RESUMEN
Epidermal growth factor receptor inhibitors (EGFRIs) frequently cause cutaneous adverse effects such as papulopustular eruptions. However, the mechanism of the reactions remains unclear. To assess the pathological mechanism of cutaneous adverse reactions caused by EGFRIs, we investigated whether EGFRIs have an influence on the innate immune response of the skin. Levels of human ß-defensins (hBDs), which serve as the first line of defence against infection by pathogenic microorganisms, in the stratum corneum samples of patients treated with EGFR. monoclonal antibodies were measured before and after starting therapy. There were no obvious trends in hBD production in patients without eruptions, whereas a significant decrease in hBD1 and hBD3 production and a nonsignficant decrease in hBD2 production were observed in patients who developed papulopustular eruptions. Our results suggest that a reduction in hBD contributes to the increased incidence of papulopustular eruptions.
Asunto(s)
Anticuerpos Monoclonales/efectos adversos , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/patología , Receptores ErbB/antagonistas & inhibidores , beta-Defensinas/efectos de los fármacos , Anciano , Anciano de 80 o más Años , Antiinfecciosos/análisis , Antiinfecciosos/metabolismo , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/uso terapéutico , Erupciones por Medicamentos/etiología , Erupciones por Medicamentos/inmunología , Erupciones por Medicamentos/microbiología , Epidermis/efectos de los fármacos , Epidermis/metabolismo , Epidermis/patología , Receptores ErbB/inmunología , Femenino , Humanos , Inmunidad Innata/efectos de los fármacos , Masculino , Persona de Mediana Edad , Infecciones Cutáneas Estafilocócicas/inducido químicamente , Infecciones Cutáneas Estafilocócicas/epidemiología , beta-Defensinas/análisisRESUMEN
AIM: The anatomy of the region between the vagina and anal canal plays an essential role when performing a proctectomy for low-lying tumours. However, the anatomical characteristics of this area remain unclear. The purpose of the present study was to clarify the configuration, and both lateral and inferior extensions, of the muscle bundles in the anorectal anterior wall in females. METHODS: Using cadaveric specimens, macroscopic anatomical and histological evaluations were conducted at the anatomy department of our institute. Macroscopic anatomical specimens were obtained from six female cadavers. Histological specimens were obtained from eight female cadavers. RESULTS: The smooth muscle fibres of the internal anal sphincter and longitudinal muscle extended anteriorly in the anorectal anterior wall of females and the muscle bundles showed a convergent structure. The anterior extending smooth muscle fibres merged into the vaginal smooth muscle layer, distributed subcutaneously in the vaginal vestibule and perineum and spread to cover the anterior surface of the external anal sphincter and the levator ani muscle. Relatively sparse space was observed in the region anterolateral to the rectum on histological analysis. CONCLUSION: Smooth muscle fibres of the rectum and vagina are intermingled in the median plane, and there is relatively sparse space in the region anterolateral to the rectum. Therefore, when detaching the anorectal canal from the vagina during proctectomy, an approach from both the lateral sides should be used.
Asunto(s)
Canal Anal/anatomía & histología , Músculo Liso/anatomía & histología , Proctectomía/métodos , Recto/anatomía & histología , Vagina/anatomía & histología , Cadáver , Femenino , HumanosRESUMEN
BACKGROUND: Allergic rhinitis, a known risk factor for asthma onset, often accompanies mouth breathing. Mouth breathing may bypass the protective function of the nose and is anecdotally considered to increase asthma morbidity. However, there is no epidemiological evidence that mouth breathing is independently associated with asthma morbidity and sensitization to allergens. In this study, we aimed to clarify the association between mouth breathing and asthma morbidity and allergic/eosinophilic inflammation, while considering the effect of allergic rhinitis. METHODS: This community-based cohort study, the Nagahama Study, contained a self-reporting questionnaire on mouth breathing and medical history, blood tests, and pulmonary function testing. We enrolled 9804 general citizens of Nagahama City in the Shiga Prefecture, Japan. RESULTS: Mouth breathing was reported by 17% of the population and was independently associated with asthma morbidity. The odds ratio for asthma morbidity was 1.85 (95% CI, 1.27-2.62) and 2.20 (95% CI, 1.72-2.80) in subjects with mouth breathing alone and allergic rhinitis alone, which additively increased to 4.09 (95% CI, 3.01-5.52) when mouth breathing and allergic rhinitis coexisted. Mouth breathing in nonasthmatics was a risk for house dust mite sensitization, higher blood eosinophil counts, and lower pulmonary function after adjusting for allergic rhinitis. CONCLUSION: Mouth breathing may increase asthma morbidity, potentially through increased sensitization to inhaled allergens, which highlights the risk of mouth-bypass breathing in the 'one airway, one disease' concept. The risk of mouth breathing should be well recognized in subjects with allergic rhinitis and in the general population.
