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Neutron-capture cross sections of neutron-rich nuclei are calculated using a Hauser-Feshbach model when direct experimental cross sections cannot be obtained. A number of codes to perform these calculations exist, and each makes different assumptions about the underlying nuclear physics. We investigated the systematic uncertainty associated with the choice of Hauser-Feshbach code used to calculate the neutron-capture cross section of a short-lived nucleus. The neutron-capture cross section for 73 Zn (n, γ ) 74 Zn was calculated using three Hauser-Feshbach statistical model codes: TALYS, CoH, and EMPIRE. The calculation was first performed without any changes to the default settings in each code. Then an experimentally obtained nuclear level density (NLD) and γ -ray strength function ( γ SF ) were included. Finally, the nuclear structure information was made consistent across the codes. The neutron-capture cross sections obtained from the three codes are in good agreement after including the experimentally obtained NLD and γ SF , accounting for differences in the underlying nuclear reaction models, and enforcing consistent approximations for unknown nuclear data. It is possible to use consistent inputs and nuclear physics to reduce the differences in the calculated neutron-capture cross section from different Hauser-Feshbach codes. However, ensuring the treatment of the input of experimental data and other nuclear physics are similar across multiple codes requires a careful investigation. For this reason, more complete documentation of the inputs and physics chosen is important. Supplementary Information: The online version contains supplementary material available at 10.1140/epja/s10050-023-00920-0.
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This corrects the article DOI: 10.1103/PhysRevLett.116.242502.
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Nuclear reactions where an exotic nucleus captures a neutron are critical for a wide variety of applications, from energy production and national security, to astrophysical processes, and nucleosynthesis. Neutron capture rates are well constrained near stable isotopes where experimental data are available; however, moving far from the valley of stability, uncertainties grow by orders of magnitude. This is due to the complete lack of experimental constraints, as the direct measurement of a neutron-capture reaction on a short-lived nucleus is extremely challenging. Here, we report on the first experimental extraction of a neutron capture reaction rate on ^{69}Ni, a nucleus that is five neutrons away from the last stable isotope of Ni. The implications of this measurement on nucleosynthesis around mass 70 are discussed, and the impact of similar future measurements on the understanding of the origin of the heavy elements in the cosmos is presented.
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The ß-decay intensity of ^{70}Co was measured for the first time using the technique of total absorption spectroscopy. The large ß-decay Q value [12.3(3) MeV] offers a rare opportunity to study ß-decay properties in a broad energy range. Two surprising features were observed in the experimental results, namely, the large fragmentation of the ß intensity at high energies, as well as the strong competition between γ rays and neutrons, up to more than 2 MeV above the neutron-separation energy. The data are compared to two theoretical calculations: the shell model and the quasiparticle random phase approximation (QRPA). Both models seem to be missing a significant strength at high excitation energies. Possible interpretations of this discrepancy are discussed. The shell model is used for a detailed nuclear structure interpretation and helps to explain the observed γ-neutron competition. The comparison to the QRPA calculations is done as a means to test a model that provides global ß-decay properties for astrophysical calculations. Our work demonstrates the importance of performing detailed comparisons to experimental results, beyond the simple half-life comparisons. A realistic and robust description of the ß-decay intensity is crucial for our understanding of nuclear structure as well as of r-process nucleosynthesis.
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Carbon based nanomaterials have emerged over the last few years as important agents for biomedical fluorescence and Raman imaging applications. These spectroscopic techniques utilize either fluorescently labelled carbon nanomaterials or the intrinsic photophysical properties of the carbon nanomaterial. In this review article we present the utilization and performance of several classes of carbon nanomaterials, namely carbon nanotubes, carbon nanohorns, carbon nanoonions, nanodiamonds and different graphene derivatives, which are currently employed for in vitro as well as in vivo imaging in biology and medicine. A variety of different approaches, imaging agents and techniques are examined and the specific properties of the various carbon based imaging agents are discussed. Some theranostic carbon nanomaterials, which combine diagnostic features (i.e. imaging) with cell specific targeting and therapeutic approaches (i.e. drug delivery or photothermal therapy), are also included in this overview.
