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PubMed1988

The primary wave of epinephrine-induced platelet aggregation represents alpha 2-adrenoceptor status.

We examined relationships between epinephrine-induced slope of primary wave of aggregation and the alpha 2-adrenoceptor status on platelets. A concentration (10(-9) to 10(-6]-dependent increase in slope of primary wave with EC50 of epinephrine at 4.5 +/- 0.4 x 10(-7) was observed. In studies on epinephrine binding to alpha 2-adrenoceptors in competition with 3H-yohimbine to platelets, (IC50) of epinephrine was 4.8 +/- 3.4 x 10(-7) M. There was a significant (P less than or equal to 0.02) correlation between EC50 of epinephrine to evoke biological response and IC50 of epinephrine to bind to alpha 2-adrenoceptors (r-0.75). There was no relationship between number of receptor sites or dissociation constant of 3H-yohimbine binding and primary wave of platelet aggregation. These data show that the slope of primary wave in response to epinephrine reflects alpha 2-adrenoceptor binding of the agonist.

Adolescent↗

PubMed1987

Binding of a thromboxane A2/prostaglandin H2 agonist [3H]U46619 to washed human platelets.

The binding characteristics of [3H]U46619 to washed human platelets were studied. [3H]U46619 binding to washed human platelets was saturable and displaceable. Kinetic studies yielded a Kd of 11 +/- 4 nM (n = 4). Scatchard analysis of equilibrium binding studies revealed one class of high affinity binding sites with a Kd of 20 +/- 7 nM and a Bmax of 9.1 +/- 2.3 fmole/10(7) platelets (550 +/- 141 binding sites per platelet) (n = 4). A number of compounds that act as either agonists or antagonists of the TXA2/PGH2 receptor were tested for their ability to inhibit the binding of [3H]U46619 to washed human platelets. The Kds of the agonists and antagonists were similar to their potencies to induce or inhibit platelet aggregation. These data provide some evidence that [3H]U46619 binds to the putative human platelet TXA2/PGH2 receptor.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

PubMed1991

Physicochemical characterization of a minor mucin component from cystic fibrosis tracheobronchial secretions.

A minor mucin glycoprotein component (HTM-2) was purified from the tracheobronchial secretions of two cystic fibrosis patients using a protocol established in our laboratory. The secretions were solubilized in 0.1 M Tris-HCl buffer (pH 7.5) containing 0.22 M potassium thiocyanate and fractionated on a Bio-Gel A-5m column, followed by digestion with DNAase, rechromatography on the same column and chromatography on hydroxyapatite which resolved the major mucin (HTM-1) from the minor mucin component (HTM-2). The mucin component HTM-2 was further purified using Superose 6 chromatography. SDS-composite gel (2% polyacrylamide + 0.5% agarose) and 6% polyacrylamide gel electrophoresis showed that the purified HTM-2 was totally free of low-molecular-weight contaminants. Equilibrium density sedimentation centrifugation of purified HTM-2 using CsCl gradients also showed the absence of proteoglycans and other low-molecular-weight proteins. Comparison of carbohydrate and amino acid compositions of the two mucin components indicated that HTM-2 was quite different from the major mucin, HTM-1, reported earlier from our laboratory (Biochemistry, 24, 7334, 1985). This suggested that HTM-2 has a different polypeptide core and is perhaps a different gene product. The effects of 6 M guanidine-HCl and different concentrations of NaCl on the molecular size of HTM-2 and its ability to form aggregates was also investigated using the technique of static light scattering. In buffer containing 6 M guanidine-HCl, HTM-2 had a weight-average molecular weight of approximately 4.5 x 10(6). However, in the presence of buffer containing 0.03, 0.10 or 0.15 M NaCl, the molecular weight of HTM-2 was estimated to be approximately 11 x 10(6). These data suggest aggregation of HTM-2 in the presence of a range of NaCl concentrations. In contrast to HTM-1, which is a more anionic glycoprotein, the apparent molecular size of HTM-2 did not decrease at the higher NaCl concentration.