Asunto(s)
Asma/epidemiología , Asma/etiología , Respiración por la Boca , Adulto , Anciano , Asma/diagnóstico , Biomarcadores , Estudios de Cohortes , Femenino , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Morbilidad , Oportunidad Relativa , Vigilancia de la Población , Pruebas de Función Respiratoria , Factores de Riesgo , AutoinformeRESUMEN
OBJECTIVE: This paper assesses the relationship between the distance between the cemento-enamel junction and alveolar crest and risk factors commonly associated with periodontitis. MATERIALS: Eighty individuals between 28 and 92 years old with known biological sex and age were analyzed from a 20th century forensic human collection from Merida, Yucatan (Mexico). METHODS: Macroscopic assessment, along with metric analysis, was employed using a probe. RESULTS: Ante-mortem tooth loss was positively correlated with the distance between the cemento-enamel junction and alveolar crest, as was the presence of root calculus in females. CONCLUSIONS: Cemento-enamel junction to alveolar crest distance is not a reliable indicator of periodontitis since it is not directly related to periodontitis-causing infectious pathogens, and since ante-mortem tooth loss can affect root exposure. SIGNIFICANCE: This study demonstrates that a purely quantitative approach to diagnosing periodontitis in archaeological and forensic human remains can be misleading. LIMITATIONS: The skeletal collection is only representative of the low socioeconomic class of Merida, and its female cohort is underrepresented. In addition, because the Xoclan collection is modern, limitations (particularly with respect to tooth wear) of the applicability of these interpretations to older archaeological remains exist. SUGGESTION FOR FURTHER RESEARCH: A combination of quantitative and qualitative characteristics of alveolar bone is needed to reliably diagnose periodontitis in skeletal populations.
Asunto(s)
Pérdida de Hueso Alveolar , Cálculos Dentales , Periodontitis , Humanos , Femenino , Persona de Mediana Edad , Anciano , Adulto , Masculino , México , Anciano de 80 o más Años , Pérdida de Hueso Alveolar/patología , Pérdida de Hueso Alveolar/historia , Periodontitis/patología , Periodontitis/historia , Cálculos Dentales/patología , Cálculos Dentales/historia , Pérdida de Diente/patología , Pérdida de Diente/historia , Paleopatología/métodos , Clase Social , Estatus Socioeconómico BajoRESUMEN
BACKGROUND: Epidemiological studies have shown that smoking increases the propensity for atopy and asthma. However, the effects of smoking on atopy and eosinophilic inflammation in asthmatics, including the elderly, remain unknown. OBJECTIVE: To determine the effects of smoking on serum immunoglobulin E (IgE) levels and eosinophilic inflammation in asthmatics of all ages. METHODS: The associations of serum IgE levels, blood eosinophil counts and fractional exhaled nitric oxide (FeNO) levels with smoking and age in steroid-naive asthmatics were cross-sectionally assessed (n = 307). Levels of sputum eosinophil and thymic stromal lymphopoietin (TSLP) that promotes Th2 inflammation were also analysed. Current smokers were excluded when analysing contributing factors of FeNO. RESULTS: Levels of serum IgE, blood eosinophil and FeNO decreased with increasing age in never-smokers, whereas decrease in serum IgE levels with increasing age was not observed in current smokers. In addition, current smoking was associated with higher blood eosinophil counts. In atopic asthmatics, age-related declines in serum IgE levels were less steep in ex-smokers than in never-smokers, and atopic ex-smokers with asthma showed higher blood eosinophil counts and higher FeNO irrespective of age. Lastly, sputum TSLP levels were associated with sputum eosinophil proportions and pack-years. Current and ex-smokers had higher TSLP levels than never-smokers. CONCLUSIONS AND CLINICAL RELEVANCE: In steroid-naive asthmatics, smoking may attenuate the age-related decrease in IgE levels and maintain eosinophilic inflammation, in which TSLP may be involved.