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Carbono , Nanoestructuras , Nanotubos de Carbono , Diagnóstico por Imagen , Sistemas de Liberación de MedicamentosRESUMEN
Carbon nanomaterials are among the most broadly discussed, researched and applied of synthetic nanomaterials. The structural diversity of these materials provides an array of unique electronic, magnetic and optical properties, which when combined with their robust chemistry and ease of manipulation, makes them attractive candidates for sensor applications. Furthermore, the biocompatibility exhibited by many carbon nanomaterials has seen them used as in vivo biosensors. Carbon nanotubes, graphene and carbon dots have come under intense scrutiny, as either discrete molecular-like sensors, or as components which can be integrated into devices. In this review we consider recent developments in the use of carbon nanoparticles and nanostructures as sensors and consider how they can be used to detect a diverse range of analytes.
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Técnicas Biosensibles , Carbono , Nanoestructuras , Materiales Biocompatibles , Células Cultivadas , Humanos , NanotecnologíaRESUMEN
A novel technique has been developed, which will open exciting new opportunities for studying the very neutron-rich nuclei involved in the r process. As a proof of principle, the γ spectra from the ß decay of ^{76}Ga have been measured with the SuN detector at the National Superconducting Cyclotron Laboratory. The nuclear level density and γ-ray strength function are extracted and used as input to Hauser-Feshbach calculations. The present technique is shown to strongly constrain the ^{75}Ge(n,γ)^{76}Ge cross section and reaction rate.
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An approach is presented to experimentally constrain previously unreachable (p, γ) reaction rates on nuclei far from stability in the astrophysical rp process. Energies of all critical resonances in the (57)Cu(p,γ)(58)Zn reaction are deduced by populating states in (58)Zn with a (d, n) reaction in inverse kinematics at 75 MeV/u, and detecting γ-ray-recoil coincidences with the state-of-the-art γ-ray tracking array GRETINA and the S800 spectrograph at the National Superconducting Cyclotron Laboratory. The results reduce the uncertainty in the (57)Cu(p,γ) reaction rate by several orders of magnitude. The effective lifetime of (56)Ni, an important waiting point in the rp process in x-ray bursts, can now be determined entirely from experimentally constrained reaction rates.
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BACKGROUND: An adequately powered, double-blind, multisite, randomised controlled trial has shown no net clinical benefit for subcutaneous ketamine over placebo in the management of cancer pain refractory to combination opioid and co-analgesic therapy. The results of the trial were disseminated widely both nationally and internationally. AIM: To determine whether the trial had impacted on clinical practice in Australasia. METHODS: Members of the Australia and New Zealand Society of Palliative Medicine were sent an online ketamine utilisation survey. RESULTS: A total of 123/392 clinicians responded (31% response rate). The majority of respondents had practised for more than 10 years in a metropolitan hospital setting. Ketamine had been prescribed by 91% of respondents, and 92% were aware of the trial. As a result, 65% of respondents had changed practice (17% no longer prescribed ketamine, 46% used less and 2% more). Thirty-five per cent had not changed practice. Reasons for change included belief in the results of the study, concerns over the toxicity reported or because there were alternatives for pain control. Of those who prescribed less, over 80% were more selective and would now only use the drug in certain clinical situations or pain types, or when all other medications had failed. CONCLUSIONS: Although two-thirds of respondents reported practice change as a result of the randomised controlled trial, a minority remained convinced of the benefit of the drug from their own observations and would require additional evidence.
