RESUMEN
PURPOSE: While some work has been done on Health-Related Quality of Life (HRQoL) in statin users, none has focused specifically on statin-associated muscle symptoms (SAMS) sufferers. The objective was to assess self-reported HRQoL, before and after statin withdrawal, in patients reporting SAMS. We hypothesized that the presence of SAMS associated with decreased self-reported physical and mental well-being. METHODS: Patients (50 men/28 women [M/W], aged 49 ± 9 years [Mean ± SD]) in primary cardiovascular prevention were recruited into three cohorts: statin users with (SAMS, 29 M/18W) or without symptoms (No SAMS, 10 M/5W) and controls (11 M/5W). The Short Form 36 Health Survey (SF-36) was used to assess HRQoL. All variables were measured before and after 2 months of statin withdrawal, and repeated measures analyses were used to verify withdrawal and group effects as well as their interaction. RESULTS: SF-36 physical and mental component scores (respectively, PCS and MCS) were lower in the SAMS group compared with other groups (both p < 0.01). Statin withdrawal led to an increase in LDL cholesterol for statin users (+69.0%, p < 0.01) and an improvement in well-being in the SAMS group, other groups showing no change. A time x category interaction (p = 0.02) was seen for PCS and post hoc analyses showed that statin withdrawal improved PCS and MCS (respectively, +12.5% [ES 0.77] and +5.1% [ES 0.27], both p < 0.05) in the SAMS group. CONCLUSION: Patients self-reporting SAMS showed improved HRQoL following drug withdrawal, but this was mirrored by a rise in LDL cholesterol. These findings should be considered by clinicians in the evaluation and follow-up of treatment with statins.
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Enfermedades Cardiovasculares , Inhibidores de Hidroximetilglutaril-CoA Reductasas , Masculino , Humanos , Femenino , Inhibidores de Hidroximetilglutaril-CoA Reductasas/efectos adversos , Calidad de Vida/psicología , LDL-Colesterol , Salud Mental , Enfermedades Cardiovasculares/prevención & controlRESUMEN
BACKGROUND: We conducted a retrospective case review of patients with mastoid cavity and active or inactive chronic otitis media (COM) who underwent cochlear implantation and ear obliteration in a single-stage procedure. The objectives of this review are to assess the rates of complications and postoperative infections and to evaluate post-implantation audiologic performance. MATERIALS AND METHODS: All patients with COM and mastoid cavity, associated or not with active disease, who undergo cochlear implantation and obliteration of the ear as a single-stage procedure from November 2004 to April 2013, were included in the review. All the complications were recorded. Open-set sentence scores were used to evaluate the audiologic gain after implantation. RESULTS: Twenty-seven patients were included in our review: Ten with active COM and seventeen with inactive COM. Overall, nine patients (9/27) presented post-operative complications (7/9 were minor): three were amongst active COM patients (30%) as compared to six amongst inactive COM patients (35%), which included the two major complications. A mean gain of 55.9% on open-set sentence scores was obtained after cochlear implantation. DISCUSSION: We found that complications rate of the one-stage cochlear implantation was higher in patients with COM than in global implant population, but most complications were minors and there was no statistical difference between active and inactive COM. In addition, these patients had audiologic scores similar to those found in patients with normal temporal bone anatomy. CONCLUSION: Cochlear implantation performed as a one-stage procedure could be considered as an option of treatment to avoid staging in patients with active and inactive COM. Although these patients need a regular follow-up, they present good post-implantation audiometric scores.
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Implantación Coclear , Implantes Cocleares , Pérdida Auditiva Conductiva/rehabilitación , Apófisis Mastoides/cirugía , Otitis Media/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Enfermedad Crónica , Implantación Coclear/métodos , Femenino , Estudios de Seguimiento , Pérdida Auditiva Conductiva/etiología , Humanos , Masculino , Persona de Mediana Edad , Otitis Media/complicaciones , Diseño de Prótesis , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
We studied the association of breast cancer with the polymorphic polyglutamine repeat of the androgen receptor (AR) in 255 incident cases of breast cancer and 461 matched controls from the Quebec City metropolitan area. Women for whom the sum of both of the AR (CAG)n-repeats alleles is 39 or less (short-allele AR genotypes) have one-half the risk of breast cancer compared with women for whom the sum of AR (CAG)n-repeats is 40 or more [odds ratio (OR), 0.5; 95% confidence interval (CI), 0.3-0.83; P = 0.007]. This association is stronger in postmenopausal women (180 cases, 297 controls; OR, 0.36; 95% CI, 0.19-0.7; P = 0.003). We also observed an interaction between the type of menopause (natural versus surgical) and the AR genotype on breast cancer risk. Alternately, when subjects were grouped according to their (CAG)n-repeat genotype [homozygous for short alleles (CAG)n < or = 20; other genotypes ("long allele")], results were similar (OR. 0.5; 95% CI, 0.27-0.82; P = 0.007). Thus, women with short-alleles AR genotypes appear to be protected against breast cancer. Short-alleles AR genotypes were observed in 16% of the general population as represented by the control group. Short polyglutamine repeats in the AR protein have been reported to be associated with an increase in the capacity of the receptor to activate transcription of reporter genes in vitro. Furthermore, androgens have been previously shown to inhibit in vitro the growth of breast cancer cell lines. This suggests that differences in the number of polyglutamines in the AR protein may influence individual risk of breast cancer, especially in postmenopausal women, and that this apparent protection could be the consequence of an increased response/sensitivity to androgens.