Asunto(s)
Eosinófilos/inmunología , Inmunoglobulina E/inmunología , Inflamación/inmunología , Fumar , Adolescente , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Asma/inmunología , Asma/metabolismo , Estudios Transversales , Citocinas/metabolismo , Espiración , Femenino , Compuestos Férricos/sangre , Humanos , Inmunoglobulina E/sangre , Inflamación/sangre , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Nitratos/sangre , Óxido Nítrico , Esputo/metabolismo , Adulto Joven , Linfopoyetina del Estroma TímicoRESUMEN
BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterised by the presence of airflow limitation caused by loss of lung elasticity and/or airway narrowing. The pathological hallmark of loss of lung elasticity is emphysema, and airway wall remodelling contributes to the airway narrowing. Using CT, these lesions can be assessed by measuring low attenuation areas (LAA) and airway wall thickness/luminal area, respectively. As previously reported, COPD can be divided into airway dominant, emphysema dominant and mixed phenotypes using CT. In this study, it is postulated that a patient's physique may be associated with the relative contribution of these lesions to airflow obstruction. METHODS: CT was used to evaluate emphysema and airway dimensions in 201 patients with COPD. Emphysema was evaluated using percentage of LAA voxels (LAA%) and airway lesion was estimated by percentage wall area (WA%). Patients were divided into four phenotypes using LAA% and WA%. RESULTS: Body mass index (BMI) was significantly lower in the higher LAA% phenotype (ie, emphysema dominant and mixed phenotypes). BMI correlated with LAA% (rho = -0.557, p<0.0001) but not with WA%. BMI was significantly lower in the emphysema dominant phenotype than in the airway dominant phenotype, while there was no difference in forced expiratory volume in 1 s %predicted between the two. CONCLUSION: A low BMI is associated with the presence of emphysema, but not with airway wall thickening, in male smokers who have COPD. These results support the concept of different COPD phenotypes and suggest that there may be different systemic manifestations of these phenotypes.
Asunto(s)
Índice de Masa Corporal , Enfermedad Pulmonar Obstructiva Crónica/patología , Anciano , Biomarcadores/sangre , Femenino , Volumen Espiratorio Forzado/fisiología , Humanos , Masculino , Persona de Mediana Edad , Fenotipo , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico por imagen , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Tomografía Computarizada por Rayos XRESUMEN
BACKGROUND: The association between gastro-oesophageal reflux disease (GORD) and chronic obstructive pulmonary disease (COPD) exacerbation has so far remained unclear. OBJECTIVE: To prospectively establish the clinical significance of GORD symptoms on exacerbation. METHODS: 82 patients with COPD and 40 age matched controls were enrolled in this study. Symptoms were evaluated by a questionnaire using the Frequency Scale for the Symptoms of GORD (FSSG). Patients with COPD were prospectively surveyed for 6 months, and episodes of exacerbation were identified using a diary based on modified Anthonisen's criteria. Exhaled breath condensate (EBC) pH was measured in both groups, and induced sputum was evaluated in patients with COPD. RESULTS: Positive GORD symptoms were reported in 22 (26.8%) patients with COPD and in five (12.5%) controls (p = 0.10). The frequency of exacerbations was significantly associated with the FSSG score (p = 0.03, r = 0.24, 95% CI 0.02 to 0.43). Multiple regression analysis revealed that GORD symptoms were significantly associated with the occurrence of exacerbations (p<0.01; relative risk 6.55, 95% CI 1.86 to 23.11). EBC pH was inversely correlated with FSSG score in both groups (p = 0.01, r = -0.37, 95% CI -0.55 to -0.14 in patients with COPD, and p<0.01, r = -0.45, 95% CI -0.67 to -0.16 in control subjects). CONCLUSIONS: GORD symptoms were identified as an important factor associated with COPD exacerbation.
Asunto(s)
Reflujo Gastroesofágico/complicaciones , Enfermedad Pulmonar Obstructiva Crónica/etiología , Anciano , Estudios de Casos y Controles , Femenino , Humanos , Concentración de Iones de Hidrógeno , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Factores de Riesgo , Encuestas y CuestionariosRESUMEN
BACKGROUND: Storage of canine packed red blood cells (pRBCs) can increase erythrocyte phosphatidylserine (PS) expression and eicosanoid concentrations. HYPOTHESIS/OBJECTIVES: To determine the effects of leukoreduction on erythrocyte PS expression and eicosanoid concentrations in stored units of canine pRBCs. Our hypothesis was that leukoreduction would decrease PS expression and eicosanoid concentrations. ANIMALS: Eight healthy dogs. METHODS: In a cross-over study, units of whole blood were leukoreduced (LR) or non-LR and stored (10 and 21 days) as pRBCs. Samples were collected at donation, and before and after a simulated transfusion. PS expression was measured by flow cytometry, and concentrations of arachidonic acid (AA), prostaglandin F2α (PGF2α ), prostaglandin E2 (PGE2 ), prostaglandin D2 (PGD2 ), thromboxane B2 (TXB2 ), 6-keto-prostaglandin F1α (6-keto-PGF1α ), and leukotriene B4 (LTB4 ) were quantified by liquid chromatography-mass spectrometry. RESULTS: There was no change in PS expression during leukoreduction, storage, and simulated transfusion for non-LR and LR units. Immediately after leukoreduction, there was a significant increase in TXB2 and PGF2α concentrations, but during storage, these eicosanoids decreased to non-LR concentrations. In both LR and non-LR units, 6-keto-PGF1α concentrations increased during storage and simulated transfusion, but there was no difference between unit type. There was no difference in AA, LTB4 , PGE2 , and PGD2 concentrations between unit types. CONCLUSIONS AND CLINICAL IMPORTANCE: Leukoreduction, storage, and simulated transfusion do not alter erythrocyte PS expression. Leukoreduction causes an immediate increase in concentrations of TXB2 and PGF2α , but concentrations decrease to non-LR concentrations with storage. Leukoreduction does not decrease the accumulation of 6-keto-PGF1α during storage.
