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1.
Minerva Med ; 106(6): 339-54, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26605556

RESUMO

Deaths from lung cancer are greater than for any other type of malignancy. Many people present with advanced stage cancer at diagnosis and survival is limited. Low radiation dose CT (LDCT) screening appears to offer part of the solution to this. The US National Lung Screening Trial (NLST) showed a 20% reduction in cancer related mortality and a 6.7% reduction in all cause mortality in patients who had LDCT compared to chest X-ray. Lung Cancer screening is now being implemented in the US using the NLST screening criteria but many questions remain about the details of the methodology of screening and its cost effectiveness. Many of these questions are being answered by ongoing European trials that are reporting their findings. In this review we objectively analyse current research evidence and explore the issues that need to be resolved before implementation, including technical considerations, selection criteria and effective nodule management protocols. We discuss the potential barriers that will be faced when beginning a national screening programme and possible solutions to them.


Assuntos
Detecção Precoce de Câncer/métodos , Neoplasias Pulmonares/diagnóstico , Radiografia Pulmonar de Massa , Programas de Rastreamento/métodos , Tomografia por Emissão de Pósitrons , Tomografia Computadorizada por Raios X , Análise Custo-Benefício , Medicina Baseada em Evidências , Saúde Global , Humanos , Neoplasias Pulmonares/economia , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/terapia , Radiografia Pulmonar de Massa/métodos , Estadiamento de Neoplasias , Seleção de Pacientes , Guias de Prática Clínica como Assunto , Valor Preditivo dos Testes , Ensaios Clínicos Controlados Aleatórios como Assunto , Medição de Risco , Fatores de Risco , Sensibilidade e Especificidade
2.
J Mol Biol ; 380(3): 504-19, 2008 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-18565342

RESUMO

It has been shown that L-731988, a potent integrase inhibitor, targets a conformation of the integrase enzyme formed when complexed to viral DNA, with the 3'-end dinucleotide already cleaved. It has also been shown that diketo acid inhibitors bind to the strand transfer complex of integrase and are competitive with the host target DNA. However, published X-ray structures of HIV integrase do not include the DNA; thus, there is a need to develop a model representing the strand transfer complex. In this study, we have constructed an active-site model of the HIV-1 integrase complexed with viral DNA using the crystal structure of DNA-bound transposase and have identified a binding mode for inhibitors. This proposed binding mechanism for integrase inhibitors involves interaction with a specific Mg(2+) in the active site, accentuated by a hydrophobic interaction in a cavity formed by a flexible loop upon DNA binding. We further validated the integrase active-site model by selectively mutating key residues predicted to play an important role in the binding of inhibitors. Thus, we have a binding model that is applicable to a wide range of potent integrase inhibitors and is consistent with the available resistant mutation data.


Assuntos
Inibidores de Integrase de HIV/metabolismo , Integrase de HIV/química , Integrase de HIV/farmacologia , HIV-1/enzimologia , Modelos Moleculares , Acetoacetatos/química , Sítios de Ligação , Cristalografia por Raios X , DNA Viral/metabolismo , Furanos/química , Integrase de HIV/genética , HIV-1/genética , Interações Hidrofóbicas e Hidrofílicas , Magnésio/metabolismo , Modelos Químicos , Mutação , Naftiridinas/química , Conformação de Ácido Nucleico , Ligação Proteica , Conformação Proteica , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Pirróis/química , Reprodutibilidade dos Testes , Triazóis/química
3.
Science ; 309(5740): 1551-6, 2005 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-16099951

RESUMO

The month-to-month variability of tropical temperatures is larger in the troposphere than at Earth's surface. This amplification behavior is similar in a range of observations and climate model simulations and is consistent with basic theory. On multidecadal time scales, tropospheric amplification of surface warming is a robust feature of model simulations, but it occurs in only one observational data set. Other observations show weak, or even negative, amplification. These results suggest either that different physical mechanisms control amplification processes on monthly and decadal time scales, and models fail to capture such behavior; or (more plausibly) that residual errors in several observational data sets used here affect their representation of long-term trends.