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Neoplasias de la Mama/genética , Péptidos/genética , Receptores Androgénicos/genética , Estudios de Casos y Controles , Femenino , Predisposición Genética a la Enfermedad , Humanos , Persona de Mediana Edad , Polimorfismo GenéticoRESUMEN
When released into the bloodstream, proinflammatory cytokines have the ability to trigger the transcription of different genes in cells of the blood-brain barrier (BBB), including members of the nuclear factor kappa B (NF-kappaB) family and cyclooxygenase-2 (COX-2), the limiting enzyme for the formation of prostaglandins (PGs). The present study investigated the possibility that interleukin-1beta (IL-1beta) plays an essential role in these events during a systemic inflammatory response. Both wild-type and IL-1beta-deficient mice were killed at different times after two different immunogenic stimuli, i.e., intraperitoneal lipopolysaccharide (LPS) injection and intramuscular turpentine injection, used here as a model of systemic localized inflammatory insult. The inhibitory factor kappaBalpha (IkappaBalpha, index of NF-kappaB activity) and COX-2 transcripts were detected throughout the brain by means of in situ hybridization. Systemic LPS injection caused a strong and rapid expression of IkappaBalpha in endothelial cells lining the BBB of large and small blood vessels and thereafter within parenchymal microglia across the brain. This treatment also provoked a transient expression of COX-2 along cells of the vascular system, and the expression pattern and intensity of the signal for both transcripts were essentially the same in wild-type and IL-1beta-deficient animals. In contrast, the induction of these genes that was quite selective to the cells of the BBB in response to intramuscularly turpentine insult was completely abolished in IL-1beta-deficient mice. Indeed, a late and prolonged expression of IkappaBalpha and COX-2 mRNAs was found along the cerebral blood vessels in response to the sterile and localized inflammation in wild-type mice, whereas such induction was absent in the brain of IL-1beta-deficient animals. These results indicate that IL-1beta has an obligatory role in the activation of NF-kappaB molecules and PGs within endothelial cells of the BBB in an experimental model of intramuscularly turpentine-induced inflammation but not during endotoxemia.
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Barrera Hematoencefálica/fisiología , Endotoxemia/fisiopatología , Inflamación/fisiopatología , Interleucina-1/fisiología , Isoenzimas/genética , FN-kappa B/metabolismo , Prostaglandina-Endoperóxido Sintasas/genética , Transcripción Genética/fisiología , Animales , Ciclooxigenasa 2 , Endotelio Vascular/metabolismo , Endotelio Vascular/patología , Endotoxemia/genética , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Inflamación/enzimología , Inflamación/genética , Inflamación/metabolismo , Inyecciones Intramusculares , Inyecciones Intraperitoneales , Interleucina-1/genética , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Noqueados/genética , Trementina/farmacologíaRESUMEN
Congenital adrenal hyperplasia is the most frequent cause of adrenal insufficiency and ambiguous genitalia in newborn children. In contrast to congenital adrenal hyperplasia due to 21-hydroxylase and 11 beta-hydroxylase deficiencies, which impair steroid formation in the adrenal cortex, exclusively, classical 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) deficiency affects steroid biosynthesis in the gonads as well as in the adrenals. The structures of the highly homologous type I and II 3 beta-HSD genes have been analyzed in three male pseudohermaphrodite 3 beta-HSD deficient patients from unrelated families in order to elucidate the molecular basis of classical 3 beta-HSD deficiency from patients exhibiting various degrees of severity of salt losing. The nucleotide sequence of DNA fragments generated by selective polymerase chain reaction amplification that span the four exons, the exon-intron boundaries, as well as the 5'-flanking region of each of the two 3 beta-HSD genes have been determined in the three male patients. The five point mutations characterized were all detected in the type II 3 beta-HSD gene, which is the gene predominantly expressed in the adrenals and gonads, while no mutation was detected in the type I 3 beta-HSD gene, predominantly expressed in the placenta and peripheral tissues. The two male patients suffering from severe salt-losing 3 beta-HSD deficiency are compound heterozygotes, one bearing the frame-shift mutation 186/insC/187 and the missense mutation Y253N, while the other bears the nonsense mutation W171X and the missense mutation E142K. The influence of the detected missense mutations on enzymatic activity was assessed by in vitro expression analysis of mutant recombinant enzymes generated by site-directed mutagenesis in heterologous mammalian cells. Recombinant mutant type II 3 beta-HSD enzymes carrying Y253N or E142K substitutions exhibit no detectable activity. On the other hand, the nonsalt-losing patient is homozygous for the missense mutation A245P. This mutation decreases 3 beta-HSD activity by approximately 90%. The present findings, describing the first missense mutations in the human type II 3 beta-HSD gene, provide unique information on the structure-activity relationships of the 3 beta-HSD superfamily. Moreover, the present findings provide a molecular explanation for the enzymatic heterogeneity responsible for the severe salt-losing form to the clinically inapparent salt-wasting form of classical 3 beta-HSD deficiency.(ABSTRACT TRUNCATED AT 400 WORDS)
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3-Hidroxiesteroide Deshidrogenasas/deficiencia , 3-Hidroxiesteroide Deshidrogenasas/genética , Hiperplasia Suprarrenal Congénita/genética , 3-Hidroxiesteroide Deshidrogenasas/química , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , ADN/química , Mutación del Sistema de Lectura , Humanos , Immunoblotting , Masculino , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Mutación , Linaje , Mutación Puntual , Reacción en Cadena de la Polimerasa , Pregnenolona/metabolismo , Progesterona/metabolismoRESUMEN
Three beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase (3 beta HSD) deficiency is a form of congenital adrenal hyperplasia characterized by severe impairment of steroid biosynthesis in the adrenals and gonads. To better understand the molecular basis of the phenotypic heterogeneity found in 3 beta HSD deficiency, we analyzed the structure of type I and II 3 beta HSD genes in a female patient with nonsalt-losing 3 beta HSD deficiency diagnosed at puberty. We directly sequenced DNA fragments generated by polymerase chain reaction amplification of the four exons, the exon-intron boundaries, and the 5'-flanking regions of each gene. No mutation was detected in the type I 3 beta HSD gene, which is the predominant species expressed in the placenta and peripheral tissues. We detected a novel missense mutation, Y254D, in one allele of the patient's type II 3 beta HSD gene, which is the almost exclusive type expressed in the adrenals and gonads. The influence of the Y254D mutation on enzymatic activity was assessed by analyzing the recombinant mutant enzyme generated by site-directed mutagenesis after its transient expression in COS-1 monkey kidney cells. Recombinant mutant type II 3 beta HSD enzyme carrying the Y254D substitution exhibits no detectable activity with C21 delta 5-steroid pregnenolone or C19 delta 5-steroid dehydroepiandrosterone used as substrate. The absence of restriction fragment length polymorphism by Southern blot analysis and the finding that all of the amplified DNA fragments possess the expected length suggest the absence of deletions, duplications, or re-arrangements in the other allele. A putative second mutation could be located farther than 1427 basepairs upstream of the initiation codon, thus potentially affecting the normal expression of this gene or within intronic regions, generating an alternative aberrant splicing site. These are possibilities that remain to be elucidated. The present findings, which describe the novel missense mutation Y254D in the human type II 3 beta HSD gene, provide useful information on the structure-activity relationships of the 3 beta HSD superfamily.