Asunto(s)
Conservación de la Sangre/veterinaria , Eicosanoides/sangre , Procedimientos de Reducción del Leucocitos/veterinaria , Fosfatidilserinas/sangre , Animales , Estudios Cruzados , Perros , Transfusión de Eritrocitos/veterinaria , Eritrocitos/metabolismo , Femenino , Citometría de Flujo/veterinaria , MasculinoRESUMEN
CLINICAL CASE: A 37-year-old man was referred because of a bilateral loss of visual acuity and metamorphopsia. On examination there was yellowish exudation bilaterally associated with serous retinal detachments, corresponding to hyperfluorescence spots in the early stage of fluorescein angiography. Photocoagulation of the leakage spots was performed in both eyes. Six months later he had recovered his visual acuity and the lesions had remitted. DISCUSSION: The clinical findings, ancillary tests and subsequent clinical course allowed a diagnosis of Best's disease to be ruled out. The presence of sub-retinal yellowish exudation in the posterior pole does not necessarily rule out the diagnosis of central serous chorioretinopathy.
Asunto(s)
Enfermedades de la Coroides/complicaciones , Enfermedades de la Coroides/diagnóstico , Enfermedades de la Retina/complicaciones , Enfermedades de la Retina/diagnóstico , Adulto , Exudados y Transudados , Humanos , MasculinoRESUMEN
The endothelium represents an important therapeutic target for containment of oxidative stress, thrombosis and inflammation involved in a plethora of acute and chronic conditions including cardiovascular and pulmonary diseases and diabetes. However, rapid blood clearance and lack of affinity to the endothelium compromise delivery to target and restrict medical utility of antioxidant enzymes (e.g., catalase) and fibrinolytics. The use of "stealth" PEG-liposomes prolongs circulation, whereas conjugation with antibodies to endothelial determinants permits targeting. Constitutive endothelial cell adhesion molecules (CAM, such as ICAM-1 and PECAM-1, which are stably expressed and functionally involved in oxidative stress and thrombosis) are candidate determinants for targeting of antioxidants and fibrinolytics. CAM antibodies and compounds conjugated with anti-CAM bind to endothelial cells and accumulate in vascularized organs (preferentially, lungs). Pathological stimuli enhance ICAM-1 expression in endothelial cells and facilitate targeting, whereas PECAM-1 expression and targeting are stable. Endothelial cells internalize 100-300 nm diameter conjugates possessing multiple copies of anti-CAM, but not monomolecular antibodies or micron conjugates. This permits size-controlled sub-cellular targeting of antioxidants into the endothelial interior and fibrinolytics to the endothelial surface. Targeting catalase to PECAM-1 or ICAM-1 protects endothelial cells against injury by oxidants in culture and alleviates vascular oxidative stress in lungs in animals. Anti-CAM/catalase conjugates are active for a few hours prior to lysosomal degradation, which can be delayed by auxiliary drugs. Conjugation of fibrinolytics to monovalent anti-ICAM permits targeting and prolonged retention on the endothelial surface. Therefore, CAM targeting of antioxidants and fibrinolytics might help to contain oxidative and thrombotic stresses, with benefits of blocking CAM. Avenues for improvement and translation of this concept into the clinical domain are discussed.
Asunto(s)
Antioxidantes/farmacología , Moléculas de Adhesión Celular/efectos de los fármacos , Endotelio Vascular/efectos de los fármacos , Fibrinolíticos/farmacología , Animales , Antioxidantes/efectos adversos , Endotelio Vascular/enzimología , Fibrinolíticos/efectos adversos , HumanosRESUMEN
We isolated a cDNA clone encoding the human prostaglandin (PG) E receptor EP4 subtype and examined the gene expression in human blood cells. Northern blot analysis revealed that the EP4 gene is expressed at a high level in peripheral blood mononuclear cells, and at lower levels in cultured human blood cell lines, THP-1 and U937 (monocytoid cell lines), MOLT-4 and Jurkat (T-cell lines), and Raji (B-cell line). To examine regulation of the EP4 gene expression in the immune system, we studied the effects of phorbol 12-myristate 13-acetate (PMA) on these cell lines. Gene expression was upregulated in THP-1, U937, and Raji cells by PMA, and was downregulated in MOLT-4 and Jurkat cells. In THP-1 cells the effects of PMA were further analyzed, and the upregulation of the EP4 gene was shown to be followed by an increase in PGE2 binding sites and in PGE2-induced cAMP accumulation. In the striking contrast, other PGE receptor subtypes (EP1, EP2 and EP3) and other prostanoid receptors (IP and DP) were shown not to be upregulated by PMA. Therefore, this is the first demonstration of a highly specific upregulation of the EP4 subtype in THP-1 cells treated with PMA, suggesting the importance of the EP4 subtype in the immune system. In the present study we also clarified that EP4 gene expression is regulated differently among human monocytoid and lymphoid lineage cells, thus leading to the better understanding of the regulatory mechanisms for the human EP4 gene expression in the immune system.
Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Linfocitos/efectos de los fármacos , Monocitos/efectos de los fármacos , Receptores de Prostaglandina E/genética , Acetato de Tetradecanoilforbol/farmacología , Linfoma de Burkitt/patología , AMP Cíclico/fisiología , ADN Complementario/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Genes , Humanos , Leucemia Monocítica Aguda/patología , Leucemia-Linfoma de Células T del Adulto/patología , Linfocitos/metabolismo , Linfoma de Células B Grandes Difuso/patología , Monocitos/metabolismo , Proteínas de Neoplasias/biosíntesis , Proteínas de Neoplasias/genética , ARN Mensajero/biosíntesis , ARN Mensajero/genética , ARN Neoplásico/biosíntesis , ARN Neoplásico/genética , Receptores de Prostaglandina E/clasificación , Transducción de Señal/fisiología , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Using specific antagonists to rat interleukin (IL)-1 alpha and IL-1 beta, the roles of these IL-1s in endotoxin-induced suppression of plasma gonadotropin levels in freely-moving rats were studied. In orchiectomized rats, recombinant rat IL-1 alpha and IL-1 beta administered into the lateral ventricles almost equipotently suppressed plasma LH levels. Twenty five micrograms of bacterial endotoxin or lipopolysaccharide (LPS) administered similarly showed a comparable effect as that of 1 microgram IL-1 alpha or IL-1 beta, and completely lowered plasma LH levels by 60 min after the injection. To examine the roles of endogenous IL-1 alpha and IL-1 beta, anti-rat IL-1 alpha antiserum (anti-IL-1 alpha) and a recombinant human IL-1 receptor antagonist (IL-1ra) were used as specific blockers for IL-1 alpha and IL-1 beta, respectively. Anti-IL-1 alpha (10 microliters) or IL-1ra (10 micrograms) administered intracerebroventricularly (icv) with 25 micrograms LPS, significantly attenuated the LPS-induced effect on plasma LH levels during the first 60 min after LPS infusion, but not during the second 60 min. LPS at a dose of 5 micrograms induced smaller but still significant changes in plasma LH levels compared with 25 micrograms LPS or 1 microgram IL-1 beta. IL-1ra (10 micrograms) completely blocked LH suppression induced by 1 microgram IL-1 beta, but did not completely reverse the changes of LH induced by 5 micrograms LPS. IL-1ra injected iv also significantly attenuated the early suppressive effect of iv administered LPS, but not its late effect on plasma LH levels. However, iv administered IL-1ra had no influence on the effects of icv administered LPS. These data indicate that at least a part of plasma LH suppression caused by icv administered LPS is mediated via IL-1 alpha and IL-1 beta synthesized within the brain, while factor(s) other than IL-1 also participate in the LPS-induced change, particularly during the later period. A similar mechanism may also work peripherally in the case of iv administered LPS-induced plasma LH suppression.
Asunto(s)
Ventrículos Cerebrales/fisiología , Interleucina-1/farmacología , Lipopolisacáridos/toxicidad , Hormona Luteinizante/sangre , Animales , Ventrículos Cerebrales/efectos de los fármacos , Escherichia coli , Sueros Inmunes , Inyecciones Intraventriculares , Interleucina-1/administración & dosificación , Interleucina-1/fisiología , Cinética , Hormona Luteinizante/metabolismo , Masculino , Orquiectomía , Ratas , Ratas Endogámicas , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/farmacología , Factores de TiempoRESUMEN
Whereas the stimulatory effect of interleukin-6 (IL-6) on the hypothalamic-pituitary-adrenal (HPA) axis is well established, its mode of action in this axis has yet to be fully elucidated. To further study the role of IL-6 in the HPA axis, we compared the expression of IL-6 messenger RNA (mRNA) in the rat hypothalamus, pituitary, and adrenal gland with that in the spleen after ip or intracerebroventricular (icv) administration of bacterial lipopolysaccharide (LPS). After either ip or icv administration, LPS induced the expression of IL-6 mRNA, which consists of 1.2 kilobases (kb) and 2.4 kb subclasses, in all these tissues of the HPA axis as well as in the spleen. Although we used 100 times less amount of LPS for the icv administration than that used for ip LPS, plasma ACTH levels in both the conditions rapidly reached comparable levels. This icv dose induced IL-6 mRNA expression in the hypothalamus faster than ip dose but also stimulated IL-6 mRNA expression in the hypothalamus, pituitary, and adrenal gland more effectively and smoothly than the ip LPS dose did. Northern blot analysis revealed that in the hypothalamus, pituitary, and adrenals, the predominant subclass of IL-6 mRNA was not 1.2 kb but 2.4 kb. In contrast, this subclass was the minor component in the spleen induced under the same circumstances. These findings indicate that IL-6-synthesizing cells in the HPA axis differ in character from those in the spleen, and that LPS applied in vivo may modulate IL-6 expression in these cells directly and/or indirectly through secondarily activated functions in the neuronal or endocrine systems.