4.
Nat Biotechnol ; 19(8): 756-9, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11479569

RESUMO

We report an approach for developing combinatorial fluorescence energy transfer (CFET) tags by tuning the tags' fluorescence emission signatures. The tags can all be excited at a single wavelength and analyzed by a simple optical system. We constructed eight CFET tags with unique fluorescence signatures, detected by a three-color capillary array electrophoresis (CAE) system with 488 nm excitation, using only three fluorescent dyes. A 1',2'-dideoxyribose phosphate spacer was used to separate the donor and acceptor to tune the energy transfer efficiency, generating unique fluorescence signatures. The spacer also served as an electrophoretic mobility tag to tune the mobility of CFET-labeled DNA for multiplex detection of single-nucleotide polymorphisms (SNPs). Six nucleotide variations were identified simultaneously using six CFET tags on synthetic DNA templates and on a PCR product from the retinoblastoma tumor suppressor gene.


Assuntos
Química/métodos , Eletroforese Capilar/métodos , Corantes Fluorescentes/farmacologia , Espectrometria de Fluorescência/métodos , Éxons , Homozigoto , Humanos , Modelos Químicos , Modelos Genéticos , Mutagênese , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único/genética , Proteína do Retinoblastoma/metabolismo
5.
Artigo em Inglês | MEDLINE | ID: mdl-10970710

RESUMO

Recognition of the autistic spectrum disorders is becoming more widespread amongst basic scientists, clinicians, and the general population. The term does not imply anything about pathology or aetiology, although it has proved to be a useful concept clinically. From Kanner's classical autism the concept has widened in scope to include milder and more subtle impairments. From a clinical perspective, there are many alternative diagnoses in an individual with autistic-like symptoms, and thorough investigation is necessary to exclude these.


Assuntos
Transtorno Autístico/classificação , Transtorno Autístico/diagnóstico , Síndrome de Asperger/diagnóstico , Síndrome de Asperger/epidemiologia , Transtorno Autístico/epidemiologia , Criança , Comorbidade , Diagnóstico Diferencial , Humanos , Transtorno Obsessivo-Compulsivo/diagnóstico , Transtorno Obsessivo-Compulsivo/epidemiologia , Transtorno da Personalidade Esquizoide/diagnóstico , Transtorno da Personalidade Esquizoide/epidemiologia
6.
Neuroendocrinology ; 71(4): 228-36, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10773742

RESUMO

These studies investigated the role of substance P (SP) in the regulation of the hypothalamic-pituitary-ovarian axis in cynomolgus monkeys with normal menstrual cycles. Plasma concentrations of SP were determined in blood samples taken every morning in normally menstruating cynomolgus monkeys throughout the menstrual cycle. There was a significant decreasing linear trend of SP during the follicular phase (cycle day -13 to day 0) and a significant inverse relationship between SP plasma values and plasma 17beta-estradiol (E(2)) values from day -13 to day 0 of the adjusted cycle. Correspondingly, SP area under the curve was significantly greater during the follicular phase than the luteal phase. In a second experiment, plasma concentrations of E(2), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and progesterone and length of cycles were measured after five daily intragastric administrations (10 mg/kg) of an NK(1) receptor (SP receptor) antagonist (RPR 100893; 10 mg/kg) initiated after serum E(2) concentrations had exceeded 125 pg/ml. There was a statistically significant reduction in the amplitude (41% of control) and the area under the curve (37% of control) of the preovulatory LH surge. In addition, there was a reduction of the duration of the LH surge (3 +/- 0.1 days in controls vs. 2.1 +/- 0.2 days in treated animals). The present results show for the first time that there are significant variations in plasma levels of SP, with a strong negative correlation with serum levels of E(2) during the follicular phase of the cynomolgus monkey, and that endogenous SP has a potentiating role in the interactive hypothalamo-anterior-pituitary mechanisms which lead to the preovulatory LH and FSH surges during the menstrual cycle in the monkey.


Assuntos
Hormônio Foliculoestimulante/sangue , Fase Folicular/fisiologia , Hormônio Luteinizante/sangue , Receptores da Neurocinina-1/fisiologia , Substância P/sangue , Animais , Estradiol/sangue , Feminino , Fase Folicular/efeitos dos fármacos , Indóis/farmacologia , Isoindóis , Macaca fascicularis , Antagonistas dos Receptores de Neurocinina-1 , Ovulação/fisiologia , Progesterona/sangue , Progesterona/metabolismo , Substância P/antagonistas & inibidores
7.
Science ; 290(5499): 2133-7, 2000 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-11118145

RESUMO

A comparison of observations with simulations of a coupled ocean-atmosphere general circulation model shows that both natural and anthropogenic factors have contributed significantly to 20th century temperature changes. The model successfully simulates global mean and large-scale land temperature variations, indicating that the climate response on these scales is strongly influenced by external factors. More than 80% of observed multidecadal-scale global mean temperature variations and more than 60% of 10- to 50-year land temperature variations are due to changes in external forcings. Anthropogenic global warming under a standard emissions scenario is predicted to continue at a rate similar to that observed in recent decades.

