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1.
Int J Cosmet Sci ; 31(1): 47-54, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19134127

RESUMO

Human body odour is important in modulating self-perception and interactions between individuals. Artificial fragrances have been used for thousands of years to manipulate personal odour, but the nature and extent of influences on person perception are relatively unexplored. Here we test the effects of a double-blind manipulation of personal odour on self-confidence and behaviour. We gave to male participants either an aerosol spray containing a formulation of fragrance and antimicrobial agents or an otherwise identical spray that lacked these active ingredients. Over several days, we found effects between treatment groups on psychometric self-confidence and self-perceived attractiveness. Furthermore, although there was no difference between groups in mean attractiveness ratings of men's photographs by a female panel, the same women judged men using the active spray as more attractive in video-clips, suggesting a behavioural difference between the groups. Attractiveness of an individual male's non-verbal behaviour, independent of structural facial features, was predicted by the men's self-reported proclivity towards the provided deodorant. Our results demonstrate the pervasive influence of personal odour on self-perception, and how this can extend to impressions on others even when these impressions are formed in the absence of odour cues.


Assuntos
Comportamento , Odorantes , Autoimagem , Desejabilidade Social , Adulto , Método Duplo-Cego , Feminino , Humanos , Estudos Longitudinais , Masculino , Inquéritos e Questionários , Adulto Jovem
2.
Neuroscience ; 135(4): 1045-54, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16165283

RESUMO

Emerging data report sex differences in how the brain responds to chronic stress. Here, we investigated the effects of chronic restraint stress (6 h/day/21 days) on hippocampal morphology and function in ovariectomized female rats. Chronic restraint stress caused CA3 apical dendritic retraction in short- and long-shafted neurons, while it reduced basal dendritic arbors in long-shafted neurons only. Chronic restraint did not affect CA1 dendritic arborization, although it increased the proportion of CA1 spine heads compared with controls. Both stressed and control animals performed well on the Y-maze, a spatial memory task. However, chronic stress enhanced Y-maze performance compared with controls, which may reflect facilitated spatial memory or reduced habituation. Y-maze performance correlated with CA1 spine head proportion. This relationship suggests that spatial ability in females may be more tightly coupled with CA1 morphology, which may override the influence of CA3 dendritic retraction. Thus, this research provides additional evidence that CA3 morphology does not always parallel spatial memory.


Assuntos
Hipocampo/patologia , Memória/fisiologia , Neurônios/patologia , Comportamento Espacial/fisiologia , Estresse Psicológico/fisiopatologia , Animais , Feminino , Aprendizagem em Labirinto/fisiologia , Ovariectomia , Ratos , Restrição Física/fisiologia , Caracteres Sexuais
3.
Int Dent J ; 53(6 Suppl 1): 404-8, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14725386

RESUMO

OBJECTIVES: Two studies were carried out to investigate whether the effect of specific oral care feed-back devices would result in better plaque removal (Study 1) and lead to changes in attitudes and views relating to oral health (Study 2). The objective of the first study was to compare the ability of a toothbrush system consisting of a new toothbrush plus plaque disclosing tablet (SIGNAL Integral) to remove supra-gingival plaque to that of a marketed toothbrush (Oral B Cross Action) following a single unsupervised brushing. METHODS: Study 1 had a cross-over design and included 21 healthy adult volunteers. Subjects refrained from any form of oral hygiene for 24 hours prior to each test session. Plaque levels (Modified Quigley-Hein Plaque index) were assessed prior to and following each unsupervised brushing. When the new toothbrush was used, subjects self-disclosed their plaque with a disclosing tablet (erythrosine) immediately prior to brushing. At each occasion, brushing time (in sec) was also recorded. Study 2 had a two-cell, parallel design (test and control group) and lasted for five days. Healthy adult volunteers were enrolled. Subjects in the test group (n = 30) were given a fluoride toothpaste and four saliva test strips to use at home. The saliva test strip is designed to change colour in the pH range of 6.5 to 10, allowing the user to verify the effect of brushing. The control group (n = 29) received no saliva test strips. Subjects completed a questionnaire after 4 days of product use. RESULTS: In Study 1, significantly more plaque was removed (p < 0.05) and brushing time was increased by more than 20% when subjects used the new toothbrush together with the disclosing tablet compared to brushing with the marketed toothbrush. In Study 2, significant increases in motivation relating to oral hygiene were found in the test group who had been using the saliva test strip. CONCLUSION: The two studies have provided significant evidence that the provision of feed-back devices in an oral care context can lead to improved removal of plaque and increased motivation.