Asunto(s)
Sistema Hipotálamo-Hipofisario/metabolismo , Interleucina-6/genética , Lipopolisacáridos/farmacología , Sistema Hipófiso-Suprarrenal/metabolismo , ARN Mensajero/metabolismo , Bazo/metabolismo , Animales , Inyecciones Intraperitoneales , Inyecciones Intraventriculares , Masculino , Ratas , Ratas WistarRESUMEN
PGE2 is known to induce uterine contraction by increasing intracellular Ca2+. In the present study, to investigate other functions of PGE2 in human uterus, two EP3 isoforms were isolated by the RT-PCR method using human uterus polyadenylated ribonucleic acid (RNA). These EP3 isoforms, named EP3-V and EP3-VI, are composed of 402 and 393 amino acid residues, respectively, which are unique compared with EP3 isoforms of other species. Their N-terminal 359 amino acid residues are identical to those of previously reported human EP3 isoforms, whereas the two isoforms contained a novel amino acid sequence in their C-terminal tails. The dissociation constant values of EP3-V and EP3-VI for PGE2 were 3.9 and 1.4 nmol/L, respectively, which were consistent with those of previously reported EP3 isoforms. Signaling experiments revealed that M&B28767, an EP3 agonist, not only inhibited forskolin-induced cAMP concentrations, but also activated mitogen-activated protein kinase in Chinese hamster ovary cells stably expressing EP3-V and EP3-VI. These responses were abolished by treatment with pertussis toxin. In addition, M&B28767 increased cAMP concentrations in EP3-VI-expressing cells, whereas it did not in EP3-V-expressing cells. M&B28767 did not stimulate phosphoinositide turnover in EP3-V or EP3-VI-expressing cells. EP3-V and EP3-VI messenger RNAs (mRNAs) were detected abundantly in human uterus, whereas weak, but substantial, bands were detected in the lung and kidney in RT-PCR specific for each mRNA. In situ hybridization revealed EP3-V and EP3-VI mRNAs in the human myometrium, but not in the endometrium. The present study suggests that EP3-V and EP3-VI are possibly involved in the proliferation of cells in human myometrium.
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Alprostadil/análogos & derivados , Receptores de Prostaglandina E/fisiología , Transducción de Señal/fisiología , Útero/fisiología , Alprostadil/farmacología , Secuencia de Aminoácidos , Animales , Células CHO , Membrana Celular/fisiología , Colforsina/farmacología , Cricetinae , AMP Cíclico/metabolismo , Femenino , Humanos , Datos de Secuencia Molecular , Isoformas de Proteínas/genética , Isoformas de Proteínas/fisiología , ARN Mensajero/genética , Receptores de Prostaglandina E/agonistas , Receptores de Prostaglandina E/genética , Subtipo EP3 de Receptores de Prostaglandina E , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Alineación de Secuencia , Transcripción Genética , TransfecciónRESUMEN
Prophet of Pit-1 (Prop-1), which is a paired-like homeodomain transcription factor, is capable of binding to sites in an early enhancer of the Pit-1 gene and regulating its expression. According to a previous report, Prop-1 messenger RNA (mRNA) is expressed in the developing pituitary gland before Pit-1 mRNA expression and maximum expression are observed at e 12.0. After e 14.5, Prop-1 mRNA expression rapidly decreases, and only trace amounts of mRNA are detectable in adult mouse pituitary. Human Pit-1 is expressed considerably, not only in normal adult pituitary but also in pituitary adenomas, so we studied human Prop-1 gene expression in adult pituitary and pituitary adenomas. We also cloned human Prop-1 complementary DNA (cDNA) and sequenced the Prop-1 cDNAs in pituitary adenomas. The amino acid sequence of human Prop-1 cDNA that we cloned was identical to that of the previously reported sequence, except Thr substituted at codon 142 instead of Ala. This amino acid substitution is considered to be a polymorphism because it did not alter transcriptional activity, and 7 of 28 alleles were Ala. Human Prop-1 transcript was detected in normal adult pituitary, by Northern blot analysis, and in all pituitary adenomas examined by RT-PCR analysis. The expression of human Prop-1 in pituitary adenomas was confirmed by in situ hybridization in one of the somatotroph adenomas. The sequence analysis of human Prop-1 cDNAs in these pituitary adenomas revealed that there were no mutations, except 5 silent nucleic acid substitutions, suggesting that mutations of Prop-1 gene do not represent a frequent mechanism of human pituitary tumorigenesis.
