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Sex difference in platelet aggregation detected by new aggregometry using light scattering.

Earlier studies in platelet aggregation have shown that females seemed to have greater aggregability than males as detected by conventional aggregometry which used light transmission (LT), but controversy still remains. This study was performed to determine whether sex difference exists in platelet aggregation by using the recently developed laser light scattering (LS) method, which can detect small aggregates (i.e., two or three platelets). Blood was drawn from healthy volunteers (10 male and 10 female in follicular phase after menstruation), and platelet aggregation was detected by either LT or LS method in platelet rich plasma. Platelet aggregation was stimulated by increasing concentration of adenosine 5'-diphosphate (ADP, 0, 0.5, 1 and 2 microM). To detect the effect of sex hormones, platelets were incubated with estradiol (10 nM) or testosterone (40 nM) for 30 min, then platelet aggregation studies were performed. LT method revealed that female had greater aggregability than male. With weak stimuli (< or = 1 microM ADP), LS method showed that females had more medium aggregates than males, and that testosterone decreased small aggregates, and that estradiol decreased all sizes of aggregates. These data suggest that the female is more conductive to platelet aggregation than the male at a physiologic concentration of ADP (< or = 1 microM), but that both estradiol (10 nM) and testosterone (40 nM) have countereffects on platelet aggregation at the same condition. Therefore, the reason why females have greater aggregability than males may partly be explained by their lack of testosterone, but the mechanism still remains to be elucidated.

Adenosine Diphosphate↗

Disease mapping and spatial regression with count data.

In this paper, we provide critical reviews of methods suggested for the analysis of aggregate count data in the context of disease mapping and spatial regression. We introduce a new method for picking prior distributions, and propose a number of refinements of previously used models. We also consider ecological bias, mutual standardization, and choice of both spatial model and prior specification. We analyze male lip cancer incidence data collected in Scotland over the period 1975-1980, and outline a number of problems with previous analyses of these data. In disease mapping studies, hierarchical models can provide robust estimation of area-level risk parameters, though care is required in the choice of covariate model, and it is important to assess the sensitivity of estimates to the spatial model chosen, and to the prior specifications on the variance parameters. Spatial ecological regression is a far more hazardous enterprise for two reasons. First, there is always the possibility of ecological bias, and this can only be alleviated by the inclusion of individual-level data. For the Scottish data, we show that the previously used mean model has limited interpretation from an individual perspective. Second, when residual spatial dependence is modeled, and if the exposure has spatial structure, then estimates of exposure association parameters will change when compared with those obtained from the independence across space model, and the data alone cannot choose the form and extent of spatial correlation that is appropriate.

Epidemiologic Methods↗

Integrating health-related data from various sources: combining surveys, records and routine data.

The paper discusses the necessity of combining data from various sources in order to enhance their usefulness for a variety of applications. As future data use can hardly be forseen in advance, major data sets in health services should fulfill several formal requirements in order to make them suitable for future linkage. These formal requirements are that there be references to defined populations, to specific persons, to defined time periods, to specific places or regions. It would be necessary for terms, definition and classification schemes to agree between data sets which are to be linked and be in wide use. Three facets of data linkage are discussed specifically namely linking data at one level of aggregation, linking different data components, and combining data sets from different sources at several levels of aggregation. Three examples are provided, describing linkages of data from various sources for epidemiological studies and a study in health services research. They show that at this point in descriptive epidemiological studies linkage on the basis of regions is of great importance. This implies that it would be desirable for large scale data collection activities in health services to provide for a uniform representation of the geographic areas. Such uniformity would greatly enhance the linkage potential of data sets and thus their usefulness for small area and regional analyses.

Adult↗

Global impairment of the ubiquitin-proteasome system by nuclear or cytoplasmic protein aggregates precedes inclusion body formation.