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Actitud del Personal de Salud , Dolor Crónico/terapia , Difusión de Innovaciones , Medicina General/métodos , Ketamina/uso terapéutico , Neoplasias/fisiopatología , Manejo del Dolor/tendencias , Cuidados Paliativos/métodos , Médicos/psicología , Pautas de la Práctica en Medicina/tendencias , Adulto , Anciano , Dolor Crónico/etiología , Ensayos Clínicos Fase III como Asunto , Utilización de Medicamentos , Femenino , Medicina General/estadística & datos numéricos , Médicos Generales/psicología , Encuestas de Atención de la Salud , Humanos , Ketamina/efectos adversos , Masculino , Persona de Mediana Edad , Manejo del Dolor/psicología , Manejo del Dolor/estadística & datos numéricos , Cuidados Paliativos/psicología , Cuidados Paliativos/tendencias , Pautas de la Práctica en Medicina/estadística & datos numéricos , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
Classical novae are expected to contribute to the 1809-keV Galactic γ-ray emission by producing its precursor 26Al, but the yield depends on the thermonuclear rate of the unmeasured 25Al(p,γ)26Si reaction. Using the ß decay of 26P to populate the key J(π)=3(+) resonance in this reaction, we report the first evidence for the observation of its exit channel via a 1741.6±0.6(stat)±0.3(syst) keV primary γ ray, where the uncertainties are statistical and systematic, respectively. By combining the measured γ-ray energy and intensity with other experimental data on 26Si, we find the center-of-mass energy and strength of the resonance to be E(r)=414.9±0.6(stat)±0.3(syst)±0.6(lit.) keV and ωγ=23±6(stat)(-10)(+11)(lit.) meV, respectively, where the last uncertainties are from adopted literature data. We use hydrodynamic nova simulations to model 26Al production showing that these measurements effectively eliminate the dominant experimental nuclear-physics uncertainty and we estimate that novae may contribute up to 30% of the Galactic 26Al.
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We have assessed whether glucose concentration and patient outcome are related in hospitalised patients when glycaemia is quantified in detail. Continuous glucose monitoring was performed on 47 consecutive subjects with an acute exacerbation of chronic obstructive pulmonary disease. Length of hospital stay increased by 10% for each mmol/L increase in mean glucose (P = 0.01). In a multivariable analysis, mean glucose was independently associated with length of hospital stay (P = 0.02). These data add weight to evidence that hyperglycaemia may adversely affect patient outcomes in hospitalised patients.
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Glucemia/metabolismo , Índice Glucémico/fisiología , Tiempo de Internación/tendencias , Enfermedad Pulmonar Obstructiva Crónica/sangre , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico , Enfermedad Aguda , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Enfermedad Pulmonar Obstructiva Crónica/terapiaRESUMEN
OBJECTIVE: To assess the efficacy of thrice daily topical 4Jointz utilizing Acteev technology (a combination of a standardized comfrey extract and a pharmaceutical grade tannic acid, 3.5 g/day) on osteoarthritic knee pain, markers of inflammation and cartilage breakdown over 12 weeks. PATIENTS AND METHODS: Adults aged 50-80 years (n = 133) with clinical knee OA were randomised to receive 4Jointz or placebo in addition to existing medications. Pain and function were measured using a visual analogue scale (VAS) and the Knee Injury and Osteoarthritis Outcome Score (KOOS) scale at baseline, 4, 8 and 12 weeks. Inflammation was measured analysing IL-6 expression and CTX-2 presence as representative for cartilage breakdown using ELISA, at baseline and 12 weeks. RESULTS: Pain scores significantly reduced in the group who received 4Jointz compared to the group who received placebo after 12 weeks using both the VAS (-9.9 mm, P = 0.034) and the KOOS pain scale (+5.7, P = 0.047). Changes in IL-6 and CTX-2 were not significant (-0.04, P = 0.5; -0.01, P = 0.68). Post-hoc analyses suggested that treatment may be most effective in women (VAS -16.8 mm, P = 0.008) and those with milder radiographic osteoarthritis (OA) (VAS -16.1 mm, P = 0.009). Rates of adverse events were similar in both groups, excepting local rash that was more common amongst participants receiving 4Jointz (21% vs 1.6%, IRR 13.2, P = 0.013), but only 26% (n = 4) of participants with rashes discontinued treatment. There were no changes in systemic blood results. CONCLUSIONS: Topical treatment using 4Jointz reduced pain but had no effect on inflammation or cartilage breakdown over 12 weeks of treatment. TRIAL REGISTRATION: Australia and New Zealand Clinical Trials registry ACTRN12610000877088.
