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1.
Pers Soc Psychol Bull ; : 1461672221133693, 2022 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-36350190

RESUMO

Issues in applied survey research, including minimizing respondent burden and ensuring measures' brevity for smartphone administration, have intensified efforts to create short measures. We conducted two studies on the psychometric properties of single-item satisfaction, love, conflict, and commitment measures. Study 1 was longitudinal, surveying college-age dating couples at three monthly waves (n =121, 84, and 68 couples at the respective waves). Partners completed single- and multi-item measures of the four constructs, along with other variables, to examine test-retest reliability and convergent, concurrent, and predictive validity. Single-item measures of satisfaction, love, and commitment exhibited impressive psychometric qualities, but our single-item conflict measure performed somewhat less strongly. Study 2, a cross-sectional online survey (n = 280), showed strong convergent validity of the single-item measures, including that of conflict.

2.
Nanoscale ; 8(9): 4984-90, 2016 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-26863903

RESUMO

Metal nanostructures have attractive electrical and thermal properties as well as structural stability, and are important for applications in flexible conductors. In this study, we have developed a method to fabricate and control novel complex platinum nanostructures with accordion-like profile using atomic layer deposition on lithographically patterned polymer templates. The template removal process results in unique structural transformation of the nanostructure profile, which has been studied and modeled. Using different template duty cycles and aspect ratios, we have demonstrated a wide variety of cross-sectional profiles from wavy geometry to pipe array patterns. These complex thin metal nanostructures can find applications in flexible/stretchable electronics, photonics and nanofluidics.

3.
J Radioanal Nucl Chem ; 304(3): 1293-1301, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26224989

RESUMO

A method for producing synthetic debris similar to the melt glass produced by nuclear surface testing is demonstrated. Melt glass from the first nuclear weapon test (commonly referred to as trinitite) is used as the benchmark for this study. These surrogates can be used to simulate a variety of scenarios and will serve as a tool for developing and validating forensic analysis methods.

4.
J Anim Physiol Anim Nutr (Berl) ; 98(5): 908-13, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24372960

RESUMO

The purpose of this study was to determine the diurnal composition and concentration of volatile fatty acids (VFA) and to determine VFA composition and concentration differences between stomach compartment 1 (C1) and caecum of alpacas fed grass and alfalfa hay. The study was divided into two experiments. In Experiment 1 (EXP 1), 10 male alpacas (3+ years old, 65 kg BW) were divided into two groups, housed in drylot pens, provided ad libitum water and fed alfalfa (AH) or grass hay (GH) for 30 days. The alpacas were slaughtered and the digestive tract collected, divided into sub-tract sections, weighed and digesta sampled for pH, dry matter (DM) and NDF. Volatile fatty acid composition and concentration were determined on C1 and caecal material. Four adult male (3+ years old, 60 kg BW), C1 fistulated alpacas were housed in metabolism crates and divided into two forage groups for Experiment 2 (EXP 2). Alpacas were fed the forages as in EXP 1. Diurnal C1 VFA samples were drawn at 1, 3, 6, 9, 12, 18 and 24 h post-feeding. There were no differences between forages for tract weight, C1 and caecum digesta DM or NDF. Differences were noted (p < 0.05) for pH between forages and sub-tract site. Volatile fatty acids concentrations were different (p < 0.05) for forage and site, and total VFA was higher for AH than GH (110.6 and 79.1 mm) and C1 than caecum (40.7 and 27.6 mm). Proportion of VFA was significant (p < 0.05) for forage and site, C1 acetate highest for GH (84.8 vs. 74.0 mm) and caecum acetate 83.7 and 76.2 mm for GH and AH respectively. These data demonstrate the level of VFA produced in C1 and the caecum of alpacas and the diurnal VFA patterns. Composition of VFA is similar to other ruminant species.