The highly conserved ubiquitin-proteasome system (UPS) controls the stability of most nuclear and cytoplasmic proteins and is therefore essential for virtually all aspects of cellular function. We have previously shown that the UPS is impaired in the presence of aggregated proteins that become deposited into cytoplasmic inclusion bodies (IBs). Here, we report that production of protein aggregates specifically targeted to either the nucleus or cytosol leads to global impairment of UPS function in both cellular compartments and is independent of sequestration of aggregates into IBs. The observation of severe UPS impairment in compartments lacking detectable aggregates or aggregation-prone protein, together with the lack of interference of protein aggregates on 26S proteasome function in vitro, suggests that UPS impairment is unlikely to be a consequence of direct choking of proteasomes by protein aggregates. These data suggest a common proteotoxic mechanism for nuclear and cytoplasmic protein aggregates in the pathogenesis of neurodegenerative disease.

Cell Line↗

Involvement of thromboxane A2 and tyrosine kinase in the synergistic interaction of platelet activating factor and calcium ionophore A23187 in human platelet aggregation.

The present study was carried out to examine the mechanisms of the synergistic interaction of PAF and A23187 mediated platelet aggregation. We found that platelet aggregation mediated by subthreshold concentrations of PAF (5 nM) and A23187 (1 mM) was inhibited by PAF receptor blocker (WEB 2086, IC50 = 0.65 mM) and calcium channel blockers, diltiazem (IC50 = 13 mM) and verapamil (IC50 = 18 mM). Pretreatment of platelets with PAF and A23187 induced rise in intracellular calcium and this effect was also blocked by verapamil. While examining the role of the down stream signaling pathways, we found that platelet aggregation induced by the co-addition of PAF and A23187 was also inhibited by low concentrations of phospholipase C (PLC) inhibitor (U73122; IC50 = 10 mM), a cyclooxygenase inhibitor (indomethacin; IC50 = 0.2 mM) and inhibitor of TLCK, herbimycin A with IC50 value of 5 mM. The effect was also inhibited by a specific TXA2 receptor antagonist, SQ 29548 with very low IC50 value of 0.05 mM. However, the inhibitors of MAP kinase, PD98059 and protein kinase C, chelerythrine had no effect on PAF and A23187-induced platelet aggregation. These data suggest that the synergism between PAF and A23187 in platelet aggregation involves activation of thromboxane and tyrosine kinase pathways.

Benzoquinones↗

Organization and representation of patient safety data: current status and issues around generalizability and scalability.

Recent reports have identified medical errors as a significant cause of morbidity and mortality among patients. A variety of approaches have been implemented to identify errors and their causes. These approaches include retrospective reporting and investigation of errors and adverse events and prospective analyses for identifying hazardous situations. The above approaches, along with other sources, contribute to data that are used to analyze patient safety risks. A variety of data structures and terminologies have been created to represent the information contained in these sources of patient safety data. Whereas many representations may be well suited to the particular safety application for which they were developed, such application-specific and often organization-specific representations limit the sharability of patient safety data. The result is that aggregation and comparison of safety data across organizations, practice domains, and applications is difficult at best. A common reference data model and a broadly applicable terminology for patient safety data are needed to aggregate safety data at the regional and national level and conduct large-scale studies of patient safety risks and interventions.

Humans↗

Studies of the regulation and reaction mechanism of the carbamyl phosphate synthetase and aspartate transcarbamylase of bakers' yeast.

Kinetic studies of the carbamyl phosphate synthetase activity (CPSase) of bakers' yeast revealed an absolute requirement for K+ ions ; KM values for two of the substrates, glutamine and bicarbonate, were found to be 5 X 10(-4) M and 3 X 10(-3) M respectively. CPSase activity of the purified enzyme aggregate (M.W. 800,000) was extremely sensitive to UTP with a Ki of 2.4 X 10(-4) M. The purine nucleotide intermediate, XMP, was a strong activator of CPSase, acting at a site different from the regulatory site at which UTP binds ; XMP activation diminished at high concentrations of the substrate Mg-ATP. Studies of the reaction mechanism of CPSase revealed that it involved the sequential addition of the substrates bicarbonate and Mg-ATP, liberation of ADP, addition of glutamine, binding of ATP and then release of ADP and the product carbamyl phosphate. Studies of the reaction mechanism of the aspartate transcarbamylase (ATCase) of the aggregate yielded data which were not compatible with any of the usual models ; whichever reaction mechanism is ultivately found to fit the data, it will probably prove applicable both to the ATCase of the aggregate and to the disaggregated ATCase subunit (MW 138,000).