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Adenoma/metabolismo , Proteínas de Homeodominio/genética , Hipófisis/metabolismo , Neoplasias Hipofisarias/metabolismo , ARN Mensajero/análisis , Adulto , Secuencia de Bases , Clonación Molecular , Humanos , Datos de Secuencia Molecular , Mutación , Transcripción GenéticaRESUMEN
Propionic acidaemia (PA) is an autosomal recessive disorder caused by mutations in either of the PCCA or PCCB genes which encode the alpha and beta subunits, respectively, of the mitochondrial enzyme propionyl-CoA carboxylase (PCC). In this work we have examined the biochemical findings and clinical outcome of 37 Spanish PA patients in relation to the mutations found in both PCCA and PCCB genes. We have detected 27 early-onset and 101 late-onset cases, showing remarkably similar biochemical features without relation to either the age of onset of the disease or the defective gene they have. Twenty-one of the patients have so far survived and three of them, now adolescents, present normal development. Different biochemical procedures allowed us to identify the defective gene in 9 PCCA deficient and 28 PCCB deficient patients. Nine putative disease-causing mutations accounting for 77.7% of mutant alleles were identified among PCCA deficient patients, each one carrying a unique genotypic combination. Of PCCB mutant alleles 98% were characterised. Four common mutations (ins/del, E168K, 1170insT and A497V) were found in 38/52 mutant chromosomes investigated, whereas the remainder of the alleles harbour 12 other different mutations. By examining the mutations identified both in PCCA and PCCB genes and the clinical evolution of patients, we have found a good correlation between certain mutations which can be considered as null with a severe phenotype, while certain missense mutations tend to be related to the late and mild forms of the disease. Expression studies, particularly of the missense mutations identified are necessary but other genetic and environmental factors probably contribute to the phenotypic variability observed in PA.
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Carboxiliasas/deficiencia , Adulto , Biotina/metabolismo , Northern Blotting , Western Blotting , Carboxiliasas/genética , Células Cultivadas , Análisis Mutacional de ADN , Femenino , Prueba de Complementación Genética , Genotipo , Humanos , Masculino , Metilmalonil-CoA Descarboxilasa , Fenotipo , España , Resultado del Tratamiento , TritioRESUMEN
By use of the signal sequence trap method, we isolated a cDNA encoding a novel aspartic protease-like protein from the mouse kidney, and termed it 'kidney-derived aspartic protease-like protein (KAP).' The protein, a 419-amino-acid polypeptide with a 16-amino-acid signal sequence, had 47% identity with mouse cathepsin D, and its overall structure was closely related to known aspartic proteases. Northern blot analysis revealed that KAP mRNA is expressed at the highest level in the kidney, at a moderate level in the lung, and at low levels in the spleen and adipose tissue. In situ hybridization analysis demonstrated that the mRNA is expressed abundantly in the proximal straight tubule and slightly, but significantly, in the proximal convoluted tubule in the kidney. This intra-renal distribution differs distinctly from those of previously reported proteases such as cathepsins B, D, and H.
Asunto(s)
Ácido Aspártico Endopeptidasas/genética , Riñón/enzimología , Secuencia de Aminoácidos , Animales , Ácido Aspártico Endopeptidasas/biosíntesis , Secuencia de Bases , Northern Blotting , Clonación Molecular , ADN Complementario , Hibridación in Situ , Ratones , Datos de Secuencia Molecular , Señales de Clasificación de Proteína/genética , ARN Mensajero/metabolismo , Homología de Secuencia de AminoácidoRESUMEN
OBJECTIVE: Prostaglandin (PG) E2, a major arachidonic acid metabolite in the kidney, acts on four receptor subtypes (EP1, EP2, EP3 and EP4). One of major causes of end-stage renal failure is hypertensive renal disease, in which enhanced renal PGE2 production has been shown. In this study, to explore the pathophysiological significance of EP subtypes in the kidney, we examined the role of EP subtypes on proliferation of mesangial cells (MCs) from stroke-prone spontaneously hypertensive rats (SHRSPs), which show faster growth than those from normotensive Wistar-Kyoto rats (WKYs). DESIGN AND METHODS: Using MCs from SHRSPs and WKYs, we investigated DNA synthesis and its upstream event, the phosphorylation of extracellular signal-regulated kinase (ERK), together with the gene expression of EP subtypes. RESULTS: Sulprostone, an EP1 agonist, dose-dependently increased DNA synthesis and the phosphorylation of ERK in MCs from both strains. The EP4 agonist, 11-deoxy-PGE1, inhibited sulprostone-induced phosphorylation of ERK in WKY-MCs. In contrast, 11-deoxy-PGE1 failed to inhibit the ERK activity in SHRSP-MCs. Interestingly, cAMP production mediated by EP4 was markedly attenuated in SHRSP-MCs as compared with that in WKY-MCs, despite the overproduction of endogenous PGE2 in SHRSP-MCs. Similar gene expressions of EP1 and EP4 and only faint expression of EP3 were detected in MCs from both strains. CONCLUSIONS: These results indicate that the PGE2/EP4 system counteracts the PGE2/EP1 system at the level of the intracellular signaling pathway. The altered EP4 signaling may play a critical role in the exaggerated mesangial growth in SHRSPs.