Amino Acids↗

PubMed1987

Automated multivariate measurement of spontaneous motor activity in mice: time course and reliabilities of the behavioral measures.

A variety of automated procedures have been developed to measure certain aspects of spontaneous motor activity in small animals. The present study used a Digiscan Animal Activity Monitor to measure six different aspects of spontaneous motor behavior in male CF-1 mice. The Digiscan system uses infrared beams and computer analysis to quantify various behavioral variables. The mice were tested for 1 hour on 2 different days of the week for 3 consecutive weeks. Both the temporal changes in the measured variables and the test-retest reliabilities were examined in a group of 30 mice. Statistical analysis of the data revealed significantly higher mean values for total movement time, average distance travelled, and horizontal activity on the first test session relative to the second session (p less than 0.01). The other 3 measures, total distance travelled, number of movements, and average speed, did not vary significantly across test sessions. All 6 behavioral variables showed good test-retest reliabilities and these could be increased by aggregating the data on a weekly basis. The present results indicate that the measures obtained from the Digiscan system are reliable and that the animals should first be habituated to the test apparatus in order to obtain stable baseline activity values.

Animals↗

PubMed2006

Case-control and case-only designs with genotype and family history data: estimating relative risk, residual familial aggregation, and cumulative risk.

In case-control studies of inherited diseases, participating subjects (probands) are often interviewed to collect detailed data about disease history and age-at-onset information in their family members. Genotype data are typically collected from the probands, but not from their relatives. In this article, we introduce an approach that combines case-control analysis of data on the probands with kin-cohort analysis of disease history data on relatives. Assuming a marginally specified multivariate survival model for joint risk of disease among family members, we describe methods for estimating relative risk, cumulative risk, and residual familial aggregation. We also describe a variation of the methodology that can be used for kin-cohort analysis of the family history data from a sample of genotyped cases only. We perform simulation studies to assess performance of the proposed methodologies with correct and mis-specified models for familial aggregation. We illustrate the proposed methodologies by estimating the risk of breast cancer from BRCA1/2 mutations using data from the Washington Ashkenazi Study.

BRCA1 Protein↗

PubMed1995

Influences of silicates and carnitine-silicate mixtures on the inhibition of aggregation of erythrocytes elicited by the presence of fibrinogen.

Carnitine and acylcarnitine derivatives have been reported to inhibit cell aggregation (Fritz and Burdzy, 1989, J. Cell. Physiol., 140:18-28). A follow-up of these observations showed that whereas the previously described effects of long-chain acylcarnitines were well replicated, those of carnitine on erythrocytes showed marked variability. The latter phenomenon was traced to the presence of silicates in carnitine solutions derived from the use of sodium hydroxide solutions stored in glass containers for the neutralization of carnitine. The present experiments have led to the discovery that oligomeric forms of silicates are powerful inhibitors of red blood cell aggregation which otherwise occurs in the presence of fibrinogen alone. The active form(s) of silicates in this assay, which appear to be generated by polymerization of silicates in metasilicate solutions on neutralization, are unstable and therefore transient under usual conditions. We estimate that the active oligomeric forms contain between 4 to 18 silicon atoms per molecule. When maintained at -18 degrees C in the presence of carnitine, but not in its absence, the active forms of oligomeric silicates remained stable for months, judging from their ability to inhibit cell aggregation. We conclude that carnitine stabilized the oligomeric form(s) of silicate, or that the species stabilized is an oligomeric silicate-carnitine complex. Comparable concentrations of choline, deoxycarnitine, or gamma-aminobutyrate were less effective in stabilizing the active silicate oligomers. The active forms of the silicate oligomers had Ki values of about 10 microM, calculated as the monomeric form, in inhibiting red blood cell aggregation. The data indicate that free carnitine does not directly inhibit erythrocyte inhibition, as previously interpreted, whereas long-chain acylcarnitine derivatives are active in the absence of silicates. Possible mechanism of actions of silicate oligomers on membranes are discussed.