8.
Hum Reprod ; 14(8): 2162-5, 1999 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10438444

RESUMO

This study assessed the behavioural and psychological profiles of children conceived by in-vitro fertilization (IVF) who are now at school age. A total of 743 IVF children born at one institution and now of school age, over 4 years old, were surveyed with Achenbach questionnaires. Follow-up telephone interviews were conducted with non-responders. The results from the study group were compared to the questionnaire control group using one-tailed t-test with statistical significance set less than 0.05. There was an 84% overall response rate. Sixty-seven per cent returned questionnaires. An additional 17% completed a telephone interview. The study group had no statistically significant increase in the rate of behavioural or psychological problems compared with the control group. There were no statistically significant differences within the study group related to sex or to multiple gestation IVF births. This large group of school-age IVF children has normal psychological development with no identified adverse effect of their status as IVF children. Determining the role, if any, of IVF in the very small number of children with behavioural and psychological problems will require additional study.


Assuntos
Desenvolvimento Infantil , Fertilização in vitro , Psicologia da Criança , Adolescente , Criança , Pré-Escolar , Feminino , Fertilização in vitro/efeitos adversos , Humanos , Masculino , Inquéritos e Questionários
9.
Bioconjug Chem ; 10(4): 629-37, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10411461

RESUMO

We synthesized three novel organoarsenicals as prototype bifunctional reagents for spatially close thiols, N-(4-arsenosophenyl) hexahydro-2-oxo-(3aS,4S,6aR)-1H-thieno[3, 4-d]imidazole-4-pentamide (1), 2-[4-[(4-arsenosophenyl)amino]-1, 4-dioxobutyl] hydrazide, (3aS,4S,6aR)-hexahydro-2-oxo- 1H-thieno[3, 4-d] imidazole-4-pentanoic acid (2), and [4-[[12-[[5-[(3aS,4S, 6aR)-hexahydro-2-oxo-1H-thieno[3, 4-d]imidazol-4-yl]-1-oxopentyl]amino]-1-oxododecyl]amino]phe nyl]-arso nous acid (3) containing both biotin and arsenic with intervening varying length spacers extending from 2 to 15 A beyond biotin bound to streptavidin. Conceptually, the arsenical group can form a stable, covalent ring structure with appropriately spaced thiols and thereby anchor the reagent to a macromolecule, while biotin allows for the detection of the reagent-macromolecule complex via avidin binding. Because the alpha-subunits of all characterized nicotinic receptors contain an easily reducible disulfide bond between adjacent cysteine residues, the reduced alpha-subunit is an attractive site for labeling. Compounds 1-3 all simultaneously bound streptavidin and dithiols, and all three decreased the number of [125I]alpha-bungarotoxin-binding sites in reduced Torpedo nicotinic receptors (IC50s 10-300 nM). Moreover, arsenylation of the receptors prevented their reoxidation with dithio-bis(nitrobenzoic acid), was reversible with 2,3-dimercaptopropanesulfonic acid, and protected the receptor from irreversible alkylation by bromoacetylcholine. However, in no case did 1-3 allow simultaneous binding to reduced nicotinic receptors and to [125I]streptavidin, although 3 alone allowed simultaneous labeling of a spatially close dithiol located in reduced antibodies.


Assuntos
Arsenicais/química , Biotina/química , Agonistas Nicotínicos/química , Receptores Nicotínicos/efeitos dos fármacos , Compostos de Sulfidrila/química , Torpedo/metabolismo , Animais , Sítios de Ligação , Ligação Competitiva/efeitos dos fármacos , Bungarotoxinas/metabolismo , Galactosidases/química , Immunoblotting , Indicadores e Reagentes , Modelos Moleculares , Agonistas Nicotínicos/farmacologia , Oxirredução , Estreptavidina/química , Estreptavidina/farmacologia
10.
J Assist Reprod Genet ; 15(9): 547-51, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9822983

