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BACKGROUND: Educational attainment is important in shaping young people's life prospects. To investigate whether being born with orofacial cleft (OFC) affects school performance, we compared school test results between children born with and without OFC. METHODS: Using record-linked datasets, we conducted a population-based cohort study of children liveborn in Western Australia 1980-2010 with a diagnosis of OFC on the Register of Developmental Anomalies, and a random sample of 6603 children born without OFC. We compared odds ratios for meeting national minimum standards in five domains (reading, numeracy, writing, spelling, grammar and punctuation), and adjusted OR (aOR) for children with cleft lip only (CLO), cleft lip and palate (CL + P) and cleft palate only (CPO) for each domain. RESULTS: Results from two testing programs (WALNA and NAPLAN) were available for 3238 (89%) children expected to participate. Most met the national minimum standards. Compared with children without OFC, children with CPO were less likely to meet minimum standards for NAPLAN reading (aOR 0.57 [95%CI 0.34, 0.96]) grammar and punctuation (aOR 0.49 [95%CI 0.32, 0.76]), WALNA writing (aOR 0.66 [95%CI 0.47, 0.92]), and WALNA and NAPLAN numeracy (aOR 0.64 [95%CI 0.43, 0.95] and aOR 0.47 [95%CI 0.28, 0.82]), respectively. Children with CL + P had significantly lower odds for reaching the spelling standard in NAPLAN tests (aOR 0.52 [95%CI 0.29, 0.94]). Children with CLO had similar odds for reaching all minimum standards. CONCLUSION: Children born with OFC, particularly children with CPO, should be monitored to identify learning difficulties early, to enable intervention to maximize school attainment.
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Labio Leporino/psicología , Fisura del Paladar/psicología , Escolaridad , Discapacidades para el Aprendizaje/etiología , Niño , Labio Leporino/epidemiología , Fisura del Paladar/epidemiología , Estudios de Cohortes , Femenino , Humanos , Discapacidades para el Aprendizaje/epidemiología , Masculino , Registro Médico Coordinado , Instituciones Académicas , Clase Social , Australia Occidental/epidemiologíaRESUMEN
BACKGROUND: Most oestrogen receptor (ER)-positive early breast cancer diagnosed today is highly curable with multimodality treatment. Systemic adjuvant treatments including endocrine therapy and chemotherapy have made a significant contribution to the increasing cure rates over the past three decades. However not all women will require chemotherapy. The IHC4+C score is a prognostic tool that integrates four immunohistochemical measures with clinicopathological features to estimate the residual risk of distant recurrence at 10 years in post-menopausal women with ER-positive breast cancer who have received 5 years of endocrine therapy. Retrospective studies indicate that the test can identify a set of women that are at such low risk of recurrence that chemotherapy can be of little benefit. METHODS: In this study, 124 patients were prospectively selected from the multidisciplinary team meeting between January 2013 and April 2014 for IHC4+C testing. Adjuvant systemic treatment recommendations by clinicians were recorded without and with the availability of the score in addition to the patient's decision. RESULTS: There was concordance in the MDT's recommendation without and with the availability of the score in 73% of cases. Clinicians recommended chemotherapy or at least its discussion to 74 (59%) patients, which fell to 32 (34%) patients after the IHC4+C score was made available, sparing one in four tested patients a chemotherapy recommendation, along with its toxicity and expense. CONCLUSION: This decision impact study shows that when used by clinicians in the multidisciplinary team meeting for adjuvant decision-making, a significant proportion of patients are spared chemotherapy recommendations.
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Neoplasias de la Mama/diagnóstico , Toma de Decisiones/fisiología , Inmunohistoquímica/métodos , Anciano , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Técnicas de Apoyo para la Decisión , Femenino , Humanos , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Pronóstico , Receptores de Estrógenos/metabolismo , Proyectos de Investigación , Estudios RetrospectivosRESUMEN
BACKGROUND: The absence of BRCA1-associated protein 1 (BAP1) expression in uveal melanoma (UM) is associated with metastatic progression and reduced survival. In this study, we examine nuclear BAP1 (nBAP1) protein expression in primary UMs (PUMs) that show both 'typical' and 'atypical' clinical courses according to their chromosome 3 status, and secondary hepatic metastatic UM (MUM), correlating the results with histological, clinical and survival data. METHODS: Nuclear BAP1 expression was immunohistochemically assessed in tissue microarrays (TMAs) of: (a) 68 PUM patients, who had been treated surgically; and (b) 13 MUM patients, with 5 cases being paired with primary tumour tissue. All cases were fully annotated. The percentage of tumour cell nuclei staining positively for BAP1 was scored by independent observers. RESULTS: Nuclear BAP1 protein expression was absent in 35 out of 68 (51%) PUM patients, correlating strongly with poor prognostic clinicopathological and genetic parameters and reduced survival (Log rank, P<0.001). Lack of nBAP1 expression importantly identified a subset of 'atypical' PUM patients with disomy of chromosome 3 but with unexpected metastatic relapse. Nuclear BAP1 expression was absent in 10 out of 13 (77%) MUM and expression was concordant in all paired PUM and MUM patients. CONCLUSIONS: Absent nBAP1 protein expression is an independent survival predictor for UM patients, easily examined using immunohistochemistry.