Assuntos
Ração Animal/análise , Camelídeos Americanos/fisiologia , Dieta/veterinária , Ácidos Graxos Voláteis/química , Medicago sativa/química , Poaceae/química , Fenômenos Fisiológicos da Nutrição Animal , Animais , Masculino
5.
Animal ; 7(3): 439-45, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23031388

RESUMO

The effect of feed restriction on gene expression of regulatory enzymes of intermediary metabolism was studied in two sheep breeds (Australian Merino and Dorper) subjected to two nutritional treatments: feed restriction (85% of daily maintenance requirements) and control (ad libitum feeding), during 42 days. The experimental animals (ram lambs) were divided into four groups, n = 5 (Australian Merino control (MC), Australian Merino Restriction (MR), Dorper control (DC) and Dorper Restriction (DR)). After the trial, animals were sacrificed and samples were taken from liver tissue to quantify glucose levels and gene expression of relevant intermediary metabolism enzymes (phosphofructokinase (PFK), pyruvate kinase (PK), phosphoenolpyruvate carboxykinase, fructose 1,6-bisphosphatase, glucose-6-phosphatase, glycogen synthase (GS), fatty acid synthase (FAS), glutamate dehydrogenase (GDH) and carbamoyl phosphate synthase (CPS)) through real-time PCR. During the experimental period, the MR animals lost 12.6% in BW compared with 5.3% lost by the Dorper lambs. MC and DC rams gained, respectively, 8.8% and 14% during the same period. Within the Dorper breed, restricted feed animals revealed a significant decrease over controls in the transcription of PFK (1.95-fold) and PK (2.26-fold), both glycolytic enzymes. The gluconeogenesis showed no change in the feed restricted animals of both breeds. DR feed group presented a significant decrease over the homologous Merino sheep group on GS. In both experimental breeds, FAS mRNA expression was decreased in restricted feed groups. GDH expression was decreased only in the DR animals (1.84-fold) indicating a reduced catabolism of amino acids in these animals. Finally, CPS was significantly (P < 0.05) higher in the Dorper sheep, indicating a facilitated urea synthesis in this breed. These results indicate a better adaptation of metabolic intermediate regulatory enzymes and hepatic glucose production of Dorper sheep to feed restriction concurring with the BW results in the experimental groups.


Assuntos
Restrição Calórica , Metabolismo Energético/fisiologia , Enzimas/metabolismo , Regulação Enzimológica da Expressão Gênica/fisiologia , Ovinos/metabolismo , Análise de Variância , Animais , Peso Corporal , Glucose/metabolismo , Fígado/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Ovinos/genética , Especificidade da Espécie , Austrália Ocidental
6.
Oncogene ; 20(44): 6300-8, 2001 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-11607832

RESUMO

Endocytosis is a regulated physiological process by which cell surface proteins are internalized along with extracellular factors such as nutrients, pathogens, peptides, toxins, etc. The process begins with the invagination of small regions of the plasma membrane which ultimately form intracellullar vesicles. These internalized vesicles may shuttle back to the plasma membrane to recycle the membrane components or they may be targeted for degradation. One role for endocytosis is in the attenuation of receptor signaling. For example, desensitization of activated membrane bound receptors such as G-protein coupled receptors (GPCRs) or receptor tyrosine kinases (RTKs) occurs, in part, through endocytosis of the activated receptor. However, accumulating evidence suggests that endocytosis also mediates intracellular signaling. In this review, we discuss the experimental data that implicate endocytosis as a critical component in cellular signal transduction, both in the initiation of a signal as well as in the termination of a signal. Furthermore, we focus our attention on a recently described adaptor protein, intersectin (ITSN), which provides a link to both the endocytic and the mitogenic machinery of a cell. Thus, ITSN functions at a crossroad in the biochemical regulation of cell function.