RESUMO

PURPOSE: This study was designed to examine changes in peripheral plasma substance-P and -K levels, their association with follicle-stimulating hormone and luteinizing hormone release in normal reproductive cycles in humans, and their correlation with plasma estradiol and progesterone. METHODS: Fourteen healthy, normally menstruating women underwent daily blood sampling (cycle day 4, 4-14 days) for measurement of estradiol, progesterone, luteinizing hormone, and follicle-stimulating hormone, substances-P and -K, and daily transvaginal ultrasounds assessing follicular growth and documentation of ovulation. RESULTS: Estradiol peaked on day 13, luteinizing hormone and follicle-stimulating hormone peaked on day 14, and progesterone began an exponential increase on about day 13. CONCLUSIONS: In contrast to other experimental designs using in vitro or in vivo rat or monkey tissue, peripheral levels of substances-P (P = 0.8391) and -K (P = 0.3205) reflected no modulation related to midcycle gonadotropin release in cycling woman.


Assuntos
Gonadotropinas/fisiologia , Ciclo Menstrual/fisiologia , Neurocinina A/fisiologia , Substância P/fisiologia , Adulto , Estradiol/sangue , Estradiol/fisiologia , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Foliculoestimulante/fisiologia , Humanos , Técnicas Imunoenzimáticas , Hormônio Luteinizante/sangue , Hormônio Luteinizante/fisiologia , Ciclo Menstrual/sangue , Microesferas , Neurocinina A/sangue , Progesterona/sangue , Progesterona/fisiologia , Radioimunoensaio , Substância P/sangue
11.
Brain Res ; 806(2): 127-40, 1998 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-9739125

RESUMO

The nucleus accumbens (NAcc) can be subdivided into 'core' and 'shell' based on anatomical connections and histochemical markers. Previous studies have demonstrated dopamine-beta-hydroxylase immunoreactive (DBH-ir) fibers in the NAcc shell, but the source of these noradrenergic (NE) afferents has not been determined. Therefore, we have investigated in detail the anatomy of NE afferents to this subregion. Dual immunohistochemistry for DBH and substance P demonstrated numerous DBH-ir fibers in the caudal NAcc shell. Neurons projecting to the NAcc were identified with Fluoro-Gold (FG) or cholera toxin B (CTb) retrograde tracing and tyrosine hydroxylase (TH) immunohistochemistry. Single- and double-labeled neurons were observed in the A2 and A1 NE cell groups following FG injections into the caudal NAcc shell. Numerous FG and CTb single-labeled neurons were found in the rostral locus coeruleus (LC), subcoeruleus and pericoerulear dendritic region, with an occasional double-labeled neuron in the LC. Few labeled neurons were seen in the brainstem after FG injections into the NAcc core, consistent with the lack of DBH-ir in this subterritory. To confirm these results, injections of Phaseolus vulgaris leucoagglutinin or biotinylated dextran amine were made into the LC or nucleus tractus solitarius (NTS). Virtually no labeled fibers were observed in the NAcc following injections into central LC. However, fibers were observed in the NAcc shell after injections in the NTS. These results indicate that the primary source(s) of NE afferents to the NAcc shell is the A2 region of the NTS, with lesser contributions from A1 and LC.


Assuntos
Mapeamento Encefálico , Neurônios Aferentes/fisiologia , Norepinefrina/fisiologia , Núcleo Accumbens/fisiologia , Estilbamidinas , Animais , Tronco Encefálico/citologia , Tronco Encefálico/metabolismo , Tronco Encefálico/fisiologia , Toxina da Cólera , Dopamina beta-Hidroxilase/metabolismo , Corantes Fluorescentes , Locus Cerúleo/fisiologia , Masculino , Fibras Nervosas/fisiologia , Ratos , Ratos Sprague-Dawley , Núcleo Solitário/fisiologia
12.
Res Commun Mol Pathol Pharmacol ; 102(1): 57-68, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9920346