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Analgésicos/uso terapéutico , Consuelda/química , Osteoartritis de la Rodilla/tratamiento farmacológico , Dolor/tratamiento farmacológico , Extractos Vegetales/uso terapéutico , Taninos/uso terapéutico , Administración Tópica , Anciano , Anciano de 80 o más Años , Método Doble Ciego , Combinación de Medicamentos , Femenino , Humanos , Articulación de la Rodilla/fisiopatología , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/complicaciones , Osteoartritis de la Rodilla/fisiopatología , Dolor/complicaciones , Dolor/fisiopatología , Manejo del Dolor , Dimensión del Dolor , Recuperación de la Función , Resultado del TratamientoRESUMEN
UNLABELLED: Thinness is a risk factor for fractures, but the effect of obesity on fracture risk is less clear. We found an association between measures of obesity and prevalence and number of vertebral deformities in women but not in men, in a cross-sectional study of 1,011 participants aged 50-80 years. INTRODUCTION: Low body weight is well recognised as a risk factor for fractures, but the association between overweight and fracture risk is less well described. This cross-sectional study describes the association between measures of obesity and vertebral deformities in 1,011 male and female participants in the Tasmanian Older Adult Cohort study. METHODS: Vertebral deformities (anterior wedging) of T4-L4 were determined by morphometric dual-emission X-ray absorptiometry. Body fat was assessed as weight, body mass index (BMI), waist-hip ratio (WHR), waist circumference and DXA measures of trunk fat (in percent) and total fat mass. RESULTS: The mean age of participants was 63 ± 7 years, and mean BMI was 28 ± 5. Prevalent thoracic vertebral deformities were associated with increasing weight [standardised ß (Sß) 0.29, p = 0.003], BMI (Sß 0.33, p < 0.001), trunk fat (Sß 0.20, p = 0.03), waist circumference (Sß 0.19, p = 0.03) and fat mass (Sß 0.23, p = 0.03), but not the WHR in women, and only with decreasing total fat mass in men. In addition, the number of vertebral deformities increased as weight, BMI or fat mass increased in women (all p < 0.05) but decreased with increasing total fat mass in men. Associations between fat mass and vertebral deformities were mainly linear, but there was some evidence of a threshold effect in women with a BMI ≥ 35. CONCLUSIONS: There is a deleterious association between increasing amounts of body fat in women but not in men and the prevalence and number of vertebral deformities, which may reflect loading of the thoracic spine.
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Obesidad/complicaciones , Osteoporosis/etiología , Curvaturas de la Columna Vertebral/etiología , Absorciometría de Fotón/métodos , Anciano , Antropometría/métodos , Índice de Masa Corporal , Densidad Ósea/fisiología , Métodos Epidemiológicos , Femenino , Humanos , Vértebras Lumbares/fisiopatología , Masculino , Persona de Mediana Edad , Obesidad/epidemiología , Obesidad/fisiopatología , Osteoporosis/epidemiología , Osteoporosis/fisiopatología , Factores de Riesgo , Distribución por Sexo , Curvaturas de la Columna Vertebral/epidemiología , Curvaturas de la Columna Vertebral/fisiopatología , Tasmania/epidemiología , Vértebras Torácicas/fisiopatologíaRESUMEN
OBJECTIVES: To estimate the prevalence and demographic distribution of treated epilepsy in a community-based population. MATERIALS & METHODS: We surveyed all residents in Tasmania, Australia, who were supplied at least one antiepileptic drug prescription between July 1, 2001 and June 30, 2002, recorded on the national prescription database. We adjusted for the effect of disease-related non-response bias by imputation methods. RESULTS: After three mail contacts, 54.0% (4072/7541) responded, with 1774 (43.6%) indicating treatment for epilepsy, representing 86.0% of the estimated total possible cases in Tasmania. The adjusted treated epilepsy prevalence was 4.36 per 1000 (95% CI 4.34, 4.39); lower in women (prevalence ratio 0.92 (95% CI 0.84, 1.00)); greater with increasing age (P < 0.001); similar in the three main geographic regions; and similar with socioeconomic status of postcode of residence. CONCLUSIONS: Although our estimates are likely to be affected by access to health services, overall treated epilepsy prevalence of 4.4 per 1000 is similar to previous studies. Our finding of high elderly prevalence has been reported in a few recent studies in developed countries and has important clinical and public health implications in populations with similar aging demographics.