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3-Hidroxiesteroide Deshidrogenasas/deficiencia , 3-Hidroxiesteroide Deshidrogenasas/genética , Hiperplasia Suprarrenal Congénita/metabolismo , Genes , Mutación , Cloruro de Sodio/metabolismo , Adolescente , Hiperplasia Suprarrenal Congénita/genética , Secuencia de Bases , Femenino , Humanos , Datos de Secuencia Molecular , Sondas de Oligonucleótidos/genética , Reacción en Cadena de la PolimerasaRESUMEN
Congenital adrenal hyperplasia (CAH) is the most frequent cause of adrenal insufficiency and ambiguous genitalia in newborn children. In contrast to CAH caused by 21 alpha-hydroxylase and 11 beta-hydroxylase deficiencies, which impairs steroid formation in the adrenal exclusively, 17 alpha-hydroxylase/17,20-lyase deficiency impairs steroid biosynthesis in the adrenals and gonads. The sequence of CYP17 gene was determined by direct sequencing of asymmetric PCR products in two French-Canadian 46,XY pseudohermaphrodite siblings suffering from combined 17 alpha-hydroxylase/17,20-lyase deficiency. The two patients are homozygous for the novel missense mutation R96W caused by a C to T transition converting codon Arg96 (CGG) into a Trp (TGG) in exon 1. The both parents are heterozygous for this missense mutation. We assessed the effect of the R96W mutation on 17 alpha-hydroxylase/17,20-lyase activity by analysis of mutant enzyme, generated by site-directed mutagenesis, expressed in COS-1 cells. The presence of R96W substitution almost completely abolished the activity of the mutant protein. The present findings provide a molecular explanation for the signs and symptoms of combined 17 alpha-hydroxylase/17,20-lyase deficiency in these two patients and provide useful information on the structure-activity relationships of the P450c17, enzyme.
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Aldehído-Liasas/genética , Sistema Enzimático del Citocromo P-450/genética , Adolescente , Adulto , Aldehído-Liasas/deficiencia , Secuencia de Bases , Sistema Enzimático del Citocromo P-450/deficiencia , Deshidroepiandrosterona/biosíntesis , Femenino , Humanos , Masculino , Datos de Secuencia Molecular , Mutación , Esteroide 17-alfa-HidroxilasaRESUMEN
Recently, the structure of two genes encoding isoenzymes responsible for 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase (3 beta HSD) activity in the human was elucidated. This activity is an essential step in the biosynthesis of all classes of steroid hormones. In the classic severe form of 3 beta HSD deficiency, patients present with adrenal insufficiency, various degrees of salt loss, and incomplete masculinization in males. Here we report the characterization of the molecular basis of congenital adrenal hyperplasia due to 3 beta HSD deficiency in a male pseudohermaphrodite born from consanguineous parents and having no clinical salt loss. To analyze the structure of the type I and II 3 beta HSD genes of the patient, DNA fragments, generated by polymerase chain reaction amplification of the four exons and the exon-intron boundaries of these genes, were directly sequenced. The patients carried a homozygous missense mutation converting Asn100 to Ser in exon 3 of his type II 3 beta HSD gene. His parents were heterozygous for the same point mutation. The absence of clinical salt loss associated with a male pseudohermaphroditism suggested that 3 beta HSD activity was impaired to different levels in the testes and adrenal. To elucidate whether this N100S missense mutation affected preferentially a steroidogenic pathway, enzymatic activity was analyzed by in vitro analysis of mutant recombinant enzyme generated by site-directed mutagenesis after its transient expression in COS-1 cells. Using homogenates from transfected cells, the N100S 3 beta HSD enzyme showed a Km value for pregnenolone of 25 +/- 3 mumol/L compared with 3.5 +/- 0.2 mumol/L for the normal human type II 3 beta HSD enzyme. Similar results were obtained using dehydroepiandrosterone as substrate. In addition to decreasing apparent affinity, the N100S mutation decreased the relative specific activity (Vmax), leading to a relative specificity (relative Vmax/Km) 2.7% and 11% that of normal type II 3 beta HSD using pregnenolone or dehydroepiandrosterone as substrate, respectively. Moreover, the mutant N100S protein had an apparent decreased affinity for NAD+, with a Km value of 650 +/- 66 mumol/L compared with 20 +/- 2 mumol/L for normal type II 3 beta HSD. Except for the hypothetical effect of local factors, these findings suggest that a very weak residual activity of the normal type II 3 beta HSD enzyme could prevent salt loss, but it was insufficient for normal male sex differentiation.(ABSTRACT TRUNCATED AT 400 WORDS)
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3-Hidroxiesteroide Deshidrogenasas/genética , Trastornos del Desarrollo Sexual/enzimología , Trastornos del Desarrollo Sexual/genética , Homocigoto , Mutación Puntual , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , Hormona Adrenocorticotrópica , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Preescolar , Consanguinidad , Análisis Mutacional de ADN , Cartilla de ADN , Trastornos del Desarrollo Sexual/fisiopatología , Femenino , Humanos , Cinética , Masculino , Mamíferos , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Linaje , Reacción en Cadena de la Polimerasa , Valores de Referencia , Homología de Secuencia de Aminoácido , Esteroides/sangreRESUMEN