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Dinoprostona/análogos & derivados , Dinoprostona/fisiología , Mesangio Glomerular/fisiopatología , Receptores de Prostaglandina E/fisiología , Animales , Células Cultivadas , AMP Cíclico/biosíntesis , ADN/biosíntesis , Dinoprostona/farmacología , Expresión Génica , Mesangio Glomerular/efectos de los fármacos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosforilación , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Receptor Cross-Talk , Receptores de Prostaglandina E/agonistas , Receptores de Prostaglandina E/antagonistas & inhibidores , Receptores de Prostaglandina E/clasificación , Receptores de Prostaglandina E/genética , Subtipo EP1 de Receptores de Prostaglandina E , Subtipo EP4 de Receptores de Prostaglandina E , Transducción de SeñalRESUMEN
We investigated the effect of aging on the extracellular matrix of the lungs by examining age-associated changes in the elastin and collagen content, and the proportion of types I and III collagen, in the whole lungs of BALB/c and SAMR1 male mice between the ages of 3 and 24 months. The elastin content was determined by the hot alkali method. The hydroxyproline content was measured and assessed to be the collagen content. The relative proportion of types I and III collagen was assessed by cyanogen bromide digestion, followed by separation of the resultant peptides by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The elastin content did not change significantly with age in either strain of mice. The total collagen content of the whole lung was significantly higher at 24 months of age, although there were no significant changes with aging in the hydroxyproline content per dry lung weight nor in the proportion of type III to type I collagen. We conclude that in terms of the extracellular matrix, the lungs of aged mice are not very different in feature from the lungs of younger mice, and this is probably the simple consequence of growth of the lungs of young mice.
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Envejecimiento/metabolismo , Colágeno/metabolismo , Elastina/metabolismo , Pulmón/metabolismo , Envejecimiento/patología , Animales , Colágeno/clasificación , Matriz Extracelular/metabolismo , Pulmón/anatomía & histología , Masculino , Ratones , Ratones Endogámicos BALB C , Tamaño de los Órganos , Especificidad de la EspecieRESUMEN
To clarify the site and mode of action of tumor necrosis factor (TNF) in the pituitary, we studied the effects, binding sites of TNF and its receptor mRNA in the two types of mouse pituitary-derived cell lines, AtT-20, ACTH-producing cells and TtT/GF, folliculo-stellate (FS)-like cells. First, we examined the expression of TNF receptor mRNA in these cells. Using Northern blot analyses with radiolabeled cDNA to murine TNF receptor p60 and p80 mRNAs as probes, we identified both types of mRNA in the poly(A)-containing RNA prepared from AtT-20 cells and p60 TNF receptor mRNA from TtT/GF. The identified mRNA were compatible in size with those detected in the immune-competent cells. Next, we studied the TNF-binding sites on these cells. Scatchard plot analysis of the significant binding of [125I]TNF revealed a single type of binding site with a Kd (dissociation constant) of 210 pM and 131 binding sites/cell on AtT-20. Similarly on TtT/GF, [125I]TNF showed 353 binding sites/cell with a Kd of 900 pM. [125I]TNF binding on both types of cells competed with TNF and lymphotoxin (TNF beta) in an equimolar fashion. Third, TNF stimulates ACTH synthesis in AtT-20 cells, while TNF increases immunoreactive interleukin (IL)-6 release from TtT/GF cells. These findings demonstrate that AtT-20 and TtT/GF cells are equipped with fully functional TNF receptor system, and suggest that ligand of the receptor, TNF alpha and/or TNF beta, can modulate ACTH synthesis and release as a direct hormonal effector on corticotrophs or indirect modulator through another paracrine mediator, such as IL-6 from FS cells.