Adenosine Triphosphate↗

Protein aggregation: folding aggregates, inclusion bodies and amyloid.

Aggregation results in the formation of inclusion bodies, amyloid fibrils and folding aggregates. Substantial data support the hypothesis that partially folded intermediates are key precursors to aggregates, that aggregation involves specific intermolecular interactions and that most aggregates involve beta sheets.

Amyloid↗

p values for pathogens: statistical inference from infectious-disease data.

Certain features of infectious-disease data-including the aggregated nature of the data, confounding variables, correlated variables, and non-linear relations-complicate the use of standard statistical procedures. Using data on a helminth infection, we review the use of three parametric tests (analysis of variance, linear regression, and logistic regression), address the complications arising from violation of the assumptions for these tests, and suggest methods of correction. We also compare the relative merits of parametric methods with equivalent non-parametric approaches, and illustrate the differences produced with results from a Kruskal-Wallis test and a t test. The value of using a resampling method-bootstrapping-is also shown. Finally, we discuss problems arising from use of a study design that requires data on the same attribute to be collected from the same individual over a period of time, and present three methods for overcoming this complication, showing that, in the example used, the mixed effect model and generalised estimating equation give similar results.

Analysis of Variance↗

Brightness of yellow fluorescent protein from coral (zFP538) depends on aggregation.

The yellow fluorescent protein from coral (zFP538) forms aggregates in water solutions. According to dynamic light scattering and gel filtration data, the aggregation number is approximately 1000-10000 at pH 8-9 and protein concentration 1 mg/mL. Gel filtration demonstrated that dissociation of the aggregates takes place upon dilution, and the molecular weight of the aggregates decreases with pH. Atomic force microscopy (AFM) and near-field scanning optical microscopy (NSOM) were used to obtain images of zFP538 in the solid state. It was shown that protein films are comprised of fluorescent ellipsoidal granules with a 50-300 nm major axis and a 30-130 nm minor axis. The dependence of zFP538 fluorescence on protein concentration between 1.2 x 10(-)(9) and 5.5 x 10(-)(7) M can be divided in two linear regions with different slopes indicating the existence of at least two different forms of zFP538. The fluorescence of zFP538 decreases with time upon acidification, and the decrease depends on pH and protein concentration. Between pH 3.5 and pH 5.5, relative residual fluorescence is higher for concentrated zFP538 solutions (about 10(-)(6) M) as compared with diluted ones (10(-)(7) M and below). Aggregation makes zFP538 more stable against fluorescence quenching upon acidification: the decrease in zFP538 fluorescence at protein concentration 1 mg/mL is completely reversible, unlike that observed for less concentrated solutions. This phenomenon may be due to the decrease in the freedom of chromophore mobility in zFP538 aggregates.

Animals↗

Regional variation in medical classification agreement: benchmarking the coding gap.

The growing use of classification and coding of patient data in medical information systems has resulted in increased dependence on the accuracy of coding practices. Information maintained on systems must be trusted by both providers and managers in order to serve as a viable tool for the delivery of healthcare in an evidence-based environment. A national survey of health information managers was employed here to assess observed levels of coder agreement with physician code selections used in classifying patient data. Findings from this survey suggest that, on a national level, the quality of coded data may suffer as a result of disagreement or inconsistent coding within healthcare provider organizations, in an era where physicians are increasingly called upon to enter and classify patient data via computerized medical records. Nineteen percent of respondents report that coder-physician classification disagreement occurred on more than 5% of all patient encounters. In some cases disagreement occurs in 20% or more instances of code selection. This phenomenon occurred to varying degrees across regions and market areas, suggesting a confounding influence when coded data is aggregated for comparative purposes. In an evidence-based healthcare environment, coded data often serves as a representation of clinical performance. Given the increasing complexity of medical information classification systems, reliance on such data may pose a risk for both practitioners and managers without consistent agreement on coding practices and procedures.