Accumulating evidence supports the existence of an innate immune response in the brain during systemic inflammation that is associated with a robust induction of proinflammatory cytokines and chemokines by specific cells of the central nervous system. The present study investigated the genetic regulation and fine cellular distribution of the monocyte chemoattractant protein-1 (MCP-1) in the brain of mice and rats in response to systemic immune insults. MCP-1 belongs to a superfamily of chemokines that have a leading role in the early chemotaxic events during inflammation. In situ hybridization histochemistry failed to detect constitutive expression of the chemokine transcript in the cerebral tissue except for the area postrema (AP) that exhibited a low signal under basal conditions. This contrasts with the strong and transient induction of the mRNA encoding MCP-1 following a single systemic bolus of lipopolysaccharide (LPS), recombinant interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF-alpha). These stimuli rapidly triggered (30 to 90 minutes) MCP-1 transcription in all the circumventricular organs (CVOs), the choroid plexus (chp), the leptomeninges, and along the cerebral blood vessels. The time-related induction and intensity of the signal differed among the challenges, route of administration and species, but MCP-1-expressing cells were always found in vascular-associated structures and those devoid of blood-brain barrier. At later times, few isolated microglia across the brain parenchyma depicted positive signal for MCP-1 mRNA. A dual-labeling procedure also provided convincing anatomical evidence that endothelial cells of the microvasculature and a few myeloid cells of the CVOs and chp were positive for the transcript during endotoxemia. This gene is under a sophisticated transcriptional regulation, as the hybridization signal returned to undetectable levels 12 to 24 hours after all the treatments in both species. Of interest are the data that only ligands that triggered nuclear factor kappa B (NF-kappa B) signaling had the ability to increase MCP-1 gene expression, because high doses of IL-6 remained without effects. These data provide the anatomical evidence that MCP-1 is expressed within specific populations of cells in response to systemic inflammatory molecules that use NF-kappa B as intracellular signaling system. This chemokine may therefore play a critical role in the cerebral innate immune response and contribute to the early chemotaxic events during chronic cerebral inflammation.
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Química Encefálica/inmunología , Quimiocina CCL2/genética , Ratones Endogámicos/inmunología , Ratas Sprague-Dawley/inmunología , Animales , Química Encefálica/efectos de los fármacos , Expresión Génica/inmunología , Inyecciones Intravenosas , Interleucina-1/farmacología , Lipopolisacáridos/farmacología , Macrófagos/inmunología , Ratones , Microglía/inmunología , FN-kappa B/inmunología , Fenotipo , ARN Mensajero/análisis , Ratas , Choque Séptico/inmunología , Activación Transcripcional/efectos de los fármacos , Activación Transcripcional/inmunología , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
The spontaneous development of autoimmune disease in MRL-lpr mice induces behavioral and endocrine changes that resemble effects of chronic stressors. To further examine the correspondence between autoimmune disease and chronic stress, we asked whether the brains of autoimmune mice show a shift in the corticotropin-releasing factor (CRF) to vasopressin (AVP) ratio. Using in situ hybridization histochemistry with 35S-labelled mouse riboprobes, the levels of mRNA transcripts encoding CRF and AVP were compared between autoimmune MRL-lpr and control MRL +/+ brains. CRF transcript levels were lower in the hypothalamic paraventricular nucleus and in the central nucleus of the amygdala in MRL-lpr mice. AVP transcript levels were higher in the paraventricular and the supraoptic nuclei in MRL-lpr mice compared to controls. CRF mRNA levels were inversely related to performance in stress-sensitive tasks and to measures of autoimmunity. As found previously for behavioral performance, immunosuppressive treatment with cyclophosphamide abolished the group difference in neuropeptide gene expression. These results indicate that an autoimmune disease process is necessary for the shift in the brain CRF:AVP ratio. Furthermore, they support the parallel between chronic stress and chronic autoimmunity/inflammation, and suggest common central mechanisms relevant to endocrine function and behavior.
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Enfermedades Autoinmunes/metabolismo , Conducta Animal/fisiología , Hormona Liberadora de Corticotropina/metabolismo , Sistema Límbico/metabolismo , Vasopresinas/metabolismo , Animales , Enfermedades Autoinmunes/inmunología , Temperatura Corporal , Química Encefálica/inmunología , Hormona Liberadora de Corticotropina/genética , ADN Complementario , Expresión Génica/inmunología , Hibridación in Situ , Sistema Límbico/inmunología , Masculino , Ratones , Ratones Endogámicos MRL lpr , Neuroinmunomodulación/inmunología , Núcleo Hipotalámico Paraventricular/inmunología , Núcleo Hipotalámico Paraventricular/metabolismo , ARN Mensajero/metabolismo , Estrés Fisiológico/inmunología , Estrés Fisiológico/metabolismo , Núcleo Supraóptico/inmunología , Núcleo Supraóptico/metabolismo , Natación , Transcripción Genética/inmunología , Vasopresinas/genéticaRESUMEN
The present study investigated the effect of serotonin depletion on the neuronal activity and transcription of corticotropin-releasing factor in the rat brain during the acute-phase response. Conscious male rats received an intraperitoneal (i.p.) injection with the immune activator lipopolysaccaride (25 microg/100 g body wt) after being treated for three consecutive days with para-chlorophenylalanine (30mg/100 g/day). This irreversible inhibitor of tryptophane-5-hydroxylase decreased hypothalamic serotonin levels by 96%. One, 3 and 6 h after a single i.p. injection of lipopolysaccharide or vehicle solution, rats were killed and their brains cut in 30-microm coronal sections. Messenger RNAs encoding c-fos, nerve-growth factor inducible-B gene, corticotropin-releasing factor and the heteronuclear RNA encoding corticotropin-releasing factor primary transcript were assayed by in situ hybridization using 35S-labeled riboprobes, whereas Fos-immunoreactive nuclei were labeled by immunocytochemistry. Lipopolysaccharide induced a wide neuronal activation indicated by the expression of both immediate-early gene transcripts and Fos protein in numerous structures of the brain. The signal for both immediate-early gene transcripts was low to moderate 1 h after lipopolysaccharide administration, maximal at 3 h and decline at 6 h post-injection, whereas at that time, Fos-immunoreactive nuclei were still detected in most of the c-fos messenger RNA-positive structures. Interestingly, the strong and widespread induction of both immediate-early gene transcripts was almost totally inhibited by para-chlorophenylalanine treatment; in the hypothalamic paraventricular nucleus for example, c-fos messenger RNA signal and the number of Fos-immunoreactive positive cells were reduced by 80 and 48%, respectively, in serotonin-depleted rats treated with the bacterial endotoxin. This blunted neuronal response was also associated with an attenuated stimulation of neuroendocrine corticotropin-releasing factor transcription and plasma corticosterone release. Indeed, lipopolysaccharide caused a selective expression of corticotropin-releasing factor primary transcript in the paraventricular nucleus of the hypothalamus and this effect was significantly reduced by treatment with the serotonin inhibitor. However, basal expression of corticotropin-releasing factor messenger RNA across the brain (bed nucleus of the stria terminalis, medial preoptic area, paraventricular nucleus of the hypothalamus, central nucleus of the amygdala, etc.) was not affected by the para-chlorophenylalanine treatment. These results suggest that the integrity of serotonin pathways plays a role in the neuronal activity triggered by the systemic endotoxin insult. The fact that serotonin depletion largely prevented activation of neurosecretory parvocellular neurons of the paraventricular nucleus of the hypothalamus and neuroendocrine corticotropin-releasing factor gene transcription in response to immunogenic challenge provides the evidence that serotonergic system is part of the brain circuitry involved in the corticotroph axis-immune interface.
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Encéfalo/metabolismo , Hormona Liberadora de Corticotropina/genética , Neuronas/metabolismo , Serotonina/fisiología , Transcripción Genética , Amígdala del Cerebelo/efectos de los fármacos , Amígdala del Cerebelo/metabolismo , Animales , Encéfalo/efectos de los fármacos , Corticosterona/sangre , Fenclonina/farmacología , Genes Inmediatos-Precoces , Genes fos , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Lipopolisacáridos/toxicidad , Masculino , Neuronas/efectos de los fármacos , Especificidad de Órganos , Proteínas Proto-Oncogénicas c-fos/genética , ARN Mensajero/análisis , Ratas , Ratas Sprague-Dawley , Serotonina/metabolismo , Factores de Tiempo , Transcripción Genética/efectos de los fármacos , Triptófano Hidroxilasa/metabolismoRESUMEN
Fluoxetine is a serotonin re-uptake blocker commonly used to treat endogenous depression. The present experiments were carried out to assess the effects of fluoxetine on c-fos induction throughout the rat brain. In addition, intron-directed in situ hybridization analysis was used to examine fluoxetine regulation of corticotropin-releasing factor heteronuclear gene transcription in the paraventricular nucleus of the hypothalamus. Because the actions of corticotropin-releasing factor are mediated by membrane-bound corticotropin-releasing factor type 1 receptors, we also evaluated the stimulation of such receptors after acute fluoxetine exposure. The immediate-early gene, c-fos, was markedly induced in several telencephalic and diencephalic brain structures. For instance, a strong hybridized signal was apparent 30 min after fluoxetine (10 mg/kg; intraperitoneal) administration in the caudate putamen, septal nucleus, bed nucleus of stria terminalis, anterodorsal preoptic area, paraventricular nucleus, supraoptic nucleus, ventromedial hypothalamus and posterior hypothalamic nucleus. In addition, c-fos-expressing neurons were also evident in discrete amygdaloid nuclei. This nuclear induction was brief in duration, as levels of the immediate-early gene were mostly undetectable 90 min after drug administration. In contrast to the extensive induction of c-fos by fluoxetine throughout the brain parenchyma, elevation of corticotropin-releasing factor heteronuclear RNA levels were confined exclusively to neurosecretory nerve cells of the paraventricular nucleus, with peak levels detected 30 min after fluoxetine exposure. Therefore, the time-course of corticotropin-releasing factor heteronuclear RNA closely paralleled that of c-fos. Significant changes in corticotropin-releasing factor type 1 receptor messenger RNA levels were also observed in the paraventricular nucleus but with a slow incremental biosynthesis of the receptor messenger RNA, as high levels were discernible only 360 min after fluoxetine treatment. Finally, we failed to detect sex-related differences in the acute response to fluoxetine, as both female and male rat brains showed a comparable induction of c-fos, corticotropin-releasing factor heteronuclear RNA and corticotropin-releasing factor type 1 receptor expression within parvocellular neurosecretory nerve cells that govern the stress response. All of these findings are discussed in terms of specific sequences of nuclear events that couple fluoxetine-based serotonin input with changes in gene expression in selective neurons.