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Neoplasias Hepáticas/metabolismo , Melanoma/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Ubiquitina Tiolesterasa/metabolismo , Neoplasias de la Úvea/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Estimación de Kaplan-Meier , Neoplasias Hepáticas/secundario , Masculino , Melanoma/mortalidad , Melanoma/secundario , Persona de Mediana Edad , Pronóstico , Modelos de Riesgos Proporcionales , Factores de Riesgo , Neoplasias de la Úvea/mortalidad , Neoplasias de la Úvea/patología , Adulto JovenRESUMEN
Detection of estrus is a key determinant of profitability of dairy herds, but estrus is increasingly difficult to observe in the modern dairy cow with shorter duration and less-intense estrus. Concurrent with the unfavorable correlation between milk yield and fertility, estrus-detection rates have declined to less than 50%. We tested ultra-wideband (UWB) radio technology (Thales Research & Technology Ltd., Reading, UK) for proof of concept that estrus could be detected in dairy cows (two 1-wk-long trials; n=16 cows, 8 in each test). The 3-dimensional positions of 12 cows with synchronized estrous cycles and 4 pregnant control cows were monitored continuously using UWB mobile units operating within a network of 8 base units for a period of 7d. In the study, 10 cows exhibited estrus as confirmed by visual observation, activity monitoring, and milk progesterone concentrations. Automated software was developed for analysis of UWB data to detect cows in estrus and report the onset of estrus in real time. The UWB technology accurately detected 9 out of 10 cows in estrus. In addition, UWB technology accurately confirmed all 6 cows not in estrus. In conclusion, UWB technology can accurately detect estrus and hence we have demonstrated proof of concept for a novel technology that has significant potential to improve estrus-detection rates.
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Detección del Estro/métodos , Estro/fisiología , Animales , Bovinos , Femenino , Fertilidad , Leche/química , Progesterona/análisis , Ondas de Radio , Factores de TiempoRESUMEN
BACKGROUND: Previously, using gene-knockdown techniques together with genome expression array analysis, we showed the gene protein Kinase C (PKC)-zeta (PRKCZ) to mediate the malignant phenotype of human prostate cancer. However, according to NCBI, the gene has undergone several major iterations. Therefore, to understand the relationship between its structure and biological activities, we have analysed its expressed sequence in prostate cancer cell lines and tissues. METHODS: Transcriptome-walking and targeted PCR were used to sequence the mRNA transcribed from PRKCZ. Hydropathy analysis was employed to analyse the hypothetical protein sequence subsequently translated and to identify an appropriate epitope to generate a specific monoclonal antibody. RESULTS: A novel sequence was identified within the 3'-terminal domain of human PRKCZ that, in prostate cancer cell lines and tissues, is expressed during transcription and thereafter translated into protein (designated PKC-ζ(-PrC)) independent of conventional PKC-ζ(-a). The monoclonal antibody detected expression of this 96 kD protein only within malignant prostatic epithelium. INTERPRETATION: Transcription and translation of this gene sequence, including previous intronic sequences, generates a novel specific biomarker of human prostate cancer. The presence of catalytic domains characteristic of classic PKC-ß and atypical PKC-ι within PKC-ζ(-PrC) provides a potential mechanism for this PRKCZ variant to modulate the malignant prostatic phenotype out-with normal cell-regulatory control.
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Biomarcadores de Tumor/biosíntesis , Biomarcadores de Tumor/genética , Neoplasias de la Próstata/enzimología , Neoplasias de la Próstata/genética , Proteína Quinasa C/biosíntesis , Proteína Quinasa C/genética , Secuencia de Aminoácidos , Secuencia de Bases , Biomarcadores de Tumor/metabolismo , Dominio Catalítico , Línea Celular , Línea Celular Tumoral , Células Epiteliales/metabolismo , Variación Genética , Humanos , Masculino , Datos de Secuencia Molecular , Fenotipo , Neoplasias de la Próstata/metabolismo , Proteína Quinasa C/metabolismo , Empalme del ARN , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Transcripción Genética , Transcriptoma/genéticaRESUMEN
Plasticity in the spinal dorsal horn is thought to underlie the development of neuropathic pain. Calcineurin (protein phosphatase 3) plays an important role in plasticity in the brain. Here we examined whether chronic constriction injury (CCI) of the sciatic nerve modifies calcineurin expression in the spinal dorsal horn. Male rats were assigned to control (uninjured), sham-operated or CCI groups. CCI animals exhibited both a shift in weight bearing and a reduction in paw withdrawal latencies as signs of pain behavior. At 3 days (3D) the pain behavior was associated with a significant increase in calcineurin gene expression, enzyme activity and content of its Aα isoform in the ipsilateral spinal dorsal horn. In contrast, while the pain behavior persisted at 7 days (7D) calcineurin gene expression returned to control levels and activity and protein content decreased. A single intrathecal injection of MK-801 15 min before the ligation attenuated both signs of pain behavior in 3D but not 7D CCI animals. The same pre-treatment also prevented the CCI-associated increases in calcineurin in these animals. These data suggested an involvement of calcineurin in CCI-elicited neuropathic pain. The time-dependent divergent changes in calcineurin expression may underlie the different phases of neuropathic pain development.