Attitude of Health Personnel↗

Examining geographic patterns of mortality: the atlas of mortality in small areas in Spain (1987-1995).

BACKGROUND: Small-area mortality atlases have been demonstrated to be a useful tool for both showing general geographical patterns in mortality data and identifying specific high-risk locations. In Spain no study has so far systematically examined geographic patterns of small-area mortality for the main causes of death. This paper presents the main features, contents and potential uses of the Spanish Atlas of Mortality in small areas (1987-1995). METHODS: Population data for 2,218 small areas were drawn from the 1991 Census. Aggregated mortality data for 14 specific causes of death for the period 1987-1995 were obtained for each small area. Empirical Bayes-model-based estimates of age-adjusted relative risk were displayed in small-area maps for each cause/gender/age group (0-64 or 65 and over) combination using the same range of values (i.e. septiles) and colour schemes. RESULTS: The 'Spanish Atlas of Mortality' includes multiple choropleth (area-shaded) small-area maps and graphs to answer different questions about the data. The atlas is divided into three main sections. Section 1 includes the methods and comments on the main maps. Section 2 presents a two-page layout for each leading cause of death by gender including 1) a large map with relative risk estimates, 2) a map that indicates high- and low-risk small areas, 3) a graph with median and interquartile range of relative risk estimates for 17 large regions of Spain, and 4) relative-risk maps for two age groups. Section 3 provides specific information on the geographical units of analysis, statistical methods and other supplemental maps. CONCLUSION: The 'Spanish Atlas of Mortality' is a useful tool for examining geographical patterns of mortality risk and identifying specific high-risk areas. Mortality patterns displayed in the atlas may have important implications for research and social/health policy planning purposes.

Adolescent↗

Relationship between platelet membrane lipid compositions and platelet aggregability in alcoholic liver disease.

We studied the relationship between changes in platelet aggregability and platelet membrane lipid in alcoholic liver disease. The maximal rate of ADP-induced platelet aggregation was significantly increased in the alcoholic liver disease group than in the control group. No significant difference was observed in the maximal rate of collagen-induced platelet aggregation. However, a lag time required for the start of platelet aggregation was significantly shortened in the alcoholic liver disease group, indicating increased platelet aggregability. Results of the platelet aggregation test suggested that alcoholic liver disease patients have their platelet aggregation affected by the abnormality of prostaglandin metabolism. The alcoholic liver disease group was further divided into two subgroups: the hyperaggregation group and the unchanged aggregation group. Both free cholesterol and phospholipid in the platelet membrane were significantly increased in the alcoholic liver disease group. In phospholipid compositions, phosphatidylserine plus phosphatidylinositol were significantly decreased in the alcoholic liver disease group, whereas a significant decrease in phosphatidylserine plus phosphatidylinositol was observed in the hyperaggregation group of alcoholic liver disease. Analysis of fatty acid compositions of platelet membrane showed significantly decreased palmitic acid in the alcoholic group. There was no significant change of arachidonic acid, which directly affects platelet aggregation. Eicosapentaenoic acid significantly decreased in the alcoholic liver disease group, but there was no difference in docosahexaenoic acid. Meanwhile, the thrombogenic index, calculated from the fatty acids of platelet membrane, showed no difference between the alcoholic liver disease group and the control group. However, the thrombogenic index was significantly increased in the hyperaggregation group than in the unchanged aggregation group. These data suggested that platelet aggregation is affected by not only a change in arachidonic acid, but also changes in fatty acid compositions of the platelet membrane.

Adult↗

Season of birth in valvular heart disease.

This study demonstrates seasonal variations of birth dates in children with congenital valvular heart disease and in adults dying from valvular heart disease. The findings are based on the 1989-98 Swiss EUROCAT data, and on 1969-94 Swiss mortality records. Seasonality was tested with aggregated monthly data using Edwards' procedure. Both data sets showed excesses between December and March, consistent in different forms of valvular disease and in both sexes. Despite the decline of rheumatic heart disease, risk factors causing season of birth effects remain relevant for congenital anomalies.

Adult↗

[Effect of urea on aggregation of immunoglobulins in water solutions].