Assuntos
Atitude Frente a Saúde , Dispositivos para o Cuidado Bucal Domiciliar , Placa Dentária/terapia , Retroalimentação , Higiene Bucal , Adulto , Cariostáticos/uso terapêutico , Estudos Cross-Over , Placa Dentária/diagnóstico , Índice de Placa Dentária , Eritrosina , Corantes Fluorescentes , Fluoretos/uso terapêutico , Humanos , Concentração de Íons de Hidrogênio , Motivação , Saúde Bucal , Fitas Reagentes , Saliva/fisiologia , Fatores de Tempo , Escovação Dentária/instrumentação , Cremes Dentais/uso terapêutico
4.
Vaccine ; 19(6): 655-60, 2000 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-11090718

RESUMO

Fragilysin, an extracellular zinc metalloprotease produced by enterotoxigenic strains of the anaerobic bacterium Bacteroides fragilis, disrupts the paracellular barrier by cleavage of the intercellular proteins between epithelial cells resulting in fluid secretion. Intranasal immunization of mice with fragilysin and co-administered ovalbumin (Ova) resulted in an Ova-specific serum IgG response that was over 18000-fold higher than Ova alone, as well as detectable levels of serum IgA. Serum IgG titers were comparable with those seen when whole cholera toxin was used as the adjuvant, although the responses obtained with fragilysin showed more variability between mice. Metalloproteases to which fragilysin is structurally related were ineffective as mucosal adjuvants. Our results and similar studies with enterotoxins that affect the paracellular barrier suggest that alteration of mucosal permeability may play an important role in the mechanisms of adjuvanticity.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Administração Intranasal , Antígenos de Bactérias/imunologia , Imunoglobulinas/sangue , Metaloendopeptidases/imunologia , Animais , Animais não Endogâmicos , Anticorpos Antibacterianos/biossíntese , Toxina da Cólera/administração & dosagem , Toxina da Cólera/imunologia , Feminino , Imunidade nas Mucosas , Metaloendopeptidases/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Vacinação
5.
Yeast ; 16(9): 811-27, 2000 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-10861905

RESUMO

In response to elevated levels of HMG-CoA reductase, an integral endoplasmic reticulum (ER) membrane protein, cells assemble novel ER arrays. These membranes provide useful models for exploration of ER structure and function, as well as general features of membrane biogenesis and turnover. Yeast express two functional HMG-CoA reductase isozymes, Hmg1p and Hmg2p, each of which induces morphologically different ER arrays. Hmg1p induces stacks of paired nuclear-associated membranes called karmellae. In contrast, Hmg2p induces peripheral ER membrane arrays and short nuclear-associated membrane stacks. In spite of their ability to induce different cellular responses, both Hmg1p and Hmg2p have similar structures, including a polytopic membrane domain containing eight predicted transmembrane helices. By examining a series of recombinant HMG-CoA reductase proteins, our laboratory previously demonstrated that the last ER-lumenal loop (Loop G) of the Hmg1p membrane domain contains a signal needed for proper karmellae assembly. Our goal was to examine the primary sequence requirements within Loop G that were critical for proper function of this signal. To this end, we randomly mutagenized the Loop G sequence, expressed the mutagenized Hmg1p in yeast, and screened for inability to generate karmellae at wild-type levels. Out of approximately 4000 strains with Loop G mutations, we isolated 57 that were unable to induce wild-type levels of karmellae assembly. Twenty-nine of these mutants contained one or more point mutations in the Loop G sequence, including nine single point mutants, four of which had severe defects in karmellae assembly. Comparison of these mutations to single point mutations that did not affect karmellae assembly did not reveal obvious patterns of sequence requirements. For example, both conservative and non-conservative changes were present in both groups and changes that altered the total charge of the Loop G region were observed in both groups. Our hypothesis is that Loop G serves as a karmellae-inducing signal by mediating protein-protein or protein-lipid interactions and that amino acids revealed by this analysis may be important for maintaining the proper secondary structure needed for these interactions.


Assuntos
Retículo Endoplasmático/metabolismo , Hidroximetilglutaril-CoA Redutases/genética , Mutação , Saccharomyces cerevisiae/enzimologia , Transdução de Sinais , Alelos , Sequência de Aminoácidos , Retículo Endoplasmático/química , Regulação Fúngica da Expressão Gênica , Hidroximetilglutaril-CoA Redutases/química , Hidroximetilglutaril-CoA Redutases/metabolismo , Hidroximetilglutaril-CoA-Redutases NADP-Dependentes , Membranas Intracelulares/química , Membranas Intracelulares/metabolismo , Isoenzimas , Dados de Sequência Molecular , Saccharomyces cerevisiae/genética , Análise de Sequência de DNA
6.
Med Sci Sports Exerc ; 31(12): 1793-8, 1999 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10613430

RESUMO

PURPOSE: African Americans have a higher incidence of hypertension than other racial groups. Furthermore, some research suggests that normotensive individuals who exhibit exaggerated blood pressure (BP) responses to exercise may be at risk for future hypertension. This study sought to determine whether normotensive African Americans exhibited exaggerated BP responses to static exercise or dynamic exercise relative to Caucasian Americans and Asian Americans. METHODS: Thirty normotensive subjects participated from each of the three racial groups (15 men and 15 women). Subjects held 30% of maximal voluntary contraction (right knee extension) for 3 min, and BP was recorded during the 3rd min. On a separate occasion, subjects cycled for six min at a power equivalent to 70% of VO2 reserve, and BP was recorded during the 6th min. RESULTS: Static exercise produced large, significant increases in both systolic and diastolic BP (35 +/- 1.5 and 29 +/- 1.3 mm Hg, respectively). Dynamic exercise produced large, significant increases in systolic BP (51 +/- 1.6 mm Hg) and moderate, yet significant, increases in diastolic BP (8 +/- 1.0 mm Hg). There were no significant differences between racial groups in BP response to either static exercise or dynamic exercise. However, during dynamic exercise, men had a higher systolic BP response than did women. CONCLUSIONS: African Americans who are normotensive at rest do not exhibit a greater BP response to static exercise or dynamic exercise than do Caucasian Americans or Asian Americans.


Assuntos
Povo Asiático , População Negra , Pressão Sanguínea/fisiologia , Exercício Físico , População Branca , Diástole , Feminino , Humanos , Masculino , Sístole
7.
Mol Biol Cell ; 10(10): 3409-23, 1999 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-10512876

RESUMO

In all cells examined, specific endoplasmic reticulum (ER) membrane arrays are induced in response to increased levels of the ER membrane protein 3-hydroxy 3-methylglutaryl coenzyme A (HMG-CoA) reductase. In yeast, expression of Hmg1p, one of two yeast HMG-CoA reductase isozymes, induces assembly of nuclear-associated ER stacks called karmellae. Understanding the features of HMG-CoA reductase that signal karmellae biogenesis would provide useful insights into the regulation of membrane biogenesis. The HMG-CoA reductase protein consists of two domains, a multitopic membrane domain and a cytosolic catalytic domain. Previous studies had indicated that the HMG-CoA reductase membrane domain was exclusively responsible for generation of ER membrane proliferations. Surprisingly, we discovered that this conclusion was incorrect: sequences at the carboxyl terminus of HMG-CoA reductase can profoundly affect karmellae biogenesis. Specifically, truncations of Hmg1p that removed or shortened the carboxyl terminus were unable to induce karmellae assembly. This result indicated that the membrane domain of Hmg1p was not sufficient to signal for karmellae assembly. Using beta-galactosidase fusions, we demonstrated that the carboxyl terminus was unlikely to simply serve as an oligomerization domain. Our working hypothesis is that a truncated or misfolded cytosolic domain prevents proper signaling for karmellae by interfering with the required tertiary structure of the membrane domain.


Assuntos
Citosol/enzimologia , Hidroximetilglutaril-CoA Redutases/metabolismo , Carbocianinas/química , Retículo Endoplasmático/metabolismo , Imunofluorescência , Hidroximetilglutaril-CoA Redutases/genética , Membranas Intracelulares/metabolismo , Microscopia Eletrônica , Mutação , Membrana Nuclear/ultraestrutura , Proteínas Recombinantes de Fusão/metabolismo , Schizosaccharomyces , beta-Galactosidase/genética
8.
Placenta ; 19(7): 517-24, 1998 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9778125

RESUMO

To determine whether the expression and activity of glucose transporters in human trophoblast are regulated by glucose, syncytiotrophoblast cells, choriocarcinoma cells, and villous fragments were incubated with a range of glucose concentrations (0-20 mM, 24 h). Expression of GLUT1 and GLUT3 glucose transporters was measured by immunoblotting, while glucose transporter activity was determined by [3H]2-deoxyglucose uptake in the cultured cells. GLUT1 expression in syncytial cells was enhanced following incubation in absence of glucose, reduced by incubation in 20 mM glucose but was not altered by incubation at 1 or 12 mM glucose. Transporter activity was inversely related to extracellular glucose over the entire range of concentrations tested (0-20 mM). Incubation of villous fragments in 20 mM glucose produced a limited suppression of GLUT1 expression, but no effects were noted following incubation at 0 or 1 mM glucose. Neither GLUT1 expression in JAr and JEG-3 choriocarcinoma cells nor transport activity in JEG-3 cells was affected by extracellular glucose concentration. Unlike syncytial cells, JAr, JEG-3 and BeWo all expressed GLUT3 protein in addition to GLUT1. These results show that while syncytiotrophoblast GLUT1 expression is altered at the extremes of extracellular glucose concentration, it is refractory to glucose alone at lower concentrations. By contrast, an inverse relationship exists between glucose transporter activity and extracellular glucose. This suggests that there are post-translational regulatory mechanisms which may respond to changes in extracellular glucose concentration.


Assuntos
Glucose/farmacologia , Proteínas de Transporte de Monossacarídeos/metabolismo , Proteínas do Tecido Nervoso , Placenta/efeitos dos fármacos , Adulto , Western Blotting , Linhagem Celular , Células Cultivadas , Coriocarcinoma/metabolismo , Desoxiglucose/metabolismo , Feminino , Transportador de Glucose Tipo 1 , Transportador de Glucose Tipo 3 , Humanos , Placenta/citologia , Placenta/metabolismo , Gravidez , Células Tumorais Cultivadas
9.
Mem Cognit ; 26(2): 402-20, 1998 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-9584445

RESUMO

Subjects exposed to members of a structured domain become sensitive to the general structure of that domain, even when they are unaware that the domain has such structure (e.g., Reber, 1993). Numerous investigators have attempted to characterize this learning as unselective in acquisition and automatic in application. However, we contend that this characterization miscasts the fundamental nature of learning. In a series of experiments, we demonstrate that what subjects learn implicitly about the structure of a domain critically depends on decisions they make about how to organize the structural components. Similarly, the application of knowledge gained implicitly is not stable, but may be selected or even created under the demands of the test task. We conclude that implicit learning, just like explicit learning, proceeds through active organization of the stimulus complex, rather than by passively absorbing any level of structure. We propose a synthesis, in which learning, with and without awareness, is understood through a common set of principles.


Assuntos
Adaptação Psicológica , Conscientização , Formação de Conceito , Rememoração Mental , Resolução de Problemas , Adulto , Tomada de Decisões , Feminino , Humanos , Masculino , Retenção Psicológica , Transferência de Experiência
10.
Infect Immun ; 66(4): 1735-9, 1998 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-9529104

RESUMO

Enterotoxigenic strains of Bacteroides fragilis produce an extracellular metalloprotease toxin (termed fragilysin) which is cytopathic to intestinal epithelial cells and induces fluid secretion and tissue damage in ligated intestinal loops. We report here that the fragilysin gene is contained within a small genetic element termed the fragilysin pathogenicity islet. The pathogenicity islet of B. fragilis VPI 13784 was defined as 6,033 bp in length and contained nearly perfect 12-bp direct repeats near its ends. Sequencing across the ends of the pathogenicity islet from two additional enterotoxigenic strains, along with PCR analysis of 20 additional enterotoxigenic strains, revealed that the islet is inserted at a specific site on the B. fragilis chromosome. The site of integration in three nontoxigenic strains contained a 17-bp GC-rich sequence which was not present in toxigenic strains and may represent a target sequence for chromosomal integration. In addition to the fragilysin gene, we identified an open reading frame encoding a predicted protein with a size and structural features similar to those of fragilysin. The deduced amino acid sequence was 28.5% identical and 56.3% similar to fragilysin and contained a nearly identical zinc-binding motif and methionine-turn region.


Assuntos
Bacteroides fragilis/patogenicidade , Enterotoxinas/genética , Metaloendopeptidases/genética , Sequência de Aminoácidos , Sequência de Bases , Metaloendopeptidases/química , Dados de Sequência Molecular , Fases de Leitura Aberta , Reação em Cadeia da Polimerase
11.
Med Sci Sports Exerc ; 29(2): 268-72, 1997 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9044233

RESUMO

Submaximal cycle ergometry is routinely used for the prediction of mode specific maximal oxygen consumption (VO2peak). Such testing is almost universally performed at a cycling cadence of 50 rpm despite the finding that a cadence of approximately 80 rpm yields greater economy of effort among individuals with cycling experience. We sought to determine if 50 or 80 rpm were superior cadences for predicting VO2peak in individuals with or without cycling experience. Thirty experienced (EXP: 16 male and 14 female) and 28 nonexperienced (NEXP: 15 male and 13 female) subjects between the ages of 18-40 yr completed two incremental exercise tests on a cycle ergometer, one at 50 rpm and one at 80 rpm. in random order. VO2peak was predicted from submaximal data according to the method of the ACSM. There was no difference between actual VO2peak at 50 rpm and at 80 rpm. Correlations between predicted and actual VO2peak were 0.79 at 50 rpm and 0.81 at 80 rpm for all subjects. No significant differences in correlations were observed between groups or between cadences. However, the predicted values of VO2peak overestimated the actual values in both groups. Therefore, we conclude that cadences of 50 and 80 rpm have similar validity in cycle ergometry prediction of VO2peak, but the ACSM protocol overestimates VO2peak.


Assuntos
Ergometria/estatística & dados numéricos , Exercício Físico/fisiologia , Aptidão Física/fisiologia , Adulto , Ciclismo , Ergometria/métodos , Feminino , Frequência Cardíaca , Humanos , Masculino , Consumo de Oxigênio
12.
J Exp Psychol Learn Mem Cogn ; 23(1): 181-200, 1997 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9028027

RESUMO

Subjects exposed to members of a structured domain become sensitive to the general structure of that domain, even when unaware that the domain has such structure. Numerous investigators have taken this as evidence for a mode of learning in which memory passively and unselectively absorbs the structure of the environment. The authors contend that this assumption miscasts the fundamental nature of learning. The authors demonstrate that even when task and stimulus structure are held constant, subjects react to variations in incidental stimulus properties by processing the stimuli in qualitatively different ways. The authors conclude that implicit learning, just like explicit learning, proceeds through active organization of the stimulus complex, rather than through passive absorption of any level of structure. The authors propose a synthesis in which learning, with and without awareness, is understood through a common set of principles.


Assuntos
Aprendizagem/fisiologia , Adaptação Fisiológica , Adulto , Feminino , Humanos , Masculino , Estimulação Luminosa
13.
FEMS Microbiol Lett ; 146(2): 279-84, 1997 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-9011050

RESUMO

The Bacteroides fragilis enterotoxin is an extracellular zinc metalloprotease that has been implicated in diarrheal disease of humans and animals. This toxin causes fluid accumulation in intestinal loops and is cytotoxic for HT-29 cells, an intestinal carcinoma cell line. Here we report the cloning and sequencing of the toxin gene (bftP). bftP is 1191 nucleotides coding for a 397 amino acid protein of 44.4 kDa. The toxin has a signal peptide of 18 amino acids that is typical of many lipoproteins followed by a 379 amino acid protoxin. The portion of the protoxin found in culture filtrates and stools begins at amino acid 212. An additional open reading frame located immediately upstream shows some sequence identity with cobra cytotoxins. If expressed, the ORF protein product could also play a role in the virulence of B. fragilis.


Assuntos
Bacteroides fragilis/genética , Enterotoxinas/genética , Metaloendopeptidases/genética , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Bacteroides fragilis/enzimologia , Sequência de Bases , Clonagem Molecular , DNA Bacteriano/análise , DNA Bacteriano/isolamento & purificação , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Análise de Sequência de DNA
14.
Anal Chem ; 69(3): 532-5, 1997 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-21639203

RESUMO

Temperature-resolved molecular emission spectroscopy is described as a thermal analysis method for the analysis of solids and liquids. The technique uses an electrically heated graphite cup to decompose and/or vaporize the sample. The vapors are carried by a stream of argon into a cool hydrogen diffusion flame. Both the quantity and the nature of the decomposed species can be determined. The technique is particularly useful for the determination of sulfur, phosphorus, or nitrogen. Calibration curves for sulfur show the expected parabolic shape, and those for phosphorus are linear. The detection limit for elemental sulfur was determined to be approximately 50 ng. The evolution of sulfur is shown to be related to the decomposition temperature which is characteristic of the sulfur-containing species. Reproducibility of the decomposition temperatures is typically ±2%.

15.
J Cell Sci ; 109 ( Pt 6): 1637-45, 1996 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8799850

RESUMO

When expressed in pituitary AtT-20 cells, parotid proline-rich proteins enter the regulated pathway. Because the short N-terminal domain of a basic proline-rich protein is necessary for efficient export from the ER, it has not been possible to evaluate the role of this polypeptide segment as a sorting signal for regulated secretion. We now show that addition of the six-amino acid propeptide of proparathyroid hormone to the proline-rich protein, and especially to a deletion mutant lacking the N-terminal domain, dramatically accelerates intracellular transport of these polypeptides. Under these conditions the chimeric deletion mutant is stored as effectively as the full-length protein in dense core granules. The propeptide does not function as a sorting signal in AtT-20 cells as it does not reroute a constitutively secreted reporter protein to the regulated pathway. During transit, the propeptide is cleaved from the chimeric polypeptides such that the original structures of the full-length and the deletion mutant proline-rich proteins are reestablished. We have also found that the percentage stimulated secretion of the proline-rich proteins increases incrementally (almost twofold) as their level of expression is elevated. The increase reflects an enrichment of these polypeptides in the granule pool and its incremental nature suggests that sorting of proline-rich proteins involves an aggregation-based process. Because we can now rule out contributions to sorting by both N- and C-terminal segments of the proline-rich protein, we deduce that the unique proline-rich domain is responsible for storage. Thus at least some of the determinants of sorting for regulated secretion are protein-specific rather than universal.


Assuntos
Glicoproteínas de Membrana , Glândula Parótida/metabolismo , Peptídeos/química , Animais , Sítios de Ligação/genética , Linhagem Celular , Retículo Endoplasmático/metabolismo , Camundongos , Estrutura Molecular , Mutagênese Sítio-Dirigida , Hormônio Paratireóideo/genética , Hormônio Paratireóideo/metabolismo , Peptídeos/genética , Peptídeos/metabolismo , Prolina/química , Domínios Proteicos Ricos em Prolina , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Processamento de Proteína Pós-Traducional , Sinais Direcionadores de Proteínas/química , Sinais Direcionadores de Proteínas/genética , Sinais Direcionadores de Proteínas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transfecção , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/metabolismo
16.
Mol Biol Cell ; 7(5): 769-89, 1996 May.
Artigo em Inglês | MEDLINE | ID: mdl-8744950