RESUMO

Reduction in whole body cytochrome P450 (CYP 450) activity is evident in humans who develop trauma and sepsis-induced multiple organ failure (MOF). It is not known whether this has any deleterious or protective effect. Intraperitoneal injection of zymosan, the cell wall of Saccharomycoses A, induces dose-dependent inflammation with concomitant MOF in rats. High dose intraperitoneal zymosan (100 mg/100 g body weight) causes mortality and organomegaly in rats; low dose zymosan (20 mg/100 g body weight) does not. To study a role for CYP 450 in zymosan-induced toxicity, we examined the effect of the non-specific CYP 450 suicide inhibitor 1-aminobenzotriazole (1-ABT)(80 mg/kg/d), on rats treated with low dose zymosan. The 90% reduction in CYP 450 content achieved by this dose of 1-ABT was associated with 58% mortality in rats treated with low dose zymosan, in contrast to no mortality in rats treated with low dose zymosan alone (p < 0.01). In survivors, liver and lung organomegaly (p < 0.01), and polymorphonuclear leukocyte accumulation in the liver (p < 0.01) were increased after zymosan administration in rats treated with 1-ABT compared to those without 1-ABT. There was no effect of treatment with 1-ABT on the increased urinary excretion of nitric oxide byproducts observed after zymosan administration. These observations are consistent with the hypothesis that the CYP 450 enzyme system is an endogenous protectant in this experimental model of inflammation-induced MOF.


Assuntos
Inibidores das Enzimas do Citocromo P-450 , Inibidores Enzimáticos/farmacologia , Triazóis/farmacologia , Zimosan/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Sinergismo Farmacológico , Inflamação/induzido quimicamente , Inflamação/enzimologia , Contagem de Leucócitos , Masculino , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/enzimologia , Neutrófilos/efeitos dos fármacos , Nitratos/urina , Nitritos/urina , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Análise de Sobrevida
13.
J Neurosci Res ; 50(1): 94-103, 1997 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-9379497

RESUMO

Utilizing a human NK1 receptor antagonist (RPR 100893), the present in vivo study was designed to test the hypothesis that endogenous substance P (SP) modulates the action of 17beta-estradiol in inducing luteinizing hormone (LH) and follicle stimulating hormone (FSH) surges in ovariectomized cynomolgus monkey. Plasma concentrations of LH and FSH as well as NK1 receptor antagonist and SP were measured during the development of the negative and positive feedback phases which follow a single administration of estradiol benzoate (50 microg/kg) to long-term ovariectomized monkeys. Daily administration by gastric intubation of 1 mg/kg or 10 mg/kg of the NK1 receptor antagonist (RPR 100893) leads to detectable levels of the antagonist in the blood of treated animals for at least 6 hr after its administration. These levels are in agreement with the experimentally determined IC50 value of the antagonist. The most striking finding of this study is that LH and FSH releases are enhanced during the descending arm of the estradiol benzoate-induced LH and FSH surges, which suggests that endogenous SP normally has an inhibitory role during this time. The enhancement of LH release is approximately 50%, regardless of the amount of the NK1 antagonist used. However, the enhanced FSH release is more important. Furthermore, blockade of the NK1 receptor with the smaller dose of the antagonist leads to a small, but significant, increase in plasma levels of SP, indicating that blockade of SP receptors leads to an increased release of SP. Collectively, these results further substantiate the link which exists between the ovarian steroid 17beta-estradiol and SP systems. Also, for the first time, these results demonstrate an inhibitory involvement of the human NK1 receptor in the 17beta-estradiol-induced pseudo-ovulatory gonadotropin surges in the ovariectomized monkey.


Assuntos
Hormônio Foliculoestimulante/sangue , Indóis/farmacologia , Hormônio Luteinizante/sangue , Antagonistas dos Receptores de Neurocinina-1 , Substância P/fisiologia , Animais , Estradiol/sangue , Estradiol/farmacologia , Feminino , Humanos , Indóis/sangue , Isoindóis , Macaca fascicularis , Ovariectomia , Radioimunoensaio , Receptores da Neurocinina-1/sangue , Substância P/antagonistas & inibidores , Substância P/sangue
14.
Hum Reprod ; 12(2): 231-5, 1997 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9070701

RESUMO

Changes in plasma concentrations of ACTH, beta-endorphin (beta-EP) and cortisol have been found to be associated during the human menstrual cycle. Changes in hypothalamic levels of gonadotrophin releasing hormone (GnRH), beta-EP and substance P (SP) have also been associated with the oestrous cycle in the rat. Therefore, an attempt was made to measure the activity of the corticotrophic axis and SP by measuring blood and follicular fluid concentrations of ACTH, beta-EP, SP and corticotrophin releasing hormone (CRH) during the hormonal ovarian stimulation phase for in-vitro fertilization (IVF), in a series of 19 patients. At the plasma level, there was no significant change over treatment days in ACTH (P = 0.1550), beta-EP (P = 0.1137), or SP concentrations (P = 0.5625). CRH was not detectable over treatment days. In addition, there was no significant change in neuropeptide over treatment days between those women who became pregnant and those who did not (P = 0.17 for all). In the follicular fluid, ACTH was not detectable, beta-EP concentration was three times higher than in the plasma, CRH was detectable, and SP concentration was similar to that of plasma. There was no apparent correlation, however, between beta-EP or SP concentrations in the plasma and follicular fluid from a given patient. In conclusion, the absence of changes in the activity of the corticotrophic axis during the hormonal ovarian stimulation suggests that there was no major stress component associated with the stimulation phase of IVF or the occurrence of a pregnancy.