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Calcineurina/biosíntesis , Neuralgia/metabolismo , Células del Asta Posterior/metabolismo , Nervio Ciático/lesiones , Animales , Constricción , Maleato de Dizocilpina/farmacología , Expresión Génica/efectos de los fármacos , Immunoblotting , Masculino , Plasticidad Neuronal/efectos de los fármacos , Plasticidad Neuronal/fisiología , Fármacos Neuroprotectores/farmacología , Células del Asta Posterior/efectos de los fármacos , ARN Mensajero/análisis , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Nervio Ciático/metabolismoRESUMEN
BACKGROUND: This study was performed to test the hypothesis that expression of small heat shock protein Hsp-27 is, at diagnosis, a reliable predictive biomarker of clinically aggressive prostate cancer. METHODS: A panel of tissue microarrays constructed from a well-characterised cohort of 553 men with conservatively managed prostate cancer was stained immunohistochemically to detect Hsp-27 protein. Hsp-27 expression was compared with a series of pathological and clinical parameters, including outcome. RESULTS: Hsp-27 staining was indicative of higher Gleason score (P<0.001). In tissue cores having a Gleason score >7, the presence of Hsp-27 retained its power to independently predict poor clinical outcome (P<0.002). Higher levels of Hsp-27 staining were almost entirely restricted to cancers lacking ERG rearrangements (chi2 trend=31.4, P<0.001), although this distribution did not have prognostic significance. INTERPRETATION: This study has confirmed that, in prostate cancers managed conservatively over a period of more than 15 years, expression of Hsp-27 is an accurate and independent predictive biomarker of aggressive disease with poor clinical outcome (P<0.001). These findings suggest that apoptotic and cell-migration pathways modulated by Hsp-27 may contain targets susceptible to the development of biologically appropriate chemotherapeutic agents that are likely to prove effective in treating aggressive prostate cancers.
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Reordenamiento Génico , Proteínas de Choque Térmico HSP27/análisis , Neoplasias de la Próstata/química , Proteínas Proto-Oncogénicas c-ets/genética , Anciano , Proteínas de Choque Térmico HSP27/fisiología , Proteínas de Choque Térmico , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Chaperonas Moleculares , Pronóstico , Neoplasias de la Próstata/patologíaRESUMEN
Prostate cancer is the most frequently diagnosed male cancer, and its clinical outcome is difficult to predict. The disease may involve the inappropriate expression of genes that normally control the proliferation of epithelial cells in the basal layer and their differentiation into luminal cells. Our aim was to identify novel basal cell markers and assess their prognostic and functional significance in prostate cancer. RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling. We identified 112 and 267 genes defining basal and luminal populations, respectively. The transcription factor TEAD1 and the ubiquitin ligase c-Cbl were identified as novel basal cell markers. Knockdown of either marker using siRNA in prostate cell lines led to decreased cell growth in PC3 and disrupted acinar formation in a 3D culture system of RWPE1. Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics. These data are consistent with basal features impacting on the development and clinical course of prostate cancers.