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Epilepsia/epidemiología , Factores de Edad , Anciano , Anciano de 80 o más Años , Anticonvulsivantes/uso terapéutico , Australia/epidemiología , Servicios de Salud Comunitaria/estadística & datos numéricos , Demografía/estadística & datos numéricos , Prescripciones de Medicamentos/estadística & datos numéricos , Epilepsia/tratamiento farmacológico , Femenino , Humanos , Masculino , Persona de Mediana Edad , Vigilancia de la Población , Prevalencia , Tasmania/epidemiología , Adulto JovenRESUMEN
Missense mutations have been identified in the coding region of the extracellular calcium-sensing receptor (CASR) gene and cause human autosomal dominant hypo- and hypercalcemic disorders. The functional effects of several of these mutations have been characterized in either Xenopus laevis oocytes or in human embryonic kidney (HEK293) cells. All of the mutations that have been examined to date, however, cause single putative amino acid substitutions. In this report, we studied a mutant CASR with an Alu-repetitive element inserted at codon 876, which was identified in affected members of families with the hypercalcemic disorders, familial hypocalciuric hypercalcemia (FHH) and neonatal severe hyperparathyroidism (NSHPT), to understand how this insertion affects CASR function. After cloning of the Alu-repetitive element into the wild-type CASR cDNA, we transiently expressed the mutant receptor in HEK293 cells. Expression of mutant and wild-type receptors was assessed by Western analysis, and the effects of the mutation on extracellular calcium (Ca2+(o)) and gadolinium (Gd3+(o)) elicited increases in the cytosolic calcium concentration (Ca2+(i)) were examined in fura-2-loaded cells using dual wavelength fluorimetry. The insertion resulted in truncated receptor species that had molecular masses some 30 kD less than that of the wild-type CASR and exhibited no Ca2+(i) responses to either Ca2+(o) or Gd3+(o). A similar result was observed with a mutated CASR truncated at residue 876. However, the Alu mutant receptor had no impact on the function of the coexpressed wild-type receptor. Interestingly, the Alu mutant receptor demonstrated decreased cell surface expression relative to the wild-type receptor, whereas the CASR (A877stop) mutant exhibited increased cell surface expression. Thus, like the missense mutations that have been characterized to date in families with FHH, the Alu insertion in this family is a loss-of-function mutation that produces hypercalcemia by reducing the number of normally functional CASRs on the surface of parathyroid and kidney cells. In vitro transcription of exon 7 of the CASR containing the Alu sequence yielded the full-length mutant product and an additional shorter product that was truncated due to stalling of the polymerase at the poly(T) tract. In vitro translation of the mutant transcript yielded three truncated protein products representing termination in all three reading frames at stop codons within the Alu insertion. Thus sequences within the Alu contribute to slippage or frameshift mutagenesis during transcription and/or translation.
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Calcio/orina , Hipercalcemia/genética , Hipercalcemia/metabolismo , Hiperparatiroidismo/genética , Hiperparatiroidismo/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Animales , Secuencia de Bases , Calcio/sangre , Línea Celular , Cartilla de ADN/genética , ADN Complementario/genética , Genes Dominantes , Glicosilación , Humanos , Inmunohistoquímica , Recién Nacido , Mutación , Receptores Sensibles al Calcio , Receptores de Superficie Celular/química , Secuencias Repetitivas de Ácidos Nucleicos , Transfección , Tunicamicina/farmacologíaRESUMEN
The calcium-sensing receptor (CaR) is a G-protein-coupled receptor that plays a key role in extracellular calcium ion homeostasis. We have engineered 11 CaR mutants that have been described in the disorders familial benign hypercalcemia (FBH), neonatal severe hyperparathyroidism (NSHPT), and autosomal dominant hypocalcaemia (ADH), and studied their function by characterizing intracellular calcium [Ca2+]i transients in response to varying concentrations of extracellular calcium [Ca2+]o or gadolinium [Gd3+]o. The wild type receptor had an EC50 for calcium (EC50[Ca2+]o) (the value of [Ca2+]o producing half of the maximal increase in [Ca2+]i) of 4.0 mM (+/- 0.1 SEM). However, five missense mutations associated with FBH or NSHPT, (P55L, N178D, P221S, R227L, and V817I) had significantly higher EC50[Ca2+]os of between 5.5 and 9.3 mM (all P < 0.01). Another FBH mutation, Y218S, had an EC50[Ca2+]o of > 50 mM but had only a mildly attenuated response to gadolinium, while the FBH mutations, R680C and P747fs, were unresponsive to either calcium or gadolinium. In contrast, three mutations associated with ADH, (F128L, T151M, and E191K), showed significantly reduced EC50[Ca2+]os of between 2.2 and 2.8 mM (all P < 0.01). These findings provide insights into the functional domains of the CaR and demonstrate that mutations which enhance or reduce the responsiveness of the CaR to [Ca2+]o cause the disorders ADH, FBH, and NSHPT, respectively.