Assuntos
Proteínas Adaptadoras de Transporte Vesicular , Endocitose , Mitógenos/metabolismo , Transdução de Sinais , Animais , Proteínas de Transporte/metabolismo , Diferenciação Celular , Humanos , Modelos Biológicos , Neoplasias/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína
7.
Sci Total Environ ; 266(1-3): 159-67, 2001 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-11258813

RESUMO

The overestimation of orthophosphate by filterable reactive phosphorus (FRP) measurement techniques has long been accepted. The aim of this study was to quantify that overestimation in 17 wetlands over time. Specifically an ultrafiltration technique was used prior to the application of the molybdenum blue phosphorus detection method to quantify orthophosphate concentrations. Samples were collected over a 6-month period and analysed for total filterable (< 0.50 microm) phosphorus (TFP), filterable reactive phosphorus (FRP < 0.50 microm) and ultrafiltered (< 10(3) Da) reactive phosphorus (PO4). FRP correlated well with PO4, however, FRP overestimated PO4 particularly with increasing colloidal phosphorus concentration. The ratio of DOC to TFP (C:P) influenced the fate of PO4 in the water, implying that DOC was forming complexes with phosphorus. The PO4 concentrations decreased with increasing C:P ratios in some of the wetlands over the 6-month monitoring period.


Assuntos
Monitoramento Ambiental/métodos , Fosfatos/análise , Poluentes Químicos da Água/análise , Coloides/química , Monitoramento Ambiental/instrumentação , Eutrofização , Filtração , Indicadores e Reagentes/química , Molibdênio
8.
Sci Total Environ ; 266(1-3): 299-307, 2001 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-11258830

RESUMO

The toxicity of heavy metals to biota in urban catchments has been regarded as a very important non-point source pollution issue. Numerous studies on heavy metal pollution in urban receiving waters have found that metal transport by surface runoff is closely correlated to the partitioning of the metal forms between dissolved and particulate phases, where sediment plays an important role in the transport process. Sediment cycling on urban streets, metal binding form, and rainfall character in the catchment area are considered to be the key factors for metal transport. A preliminary model is developed based on these considerations. Starting from classical build-up and wash-off processes for the suspended sediment (SS) on the urban impervious surface, the model links the transport of suspended sediment to the transport of metal species. Monitoring data from a small highway catchment were used in the model development. A total of 47 rain events over 1 year were monitored intensively at short time intervals (5-10 min) for hydrological data, rainfall intensity, and stormwater quality. In developing the model, lead was used for the metal load prediction, as it has been a common fuel additive for urban transportation. Agreement between model results and monitoring data indicates that the model can be used in predicting metal load from impervious urban areas, such as streets and roadways, on a long-term basis.


Assuntos
Metais Pesados/análise , Modelos Teóricos , Chuva , Poluentes Químicos da Água/análise , Animais , Monitoramento Ambiental , Previsões , Humanos , Metais Pesados/efeitos adversos , Veículos Automotores , Meios de Transporte , População Urbana , Poluentes Químicos da Água/efeitos adversos
9.
J Cardiovasc Pharmacol ; 35(6): 871-80, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10836720

RESUMO

Formation of mature active neuropeptides such as substance P (SP) from their glycine extended precursors entails alpha-amidation of peptide precursors by the sequential enzymatic action of peptidylglycine alpha-monooxygenase (PAM) and peptidylamidoglycolate lyase (PGL). We reported that these two enzymes that can produce mature active neuropeptides are present in cultured bovine aortic endothelial cells (BAECs). We hypothesize that alpha-amidation of peptides occurs in endothelial cells and that these peptides are critically involved in the overall regulation of cardiovascular function. In this study, this hypothesis was tested using specific amidation inhibitors to determine their effects on the actions of SP and its glycine-extended precursor (SP-Gly). We have found that SP and SP-Gly are equipotent in stimulating nitric oxide (NO) release by BAECs. At 10(-5) M, the specific inhibitors of PAM (4-phenyl-3-butenoic acid; PBA) and PGL (5-acetamido-2,4-diketo-6-phenyl-hexanoic acid and its methyl ester) reduced NO basal release by 40, 34, and 45%, respectively. They also reduced the production of NO induced by SP-Gly by 63, 68, and 69%, respectively, but had no effect on NO production in response to either SP or acetylcholine. SP and SP-Gly also were equipotent in relaxing rat aortic segments. The vasorelaxation to SP-Gly was endothelium dependent and inhibited by the NOS antagonist L-nitroarginine methyl ester (L-NAME), but it was not affected by inhibition of prostaglandin synthesis. Inhibitors of both PAM and PGL significantly reduced the vasorelaxing actions of SP-Gly, whereas responses to SP were not affected. A cumulative infusion of PBA into the femoral artery of rabbits, at final concentrations of 2.4, 24, and 240 microM for 20 min each, increased the vascular resistance (VR), indicating the tonic production of vasodilating amidated peptide(s). This effect was maximum at 60 min after infusion (20.5 +/- 4.7 vs. 8.2 +/- 0.7 mm Hg/ml/min; p < 0.05). These results suggest that endothelial cells can produce mature SP from its SP-Gly precursor and that a product of peptide alpha-amidation tonically stimulates endothelial cell NO release to control vascular tone.


Assuntos
Endotélio Vascular/efeitos dos fármacos , Complexos Multienzimáticos , Substância P/farmacologia , Vasodilatação/efeitos dos fármacos , Amidina-Liases/metabolismo , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/fisiologia , Bovinos , Células Cultivadas , Relação Dose-Resposta a Droga , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Inibidores Enzimáticos/farmacologia , Ácidos Graxos Monoinsaturados/farmacologia , Técnicas In Vitro , Masculino , Oxigenases de Função Mista/metabolismo , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/metabolismo , Coelhos , Ratos , Ratos Sprague-Dawley , Substância P/análogos & derivados
10.
Am J Clin Nutr ; 71(4): 944-9, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10731501

RESUMO

BACKGROUND: Soluble silica, a ubiquitous component of the diet, may be the natural ligand for dietary aluminum and may prevent its accumulation and toxicity in animals. However, previous studies on the inhibition of aluminum absorption and toxicity by soluble silica have produced conflicting results. We recently identified a soluble silica polymer, oligomeric silica, that has a much higher affinity for aluminum than does monomeric silica and that may be involved in the sequestration of aluminum. OBJECTIVE: By using (26)Al as a tracer, we investigated the effects of oligomeric and monomeric silica on the bioavailability of aluminum (study 1) and compared the availability of silicon from oligomeric and monomeric silica in the human gastrointestinal tract (study 2). DESIGN: In study 1, three healthy volunteers each ingested aluminum alone (control), aluminum with oligomeric silica (17 mg), and aluminum with monomeric silica (17 mg). In study 2, five healthy volunteers ingested both the oligomeric and monomeric forms of silica (34 mg). Serum and urine samples were analyzed for aluminum and silicon. RESULTS: Oligomeric silica reduced the availability of aluminum by 67% (P = 0.01) compared with the control, whereas monomeric silica had no effect (P = 0.40). Monomeric silica was readily taken up from the gastrointestinal tract and then excreted in urine (53%), whereas oligomeric silica was not detectably absorbed or excreted. CONCLUSIONS: The oligomeric, high-aluminum-affinity form of soluble silica reduces aluminum availability from the human gastrointestinal tract. Its potential role in the amelioration of aluminum toxicity in other biological systems requires attention.


Assuntos
Alumínio/farmacocinética , Absorção Intestinal/efeitos dos fármacos , Dióxido de Silício/química , Dióxido de Silício/farmacologia , Adulto , Alumínio/sangue , Alumínio/urina , Disponibilidade Biológica , Feminino , Humanos , Cinética , Substâncias Macromoleculares , Masculino , Radioisótopos , Silício/sangue , Silício/urina , Dióxido de Silício/administração & dosagem , Relação Estrutura-Atividade
11.
J Orthop Res ; 18(6): 882-90, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11192247

RESUMO

Dedifferentiated human articular chondrocytes exhibited a wide variation in their capacity to proliferate and redifferentiate in an alginate suspension culture system. The greatest extent of proliferation and redifferentiation was seen to be dependent on the formation of clonal populations of chondrocytes and correlated inversely with the initial cell seeding density. Redifferentiating chondrocytes seeded at low density (1 x 10(4) cells/ml alginate) compared with chondrocytes that were seeded at high density (1 x 10(6) cells/ml alginate) showed a nearly 3-fold higher median increase in cell number. a 19-fold greater level of type-II collagen mRNA expression, a 4-fold greater level of aggrecan mRNA expression, and a 6-fold greater level of sulfated glycosaminoglycan deposition at 4 weeks of culture. Matrix molecules from low-density cultures were assembled into chondrocyte-encapsulated, spherical extracellular matrices that were readily visualized in sections from 12-week cultures stained with antibodies against types I and II collagen and aggrecan. Ultrastructural analysis of 12-week low-density cultures confirmed the presence of thin collagen fibrils throughout the matrix.


Assuntos
Alginatos/farmacologia , Cartilagem Articular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Meios de Cultura/farmacologia , Proteínas da Matriz Extracelular , Adulto , Agrecanas , Cartilagem Articular/crescimento & desenvolvimento , Cartilagem Articular/ultraestrutura , Técnicas de Cultura de Células , Diferenciação Celular/fisiologia , Divisão Celular/fisiologia , Condrócitos/metabolismo , Condrócitos/ultraestrutura , Colágeno/genética , Colágeno/metabolismo , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Matriz Extracelular/ultraestrutura , Feminino , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Glicosaminoglicanos/metabolismo , Humanos , Lectinas Tipo C , Masculino , Pessoa de Meia-Idade , Proteoglicanas/genética , Proteoglicanas/metabolismo , RNA Mensageiro/metabolismo , Sulfatos/metabolismo
12.
Analyst ; 122(10): 1049-55, 1997 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9463954

RESUMO

Studies of the biological chemistry of aluminium can gain significantly from the use of the long-lived isotope 26Al as a tracer, although the cost of the isotope often precludes its determination by radiochemical counting techniques. Accelerator mass spectrometry (AMS) provides an ultra-sensitive method of determination, free from isobaric interference from atomic (26Mg) or molecular species. The source materials for AMS can be aluminium oxide or phosphate, both of which can be readily prepared at a sufficient level of purity from biological substrates. Natural aluminium (27Al, 100%) is added to the preparations as a chemical yield monitor and to provide the reference for the isotope ratio measurement. 26Al/27Al ratios can be determined over the range 10(-14)-10(-7), implying a limit of detection for 26Al of around 10(-18) g. The precision of measurement and long-term reproducibility are < 5% and < 7% (RSD), respectively. Chemical methodologies for routine measurements on blood and urine samples have been developed.


Assuntos
Alumínio/análise , Radioisótopos/análise , Alumínio/urina , Espectrometria de Massas , Aceleradores de Partículas , Radioisótopos/urina
13.
Am J Physiol ; 273(6): C1908-14, 1997 12.
Artigo em Inglês | MEDLINE | ID: mdl-9435496

RESUMO

Substance P (SP), an amidated peptide present in many sensory nerves, is known to affect cardiovascular function, and exogenously supplied SP has been shown to activate nitric oxide synthase (NOS) in endothelial cells. We now report that SP-Gly, the glycine-extended biosynthetic precursor of SP (which is enzymatically processed to the mature amidated SP), causes relaxation of rat aortic strips with an efficacy and potency comparable to that of SP itself. Pretreatment of the aortic strips with 4-phenyl-3-butenoic acid (PBA), an irreversible amidating enzyme inactivator, results in marked inhibition of the vasodilation activity induced by SP-Gly but not of that induced by SP itself. Isolated endothelial cell basal NOS activity is also decreased by pretreatment with PBA, with no evidence of cell death or direct action of PBA on NOS activity. Both bifunctional and monofunctional forms of amidating enzymes are present in endothelial cells, as evidenced by affinity chromatography and Western blot analysis. These results provide evidence for a link between amidative peptide processing, NOS activation in endothelial cells, and vasodilation and suggest that a product of amidative processing provides intrinsic basal activation of NOS in endothelial cells.


Assuntos
Aorta Torácica/fisiologia , Endotélio Vascular/fisiologia , Oxigenases de Função Mista/metabolismo , Complexos Multienzimáticos , Músculo Liso Vascular/fisiologia , Óxido Nítrico Sintase/metabolismo , Fragmentos de Peptídeos/farmacologia , Substância P/análogos & derivados , Substância P/farmacologia , Sequência de Aminoácidos , Animais , Aorta Torácica/efeitos dos fármacos , Bovinos , Células Cultivadas , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/enzimologia , Técnicas In Vitro , Cinética , Masculino , Dados de Sequência Molecular , Relaxamento Muscular/efeitos dos fármacos , Relaxamento Muscular/fisiologia , Músculo Liso Vascular/efeitos dos fármacos , Fragmentos de Peptídeos/síntese química , Ratos , Ratos Sprague-Dawley , Substância P/química
14.
J Trauma ; 37(3): 439-41, 1994 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8083906

RESUMO

Diagnostic peritoneal lavage (DPL) and computed tomography (CT) are the primary diagnostic modalities in the evaluation of patients with suspected blunt abdominal trauma (BAT). Diagnostic peritoneal lavage is fast and accurate but associated with complications. Computed tomography is also accurate, yet requires that patients be stable and transportable. A prospective study was designed to determine the utility of emergency ultrasound (US) studies in the initial assessment of BAT. Two hundred acutely injured patients with suspected BAT were evaluated with US. Patients were eligible for the study if they met trauma criteria and had suspected BAT. Subsequently, without knowledge of the US results, DPL or CT was performed. Ultrasound showed a sensitivity of 83%, a specificity of 100%, and an accuracy of 97% in detecting intra-abdominal injuries. Six injuries were missed but only one was felt to be significant. If US had been used in all 200 patients, 199 would have had appropriate care. We conclude US is reliable in the detection of free intraperitoneal fluid and may be used in place of DPL or CT.


Assuntos
Traumatismos Abdominais/diagnóstico , Lavagem Peritoneal , Ferimentos não Penetrantes/diagnóstico por imagem , Ferimentos não Penetrantes/diagnóstico , Traumatismos Abdominais/diagnóstico por imagem , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Sensibilidade e Especificidade , Tomografia Computadorizada por Raios X , Ultrassonografia
15.
Biochem J ; 300 ( Pt 1): 31-6, 1994 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-8198547

RESUMO

C-terminal alpha-amidation is a structural feature essential to the biological activity of many peptide hormones. Peptidylglycine alpha-amidating mono-oxygenase (PAM; EC 1.14.17.3) catalyses conversion of glycine-extended peptide hormone precursors into their corresponding alpha-hydroxyglycine derivatives. This reaction is the first step in the C-terminal amidation process. We report here that in the presence of molecular O2, copper and PAM substrate, NN-dimethyl-1,4-phenylenediamine (DMPD) serves as the requisite electron donor for the mono-oxygenase, being oxidized in the process to a stable and highly chromophoric cation radical. By monitoring the rate of increase in absorbance at 515 nm, PAM activity can be easily followed. This provides a spectrophotometric assay for PAM, which represents the first continuous assay reported for this enzyme. DMPD-supported PAM-catalysed mono-oxygenation exhibits normal Michaelis-Menten kinetic behaviour. Steady-state kinetic studies established that both the ascorbate-supported and DMPD-supported PAM reactions exhibit apparent 'Ping Pong' kinetics. In addition, both electron donors give rise to similar pH profiles and identical inhibition patterns towards known competitive inhibitors of PAM. The stoichiometry between formation of the DMPD cation radical and the alpha-hydroxyglycine PAM product was determined to be 2:1, the value expected for a monooxygenase-catalysed reaction. The optimum pH for the DMPD-supported continuous PAM assay was found to be about 5.5. The major advantage of this assay over all previously reported methods is that it is continuous; thus accurate initial rates are easily obtained. Moreover, unlike previous assay methods, 125I-labelled or chromophorically modified substrates are not required. Kinetic parameters for a broad range of PAM substrates and inhibitors have been successfully obtained using this assay.