Assuntos
Hormônio Adrenocorticotrópico/sangue , Hormônio Liberador da Corticotropina/sangue , Fertilização in vitro , Ciclo Menstrual/sangue , Substância P/sangue , beta-Endorfina/sangue , Adulto , Animais , Gonadotropina Coriônica/administração & dosagem , Feminino , Humanos , Ciclo Menstrual/efeitos dos fármacos , Gravidez , Ratos
15.
J Pharmacol Exp Ther ; 276(1): 169-77, 1996 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8558427

RESUMO

Nereistoxin (100 microM, 2-10 min) blocks nicotinic receptors in the intact chick ciliary ganglion. This effect mimics blockade by the reducing agent dithiothreitol (2 mM, 20 min), which is not reversed until oxidation with dithiobisnitrobenzoic acid (1 mM, 5 min). After treating intact ganglia with either nereistoxin or dithiothreitol, the affinity alkylating agent bromoacetylcholine causes irreversible blockade that cannot be reversed by dithiobisnitrobenzoic acid. These data suggest that nereistoxin, or a metabolite, acts to reduce nicotinic receptors, although nereistoxin differs from dithiothreitol in that agonists only partially protect against nereistoxin reduction. In studies on chick retina, we previously proposed that a metabolite of nereistoxin (such as dihydronereistoxin) is the actual reducing agent for neuronal nicotinic receptors. Current findings in chick ciliary ganglion supporting this hypothesis include: 1) changing pH alters the minimal nereistoxin concentration needed for blockade in intact ganglia, but has little effect on the minimal concentration needed for dithiothreitol, 2) application of a quaternary analog of nereistoxin has little effect on intact ganglion, but a quaternary analog of dihydronereistoxin blocks nicotinic receptors by reduction, 3) nereistoxin weakly oxidizes rather than reduces immunoprecipitated receptors from chick brain and 4) in whole-cell patch-clamp studies, nereistoxin clearly does not reduce receptors on chick ciliary neurons, although dihydronereistoxin mimics receptor blockade by dithiothreitol, and requires oxidation by dithiobisnitrobenzoic acid for reactivation. Together, these data suggest that nereistoxin is not a direct reducing agent for neuronal nicotinic receptors.


Assuntos
Gânglios Parassimpáticos/efeitos dos fármacos , Gânglios Parassimpáticos/ultraestrutura , Toxinas Marinhas/farmacologia , Neurônios/efeitos dos fármacos , Neurônios/ultraestrutura , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/metabolismo , Acetilcolina/análogos & derivados , Acetilcolina/farmacologia , Animais , Células Cultivadas , Galinhas , Gânglios Parassimpáticos/metabolismo , Concentração de Íons de Hidrogênio , Toxinas Marinhas/metabolismo , Neurônios/metabolismo , Oxirredução , Testes de Precipitina , Compostos de Amônio Quaternário/farmacologia , Compostos de Sulfidrila/farmacologia , Tiofenos/farmacologia
16.
J Hypertens ; 14(1): 19-29, 1996 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12013490

RESUMO

OBJECTIVE: To describe several emerging concepts regarding the biosynthesis, metabolism and biological roles of adenosine and to illustrate the possible significance of these ideas to vascular biology by proposing a hypothesis pertaining to the role of erythrocyte-derived adenosine deaminase in vaso-occlusive diseases associated with damaged erythrocytes. MULTIPLE PATHWAYS FOR ADENOSINE FORMATION: Three pathways of adenosine biosynthesis have been well established: the intracellular ATP pathway, the extracellular ATP pathway and the transmethylation pathway. A fourth pathway that has received relatively little attention, but could be particularly important in vascular smooth muscle, is the cyclic AMP-adenosine pathway. This pathway involves the extracellular or intracellular metabolism, or both, of cyclic AMP to AMP and hence to adenosine. Our recent experiments in cultured vascular smooth muscle cells, isolated vascular beds and intact animals support the existence of the cyclic AMP-adenosine pathway. Together these four pathways of adenosine formation should assure pharmacologically active levels of vascular adenosine. ANTIVASO-OCCLUSIVE ACTIONS OF ADENOSINE: The overall pharmacologic profile of adenosine suggests that this nucleoside functions to protect vascular beds from vaso-occlusive events. In this regard, some well known effects of adenosine include vasodilation, release of nitric oxide from vascular endothelial cells and inhibition of platelet aggregation, platelet adhesion, neutrophil-endothelial interactions, renin release and noradrenergic neurotransmission. Moreover, studies indicate that adenosine also releases nitric oxide from cultured vascular smooth muscle cells and inhibits vascular smooth muscle cell growth. Thus, any condition that reduces the levels of adenosine in the blood vessel wall or blood vessel-blood interface might predispose toward vaso-occlusive events. DAMAGE TO ERYTHROCYTES REDUCES ADENOSINE LEVELS: Adenosine deaminase rapidly metabolizes adenosine to inosine, which lacks antivaso-occlusive properties. Because erythrocytes are generously endowed with adenosine deaminase, any condition that damages erythrocytes will cause leakage of adenosine deaminase from erythrocytes directly onto the blood vessel wall, thus diminishing local vascular levels of adenosine. Experiments using dimethyl sulfoxide-induced hemolysis have confirmed the hypothesis that erythrocyte-derived adenosine deaminase can reduce adenosine levels in vivo. HYPOTHESIS: THE ROLE OF ERYTHROCYTE-DERIVED ADENOSINE DEAMINASE IN VASO-OCCLUSIVE DISEASES: Because multiple biosynthetic pathways maintain pharmacologically active levels of adenosine within the blood vessel wall and adenosine exerts a number of antivaso-occlusive effects, release of adenosine deaminase from erythrocytes may increase the risk for vaso-occlusive events.


Assuntos
Adenosina Desaminase/fisiologia , Arteriopatias Oclusivas/fisiopatologia , Doenças Vasculares/fisiopatologia , Adenosina Desaminase/farmacologia , Animais , Arteriopatias Oclusivas/metabolismo , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Humanos , Músculo Liso Vascular/citologia , Músculo Liso Vascular/metabolismo
17.
J Assist Reprod Genet ; 12(9): 581-4, 1995 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-8580653

RESUMO

PURPOSE: To assess the impact of assisted hatching on in vitro fertilization (IVF) outcome in women age 40 and older. METHODS: A retrospective analysis was performed to compare 28 cycles of IVF without assisted hatching to 38 cycles of IVF with assisted hatching. All patients in both groups were age 40 or older and the mean age was similar. RESULTS: The delivery rate per oocyte retrieval was significantly higher in the assisted hatching group (18/38; 48%) compared to the nonhatched controls (3/28; 11%, P = 0.0003). The implantation rate of hatched embryos (40/175; 22%) was clearly enhanced, compared to the nonhatched embryos (7/126; 6%, P < 0.001). The fertilization rate, number of oocytes and the number of embryos per patient were comparable in the two groups. CONCLUSIONS: Assisted hatching dramatically improves embryonic implantation and term pregnancy rates in women age 40 and older undergoing IVF.


Assuntos
Envelhecimento/fisiologia , Fertilidade/fisiologia , Fertilização in vitro/normas , Adulto , Desenvolvimento Embrionário e Fetal/fisiologia , Feminino , Fertilização in vitro/métodos , Humanos , Pessoa de Meia-Idade , Oócitos/fisiologia , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Estudos Retrospectivos
18.
J Neurosci Res ; 42(2): 228-35, 1995 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-8568923