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Biomarcadores de Tumor/análisis , Proteínas de Unión al ADN/biosíntesis , Proteínas Nucleares/biosíntesis , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Proteínas Proto-Oncogénicas c-cbl/biosíntesis , Factores de Transcripción/biosíntesis , Adulto , Anciano , Anciano de 80 o más Años , Western Blotting , Células Epiteliales/metabolismo , Células Epiteliales/patología , Técnica del Anticuerpo Fluorescente , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Masculino , Microdisección , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Pronóstico , Neoplasias de la Próstata/mortalidad , ARN Interferente Pequeño , Factores de Transcripción de Dominio TEA , Análisis de Matrices Tisulares , TransfecciónRESUMEN
BACKGROUND: Previously, proteomic methods were applied to characterise differentially expressed proteins in microdissected pancreatic ductal adenocarcinoma cells. AIMS: To report that CapG and a related protein, gelsolin, which have established roles in cell motility, are overexpressed in metastatic pancreatic cancer; and to describe their pattern of expression in pancreatic cancer tissue and their effect on cell motility in pancreatic cancer cell lines. METHODS: CapG was identified by mass spectrometry and immunoblotting. CapG and gelsolin expression was assessed by immunohistochemical analysis on a pancreatic cancer tissue microarray and correlated with clinical and pathological parameters. CapG and gelsolin levels were reduced using RNA interface in Suit-2, Panc-1 and MiaPaCa-2 cells. Cell motility was assessed using modified Boyden chamber or wound-healing assays. RESULTS: Multiple isoforms of CapG were detected in pancreatic cancer tissue and cell lines. Immunohistochemical analysis of benign (n = 44 patients) and malignant (n = 69) pancreatic ductal cells showed significantly higher CapG staining intensity in nuclear (p<0.001) and cytoplasmic (p<0.001) compartments of malignant cells. Similarly, gelsolin immunostaining of benign (n = 24 patients) and malignant (n = 68 patients) pancreatic ductal cells showed higher expression in both compartments (both p<0.001). High nuclear CapG was associated with increased tumour size (p = 0.001). High nuclear gelsolin was associated with reduced survival (p = 0.01). Reduction of CapG or gelsolin expression in cell lines by RNAi was accompanied by significantly impaired motility. CONCLUSIONS: Up regulation of these actin-capping proteins in pancreatic cancer and their ability to modulate cell motility in vitro suggest their potentially important role in pancreatic cancer cell motility and consequently dissemination.
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Movimiento Celular/fisiología , Gelsolina/análisis , Proteínas de Microfilamentos/análisis , Proteínas de Neoplasias/análisis , Proteínas Nucleares/análisis , Neoplasias Pancreáticas/química , Adenocarcinoma/química , Adenocarcinoma/genética , Adenocarcinoma/patología , Línea Celular Tumoral , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Femenino , Humanos , Isomerismo , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Páncreas/metabolismo , Páncreas/patología , Conductos Pancreáticos/metabolismo , Conductos Pancreáticos/patología , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , Interferencia de ARN/fisiología , ARN Neoplásico/metabolismo , Regulación hacia ArribaRESUMEN
Researchers investigating climate change have used historical tide-gauge measurements from all over the world to investigate the changes in sea-level that have occurred over the last century or so. However, such estimates are a combination of any true sea-level variations and any vertical movements of the land at the specific tide-gauge. For a tide- gauge record to be used to determine the climate related component of changes in sea-level, it is therefore necessary to correct for the vertical land movement component of the observed change in sea-level.In 1990, the Institute of Engineering Surveying and Space Geodesy and Proudman Oceanographic Laboratory started developing techniques based on the Global Positioning System (GPS) for measuring vertical land movements (VLM) at tide-gauges in the UK. This paper provides brief details of these early developments and shows how they led to the establishment of continuous GPS (CGPS) stations at a number of tide-gauges. The paper then goes on to discuss the use of absolute gravity (AG), as an independent technique for measuring VLM at tide-gauges. The most recent results, from CGPS time-series dating back to 1997 and AG time-series dating back to 1995/1996, are then used to demonstrate the complementarity of these two techniques and their potential for providing site-specific estimates of VLM at tide-gauges in the UK.
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Geografía , Gravitación , Agua de Mar , Clima , Europa (Continente) , Sistemas de Información Geográfica , Formulación de Políticas , Comunicaciones por Satélite , Factores de Tiempo , Reino Unido , Movimientos del AguaRESUMEN
BACKGROUND AND AIMS: Previous chromatographic analysis of colonic mucins from monozygotic twins with inflammatory bowel disease (IBD) suggested a genetic mucin alteration in ulcerative colitis (UC). This study explores this further by assessing mucosal expression of the oncofetal carbohydrate antigen TF (galactose beta1, 3 N-acetylgalactosamine alpha-), among the same IBD twins. MATERIALS AND METHODS: Formalin fixed paraffin embedded rectal biopsies were studied from 22 monozygotic twin pairs with IBD. These included eight UC twin pairs and 14 Crohn's disease (CD) twin pairs, with six pairs concordant for disease and 16 unaffected twin siblings. Closely adjacent sections were assessed by peanut lectin histochemistry for TF expression and immunohistochemically for nuclear factor kappaB (NFkappaB) activation with investigators blinded to the diagnosis. RESULTS: Unaffected twins were almost all TF positive (15/16) compared with 5/29 histologically normal controls (p<0.0001). Unaffected UC (7/8) and CD twins (8/8) were similarly TF positive. TF positivity was confined mainly to the superficial epithelium and absent from the stem cell compartment of the lower crypts, suggesting that glycosylation changes are acquired rather than genetically determined. Activated NFkappaB was present in the surface epithelium of mucosal biopsies from 13/14 unaffected IBD twins but in only 6/22 histologically normal controls (p=0.0004). All 22 affected IBD twins were TF positive and 18 were positive for activated NFkappaB. CONCLUSIONS: Altered mucosal glycosylation in unaffected identical twins of IBD patients was confirmed in this study. This occurred in both UC and CD twins. The changes are probably acquired rather than congenital and may reflect "preinflammatory" NFkappaB activation.