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Calcio/metabolismo , Hipercalcemia/etiología , Hiperparatiroidismo/etiología , Hipocalcemia/etiología , Riñón/metabolismo , Receptores de Superficie Celular/fisiología , Células Cultivadas , Humanos , Riñón/embriología , Mutagénesis Sitio-Dirigida , Receptores de Superficie Celular/análisis , Receptores de Superficie Celular/genéticaRESUMEN
There is a need for a validated endotracheal intubation trainer that has variable difficulty settings for the training and assessment of medical practitioners. In this study three anatomical modifications were retrofitted to a commercial manikin and then validated. These modifications included restricted movements of the mandible as well as changes to the upper incisors. A total of 130 participants comprising specialists, trainees and medical students volunteered for this study. Validity was tested using randomised between-groups comparison of the time taken to intubate the manikin on all settings. Overall, and at each setting, there was a significant difference between the times to intubation among the three levels of experience (P <0.001). Novices were more than 12 times more likely to fail than experts (odds ratio [OR] 12.4, 95% confidence intervals [CI] 3.8, 41.8, P <0.001). The median time to intubation for all three groups changed significantly between settings 1 (easiest) and 4 (most difficult), novice 18 seconds (CI 8.9, 27.1, P <0.001), intermediate 15 seconds (CI 6.5, 23.5, P=0.001), and expert 9 seconds (CI 0.4, 17.6, P=0.04). The novice group was significantly different from the expert group at all attempts (P <0.002), and from the intermediates at all attempts apart from the third (P=0.055). The time for the novice and intermediate groups improved significantly by the fourth attempt, novice 15 seconds (CI 5.4, 24.6, P=0.002) and intermediate 10 seconds (CI 1.0, 19.0, P=0.03). Other aspects of validity were also satisfied during this study. A high degree of validity was established for these modifications, which can be retrofitted to an existing manikin and then used for teaching or assessment.
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Anestesiología/educación , Intubación Intratraqueal , Humanos , Modelos Logísticos , Maniquíes , Factores de TiempoRESUMEN
The extracellular Ca(2+)-sensing receptor (CaR) is an unusual member of the diverse superfamily of seven-transmembrane domain G-protein-coupled receptors. Originally identified as the receptor providing the calciostat for extracellular ionized Ca(2+) ¿[Ca(2+)](o)¿, the CaR corrects small changes in [Ca(2+)](o) by regulating the secretion of the hormone that controls Ca(2+) fluxes between the blood and Ca(2+) stores in bone, and between blood and the urine. Now, research is beginning to reveal the structure and function of its unusually large N-terminal head. In addition to its role as a divalent and polyvalent cation sensor, recent studies indicate that the receptor also responds sensitively to changes in ionic strength and pH. Furthermore, new work indicates that the CaR is subject to allosteric activation by L-amino acids.