Carnitine↗

PubMedDate not supplied

Modifications of platelet shape change and ATP release during cardiopulmonary bypass.

Platelet activation is accompanied by characteristic morphological changes: smooth-disc platelets become more spherical in shape and develop psudopods. The purpose of this study is to investigate whether platelets change after extracorporeal bypass. Twenty-two patients undergoing cardiopulmonary bypass (CPB) were studied prior to anesthesia and immediately after the operation. Platelets activated by different agonists were monitored simultaneously for morphological changes, ATP release and aggregation. While shape change measured before surgery was large, it was significantly reduced after bypass surgery (p < 0.01); morphological changes were quicker postoperatively. Several other parameters also changed: the time lapse between administration of the agonist and the start of ATP secretion decrease significantly (p < 0.01). After activation with high concentrations of ADP, ATP release was significantly increased (p < 0.01). On the other hand, less ATP was released after platelet activation with collagen and arachidonic acid, suggesting a change in platelet adhesion or a downregulation of endoperoxide synthesis. In our study, the importance of preactivation change in shape, estimated quantitatively by percent loss in ability of changing shape, can be compared using various agonists, with proportional defects in release and aggregation. These data provide evidence for different intrinsic levels in platelet defects after CPB.

Adenosine Diphosphate↗

PubMed2005

The pre-vaccination regional epidemiological landscape of measles in Italy: contact patterns, effort needed for eradication, and comparison with other regions of Europe.

BACKGROUND: Strong regional heterogeneity and generally sub-optimal rates of measles vaccination in Italy have, to date, hampered attainment of WHO targets for measles elimination, and have generated the need for the new Italian National Measles Elimination Plan. Crucial to success of the plan is the identification of intervention priorities based upon a clear picture of the regional epidemiology of measles derived from the use of data to estimate basic parameters. Previous estimates of measles force of infection for Italy have appeared anomalously low. It has been argued elsewhere that this results from Italian selective under-reporting by age of cases and that the true measles force of infection in Italy is probably similar to that of other European countries. A deeper examination of the evidence for this conjecture is undertaken in the present paper. METHODS: Using monthly regional case notifications data from 1949 to the start of vaccination in 1976 and notifications by age from 1971-76, summary equilibrium parameters (force of infection (FOI), basic reproductive ratio (R0) and critical vaccination coverage (pc)) are calculated for each region and for each of 5 plausible contact patterns. An analysis of the spectra of incidence profiles is also carried out. Finally a transmission dynamics model is employed to explore the correspondence between projections using different estimates of force of infection and data on seroprevalence in Italy. RESULTS: FOI estimates are lower than comparable European FOIs and there is substantial regional heterogeneity in basic reproductive ratios; certain patterns of contact matrices are demonstrated to be unfeasible. Most regions show evidence of 3-year epidemic cycles or longer, and compared with England & Wales there appears to be little synchronisation between regions. Modelling results suggest that the lower FOI estimated from corrected aggregate national data matches serological data more closely than that estimated from typical European data. CONCLUSION: Results suggest forces of infection in Italy, though everywhere remaining below the typical European level, are historically higher in the South where currently vaccination coverage is lowest. There appears to be little evidence to support the suggestion that a higher true force of infection is masked by age bias in reporting.

Journal Article↗

PubMed2002

Sinus headache or migraine? Considerations in making a differential diagnosis.

Sinus headache is commonly diagnosed, and patients with headache often cite sinus pain and pressure as a cause of their headaches. A high frequency of diagnosis of sinus headache, which specialists consider to be relatively rare, among patients meeting International Headache Society (IHS) diagnostic criteria for migraine raises the possibility that migraine and perhaps other headache types are sometimes mistaken for sinus headache. This article considers clinical, epidemiologic, and pathophysiologic relationships between sinus headache and migraine and discusses the implications for clinical management of headache. Both historic and new data show that nasal symptoms frequently accompany migraine, although these symptoms are not part of the IHS diagnostic criteria for migraine. Parasympathetic activation, as well as the hypothesized mechanism of neurogenic or immunogenic switching (i.e., crossover interactions of neurogenic and immunogenic inflammation), may account for both the frequent occurrence of nasal symptoms in migraine and the possibility that sinus inflammation can sometimes act as a migraine trigger. Considered in aggregate, the data show that the occurrence of nasal symptoms associated with a headache should neither trigger a diagnosis of sinus disease nor exclude a diagnosis of migraine. It should, in fact, prompt diagnostic consideration of both conditions.