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Antígenos de Carbohidratos Asociados a Tumores/metabolismo , Colon/metabolismo , Enfermedades Inflamatorias del Intestino/genética , Antígenos de Carbohidratos Asociados a Tumores/análisis , Estudios de Casos y Controles , Colitis Ulcerosa/metabolismo , Enfermedad de Crohn/metabolismo , Glicoproteínas/análisis , Glicoproteínas/metabolismo , Histocitoquímica/métodos , Humanos , Enfermedades Inflamatorias del Intestino/metabolismo , FN-kappa B/análisis , Aglutinina de Mani , Estadísticas no Paramétricas , Gemelos MonocigóticosRESUMEN
In a strategy aimed at identifying novel markers of human prostate cancer, we performed expression analysis using microarrays of clones randomly selected from a cDNA library prepared from the LNCaP prostate cancer cell line. Comparisons of expression profiles in primary human prostate cancer, adjacent normal prostate tissue, and a selection of other (nonprostate) normal human tissues, led to the identification of a set of clones that were judged as the best candidate markers of normal and/or malignant prostate tissue. DNA sequencing of the selected clones revealed that they included 10 genes that had previously been established as prostate markers: NKX3.1, KLK2, KLK3 (PSA), FOLH1 (PSMA), STEAP2, PSGR, PRAC, RDH11, Prostein and FASN. Following analysis of the expression patterns of all selected and sequenced genes through interrogation of SAGE databases, a further three genes from our clone set, HOXB13, SPON2 and NCAM2, emerged as additional candidate markers of human prostate cancer. Quantitative RT-PCR demonstrated the specificity of expression of HOXB13 in prostate tissue and revealed its ubiquitous expression in a series of 37 primary prostate cancers and 20 normal prostates. These results demonstrate the utility of this expression-microarray approach in hunting for new markers of individual human cancer types.
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Biomarcadores de Tumor/análisis , Proteínas de Homeodominio/genética , Análisis de Secuencia por Matrices de Oligonucleótidos , Neoplasias de la Próstata/genética , Línea Celular Tumoral , ADN Complementario/análisis , Proteínas de Homeodominio/biosíntesis , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Próstata/metabolismo , Neoplasias de la Próstata/metabolismoRESUMEN
Heat shock protein 27 (hsp-27) is a regulator of oestrogen receptor (ER) expression and a modulator of intracellular homeostasis. In this laboratory, Shaaban et al demonstrated the importance of ER-alpha, together with Ki67, in enhancing the progression of benign breast lesions of defined morphological types. To better understand the mechanisms by which ER-alpha promotes breast neoplasia, this study was performed to test the hypothesis that the roles of ER-alpha and hsp-27 may be defined by their quantitative expression in proliferative breast lesions of varying histological risk. The expression of hsp-27 was identified using a specific monoclonal antibody and analysed to assess the proportion of positive epithelial cells using digitised morphometric image analysis. The expression of ER-alpha was analysed by immunohistochemistry and Western blotting in a variety of benign (HUMA121) and malignant mammary cell lines, including ER-alpha(+) (MCF7, ZR-75, T47D) and ER-alpha(-) (MDA-MB 231) breast cancer cell lines. The data confirm that, during progression from normal through proliferative breast lesions to in situ cancer, there was a significant increase in both the proportion and the optical density of the epithelial cells expressing hsp-27. The mean levels of expression ranged from 7.4% of the total number of epithelial cells in normal lobules to 25.17% of epithelial cells in hyperplasias of usual type (HUT) to 61.1% of epithelial cells in ductal carcinoma in situ (P<0.001). The study has confirmed the expression of hsp-27 to be closely associated with ER-alpha(+) expression, and that its regulated expression occurs early along the mammary oncogenic pathway, supporting the initial hypothesis. It is our proposal that the differential expression of hsp-27 modulates the phenotypic behaviour of morphologically benign epithelial cells and hence may be an important determinant in initiating, or promoting, a population of human mammary cancers.