Asunto(s)
Química Encefálica/efectos de los fármacos , Hormona Liberadora de Corticotropina/biosíntesis , Fluoxetina/farmacología , Genes fos/genética , Receptores de Hormona Liberadora de Corticotropina/biosíntesis , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Transcripción Genética/efectos de los fármacos , Animales , Hormona Liberadora de Corticotropina/genética , Femenino , Hipotálamo/metabolismo , Hibridación in Situ , Masculino , Sondas ARN , ARN Mensajero/biosíntesis , Ratas , Receptores de Hormona Liberadora de Corticotropina/genética , Caracteres SexualesRESUMEN
1. The present study investigated the effect of intraperitoneal (i.p.) administration of the indirect 5-hydroxytryptamine (5-HT) receptor agonist, dexfenfluramine, on the transcriptional activity of corticotropin-releasing factor (CRF) and its type 1 receptor in the brains of conscious male Sprague-Dawley rats via in situ hybridization histochemistry (ISHH) using both intronic and exonic probe technology. 2. The immediate early gene (IEG) c-fos mRNA was also used as index of cellular activity, whereas localization between CRF-immunoreactive (ir) perikarya and the IEG was accomplished to determine the site of CRF neuronal activation in the brain of dexfenfluramine-treated rats. 3. Thirty minutes, 1, 3, and 6 h after a single injection of either dexfenfluramine (10 mg kg-1) or the vehicle solution, adult male rats (230-260 g) were deeply anaesthetized and rapidly perfused with a 4% paraformaldehyde-borax solution (PF). The brains were removed from the skull, postfixed, and placed in a solution of 4% PF-10% sucrose overnight at 4 degrees C. Frozen brains were mounted on a microtome and cut from the olfactory bulb to the medulla in 30-microns coronal sections. 4. Dexfenfluramine induced a general neuronal activation as indicated by the strong signal of c-fos mRNA in several structures of the brain, including the parietal cortex, caudate putamen, circumventricular organs, medial preoptic area, bed nucleus of the stria terminalis, choroid plexus, choroidal fissure, supraoptic nucleus, paraventricular nucleus of the hypothalamus (PVN), paraventricular nucleus of the thalamus, central nucleus of the amygdala, dorsomedial nucleus of the hypothalamus, laterodorsal tegmental nucleus, locus coeruleus, and several subdivisions of the dorsal vagal complex. In most of these structures, the signal was maximal at 30 min, still strong and positive at 60 min, largely decreased at 3 h, and had completely disappeared 6 h after injection. 5. In the parvocellular division of the PVN, the large majority of CRF-ir perikarya displayed a positive signal for the mRNA encoding c-fos, indicating a profound CRFergic activation within this neuroendocrine nucleus after dexfenfluramine administration. 6. Colocalization between CRF-ir neurones and c-fos positive cells was not detected in any other regions. This selective activation of PVN CRF neurones was also confirmed by the presence of CRF primary transcript; 30 min after i.p. injection of the indirect 5-HT agonist, a positive signal for CRF hnRNA was observed, specifically in the parvocellular PVN. 7. Transcription of the gene encoding the type 1 receptor for CRF was highly stimulated in the PVN following 5-HT activation. Although this hypothalamic nucleus exhibited a barely detectable signal under basal conditions, dexfenfluramine induced a strong signal of CRF1 receptor mRNA in the parvocellular PVN. Interestingly, CRF-ir neurones displayed a positive signal for the mRNA encoding the CRF1 receptor, 3 and 6 h after systemic treatment with dexfenfluramine. 8. These results indicate that although dexfenfluramine can generate a wide neuronal activation throughout the brain, this 5-HT agonist triggers the activity of CRF neurones selectively in the parvocellular division of the PVN, a mechanism possibly related to the activity of hypothalamic-pituitary-adrenal axis. Induction of CRF1 receptor mRNA in CRF cells of the PVN indicates that neuroendocrine CRF neurones can be targeted by CNS CRF under 5-HT stimulation.
Asunto(s)
Hormona Liberadora de Corticotropina/genética , Fenfluramina/farmacología , Núcleo Hipotalámico Paraventricular/efectos de los fármacos , Receptores de Hormona Liberadora de Corticotropina/genética , Agonistas de Receptores de Serotonina/farmacología , Transcripción Genética/efectos de los fármacos , Animales , Inmunohistoquímica , Hibridación in Situ , Masculino , Núcleo Hipotalámico Paraventricular/metabolismo , Núcleo Hipotalámico Paraventricular/ultraestructura , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Proteínas Proto-Oncogénicas c-fos/genética , ARN Mensajero/análisis , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/biosíntesis , Factores de TiempoRESUMEN
The purpose of this study was to investigate the effect of the immune activator lipopolysaccharide (LPS) on the expression of corticotropin-releasing factor (CRF) and glucocorticoid receptor (GR) mRNA in the paraventricular nucleus (PVN) of transgenic mice with impaired GR function caused by endogenous expression of GR antisense RNA. At 3 and 8 wk of age, control and transgenic mice were sacrificed 4.5 h after a single ip administration of LPS (100 micrograms/100 g of body wt) or vehicle. Frozen brains were mounted on a microtome and cut in 20-microns sections. mRNAs encoding CRF and GR were assayed by in situ hybridization histochemistry using 35S-labeled riboprobes, and localization of Fos-immunoreactive (Fos-ir) nuclei was determined by immunocytochemistry. Basal expression of CRF mRNA in the PVN, central nucleus of the amygdala (CeA), and geniculate complex (GN) was similar in the control and transgenic mice. LPS induced a comparable neuronal activation in the PVN of control and transgenic mice as revealed by the number of Fos-ir neurons. Moreover, the endotoxin caused a significant increase in the CRF mRNA levels within the PVN and CeA, an effect observed in both animal models. The endotoxin did not notably modulate CRF expression in other regions, such as GN. Although GR mRNA was expressed in the PVN of control mice under basal conditions, this transcript was not detected in this hypothalamic structure in LPS-treated and transgenic animals. This indicated that endogenous Type II GR mRNA is decreased in the PVN of mice expressing Type II GR antisense RNA and that gene is downregulated by LPS. Hybridization signal for CRF and GR transcripts was not notably altered by the age of mice. These results provide evidence that the basal expression of CRF and the increase of neuroendocrine CRF transcription in response to immunogenic challenges are not significantly affected by impairment of the Type II GR function.