Diagnosis, Differential↗

PubMed1981

Modulation of platelet aggregation by native DNA - initial description of platelet receptor type, number and discrimination for native DNA.

Native DNA (dsDNA) induces the aggregation of isolated human platelets. Using isotopically labeled dsDNA (125I-dsDNA) and Scatchard analysis, a single class of platelet receptor was detected with a KD = 190 pM and numbering approximately 275/platelet. This receptor was discriminatory in that heat denatured dsDNA, poly A, poly C, poly C x I and poly C x poly I failed to substantially inhibit either the platelet binding of, or platelet aggregation induced by, dsDNA; by themselves, these polynucleotides were ineffective as platelet agonists. However, poly G, poly I and poly G x I effectively and competitively inhibited platelet binding of the radioligand, independently activated the platelet and when used at a sub-activating concentration decreased the extent of dsDNA stimulated platelet aggregation. These data depict a receptor on human platelets for dsDNA and perhaps certain additional polynucleotides and relate receptor-ligand interactions to a physiologic platelet function.

Animals↗

PubMed1997

Binding of a thrombin receptor tethered ligand analogue to human platelet thrombin receptor.

A thrombin receptor-radioligand binding assay was developed using [3H]A(pF-F)R(ChA)(hR)Y-NH2 ([3H]haTRAP), a high affinity thrombin receptor-activating peptide (TRAP), and human platelet membranes. Scatchard analysis of saturation binding data indicated that [3H]haTRAP bound to platelet membranes with a Kd of 15 nM and a Bmax of 5.2 pmol/mg of protein. The binding was reduced by GPPNHP, a nonmetabolizable GTP analogue. Various TRAPs and a TRAP antagonist, but not other receptor agonists, displaced [3H]haTRAP from the binding sites. SFLLRN-NH2, a thrombin receptor-tethered ligand analogue, and [3H]haTRAP exhibited competitive binding for the same binding sites. The relative affinity of these peptides for the binding site paralleled their EC50 or IC50 values for platelet aggregation. These data indicate that [3H]haTRAP binds specifically and saturably to the functioning G protein-linked thrombin (tethered ligand) receptor in human platelet membranes.

Dose-Response Relationship, Drug↗

PubMed2026

Likelihood-based optimization enables accurate copy number estimation for paralogous genes using exome data.

MOTIVATION: Exome sequencing is widely used for genetic studies; however, accurate detection of copy number variants (CNV) in paralogous genes is challenging due to short-read mapping ambiguity and extensive copy-number variation. The human genome contains several hundred paralogous genes, many of which are known to harbor disease-associated CNVs. Existing exome CNV callers are primarily designed for rare CNV detection in uniquely mappable regions and are not well-suited for paralogous genes. METHODS: We describe a computational method (EdgeCopy) for copy number profiling of paralogous genes using whole-exome sequence data. EdgeCopy aggregates reads mapped to all copies of paralogous genes and relates observed read depth to copy number for multiple exome samples using an approximate composite likelihood function. The likelihood function is optimized using numerical optimization to obtain gene-level fractional copy number estimates that are discretized and refined using a Hidden Markov Model to obtain exon-level copy number estimates. RESULTS: Benchmarking of Edgecopy using experimental copy number data showed high concordance (mean = 0.973) for six disease-associated paralogous genes. We evaluated performance using whole-exome data from approximately 2400 samples across five continental populations from the 1000 Genomes Project. EdgeCopy shows robust concordance with whole-genome sequencing based estimates (0.974-0.982) across populations and 130 paralogous genes spanning a wide range of copy-number variation. In comparison, copy number analysis using a state-of-the-art exome CNV caller failed to estimate copy number for paralogous genes with very high mapping ambiguity and showed much lower concordance (0.565) for CNV events compared to EdgeCopy (0.908). AVAILABILITY: EdgeCopy is freely available at https://github.com/vibansal-lab/edgecopy.