RESUMO

The present work describes time-dependent changes in the content of corticotropin-releasing hormone (CRH), adrenocorticotropin (ACTH), and beta-endorphin (beta-EP) in the hypothalamus (HT) and anterior pituitary (AP) and in the concentration of ACTH and beta-EP in the plasma during the 17 beta estradiol (E2) benzoate (E2B)-induced luteinizing hormone (LH) surge in ovariectomized cynomolgus monkeys. Monkeys were euthanized at 0, 30, 48, 72, and 96 hr post-E2B. HT and AP were rapidly dissected, extracted in 2 N acetic acid containing 1 mM phenylmethane sulfonyl fluoride at 4 degrees C, and centrifuged at 18,000g for 30 min. Peptide concentrations were measured in the supernatant by specific radioimmunoassays (RIAs). In the HT, there were significant (P < 0.05) decreases in ACTH and beta-EP content by 30 hr post-E2B and a significant (P < 0.05) decrease in HT CRH content 48 hr post-E2B. Thereafter, CRH, ACTH, and beta-EP content increased up to 72 hr post-E2B. In the AP, there was an almost linear decrease in the CRH content through 48 hr post-E2B followed by a marked 20-fold (P < 0.01) increase in the AP CRH content at 72 hr post-E2B, which corresponds to the time of the descending arm of the LH surge. The patterns of ACTH and beta-EP content were very similar in the AP, while that of CRH differed markedly. In contrast, in the HT CRH, ACTH, and beta-EP profiles were very similar. Significant (P < 0.05) increases in circulating levels of ACTH, beta-EP, and cortisol were evident at 30 hr (all 3 hormones), 48 hr (beta-EP and cortisol), and 72 hr (cortisol) post-E2B, which corresponds with the time of decreased hypothalamic content of CRH, ACTH, and beta-EP. These results suggest that there maybe a marked activation of the hypothalamo-anterior pituitary-adrenal axis during the negative and positive feedback phases of the E2B-induced LH surge in the ovariectomized monkey.


Assuntos
Hormônio Adrenocorticotrópico/biossíntese , Hormônio Liberador da Corticotropina/biossíntese , Estradiol/farmacologia , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Hormônio Luteinizante/metabolismo , beta-Endorfina/biossíntese , Hormônio Adrenocorticotrópico/genética , Animais , Hormônio Liberador da Corticotropina/genética , Estro , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Hidrocortisona/biossíntese , Sistema Hipotálamo-Hipofisário/metabolismo , Macaca fascicularis , Ovariectomia , beta-Endorfina/genética
19.
Fertil Steril ; 62(3): 551-4, 1994 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8062951

RESUMO

OBJECTIVE: To access the effect of augmenting IVF with assisted hatching in the treatment of poor-prognosis patients. DESIGN: Thirty-three poor-prognosis IVF patients were treated with assisted hatching and were compared with 43 control subjects without assisted hatching. SETTING: Center for Reproductive Medicine, Swedish Medical Center, Englewood, Colorado. PARTICIPANTS: Seventy-six women undergoing IVF with a poor prognosis for pregnancy. Poor prognosis was defined as Elevated day 3 FSH level; age > or = 39 years; and multiple prior IVF failures. MAIN OUTCOME MEASURES: Pregnancy and implantation rates per embryo. RESULTS: The incidence of ongoing pregnancy in the assisted hatching group was 64% compared with 19% in the control group. Implantation rate per embryo transferred was 33% in the assisted hatching group versus 6.5% in the control group. CONCLUSIONS: These results demonstrate that assisted hatching, when applied to poor-prognosis patients, improves embryonic implantation and pregnancy rates.


Assuntos
Embrião de Mamíferos , Fertilização in vitro , Micromanipulação , Técnicas Reprodutivas , Adulto , Implantação do Embrião , Feminino , Humanos , Gravidez , Prognóstico
20.
J Biomed Mater Res ; 28(2): 233-40, 1994 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8207036

RESUMO

An In vitro comparison of the corrosion response of 316LVM stainless steel and MP35N (a CoNiCrMo alloy) electrodes under conditions appropriate to applications in functional electrical stimulation (FES) was made. Electrodes of both alloys were subjected to a cathodic 40 microC/cm2 charge injection protocol and the potential transient response was recorded over a 96 h period. The transient responses were compared with potentiodynamic polarization data used to establish the quasiequilibrium response of the alloys in the carbonate and phosphate-buffered saline electrolyte used in the study. The MP35N electrodes exhibited extensive pitting corrosion during charge injection, whereas little corrosion was observed on 316LVM electrodes. An explanation for the susceptibility of MP35N to corrosion during charge injection is found in the potentiodynamic polarization data, which reveal a breakdown potential (critical pitting potential) of 0.45 V (SCE) for MP35N compared with 1.05 V (SCE) for 316LVM. Factors that may influence corrosion response during charge injection from alloys exhibiting active-passive behavior are discussed.


Assuntos
Ligas de Cromo , Eletrodos Implantados/normas , Teste de Materiais , Aço Inoxidável , Corrosão , Estimulação Elétrica , Microscopia Eletrônica de Varredura
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