Asunto(s)
Hormona Liberadora de Corticotropina/genética , Expresión Génica , Núcleo Hipotalámico Paraventricular/metabolismo , ARN sin Sentido/biosíntesis , Receptores de Glucocorticoides/genética , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Humanos , Hibridación in Situ , Lipopolisacáridos/farmacología , Ratones , Ratones Transgénicos , Núcleo Hipotalámico Paraventricular/inmunología , Proteínas Proto-Oncogénicas c-fos/genética , Proteínas Proto-Oncogénicas c-fos/metabolismo , ARN sin Sentido/metabolismo , ARN Mensajero/metabolismo , RatasRESUMEN
The present study investigated the effect of the acute-phase response of a systemic immune activation on the transcription of various immediate early genes (IEGs) and neuropeptides in the brain of conscious rats. One, 3, 6, 9, and 12 h after a single intraperitoneal (i.p.) administration of either the immune activator lipopolysaccharide (LPS) or the vehicle solution, adult male rats were sacrificed and their brains cut in 30-microns coronal sections. mRNA encoding the IEGs c-fos and nerve growth factor inducible-B (NGFI-B), and neuropeptides corticotropin-releasing factor (CRF), oxytocin (OT), and vasopressin (AVP) were assayed by in situ hybridization histochemistry using a 35S-labeled riboprobes. The primary transcripts [heteronuclear (hn)RNA] for these neuropeptides were also detected using intronic probe technology, and colocalization of c-fos mRNA within CRF, AVP, and OT neurons was determined by means of a combination of immunocytochemistry and in situ hybridization techniques on same the brain sections. One h after LPS treatment, both c-fos and NGFI-B genes were expressed in the parvocellular division of the paraventricular nucleus (PVN) of the hypothalamus. The medial preoptic area/organum vasculosum of the lamina terminalis, the supraoptic nucleus (SON), the magnocellular division of the PVN, the arcurate nucleus/median eminence, the locus coeruleus, the nucleus of the solitary tract, and the area postrema also exhibited a strong signal for these two transcripts 3 h after endotoxin administration. A smaller but a significant c-fos expression was observed in various structures, including the dorsomedial hypothalamic area, the central nucleus of the amygdala, the ventral part of the tuberomammillary nucleus, the laterodorsal tegmental nucleus, the external lateral part of the parabrachial nucleus, the dorsal division of the ambiguus nucleus, and the lateral reticular nucleus of LPS-injected rats. The signal for c-fos and NGFI-B mRNA in most of these brain nuclei reached a maximum at 3 h postinjection, declined at 6 h, and vanished 9 to 12 h after LPS treatment. In the parvocellular nucleus of the PVN, c-fos was largely expressed in CRF-immunoreactive (ir) neurons, whereas in the magnocellular part of that nucleus and in the SON, this transcript was colocalized in numerous OT-ir and few AVP-ir neurons. Relative levels of CRF mRNA in the parvocellular PVN were also significantly increased 6 h following LPS, but endotoxin did not alter the genetic expression of this stress-related neuropeptide in other brain regions.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Reacción de Fase Aguda/inmunología , Encéfalo/metabolismo , Inmunización , Neuronas/inmunología , Neuropéptidos/genética , Transcripción Genética , Reacción de Fase Aguda/metabolismo , Animales , Encéfalo/patología , Genes Inmediatos-Precoces , Histocitoquímica , Sistema Hipotálamo-Hipofisario/fisiología , Hibridación in Situ , Masculino , Sistema Hipófiso-Suprarrenal/fisiología , Proteínas Proto-Oncogénicas c-fos/genética , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVES: The association between job strain and ambulatory blood pressure was studied among female white-collar workers. METHODS: This cross-sectional investigation studied 210 women in high- or low-strain jobs randomly selected from 3183 women of all ages, employed as white-collar workers. The women wore an ambulatory blood pressure monitor for 24 hours during a workday. Mean blood pressures were calculated. Psychological demands and decisional latitude were measured twice (14 months before and 7 days before the blood pressure measurement) with 2 scales recommended by Karasek. RESULTS: Significant differences in blood pressure were found according to current job strain among the women holding a university degree. Their mean blood pressures during work were significantly higher [8.0 mm Hg (1.1 kPa) systolic and 6.4 mm Hg (0.8 kPa) diastolic blood pressure] in the high-strain group than in the low-strain group. Statistically significant elevations in blood pressure over the 24-hour period were also found for women with a university degree. Cumulative exposure to high strain over 14 months was also significantly associated with high systolic blood pressure at work, in the evening, and over a 24-hour period irrespective of other factors related to blood pressure. Among the women without a university degree, the blood pressure differences observed between the job strain groups were less than 1 mm Hg (0.1 kPa) and not statistically significant. CONCLUSIONS: These results provide support for the effect of job strain on ambulatory blood pressure only among female white-collar workers holding a university degree.
Asunto(s)
Presión Sanguínea , Ocupaciones , Estrés Psicológico , Mujeres Trabajadoras/psicología , Adolescente , Adulto , Monitoreo Ambulatorio de la Presión Arterial , Índice de Masa Corporal , Estudios Transversales , Escolaridad , Femenino , Humanos , Persona de Mediana Edad , Quebec , Encuestas y Cuestionarios , Lugar de TrabajoRESUMEN
A questionnaire was mailed to all vaccinators in Quebec in 1998. The objective of this survey was to document vaccinators' attitudes, knowledge, and practices related to vaccination. Vaccinators generally believe in the security, efficacy and usefulness of vaccines given to young children. However, 41% of nurses do not fully agree with these opinions. More than 94% of pediatricians completely disagree that "certain practices (homeopathy, good eating habits and a healthy lifestyle) can eliminate the need for vaccination", compared with 85% of general practitioners and only 60% of nurses. Less than 25% of doctors recall children who are late in getting their immunizations; approximately 45% of vaccinators are in complete agreement with simultaneous injections of two vaccines; many circumstances are incorrectly seen as contra indications for vaccination. Public health authorities should target systematic interventions towards vaccinators to improve this situation and to increase nurses' conviction regarding the benefits of vaccination.