Humans↗

PubMed1999

Effect of a reorganized after-hours family practice service on frequent attenders.

BACKGROUND AND OBJECTIVES: A governmental reorganization of the after-hours general practice service in Denmark was launched in January 1992. The biggest change was the introduction of mandatory county-wide telephone triage systems staffed by general practitioners. This study assesses the effect of this reorganization on the use of services by frequent attenders (FAs). METHODS: From 1990 to 1994, methods of contact and annual costs per attender were analyzed in an ecological time-trend study based on aggregated administrative data collected from the database of the Public Health Insurance, Aarhus County, Denmark (600,000 inhabitants). The study only included attenders ages 18 and over. FAs were defined as the group that, within each calendar year (12 months), had 4 or more contacts with the after-hours family practice service. RESULTS: FAs made up 9.5% of the attenders and accounted for more than 40% of the contacts and the aggregate costs. The effect of the reorganization was a 12% decrease in the number of attenders, a 16% decrease in the number of contacts, and a 29% decrease in the costs. Reorganization had a significantly bigger effect on FA attendance than on non-FA attendance, and more than half of the overall reduction in use of services could be ascribed to changes in FA behavior. CONCLUSIONS: The reorganization of the after-hours service produced a significant fall in attendance and costs, especially with respect to adult FAs.

Adolescent↗

PubMed1992

The effects of 7.5% NaCl/6% dextran 70 on coagulation and platelet aggregation in humans.

The combination solution of 7.5% NaCl/6% dextran 70 (HSD) administered IV gives hemodynamic improvement in the treatment of hemorrhagic hypotension. Since earlier dextran solutions were reported to interfere with blood coagulation, the effects of HSD on the prothrombin time (PT), the activated partial thromboplastin time (APTT), platelet aggregation, and platelet concentration were studied. The HSD mixed with human plasma (1:5 and 1:10) slightly prolonged PT, but had no effect on the APTT, compared with saline controls. The HSD also decreased human platelet aggregation at the 1:5 dilution. In separate mixing studies, the hypertonic saline component of HSD was associated with the prolongation of PT and decreased platelet aggregation. The data from these studies indicate that at its proposed therapeutic dose, HSD is expected to have minimal effect on blood coagulation.

Adult↗

PubMed2000

Employment-seeking experiences of residents in 1996: a window into the neurology marketplace.

OBJECTIVE: To assess career choice and employment-seeking experience of senior neurology residents in 1996. METHODS: Graduating residents in adult and pediatric neurology (n = 573) were surveyed to obtain career plans, initial job selection, health care attitudes, and demographic information. Results were compared with 1996 data on all United States neurologists and data from an American Medical Association (AMA) resident survey regarding the employment status of new physicians. RESULTS: Survey response rate was 71%. There was a significant increase in international medical graduates and women entering neurology compared with the current workforce. Seventy-four percent of graduates planned to enter a fellowship position; 19%, private practice; 5%, an academic position; and 2%, a career outside of clinical medicine. Neurology residents differ from aggregate national data because only 28% of residents responding to an AMA survey across all specialties applied to fellowships in 1996. Overall, 44% of neurology graduates planned an academic career. CONCLUSIONS: Changing demographics and career choice of recent graduates may require continued monitoring and could be important in neurology workforce planning. The high rate of fellowship training and plans for academic careers in 1996 is of interest and may reflect both resident response to new demands in the changing health care market and a need to modify residency programs to enhance academic training and competitiveness of neurology graduates.

Adult↗

PubMed2003

The effect of hypercholesterolemia on platelet soluble guanylyl cyclase.