The methods of molecular light scattering and of gel filtration were used to study the degree of urea-induced desaggregation of immunoglobulin (IgG) molecules preaggregated by heating. Urea was shown to desaggregate considerably (70--80%) protein IgG aggregates. The data obtained suggest an important role of hydrophobic interactions in the immunoglobulin aggregation.

Immunoglobulin G↗

Photodynamic inactivation of cultured bladder tumor cells: a preliminary study of the effects of porphyrin aggregation.

This study was designed to determine the influence, if any, of intracellular porphyrin aggregation upon in vitro photosensitivity. HpD uptake conditions were manipulated in such a way as to yield three experimental groups, each containing equal levels of cellular porphyrins but having different porphyrin aggregation status. Aggregation status was determined through comparison of the spectral properties of intact cells. Subsequent exposure to red light resulted in a family of survival curves which generally corresponded to cellular HpD fluorescence yields and optical densities (indicators of aggregation status). These data suggest that porphyrin aggregation within the cell may be partially responsible for the observed differential photosensitivity following various uptake conditions.

Biological Transport↗

Purification and functional characterization of homodimeric gamma B-gamma B fibrinogen from rat plasma.

Heterogeneity in the human and rat plasma fibrinogen (FBG) gamma chains is due to differential splicing of the primary gamma chain mRNA transcript. The subunit composition of FBG containing the predominant form of the gamma chain is homodimeric (human: A alpha 2, B beta 2, gamma 50-gamma 50; rat: A alpha 2, B beta 2, gamma A-gamma A). The subunit composition of FBG containing the longer, minor form of the gamma chain is heterodimeric (human: A alpha 2, B beta 2, gamma 50-gamma 57.5; rat: A alpha 2, B beta 2, gamma A-gamma B). The variant gamma chain-FBGs comprise about 10% of the total plasma FBG in the human and 30% in the rat. Although the presence of homodimeric gamma B-gamma B FBG has been speculated, proof of its existence has been difficult to obtain experimentally. We now show that 5% to 6% of rat plasma FBG is found as homodimeric FBG with a subunit composition of A alpha 2, B beta 2, gamma B-gamma B. Commercially purified rat FBG was further purified by diethylaminoethyl-Sephacel chromatography with a combined pH and ionic strength gradient. The enriched gamma B-gamma B FBG fraction eluted at the lowest pH (6.3) and highest ionic strength (4.5 mmho) due to its increased net negative charge. To further purify gamma B-gamma B FBG, a Mono Q column with an NaCl gradient was used. The subunit composition of the purified gamma B-gamma B FBG was confirmed by its electrophoretic mobility under reducing and denaturing conditions both as FBG, and clotted and cross-linked fibrin. Homodimeric gamma B-gamma B FBG was unable to support adenosine diphosphate (ADP)-induced platelet aggregation, whereas heterodimeric gamma A-gamma B FBG was able to support ADP-induced platelet aggregation at 40% of that achieved with homodimeric gamma A-gamma A FBG, similar to previous observations with human FBGs. Taken together, these results support the hypothesis that the A alpha-RGD sites alone are not sufficient for dimeric FBG support of platelet aggregation. Furthermore, the data suggest that at least one intact platelet recognition site at the carboxyterminus of the gamma A or gamma 50 chain is required.

Amino Acid Sequence↗

Aggregation of colon cancer in family data.

Although only a small proportion of common cancers show familial aggregation, studying such families can elucidate the roles of shared environment and genes in the development of neoplasia. We report an analysis of nine colon cancer pedigrees using new nonparametric objective methods to measure familial aggregation as a means of determining the existence of heterogeneity in the data. Each family was selected through a proband with nonpolyposis colon cancer who had a first-degree relative with documented colon cancer. To assess the aggregation of different cancers in these families we employ a method which evaluates both excess number of cases as well as distribution by risk in family members. We find that eight of the nine families exhibit significant aggregation of colon cancer: endometrial cancer aggregates in three families, breast in none, kidney in one, and all sites in eight. In this way, we show that two families fit the criteria for Cancer Family Syndrome, and that one is not a high-risk cancer family.

Biometry↗