patients with BD produced higher levels of IL-4, IL-10, and IL-13 (Th2 cell response), almost normal levels of IL-2 (Th1 cell response), but highly deficient IFN-gamma and IL-12 (Th1 cell response). The additions of exogenous recombinant IFN-gamma to the anti-CD3 stimulated PBMC dramatically changed this pattern with enhanced IL-12 and concomitantly decreased IL-4 and IL-10 production. CONCLUSION: The immune system in BD may be characterized by a divergent cytokine production profile of mixed Th1/Th2 (Th0) cell type; IFN-gamma is critical in modulating the IL-4, IL-10, and IL-12 cytokine network pathway in this disease.