We sought to determine whether hypercholesterolemia impacts on the NO-stimulated activity of platelet soluble guanylyl cyclase (sGC). We investigated two groups of nine New Zealand white rabbits receiving either a standard (NC) or a cholesterol chow (HC, 0.75%) for 15 weeks. The plasma content of cGMP and the specific activity of sGC in intact platelets were measured by a cGMP-specific radioimmunoassay. In HC, 47.9+/-3.1% of the aortic intimal area was covered with atherosclerotic lesions, and plasma cGMP levels (pmol/ml) were increased from 12.6+/-1.2 to 27.9+/-3.5 (P<.0001). In striking contrast, hypercholesterolemia had no effect on sGC activity stimulated by the NO donor SNAP. At 100 microM SNAP, the specific activities of sGC (pmol/10(9) platelets/min) were 81.8+/-14.5 in NC and 86.2+/-8.1 in HC. Basal sGC activity (pmol/10(9) platelets/min) was also similar in NC (0.21+/-0.04) and HC (0.460+/-0.11, P=.7813). In accordance, washed platelets from both groups showed a similar SNAP-induced inhibition of aggregation. These data suggest that an impaired response of platelets to NO is most likely not involved in platelet hyperreactivity in hypercholesterolemia.

Animals↗

PubMed2000

Outlet density and criminal violence in Norway, 1960-1995.

OBJECTIVE: To analyze the relationship between criminal violence and number of outlets for on-premise sales of alcohol. METHOD: The data comprise aggregate time series for Norway for the period 1960-95. Two crime indicators were used: (1) crimes of violence investigated by the police per 100,000 inhabitants aged 15 years and above (15+) and (2) convictions for criminal violence per 100,000 inhabitants (15+). Outlet density was measured as number of public drinking places per 10,000 inhabitants (15+). RESULTS: On the basis of autoregressive integrated moving average (ARIMA) analyses of differenced data, a positive and statistically significant relationship (p = .03) was found between outlet density and crimes of violence investigated by the police. This replicates the findings that are reported from cross-sectional studies. The relationship was also positive, but of borderline significance (p = .06), when convictions for criminal violence were used as outcome. CONCLUSIONS: The findings suggest that increases in the number of public drinking places are associated with increases in criminal violence. Although other kinds of data support the findings, these are the first to be based on longitudinal data. To test for robustness and cultural specificity it, therefore, seems warranted to replicate the study on data from other drinking cultures.

Alcoholic Beverages↗

PubMed2006

Bifenthrin activates homotypic aggregation in human T-cell lines.

BACKGROUND: Here, we addressed the concern that, despite the lack of overt toxicity, exposure to low levels of the common household pyrethroid pesticide, bifenthrin, could cause harm to the immune system. To do this, we measure the effect of bifenthrin on phytohemagglutinin (PHA) activation of homotypic aggregation in human T-cell lines. MATERIAL/METHODS: The human CD4+ H9, and Jurkat cell lines and the human promonocyte U937 cell line, were exposed to varying concentrations of bifenthrin. Cell viability was determined using the AlmarBlue Toxicity Assay. Concentrations of bifenthrin which did not reduce cell viability were determined and these concentrations were tested for the effect of bifenthrin on PHA-mediated homotypic aggregation. Blocking antibodies to ICAM and LFA-1 were used to disrupt aggregation and a nonspecific IgG was used as a control. RESULTS: Bifenthrin was found to be nontoxic at concentrations ranging from 10(-4) to 10(-13) M. Bifenthrin did not inhibit PHA induced cell aggregation in all cell lines tested. However, at 10(-4) M, bifenthrin to form aggregates stimulated homotypic aggregation in the H9 and Jurkat T-cell lines. The bifenthrin-induced aggregate formation, like that seen with PHA, was blocked by treating the cells with antibodies to either LFA-1 or ICAM. CONCLUSIONS: The results here show that bifenthrin activates T-cell function by stimulating ICAM/LFA-1 mediated homotypic aggregation. This data suggests that exposure to bifenthrin, even at "acceptable" limits, can increase the risk for and frequency of inflammatory responses and diseases such as asthma.

Animals↗