Assuntos
Oxigenases de Função Mista/metabolismo , Complexos Multienzimáticos , Fenilenodiaminas/química , Sequência de Aminoácidos , Animais , Catálise , Bovinos , Cromatografia Líquida de Alta Pressão , Cinética , Oxigenases de Função Mista/química , Dados de Sequência Molecular , Oxirredução , Análise Espectral
16.
Biochem Biophys Res Commun ; 184(1): 323-9, 1992 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-1567439

RESUMO

Carboxy-terminal amidation is a prevalent post-translational modification necessary for the bioactivity of many peptides. We now report that the two enzymes essential for amidation, peptidylglycine alpha-monooxygenase (PAM) and peptidylamidoglycolate lyase (PGL), are present in both the cytosol and membrane fractions of cultured bovine aortic endothelial cells. Endothelial PAM exhibits ascorbate-dependent turnover and is inactivated by the mechanism-based inactivator, 4-phenyl-3-butenoic acid (PBA), whereas PGL activity is independent of ascorbate and is not affected by PBA. These enzymological characteristics correspond to those of amidating enzymes from other tissues. These results suggest a heretofore unrecognized role for alpha-amidated peptides in cardiovascular function.


Assuntos
Amidina-Liases , Endotélio Vascular/enzimologia , Liases/metabolismo , Oxigenases de Função Mista/metabolismo , Complexos Multienzimáticos , Animais , Aorta , Ácido Ascórbico/farmacologia , Bovinos , Membrana Celular/enzimologia , Células Cultivadas , Ácidos Graxos Monoinsaturados/farmacologia , Cinética
17.
J Reprod Fertil ; 84(1): 313-24, 1988 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-3184049

RESUMO

The effects of season and of oestradiol and progesterone on the tonic secretion of LH were studied in ovariectomized Merino and Suffolk ewes, two breeds which differ markedly in the seasonal pattern of their reproductive activity. In the absence of exogenous steroids, the frequency of LH pulses was lower and the amplitude of the pulses was higher in anoestrus than in the breeding season for Merino and Suffolk ewes 30 days after ovariectomy. In long-term (190 days) ovariectomized ewes, this seasonal change in LH secretion was observed in Suffolk ewes only. During seasonal anoestrus, treatment of ewes with subcutaneous oestradiol-17 beta implants (3, 6 or 12 mm in length) decreased the frequency of LH pulses in a dose-dependent manner, with Suffolk ewes being far more sensitive to the inhibitory effects of oestradiol than Merino ewes. The lowest dose of oestradiol (3 mm) had no effect on the secretion of LH in Merino ewes, but reduced secretion in Suffolk ewes. Treatment of ewes with the highest dose of oestradiol (12 mm) completely abolished LH pulses in Suffolk ewes, whereas infrequent pulses remained evident in Merino ewes. During the breeding season, oestradiol alone had no effect on the pulsatile release of LH in either breed, but in combination with progesterone there was a significant reduction in LH pulse frequency. Progesterone effectively decreased LH secretion in both breeds in both seasons. It was concluded that differences between breeds in the 'depth' of anoestrus could be related to differences in the sensitivity of the hypothalamus to both negative feedback by oestradiol and the direct effects of photoperiod.