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Aminoácidos/metabolismo , Calcio/metabolismo , Homeostasis/fisiología , Receptores de Superficie Celular/fisiología , Animales , Humanos , Concentración de Iones de Hidrógeno , Concentración Osmolar , Enfermedades de las Paratiroides/tratamiento farmacológico , Enfermedades de las Paratiroides/metabolismo , Receptores Sensibles al Calcio , Receptores de Superficie Celular/química , Receptores de Superficie Celular/uso terapéuticoRESUMEN
A 120 kDa, G protein-coupled calcium-sensing receptor (CaR) was recently identified and cloned from bovine parathyroid and rat kidney. We report here that a similar calcium-sensing receptor is also present in rat and mouse pituitary as well as in the mouse pituitary cell line, AtT-20. Fragments (383-bp) of the extracellular domain of the calcium-sensing receptor from the AtT-20 cells and mouse pituitary were amplified by RT-PCR, sequenced, and found to be identical. By Northern blot analysis, AtT-20 cells expressed a major CaR mRNA transcript of 7.5 kb and three minor transcripts of 9.5, 4.0, and 1.5 kb. Except for the 9.5-kb species, these CaR transcripts were also found to be present in mouse kidney, where the 7.5-kb transcript was again the predominant form. The presence of the CaR protein in AtT-20 cells was documented directly by fluorescence immunocytochemistry using an antibody directed against the extracellular domain of the CaR. Exposure of AtT-20 cells to increasing extracellular calcium concentrations from 0.3 t 3 mM for 24 h resulted in a 2- to 4-fold increase in the levels of CaR mRNA, but not of the RNAs for beta-actin or POMC. The CaR appeared to be functional in AtT-20 cells, since acute increases in extracellular calcium between 2 and 5 mM induced increases in the cellular content of total inositol phosphates, cytosolic calcium, and cAMP. This report suggests that pituitary cells respond to changes in extracellular calcium via a G protein-coupled CaR.
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Calcio/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Hipófisis/metabolismo , Receptores de Superficie Celular/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Northern Blotting , Calcio/metabolismo , Bovinos , Línea Celular , Proteínas de Unión al GTP/fisiología , Humanos , Ratones , Datos de Secuencia Molecular , Hipófisis/química , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , ADN Polimerasa Dirigida por ARN , Ratas , Receptores Sensibles al Calcio , Receptores de Superficie Celular/química , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
Previous studies have yielded conflicting results concerning the effects of angiotensin-II (Ang II) on Ca2+ fluxes in adrenal zona glomerulosa (ZG) cells. The present study was designed to investigate the kinetics and dose-dependency of Ang II-mediated changes in Ca2+ influx and efflux in cultured bovine ZG cells. At a high (10 nM) Ang II concentration, cytosolic Ca2+ (Cai) shows a peak-plateau response for the first 15 min, with small Cai transients commonly observed with longer stimulations. At 50 pM Ang II, more sustained Cai changes were elicited, typically consisting of Cai oscillations. The underlying changes in Ca2+ influx and efflux were studied. The early modifications of Ca2+ influx after 2 min of agonist stimulation were biphasic, with uptake increased by 90% between 1-100 pM Ang II and inhibited by 30% at 10 nM Ang II. Furthermore, high (10 nM) Ang II doses inhibited extracellular K(+)-stimulated Ca2+ influx. After 30 min of Ang II stimulation, the later dose response of Ca2+ influx was of similar magnitude but shifted to the left, showing a maximal influx at 10 pM Ang II and a modest enhancement at 10 nM. Basal Ca2+ efflux followed a two-compartment exponential decay, reflecting rapid Ca2+ displacement from extracellular sites (k1) and active Ca2+ transport (k2). A high (10 nM) Ang II concentration induced a transient large increase (130%) in k2 during the initial phase of Ang II stimulation, which returned to basal values within 10 min. A low (50 pM) Ang II concentration induced a small sustained increase (30%) in k2. A 10-nM Ang II concentration markedly reduced the exchangeable Ca2+ pool, as Ca2+ mobilized from intracellular stores into the cytosol was rapidly extruded, while Ca2+ influx was inhibited. A more physiological (50 pM) concentration of Ang II did not significantly alter the total exchangeable Ca2+ pool due to modest stimulation of both Ca2+ efflux and influx. In summary, the initial transient Cai response to high Ang II results from a large Ca2+ mobilization combined with inhibition of Ca2+ influx, which does not allow for the refilling of Ca2+ stores. At later times, small increases in Ca2+ influx allow for the eventual recovery of exchangeable cell Ca2+ and an enhanced elevation in Cai. At low Ang II concentrations, stimulation of both Ca2+ influx and efflux are concurrent and maintained, allowing for a sustained increase in Cai with little change in exchangeable cell Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS)