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‣ Measures of degeneracy and redundancy in biological networks
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Publicado em 16/03/1999
Português
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Degeneracy, the ability of elements that are structurally different to perform the same function, is a prominent property of many biological systems ranging from genes to neural networks to evolution itself. Because structurally different elements may produce different outputs in different contexts, degeneracy should be distinguished from redundancy, which occurs when the same function is performed by identical elements. However, because of ambiguities in the distinction between structure and function and because of the lack of a theoretical treatment, these two notions often are conflated. By using information theoretical concepts, we develop here functional measures of the degeneracy and redundancy of a system with respect to a set of outputs. These measures help to distinguish the concept of degeneracy from that of redundancy and make it operationally useful. Through computer simulations of neural systems differing in connectivity, we show that degeneracy is low both for systems in which each element affects the output independently and for redundant systems in which many elements can affect the output in a similar way but do not have independent effects. By contrast, degeneracy is high for systems in which many different elements can affect the output in a similar way and at the same time can have independent effects. We demonstrate that networks that have been selected for degeneracy have high values of complexity...
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‣ Separation of phylogenetic and functional associations in biological sequences by using the parametric bootstrap
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Quantitative analyses of biological sequences generally proceed under the assumption that individual DNA or protein sequence elements vary independently. However, this assumption is not biologically realistic because sequence elements often vary in a concerted manner resulting from common ancestry and structural or functional constraints. We calculated intersite associations among aligned protein sequences by using mutual information. To discriminate associations resulting from common ancestry from those resulting from structural or functional constraints, we used a parametric bootstrap algorithm to construct replicate data sets. These data are expected to have intersite associations resulting solely from phylogeny. By comparing the distribution of our association statistic for the replicate data against that calculated for empirical data, we were able to assign a probability that two sites covaried resulting from structural or functional constraint rather than phylogeny. We tested our method by using an alignment of 237 basic helix–loop–helix (bHLH) protein domains. Comparison of our results against a solved three-dimensional structure confirmed the identification of several sites important to function and structure of the bHLH domain. This analytical procedure has broad utility as a first step in the identification of sites that are important to biological macromolecular structure and function when a solved structure is unavailable.
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‣ A biological function for cadmium in marine diatoms
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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The oceanic distribution of cadmium follows closely that of major algal nutrients such as phosphate. The reasons for this “nutrient-like” distribution are unclear, however, because cadmium is not generally believed to have a biological function. Herein, we provide evidence of a biological role for Cd in the marine diatom Thalassiosira weissflogii under conditions of low zinc, typical of the marine environment. Addition of Cd to Zn-limited cultures enhances the growth rate of T. weissflogii, particularly at low pCO2. This increase in growth rate is reflected in increased levels of cellular carbonic anhydrase (CA) activity, although the levels of TWCA1, the major intracellular Zn-requiring isoform of CA in T. weissflogii, remain low. 109Cd label comigrates with a protein band that shows CA activity and is distinct from TWCA1 on native PAGE of radiolabeled T. weissflogii cell lysates. The levels of the Cd protein are modulated by CO2 in a manner that is consistent with a role for this enzyme in carbon acquisition. Purification of the CA-active fraction leads to the isolation of a Cd-containing protein of 43 kDa. It is now clear that T. weissflogii expresses a Cd-specific CA, which, particularly under conditions of Zn limitation, can replace the Zn enzyme TWCA1 in its carbon-concentrating mechanism.
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‣ Accelerated reaction of nitric oxide with O2 within the hydrophobic interior of biological membranes
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Publicado em 03/03/1998
Português
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We demonstrate herein dramatic acceleration of aqueous nitric oxide (NO) reaction with O2 within the hydrophobic region of either phospholipid or biological membranes or detergent micelles and demonstrate that the presence of a distinct hydrophobic phase is required. Per unit volume, at low amounts of hydrophobic phase, the reaction of NO with O2 within the membranes is approximately 300 times more rapid than in the surrounding aqueous medium. In tissue, even though the membrane represents only 3% of the total volume, we calculate that 90% of NO reaction with O2 will occur there. We conclude that biological membranes and other tissue hydrophobic compartments are important sites for disappearance of NO and for formation of NO-derived reactive species and that attenuation of these potentially damaging reactions is an important protective action of lipid-soluble antioxidants such as vitamin E.
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‣ Iron-sulfur cluster disassembly in the FNR protein of Escherichia coli by O2: [4Fe-4S] to [2Fe-2S] conversion with loss of biological activity
Fonte: The National Academy of Sciences of the USA
Publicador: The National Academy of Sciences of the USA
Tipo: Artigo de Revista Científica
Publicado em 10/06/1997
Português
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The transcription factor FNR (fumarate nitrate reduction) requires the presence of an iron-sulfur (Fe-S) cluster for its function as a global transcription regulator in Escherichia coli when oxygen becomes scarce. To define the oxidation state and type of Fe-S cluster present in the active form of FNR, we have studied anaerobically purified FNR with Mössbauer spectroscopy. Our data showed that this form of FNR contained a [4Fe-4S]2+ cluster (δ = 0.45 mm/s; ΔEQ = 1.22 mm/s) and that the [4Fe-4S]2+ cluster was rapidly destroyed on exposure of FNR to air. Under these conditions, the yellow–green active form of FNR turned deep red; analysis of sulfide indicated that 70% of the labile sulfide was still present, suggesting that the Fe-S cluster had been converted into a different form. Little [3Fe-4S] cluster was, however, detected by EPR. According to Mössbauer spectroscopy, the [4Fe-4S]2+ cluster was converted in about 60% yield to a [2Fe-2S]2+ cluster (δ = 0.28 mm/s; ΔEQ = 0.58 mm/s) following 17 min of exposure to air. The [2Fe-2S]2+ cluster form of FNR was much more stable to oxygen, but was unable to sustain biological activity (e.g., DNA binding). However, DNA binding and the absorption spectrum characteristic of the [4Fe-4S]2+ cluster could be largely restored from the [2Fe-2S]2+ form when Cys...
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‣ Control of time-dependent biological processes by temporally patterned input
Fonte: The National Academy of Sciences of the USA
Publicador: The National Academy of Sciences of the USA
Tipo: Artigo de Revista Científica
Publicado em 16/09/1997
Português
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Temporal patterning of biological variables, in the form of
oscillations and rhythms on many time scales, is ubiquitous. Altering
the temporal pattern of an input variable greatly affects the output of
many biological processes. We develop here a conceptual framework for a
quantitative understanding of such pattern dependence, focusing
particularly on nonlinear, saturable, time-dependent processes that
abound in biophysics, biochemistry, and physiology. We show
theoretically that pattern dependence is governed by the nonlinearity
of the input–output transformation as well as its time constant. As a
result, only patterns on certain time scales permit the expression of
pattern dependence, and processes with different time constants can
respond preferentially to different patterns. This has implications for
temporal coding and decoding, and allows differential control of
processes through pattern. We show how pattern dependence can be
quantitatively predicted using only information from steady,
unpatterned input. To apply our ideas, we analyze, in an experimental
example, how muscle contraction depends on the pattern of motorneuron
firing.
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‣ Susceptibility to the biological effects of polyaromatic hydrocarbons is influenced by genes of the major histocompatibility complex
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Publicado em 08/12/1998
Português
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Polyaromatic hydrocarbons are ubiquitous environmental chemicals that are important mutagens and carcinogens. The purpose of this study was to determine whether genes within the major histocompatibility complex (MHC) influence their biological activities. Cell-mediated immunity to dimethylbenz(a)anthracene (DMBA) was investigated in congenic strains of mice. On three different backgrounds, H-2k and H-2a haplotype mice developed significantly greater contact-hypersensitivity responses to DMBA than H-2b, H-2d, and H-2s mice. In B10.A(R1) mice, which are Kk and Id, a vigorous contact-hypersensitivity response was present, indicating that the response was governed by class I, rather than class II, MHC genes. C3H/HeN (H-2k) and C3H.SW (H-2s) strains were also compared for the development of skin tumors and the persistence of DMBA–DNA adducts. When subjected to a DMBA initiation, phorbol 12-tetradecanoate 13-acetate (TPA)-promotion skin-tumorigenesis protocol, C3H/HeN mice, (which develop cell-mediated immunity to DMBA) were found to have significantly fewer tumors than C3H.SW mice (a strain that failed to develop a cell-mediated immune response to DMBA). DMBA–DNA adducts were removed more rapidly in C3H/HeN than in C3H.SW mice. The results indicate that genes within the MHC play an important role in several of the biological activities of carcinogenic polyaromatic hydrocarbons. The observations are consistent with the hypothesis that cell-mediated immunity to chemical carcinogens serves to protect individuals by removing mutant cells before they can evolve into clinically apparent neoplasms.
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‣ Digital in-line holography for biological applications
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Publicado em 25/09/2001
Português
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Digital in-line holography with numerical reconstruction has
been developed into a new tool, specifically for biological
applications, that routinely achieves both lateral and depth
resolution, at least at the micron level, in three-dimensional imaging.
The experimental and numerical procedures have been incorporated into a
program package with a very fast reconstruction algorithm that is now
capable of real-time reconstruction. This capability is demonstrated
for diverse objects, such as suspension of microspheres and biological
samples (diatom, the head of Drosophila melanogaster), and
the advantages are discussed by comparing holographic reconstructions
with images taken by using conventional compound light microscopy.
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‣ Mutation frequency and biological cost of antibiotic resistance in Helicobacter pylori
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Among the several factors that affect the appearance and
spread of acquired antibiotic resistance, the mutation frequency and
the biological cost of resistance are of special importance.
Measurements of the mutation frequency to rifampicin resistance in
Helicobacter pylori strains isolated from dyspeptic
patients showed that ≈1/4 of the isolates had higher mutation
frequencies than Enterobacteriaceae mismatch-repair
defective mutants. This high mutation frequency could explain why
resistance is so frequently acquired during antibiotic treatment of
H. pylori infections. Inactivation of the
mutS gene had no substantial effect on the mutation
frequency, suggesting that MutS-dependent mismatch repair is absent in
this bacterium. Furthermore, clarithromycin resistance conferred a
biological cost, as measured by a decreased competitive ability of the
resistant mutants in mice. In clinical isolates this cost could be
reduced, indicating that compensation is a clinically relevant
phenomenon that could act to stabilize resistant bacteria in a
population.
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‣ ADP ribosylation of human neutrophil peptide-1 regulates its biological properties
Fonte: The National Academy of Sciences
Publicador: The National Academy of Sciences
Tipo: Artigo de Revista Científica
Publicado em 11/06/2002
Português
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In human airways, epithelial cells lining the lumen and intraluminal cells (e.g., polymorphonuclear cells) participate in the innate immune response. These cells secrete or express on their surfaces arginine-specific ADP ribosyltransferases. Defensins, antimicrobial proteins secreted by immune cells, are arginine-rich, leading us to hypothesize that ADP ribosylation could modify their biological activities. We found that an arginine-specific ADP ribosyltransferase-1 present on airway epithelial cells modifies Arg-14 of α defensin-1. ADP-ribosylated defensin-1 had decreased antimicrobial and cytotoxic activities but still stimulated T cell chemotaxis and IL-8 release from A549 cells. Further, ADP-ribosylated defensin-1 inhibited cytotoxic and antimicrobial activities of unmodified defensin-1. We identified ADP-ribosylated defensin-1 in bronchoalveolar lavage fluid from smokers but not from nonsmokers, confirming its existence in vivo. Thus, airway mono-ADP-ribosyltransferases could have an important regulatory role in the innate immune response through modification of α defensin-1 and perhaps other basic molecules, with alteration of their biological properties.
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‣ Biological models and genes of tumor reversion: Cellular reprogramming through tpt1/TCTP and SIAH-1
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Tumor reversion is the process by which some cancer cells lose their malignant phenotype. This study was aimed at defining some of the molecular and phenotypic properties of this process. Biological models of tumor reversion were isolated from human leukemia and breast cancer cell lines by using the H-1 parvovirus as a selective agent. Differential gene expression analysis was performed between the parental malignant cells and their revertants or alternatively between these parental cells and their SIAH-1 transfectant counterparts. These SIAH-1 transfectants have a suppressed malignant phenotype and were used as a control for a viral-free system. Two hundred sixty-three genes were found to be either activated or inhibited during the reversion process, as confirmed by Northern blot analysis or quantitative PCR. Of these, 32% were differentially expressed in all systems, irrespective of whether parvovirus-selected, SIAH-1 overexpressing, or p53 mutant or wild-type cell lines were used, suggesting the existence of a universal mechanism underlying tumor reversion. Translationally Controlled Tumor Protein (tpt1/TCTP) has the strongest differential expression, down-regulated in the reversion of U937- and SIAH-1-overexpressing cells. Inhibition of TCTP expression by anti-sense cDNA or small interfering RNA molecules results in suppression of the malignant phenotype and in cellular reorganization...
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‣ Demasking biological oscillators: Properties and principles of entrainment exemplified by the Neurospora circadian clock
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Oscillations are found throughout the physical and biological worlds. Their interactions can result in a systematic process of synchronization called entrainment, which is distinct from a simple stimulus-response pattern. Oscillators respond to stimuli at some times in their cycle and may not respond at others. Oscillators can also be driven if the stimulus is strong (or if the oscillator is weak); i.e., they restart their cycle every time they receive a stimulus. Stimuli can also directly affect rhythms without entraining the underlying oscillator (masking): Drivenness and masking are often difficult to distinguish. Here we use the circadian biological clock to explore properties of entrainment. We confirm previous results showing that the residual circadian system in Neurospora can be entrained in a mutant of the clock gene frequency (frq9, a strain deficient in producing a functional FRQ protein). This finding has implications for understanding the evolution of circadian programs. By comparing data sets from independent studies, we develop a template for analyzing, modeling, and dissecting the interactions of entrained and masked components. These insights can be applied to oscillators of all periodicities.
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‣ Effects of threshold choice on biological conclusions reached during analysis of gene expression by DNA microarrays
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Global analysis of gene expression by using DNA microarrays is employed increasingly to search for differences in biological properties between normal and diseased tissue. In such studies, expression that deviates from defined thresholds commonly is used for creating genetic signatures that characterize disease vs. normality. Although it is axiomatic that the threshold parameters applied to microarray analysis will alter the contents of such genetic signatures, the extent to which threshold choice can affect the fundamental conclusions made from microarray-based studies has not been elucidated. We used gabriel (Genetic Analysis By Rules Incorporating Expert Logic), a platform of knowledge-based algorithms for the global analysis of gene expression, together with conventional statistical approaches, to examine the sensitivity of conclusions to threshold choice in recently published microarray-based studies. An analysis of the effects of threshold decisions in one of these studies [Ramaswamy, S., Ross, K. N., Lander, E. S. & Golub, T. R. (2003) Nat. Genet. 33, 49–54], which arrived at the important conclusion that the metastatic potential of primary tumors is encoded by the bulk of cells in the tumor, is the focus of this article. We discovered that support for this conclusion highly depends on the threshold used to create gene expression signatures. We also found that threshold choice dramatically affected the gene function categories represented nonrandomly in signatures. Our results suggest that the robustness of biological conclusions made by using microarray analysis should be routinely assessed by examining the validity of the conclusions by using a range of threshold parameters.
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‣ High biological species diversity in the arctic flora
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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The arctic flora is considered to be impoverished, but estimates of species diversity are based on morphological assessments, which may not provide accurate counts of biological species. Here we report on crossing relationships within three diploid circumpolar plant species in the genus Draba (Brassicaceae). Although 99% of parental individuals were fully fertile, the fertility of intraspecific crosses was surprisingly low. Hybrids from crosses within populations were mostly fertile (63%), but only 8% of the hybrids from crosses within and among geographic regions (Alaska, Greenland, Svalbard, and Norway) were fertile. The frequent occurrence of intraspecific crossing barriers is not accompanied by significant morphological or ecological differentiation, indicating that numerous cryptic biological species have arisen within each taxonomic species despite their recent (Pleistocene) origin.
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‣ RabGEF1 regulates stem cell factor/c-Kit-mediated signaling events and biological responses in mast cells
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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We recently reported that RabGEF1 is a negative regulator of high-affinity Fc receptor for IgE (FcεRI)-dependent mast cell activation and that mice lacking RabGEF1 develop severe skin inflammation and increased numbers of dermal mast cells. To better understand how RabGEF1 can regulate signaling events and biological responses in mast cells, we examined the responses of bone marrow-derived cultured mast cells (BMCMCs) from wild-type (+/+) and Rabgef1 knockout (−/−) mice after stimulation with the c-Kit ligand, stem cell factor (SCF), an important regulator of mast cell development, survival, proliferation, and activation. We found that RabGEF1-deficient mast cells exhibited enhanced and prolonged activation of Ras and extracellular regulated kinase, and significantly elevated IL-6 secretion, after stimulation with SCF. SCF-induced activation of c-Jun N-terminal kinase was increased in Rabgef1−/− BMCMCs, but without corresponding significant increases in SCF-induced migration or adhesion. SCF-mediated activation of the survival-enhancing kinase, Akt, also was increased in Rabgef1−/− BMCMCs, and these cells had a survival advantage over their +/+ counterparts in vitro. Despite enhanced Ras activation in the absence of RabGEF1...
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‣ A chemical approach to unraveling the biological function of the glycosylphosphatidylinositol anchor
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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The glycosylphosphatidylinositol (GPI) anchor is a C-terminal posttranslational modification found on many eukaryotic proteins that reside in the outer leaflet of the cell membrane. The complex and diverse structures of GPI anchors suggest a rich spectrum of biological functions, but few have been confirmed experimentally because of the lack of appropriate techniques that allow for structural perturbation in a cellular context. We previously synthesized a series of GPI anchor analogs with systematic deletions within the glycan core and coupled them to the GFP by a combination of expressed protein ligation and native chemical ligation [Paulick MG, Wise AR, Forstner MB, Groves JT, Bertozzi CR (2007) J Am Chem Soc 129:11543–11550]. Here we investigate the behavior of these GPI-protein analogs in living cells. These modified proteins integrated into the plasma membranes of a variety of mammalian cells and were internalized and directed to recycling endosomes similarly to GFP bearing a native GPI anchor. The GPI-protein analogs also diffused freely in cellular membranes. However, changes in the glycan structure significantly affected membrane mobility, with the loss of monosaccharide units correlating to decreased diffusion. Thus, this cellular system provides a platform for dissecting the contributions of various GPI anchor components to their biological function.