Assuntos
Estradiol/farmacologia , Estro , Hipotálamo/efeitos dos fármacos , Hormônio Luteinizante/metabolismo , Progesterona/farmacologia , Animais , Relação Dose-Resposta a Droga , Retroalimentação , Feminino , Luz , Ovariectomia , Taxa Secretória/efeitos dos fármacos , Ovinos
18.
J Reprod Fertil ; 84(1): 333-9, 1988 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-3184051

RESUMO

In Exp. 1, 4 groups of 50 recently weaned ewes were exposed to various degrees of contact with rams for 65 days, followed by exposure to novel rams for 4 days. Ovarian activity in the ewes was determined by laparoscopy on Days 29, 65 and 69 of treatment. There were no treatment differences in the percentage of ewes ovulating on Day 4 whereas by Day 29 more ewes in clear fenceline and full ram contact were ovulating compared to controls (P less than 0.05, P less than 0.001). After 65 days ovarian activity was significant only in those ewes in full contact with rams (P less than 0.001). Between 89 and 95% of ewes remaining anovulatory after 65 days ovulated after 4 days of full contact with novel rams. In Exp. 2, 4 groups of about 30 anovulatory ewes were exposed to various degrees of contact with rams for 5 days. Ovarian activity was assessed before and after treatment by laparoscopy. After 5 days, more ewes were ovulating in response to full ram contact than in any other treatment (P less than 0.05) and more ewes in fenceline contact with rams or with rams plus ewes were ovulating than in the isolated control treatment (P less than 0.01). In Exp. 3, 6 groups of about 40 anovulatory ewes were exposed to face masks with and without rams' wool and/or various degrees of contact with rams for 5 days. More ewes were ovulating after 5 days in the group in full physical contact with rams than in any other group (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Ovário/fisiologia , Indução da Ovulação , Reprodução , Estações do Ano , Ovinos/fisiologia , Animais , Feminino , Masculino
19.
J Reprod Fertil ; 84(1): 43-50, 1988 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-3184059

RESUMO

Ewes that were untreated, fed lupins or fed lupins and immunized against androstenedione were artificially inseminated. The percentage of ewes pregnant at 36-45 days after insemination (fertility) was 8% higher in ewes that had more than one ovulation than in those that had only one ovulation. Maximum fertility was achieved with 50 x 10(6) spermatozoa and this did not vary with the number of ovulations that ewes had. Among the pregnant, twin-ovulating ewes, embryo survival increased as the number of spermatozoa inseminated increased from 25 x 10(6) to 400 x 10(6). Immunization of ewes against androstenedione increased ovulation rate but reduced fertility, and reduced embryo survival among twin-ovulation ewes.


Assuntos
Androstenodiona/fisiologia , Fertilidade , Inseminação Artificial/veterinária , Ovulação , Animais , Feminino , Morte Fetal , Masculino , Gravidez , Ovinos , Espermatozoides/fisiologia
20.
Aust J Biol Sci ; 41(1): 23-6, 1988.
Artigo em Inglês | MEDLINE | ID: mdl-3269698

RESUMO

The pattern of ovulation of Merino ewes was studied by repeated laparoscopy each 14 days in the anoestrous (n = 97) and breeding (n = 87) seasons. In the anoestrous season the proportion of ewes ovulating did not decrease below 11%, 42% of ewes never ovulated and the remainder fluctuated between the two states. On 20 occasions a clear anovulatory period was interrupted by an isolated spontaneous ovulation. In the breeding season the overall mean proportion of ewes with corpora lutea or albicantia at laparoscopy was 87%, 54% of ewes ovulated regularly throughout while in another 31% absence of corpora lutea or albicantia coincided with the follicular phase of an oestrous cycle as evidenced by an appropriately aged corpora lutea at the next laparoscopy. Of the remaining 15% of the flock 3% had anovulatory periods greater than 14 days while the remainder experienced irregular ovulatory cycles--the majority due to short periods of anovulation but some ewes retained corpora lutea for longer than 14 days while others ovulated twice between successive laparoscopies.


Assuntos
Anestro/fisiologia , Cruzamento , Estro/fisiologia , Ovulação/fisiologia , Estações do Ano , Ovinos , Animais , Feminino
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