RESUMO

In all eucaryotic cell types analyzed, proliferations of the endoplasmic reticulum (ER) can be induced by increasing the levels of certain integral ER proteins. One of the best characterized of these proteins is HMG-CoA reductase, which catalyzes the rate-limiting step in sterol biosynthesis. We have investigated the subcellular distributions of the two HMG-CoA reductase isozymes in Saccharomyces cerevisiae and the types of ER proliferations that arise in response to elevated levels of each isozyme. At endogenous expression levels, Hmg1p and Hmg2p were both primarily localized in the nuclear envelope. However, at increased levels, the isozymes displayed distinct subcellular localization patterns in which each isozyme was predominantly localized in a different region of the ER. Specifically, increased levels of Hmg1p were concentrated in the nuclear envelope, whereas increased levels of Hmg2p were concentrated in the peripheral ER. In addition, an Hmg2p chimeric protein containing a 77-amino acid lumenal segment from Hmg1p was localized in a pattern that resembled that of Hmg1p when expressed at increased levels. Reflecting their different subcellular distributions, elevated levels of Hmg1p and Hmg2p induced sets of ER membrane proliferations with distinct morphologies. The ER membrane protein, Sec61p, was localized in the membranes induced by both Hmg1p and Hmg2p green fluorescent protein (GFP) fusions. In contrast, the lumenal ER protein, Kar2p, was present in Hmg1p:GFP membranes, but only rarely in Hmg2p:GFP membranes. These results indicated that the membranes synthesized in response to Hmg1p and Hmg2p were derived from the ER, but that the membranes were not identical in protein composition. We determined that the different types of ER proliferations were not simply due to quantitative differences in protein amounts or to the different half-lives of the two isozymes. It is possible that the specific distributions of the two yeast HMG-CoA reductase isozymes and their corresponding membrane proliferations may reveal regions of the ER that are specialized for certain branches of the sterol biosynthetic pathway.


Assuntos
Retículo Endoplasmático/enzimologia , Retículo Endoplasmático/fisiologia , Hidroximetilglutaril-CoA Redutases/metabolismo , Isoenzimas/metabolismo , Saccharomyces cerevisiae/enzimologia , Retículo Endoplasmático/ultraestrutura , Hidroximetilglutaril-CoA Redutases/ultraestrutura , Imuno-Histoquímica , Membranas Intracelulares/efeitos dos fármacos , Membranas Intracelulares/enzimologia , Membranas Intracelulares/ultraestrutura , Isoenzimas/ultraestrutura , Lovastatina/farmacologia , Microscopia Confocal , Microscopia Eletrônica , Plasmídeos , Saccharomyces cerevisiae/ultraestrutura
17.
FEBS Lett ; 384(3): 235-9, 1996 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-8617361

RESUMO

Abstract A gene, CYP119, for a potential cytochrome P450 has been isolated and sequenced from the extreme acidothermophilic archaeon Sulfolobus solfataricus. The gene predicts a polypeptide of 368 amino acids containing the consensus heme-binding sequence Phe-Gly-Xaa-Gly-Xaa-His-Xaa-Cys-Xaa-Gly- Xaa3-Ala-Arg-Xaa-Glu. It most closely resembles the cytochrome P450s found in the bacterium Bacillus subtilis, with which it shares 129 identical amino acid residues (35%). This first sequence of a potential archaeal cytochrome P450 represents an important step in tracing the complex evolutionary history of this biologically important enzyme family.


Assuntos
Sistema Enzimático do Citocromo P-450/genética , Sulfolobus/enzimologia , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Sequência Conservada , Sistema Enzimático do Citocromo P-450/química , Evolução Molecular , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Homologia de Sequência de Aminoácidos , Sulfolobus/genética
18.
Arch Toxicol ; 70(8): 508-9, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8783815

RESUMO

The high-temperature combustion of synthetic ester turbine engine lubricants has been performed by diluting the lubricant 5, 15, or 25% in diesel fuel and burning the mixture in a pilot-scale boiler facility. The effluent gas from this combustion system was carefully monitored for the formation of a potent neurotoxin, trimethylolpropane phosphate (TMPP). Although TMPP was not detected in the gaseous effluent, elevated levels of the neurotoxin were found in scrapings from the inside of the boiler system. Because of the extreme toxicity of this compound, significant dermal exposure could be a potential risk to incinerator operation and maintenance personnel.


Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/química , Poluentes Ambientais , Óleos Combustíveis , Neurotoxinas/química , Fosfitos/química , Centrais Elétricas , Propilenoglicóis/química
19.
J Cell Sci ; 108 ( Pt 12): 3827-37, 1995 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8719889

RESUMO

In pituitary-derived AtT-20 cells, recombinant fibronectin containing the N-terminal matrix assembly domain and the C-terminal half of fibronectin does not follow the regulated secretory pathway but instead concentrates in distinct organelles prior to secretion. These organelles are larger than the dense-core granules and localize to the cell body at sites that differ from lysosomes, endosomes and endoplasmic reticulum. Unlike the dense-core granules, their discharge is not stimulated by 8-bromo-cyclic-AMP or phorbol esters. The kinetics of intracellular transport and secretion of the recombinant fibronectin suggest that it is present in a post-Golgi pool that turns over more slowly than constitutive vesicles. Indeed, the fibronectin-containing organelles disappear with a half-time of 3 hours after inhibiting protein synthesis. Presence of the organelles correlates with intracellular aggregation of dimeric fibronectin polypeptides. The organelles are absent in cells expressing monomeric recombinant fibronectin (lacking C-terminal dimerization sites) or the C-terminal half of fibronectin (which dimerizes but lacks the N-terminal matrix assembly domain), both of which aggregate less efficiently than dimeric fibronectin. Instead, the latter polypeptides enter the dense-core granules. Thus while the formation of the fibronectin-containing organelles may require efficient aggregation, it may not require a specific structural signal. Moreover, efficient aggregation is not necessarily a prerequisite for following the regulated pathway.


Assuntos
Fibronectinas/metabolismo , Hipófise/metabolismo , Transdução de Sinais/fisiologia , Hormônio Adrenocorticotrópico/metabolismo , Sequência de Aminoácidos , Animais , Transporte Biológico/fisiologia , Agregação Celular/fisiologia , Linhagem Celular , Camundongos , Dados de Sequência Molecular , Organelas/metabolismo , Hipófise/citologia , Proteínas Recombinantes/metabolismo , beta-Endorfina/metabolismo
20.
Mol Biol Cell ; 6(11): 1535-47, 1995 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8589454

RESUMO

In all eukaryotic cells that have been examined, specific membrane arrays are induced in response to increased levels of the ER membrane protein, HMG-CoA reductase. Analysis of these inducible membranes has the potential to reveal basic insights into general membrane assembly. Yeast express two HMG-CoA reductase isozymes, and each isozyme induces a morphologically distinct proliferation of the endoplasmic reticulum. The isozyme encoded by HMG1 induces karmellae, which are long stacks of membranes that partially enclose the nucleus. In contrast, the isozyme encoded by HMG2 induces short stacks of membrane that may be associated with the nucleus, but are frequently present at the cell periphery. To understand the molecular nature of the different cellular responses to Hmg1p and Hmg2p, we mapped the region of Hmg1p that is needed for karmellae assembly. For this analysis, a series of exchange alleles was examined in which a portion of the Hmg2p membrane domain was replaced with the corresponding Hmg1p sequences. Results of this analysis indicated that the ER lumenal loop between predicted transmembrane domains 6 and 7 was both necessary and sufficient for karmellae assembly, when present in the context of an HMG-CoA reductase membrane domain. Immunoblotting experiments ruled out the simple possibility that differences in the amounts of the various chimeric HMG-CoA reductase proteins was responsible for the altered cellular responses. Our results are consistent with the hypothesis that each yeast isozyme induces or organizes a qualitatively different organization of ER membrane.


Assuntos
Retículo Endoplasmático/ultraestrutura , Hidroximetilglutaril-CoA Redutases/química , Hidroximetilglutaril-CoA Redutases/metabolismo , Isoenzimas/metabolismo , Membrana Nuclear/ultraestrutura , Estrutura Secundária de Proteína , Saccharomyces cerevisiae/ultraestrutura , Sequência de Aminoácidos , Eletroforese em Gel de Poliacrilamida , Retículo Endoplasmático/enzimologia , Hidroximetilglutaril-CoA Redutases/biossíntese , Immunoblotting , Isoenzimas/biossíntese , Isoenzimas/química , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/enzimologia , Homologia de Sequência de Aminoácidos , Especificidade da Espécie
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