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Biomarcadores de Tumor/análisis , Enfermedades de la Mama/genética , Enfermedades de la Mama/fisiopatología , Neoplasias de la Mama/genética , Neoplasias de la Mama/fisiopatología , Transformación Celular Neoplásica , Proteínas de Choque Térmico/biosíntesis , Receptores de Estrógenos/biosíntesis , Western Blotting , Células Epiteliales/fisiología , Receptor alfa de Estrógeno , Femenino , Proteínas de Choque Térmico/análisis , Humanos , Inmunohistoquímica , Fenotipo , Receptores de Estrógenos/fisiología , Factores de RiesgoRESUMEN
Serotonin (5-hydroxytryptamine, 5-HT) is synthesized and released in the airways by pulmonary neuroendocrine cells located in the vicinity of airway smooth muscle (ASM). The aim of this study was to determine whether ASM cells contribute to the inactivation of serotonin, and investigate the role of the serotonin transporter (SERT) and monoamine oxidase (MAO) in this process. Cultured guinea pig tracheal smooth muscle cells, maintained in culture medium containing serotonin for 1-4 days, induced a decrease in 5-HT and increase in 5-HIAA in the culture medium. Changes in indole concentrations were prevented by fluvoxamine and iproniazid. Na+-sensitive [3H]-serotonin uptake into cultured ASM cells was time- and concentration-dependent (Km, 561 nM; Vmax, 1.06 pmol/mg protein/min), and inhibited by clomipramine (IC50, 13.7 nM), fluvoxamine (IC50, 0.16 microM) and fluoxetine (IC50, 0.32 microM). Western blot analysis with an anti-SERT antibody revealed a single 115 kDa immunoreactive band in ASM cell lysates. The results of this study suggest that ASM contributes to the uptake and metabolism of serotonin via SERT and MAO, respectively, and may therefore play a role in the inactivation of endogenous serotonin generated within the airway wall.
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Músculo Liso/metabolismo , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Serotonina/metabolismo , Animales , Células Cultivadas , Clomipramina/farmacología , Fluoxetina/farmacología , Fluvoxamina/farmacología , Cobayas , Músculo Liso/efectos de los fármacos , Tráquea/efectos de los fármacos , Tráquea/metabolismoRESUMEN
Multipotent cells within the epithelial compartment, together with phenotypically 'plastic' mesenchyma cells (stromal stem cells), provide a repository of protected genetic information from which the structure, stability and functionality of the prostate gland can be maintained. However, mere preservation of cells in a non-dividing state is insufficient to provide the necessary reservoir of information from which the structure and function of the prostate gland can be retained or recreated. Rather, there is a constant dynamic interaction, at the level of information exchange, between stem cells (whether epithelial or mesenchymal) and their surrounding environment (both humoral and physical). Thus, with respect to epithelial stem cells, these reside within environmental 'niches' which allow their controlled and limited proliferation while preserving genomic integrity. Similar 'mesenchymal niches' are also predicted to occur, although not yet identified, thus providing the multipotent source from which the full spectrum of stromal phenotypes might be regenerated. Recent data from studies of the haematopoietic and hepato-biliary systems indicate that the potential scope of stem cells far exceeds the immediate phenotypic complement of those tissues within which they originate, being dependent upon their precise environment as well as their genomic integrity.
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Próstata/citología , Células Madre/citología , Diferenciación Celular , División Celular , Células Epiteliales , Humanos , Masculino , Próstata/crecimiento & desarrollo , Células del Estroma/citologíaRESUMEN
Tubers of eleven cultivars of potato were baked and the flavour compounds from the flesh were isolated by headspace adsorption onto Tenax and analysed by gas chromatography-mass spectrometry (GC-MS). Lipid degradation and the Maillard reaction were the main sources of flavour compounds, accounting for 22-69% and 28-77%, respectively, of the total yields. Various sulfur compounds, methoxypyrazines and terpenes were also identified at lower levels. Relative aroma impact values (RAVs) were calculated by dividing compound yields by the odour threshold value. Compounds contributing most to aroma (RAV > 10,000 in at least one cultivar) were 2-isobutyl-3-methoxypyrazine, 2-isopropyl-3-methoxypyrazine, beta-damascenone, dimethyl trisulfide, decanal and 3-methylbutanal. The observed differences in yields and RAVs for compounds among cultivars would be expected to result in differences in perceived flavour.
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Extractos Vegetales/análisis , Solanum tuberosum/química , Gusto , Culinaria , Manipulación de Alimentos , Cromatografía de Gases y Espectrometría de Masas , Lípidos/química , Reacción de Maillard , Odorantes , Oxidación-Reducción , VolatilizaciónRESUMEN
1. The effect of bradykinin on the Na+-K+ pump of airway smooth muscle was investigated by measuring ouabain-sensitive (86)Rb(+) uptake in cultured guinea-pig tracheal smooth muscle cells. 2. Bradykinin induced a concentration-dependent increase in ouabain-sensitive (86)Rb(+) uptake, with an EC(50) of 3 nM (pD(2) = 8.50+/-0.10). Stimulation was not affected by indomethacin (1 microM) suggesting that it is not mediated by cycloxygenase products of arachidonic acid. 3. The B(1) receptor agonists Lys-des-Arg(9)-bradykinin and des-Arg(9)-bradykinin had no effect on ouabain-sensitive (86)Rb(+) uptake. In contrast, the B(1) and B(2) receptor agonist Lys-bradykinin induced a concentration-dependent increase in ouabain-sensitive (86)Rb(+) uptake with an EC(50) of 6 nM (pD(2) = 8.21 +/- 0.20). 4. The B(1) receptor antagonist des-Arg(10)-HOE 140 (1 microM) had no effect on bradykinin-stimulated ouabain-sensitive (86)Rb(+) uptake. The B(2) receptor antagonists HOE 140 and WIN 64338 antagonized bradykinin-stimulated ouabain-sensitive (86)Rb(+) uptake with pK(B) values (-log M) of 8.20 +/- 0.08 and 8.11 +/- 0.20 respectively. 5. Reducing extracellular Na+ from 146 mM to 11 mM caused a 53.5% decrease in basal ouabain-sensitive (86)Rb+ uptake and abolished bradykinin-induced uptake. Two inhibitors of the Na(+)-H(+) exchanger, methylisobutyl-amiloride (MIA; 1 - 100 microM) and ethylisopropyl-amiloride (EIPA; 0.1 - 10 microM), inhibited bradykinin-stimulated ouabain-sensitive (86)Rb(+) uptake without affecting basal uptake. 6. These results suggest that bradykinin increases Na+-K+ pump activity of guinea-pig tracheal smooth muscle via stimulation of B(2) receptors and activation of the Na+-H+ exchanger.
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Bradiquinina/farmacología , Músculo Liso/efectos de los fármacos , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Tráquea , Animales , Bradiquinina/agonistas , Bradiquinina/antagonistas & inhibidores , Antagonistas de los Receptores de Bradiquinina , Células Cultivadas , Cobayas , Masculino , Monensina/farmacología , Músculo Liso/citología , Músculo Liso/metabolismo , Ouabaína/farmacología , Receptor de Bradiquinina B2 , Receptores de Bradiquinina/agonistas , Receptores de Bradiquinina/metabolismo , Rubidio/metabolismo , Sodio/metabolismo , Intercambiadores de Sodio-Hidrógeno/antagonistas & inhibidores , Intercambiadores de Sodio-Hidrógeno/metabolismoRESUMEN
BACKGROUND: The aim of this study was to evaluate expression of the bcl-2 family of apoptosis regulating proteins in normal and diseased human pancreatic tissues. METHOD: Expression of bcl-2, bax, bcl-x, bak and p53 was determined in formalin-fixed paraffin wax-embedded archival specimens of normal pancreatic tissue (n = 7), chronic pancreatitis (n = 7), pancreatic ductal adenocarcinoma (n = 23) and ampullary cancer (n = 7) by immunohistochemistry using specific antibodies. RESULTS: In normal pancreas and chronic pancreatitis tissues, bcl-2, bax and bcl-x were predominantly expressed in ductal epithelial cells while p53 was not detected. In pancreatic ductal adenocarcinoma and ampullary cancer, bcl-2 was not detected compared with expression seen in normal acini (p < 0.01), minor (p < 0.001) and major ducts (p < 0.01), bax expression was reduced with respect to minor ducts (p < 0.01) but no different from normal acini or major ducts. bak and bcl-x were more strongly expressed in malignant epithelia compared with acini and major ducts but reduced when compared with minor ducts (p < 0.01). Overexpression of p53 was identified in 11 (48%) of 23 pancreatic adenocarcinomas and 4 (57%) of 7 ampullary cancers. Differential survival of individual patients was predicted by the relative level of bcl-x expression but not bax or bak, such that strong expression of bcl-x was associated with a median postoperative survival of 171 days when compared with 912 days for diminished expression (p < 0.001) of bcl-x. CONCLUSION: Pancreatic and ampullary cancer are associated with absent bcl-2 expression. bax, bak and bcl-x expression was reduced compared with normal minor ducts whilst bak and bcl-x expression was increased when compared with major ducts. bcl-x expression correlates with survival following resection and may represent a potential prognosis marker.