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‣ Regulated programmed lysis of recombinant Salmonella in host tissues to release protective antigens and confer biological containment
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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We have devised and constructed a biological containment system designed to cause programmed bacterial cell lysis with no survivors. We have validated this system, using Salmonella enterica serovar Typhimurium vaccines for antigen delivery after colonization of host lymphoid tissues. The system is composed of two parts. The first component is Salmonella typhimurium strain χ8937, with deletions of asdA and arabinose-regulated expression of murA, two genes required for peptidoglycan synthesis and additional mutations to enhance complete lysis and antigen delivery. The second component is plasmid pYA3681, which encodes arabinose-regulated murA and asdA expression and C2-regulated synthesis of antisense asdA and murA mRNA transcribed from the P22 PR promoter. An arabinose-regulated c2 gene is present in the chromosome. χ8937(pYA3681) exhibits arabinose-dependent growth. Upon invasion of host tissues, an arabinose-free environment, transcription of asdA, murA, and c2 ceases, and concentrations of their gene products decrease because of cell division. The drop in C2 concentration results in activation of PR, driving synthesis of antisense mRNA to block translation of any residual asdA and murA mRNA. A highly antigenic α-helical domain of Streptococcus pneumoniae Rx1 PspA was cloned into pYA3681...
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‣ Protein contents in biological membranes can explain abnormal solvation of charged and polar residues
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Transmembrane helices are generally believed to insert into membranes based on their hydrophobicity. Nevertheless, there are important exceptions where polar residues have great functional importance, for instance the S4 helix of voltage-gated ion channels. It has been shown experimentally that insertion can be accomplished by hydrophobic counterbalance, predicting an arginine insertion cost of only 2.5 kcal/mol, compared with 14.9 kcal/mol in cyclohexane. Previous simulations of pure bilayers have produced values close to the pure hydrocarbon, which has lead to spirited discussion about the experimental conditions. Here, we have performed computer simulations of models better mimicking biological membranes by explicitly including protein helices at mass fractions from 15% to 55%, as well as an actual translocon. This has a striking effect on the solvation free energy of arginine. With some polar residues present, the solvation cost comes close to experimental observation at approximately 30% mass fraction, and negligible at 40%. In the presence of a translocon in the membrane, the cost of inserting arginine next to the lateral gate can be as low as 3–5 kcal/mol. The effect is mainly due to the extra helices making it easier to retain hydration water. These results offer a possible explanation for the discrepancy between the in vivo hydrophobicity scale and computer simulations and highlight the importance of the high protein contents in membranes. Although many membrane proteins are stable in pure bilayers...
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‣ Dissociable contribution of prefrontal and striatal dopaminergic genes to learning in economic games
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Game theory is used throughout the social and biological sciences to study behavior in social interactions. Recent research suggests an important role for the dopamine neurotransmitter system in these types of decisions. This study used a competitive game to study how people varied in their decision-making processes and related these differences in the set of genes that carry out biological functions required for dopaminergic functioning. We found that genes differentially expressed in separate brain regions influenced distinct components of people’s decision-making processes and that a surprising degree of consistency exists with what is known at the brain level about how people make decisions in social interactions.
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‣ Allostatic load as a marker of cumulative biological risk: MacArthur studies of successful aging
Fonte: National Academy of Sciences
Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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Allostatic load (AL) has been proposed as a new conceptualization
of cumulative biological burden exacted on the body through attempts to
adapt to life's demands. Using a multisystem summary measure of AL, we
evaluated its capacity to predict four categories of health outcomes, 7
years after a baseline survey of 1,189 men and women age 70–79. Higher
baseline AL scores were associated with significantly increased risk
for 7-year mortality as well as declines in cognitive and physical
functioning and were marginally associated with incident cardiovascular
disease events, independent of standard socio-demographic
characteristics and baseline health status. The summary AL measure was
based on 10 parameters of biological functioning, four of which are
primary mediators in the cascade from perceived challenges to
downstream health outcomes. Six of the components are secondary
mediators reflecting primarily components of the metabolic syndrome
(syndrome X). AL was a better predictor of mortality and decline in
physical functioning than either the syndrome X or primary mediator
components alone. The findings support the concept of AL as a measure
of cumulative biological burden.
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