Asunto(s)
Actitud del Personal de Salud , Actitud Frente a la Salud , Conocimientos, Actitudes y Práctica en Salud , Enfermeras y Enfermeros/psicología , Médicos de Familia/psicología , Vacunación/normas , Adulto , Competencia Clínica/normas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pediatría/educación , Pediatría/estadística & datos numéricos , Médicos de Familia/educación , Médicos de Familia/estadística & datos numéricos , Pautas de la Práctica en Medicina/estadística & datos numéricos , Quebec , Encuestas y Cuestionarios , Vacunación/efectos adversos , Vacunación/estadística & datos numéricosRESUMEN
Inherited deficiency of medium-chain acyl-CoA dehydrogenase (MCAD) is a severe, sometimes fatal disorder. A single mutation in the MCAD gene, 985A>G, is involved in approximately 90% of cases. To evaluate the relevance of implementing a systematic population-based screening program in the province of Quebec using a biochemical test, we measured the prevalence of this mutation in a set of anonymous newborn samples from the Quebec City area, a region where the majority of its inhabitants are French-Canadians. An allele-specific polymerase chain reaction assay was designed and used to detect the mutation in 7143 DNA samples obtained from consecutive anonymous newborns. Pools of eight DNA samples were genotyped in parallel for the same mutation to validate this pooling strategy. The allelic frequency of the MCAD 985A>G mutation was found to be 0.71% and the carrier frequency 1:71 (95% confidence interval 1:55 to 1:98). This estimate predicts a homozygous frequency of 1:19,837. Ninety-nine heterozygous carriers and one homozygous individual were identified out of 7143 samples. There was 100% concordance between the individual and pooled analyses, and the pooling strategy reduced the total genotyping costs by approximately 70%. The carrier frequency estimated for this population is similar to other northwestern European populations and would support implementation of systematic newborn screening (such as tandem mass spectrometry screening) for this disease. Pooling DNA samples followed by genotyping appears to be cost-effective for estimating prevalence of rare mutations.
Asunto(s)
Acil-CoA Deshidrogenasa/genética , Errores Innatos del Metabolismo Lipídico/genética , Mutación Puntual , Reacción en Cadena de la Polimerasa/métodos , Acil-CoA Deshidrogenasa/deficiencia , Alelos , Análisis Mutacional de ADN , Francia/etnología , Frecuencia de los Genes , Pruebas Genéticas/economía , Pruebas Genéticas/métodos , Genotipo , Humanos , Recién Nacido , Errores Innatos del Metabolismo Lipídico/enzimología , Errores Innatos del Metabolismo Lipídico/epidemiología , Quebec/epidemiología , Reproducibilidad de los ResultadosRESUMEN
Screening for hereditary hemochromatosis, although largely discussed, is not yet implemented in clinical practice. We evaluated the cost-effectiveness of 165 hemochromatosis population-screening algorithms involving two or three of several screening tests by developing a computer program that simulates all possible screening scenarios. Input data comprised government estimates of health services data and costs and a virtual population with user-defined demographic characteristics (including variable HFE mutation frequencies and penetrance values). We show that when C282Y homozygote prevalence is set at 3:1000, population screening appears cost-effective when penetrance of the biochemical phenotype is >0.70. When only hepatocellular carcinoma and cirrhosis are considered as the cost-driving complications, population-based screening is not significantly more cost-efficient than no screening, but life expectancy of individuals identified with hereditary hemochromatosis and treated is still improved by 7 years. Among the 165 screening algorithms tested in 91 different virtual populations of one million individuals, biochemical tests usually perform better as the initial test than genetic testing. Indeed, the genetic testing is most cost-effective as the final confirmatory test. Finally, for most combinations of prevalence and penetrance of HFE, one screening algorithm--unbound iron-binding capacity + transferrin saturation--appeared robust enough to be always within the top 5 most cost-effective strategies.
Asunto(s)
Algoritmos , Predisposición Genética a la Enfermedad/genética , Pruebas Genéticas/economía , Hemocromatosis/epidemiología , Hemocromatosis/genética , Mutación/genética , Simulación por Computador , Análisis Costo-Beneficio , Pruebas Genéticas/métodos , Humanos , PrevalenciaRESUMEN
Expression of the inhibitory factor kappaB alpha (IkappaB alpha) reflects the activity of nuclear factor kappaB(NF-kappaB) and is a powerful tool to investigate the regulation of the transcription factor within the CNS. IkappaB alpha mRNA was evaluated in the rat brain by means of in situ hybridization following different immunogenic stimuli; i.e., intraperitoneal (i.p.) and intravenous (i.v.) lipopolysaccharide (LPS), i.v. recombinant rat interleukin (IL) 1beta, IL-6, or tumor necrosis factor-alpha (TNF-alpha), and intramuscular (i.m.) turpentine injection, used here as a model of systemic localized inflammatory insult. Systemic LPS, IL-1beta, and TNF-alpha caused a rapid and transient transcriptional activation of IkappaB alpha along the blood vessels of the entire brain; the signal was very intense 30-60 min after the i.v. injections and returned to undetectable levels from 2 to 12 h depending on the challenge. Double-labeling procedure provided the anatomical evidence that IkappaB alpha-expressing cells within the microvasculature were essentially of the endothelial type, as they were immunoreactive to the von Willebrand factor. Scattered small cells were also found across the brain of LPS-, IL-1beta-, and TNF-alpha-injected rats at time 1-3 h, and microglial (OX-42)-immunoreactive cells were positive for the transcript. Such expression within parenchymal microglia was nevertheless not observed in the brain following a localized and sterile inflammatory insult. Indeed, i.m. turpentine administration stimulated IkappaB alpha transcription quite uniquely within the endothelium of the brain capillaries, an effect that paralleled the swelling of the injection site and lasted up to 24 h after the aggression. In contrast to these immunogenic challenges, i.v. IL-6 injection failed to activate the gene encoding IkappaB alpha in the rat brain. These results indicate that NF-kappaB may play a crucial role in specific cellular populations of the CNS to trigger transcription of immune-related genes and that IkappaB alpha resynthesis may act as a dynamic intracellular inhibitory feedback to avoid exaggeration of the response. It is possible that IkappaB alpha expression in cells of the blood-brain barrier is a general mechanism that takes place during systemic inflammation, whereas the participation of NF-kappaB-related molecules within parenchymal cells of the CNS is solicited during more severe conditions such as blood sepsis and endotoxemia.