Asunto(s)
Carcinoma Ductal Pancreático/patología , Proteínas de la Membrana/análisis , Páncreas/citología , Neoplasias Pancreáticas/patología , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Adenocarcinoma/mortalidad , Adenocarcinoma/patología , Apoptosis , Carcinoma Ductal Pancreático/mortalidad , Humanos , Inmunohistoquímica , Páncreas/fisiología , Neoplasias Pancreáticas/mortalidad , Tasa de Supervivencia , Factores de Tiempo , Proteína Destructora del Antagonista Homólogo bcl-2 , Proteína bcl-XRESUMEN
The effect of 5-hydroxytryptamine (5-HT) or serotonin on Na(+)-K(+) pump activity of airway smooth muscle was investigated by measuring (86)Rb(+) uptake in cultured guinea pig tracheal smooth muscle cells. (86)Rb(+) uptake consisted of three distinct components, one sensitive to ouabain, one to bumetanide, and one insensitive to either inhibitor. 5-HT induced a concentration-dependent increase in ouabain-sensitive (86)Rb(+) uptake (EC(50) = 21 nM) but had no effect on bumetanide-sensitive uptake, suggesting that it stimulates the Na(+)-K(+) pump but not the Na(+)-K(+)-Cl(-) cotransporter. Ouabain-sensitive uptake also was stimulated by the 5-HT(2A/2C) agonists 2,5-dimethoxy-4-iodoamphetamine and alpha-methyl-5-HT, but not by the 5-HT(1) agonist 5-carboxamidotryptamine, the 5-HT(1A/1B/2C) agonist 1-(3-chlorophenyl)piperazine, or the 5-HT(3) agonist 1-(3-chlorophenyl)biguanide. 5-HT-stimulated (86)Rb(+) uptake was inhibited by the 5-HT(2A) antagonists ketanserin and spiperone, but not by the 5-HT(1A) antagonist NAN 190 or the 5-HT(3) antagonist Y25310. 5-HT-stimulated (86)Rb(+) uptake was inhibited by reducing extracellular Na(+) concentration and by the Na(+)-H(+) exchange inhibitors dimethylamiloride and 5-(N-methyl-N-isobutyl)-amiloride. These observations suggest that 5-HT stimulates the Na(+)-K(+) pump of airway smooth muscle via 5-HT(2A) receptors by a mechanism dependent on Na(+) influx, possibly through the Na(+)-H(+) exchanger. Because stimulation of the Na(+)-K(+) pump produces hyperpolarization, this may represent a negative-feedback mechanism that opposes contraction in response to 5-HT.
Asunto(s)
Músculo Liso/enzimología , Serotonina/farmacología , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Tráquea/enzimología , Animales , Células Cultivadas , Retroalimentación/efectos de los fármacos , Retroalimentación/fisiología , Cobayas , Técnicas In Vitro , Masculino , Contracción Muscular/efectos de los fármacos , Relajación Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Ouabaína/farmacología , Radioisótopos de Rubidio , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacología , Sodio/metabolismo , Intercambiadores de Sodio-Hidrógeno/efectos de los fármacos , Intercambiadores de Sodio-Hidrógeno/metabolismo , Estimulación Química , Tráquea/efectos de los fármacosRESUMEN
Heat shock proteins (hsps) occupy a central role in the regulation of intracellular homeostasis, and differential expression of individual hsps occurs in a broad range of neoplastic processes. This study was performed to test the hypothesis that the particular patterns by which individual hsps become specifically modulated in human prostate cancers are correlated with behavioral phenotype and hence may be of value in determining the most appropriate clinical management of individual patients. Monoclonal antibodies specific for each hsp protein were used to assess expression of hsp27, hsp60, and hsp70 in formalin-fixed, paraffin wax-embedded, archival tissue specimens of early prostatic adenocarcinomas (pT1-2N0M0) removed at radical prostatectomy (n = 25) and in advanced cancers (n = 95) identified at transurethral resection of prostate (TURP). These findings were compared with similar data from control prostates (n = 10) removed at primary cystectomy for urinary bladder neoplasia not involving the prostate and also at TURP for benign prostatic hyperplasia (n = 50). Western blotting of whole cell lysates derived from established human prostatic epithelial cell lines PNT2, LNCaP, DU145, and PC3 was compared with expression of hsps by the primary human tissues. This study found that early in situ neoplastic transformation of normal prostatic epithelium was consistently associated with loss of hsp27 expression and that the level of hsp27 expression by individual prostate cancers was correlated with their Gleason grade. In advanced cancers, hsp27 expression was invariably associated with poor clinical outcome (P = 0.0001). Data from cell lines supported the primary tissue findings, with elevated hsp27 expression only in aggressive malignant cell lines and androgen-insensitive cell lines. Expression of hsp60 was significantly increased in both early and advanced prostate cancer when compared with nonneoplastic prostatic epithelium (P < 0.0001), as well as in malignant prostate cancer cell lines. Expression of hsp70 was unaltered in early prostate cancers when compared with nonneoplastic prostatic epithelium but showed a diminished expression in morphologically advanced cancers (P = 0.0029). No consistent correlation was found between levels of hsp60 or hsp70 expression and phenotypic behavior of individual primary prostatic cancers. Thus, patterns of hsp expression have been confirmed to be specifically and consistently modulated in both early and advanced human prostate cancers. Whereas absence of hsp27 is a reliable objective marker of early prostatic neoplasia, reexpression of this protein by an individual invasive prostatic carcinoma invariably heralds poor clinical prognosis. Because this protein has been shown to alter the balance between proliferation and apoptosis, understanding the mechanism(s) by which individual hsps regulate intracellular homeostasis may assist in explaining some key processes that occur during evolution of human prostate cancers. We suggest that hsp27 expression provides novel diagnostic and prognostic information on individual patient survival which, if obtained at the time of primary diagnosis, would assist in determining tumor-specific management strategies. Development of techniques to therapeutically modulate hsp27 expression raises the possibility of novel targeted approaches to regulate this homeostatic mechanism, thus allowing better control over tumor cell proliferation and hence patient survival.