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Aboriginal new world epidemiolgy and medical care, and the impact of Old World disease imports.

Various workers, including T. D. Stewart, claim that the aboriginal Americas were relatively disease-free because of the bering Strait cold-screen, eliminating many pathogens, and the paucity of zoonotic infections because of few domestic animals. Evidence of varying validity suggests that precontact Americns had their own strains of treponemic infections, bacillary and amoebic dysenteries, influenza and viral penumonia and other respiratory diseases, salmonellosis and perhaps other food poisoning, various arthritides, some endoparasites such as the ascarids, and several geographically circumscribed diseases such as the rickettsial verruca (Carrion's disease) and New World leishmaniasis and trypanosomiasis. Questionably aboriginal are tuberculosis and typhus. Accordingly, virtually all the "crowd-type" ecopathogenic diseases such as smallpox, yellow fever, typhoid, malaria, measles, pertussis, polio, etc., appear to have been absent from the New World, and were only brought in by White conquerors and their Black slaves. My hypothesis is that native American medical care systems--especially in the more culturally advanced areas--were sufficiently sophisticated to deal with native disease entities with reasonable competence. But native medical systems could not cope with the "crowd-type" disease imports that struck Indian and Eskimos as "virgin-field" populations. Reanalysis of native population losses through a genocidal combination of diease, war, slavery and attendant cultural disruption by Dobyns, Cook and others strongly suggest that traditiona estimates underplayed the death toll by a factor of the general order of ten. This would make for an immediately pre-contact Indian population of some 90-111 million instead of the tradition 8-11 million. Evidence is growing that Indians may have been no more susceptible to new pathogens that are other "virgin soil" populations, and thus their immune systems need not be considered less effective than those in other people. Present-day high mortality rates in Indians of both continents from infectious disease imports may be more socioeconomic than anything else.

Delivery of Health Care↗

Further morphometric studies of the ulna from the Omo Basin, Ethiopia.

A reanalysis of the L40-19 fossil ulna from the Omo Basin, Ethiopia is presented. Using covariance adjustment rather than ratios to correct metric data for variations due to body size, a comparative sample reflecting 21 contemporary Anthropoid taxa, and both distribution-free and multivariate statistical procedures, this study indicates that the earlier conclusion drawn by McHenry et al. ('76), viz., that the fossil is "unique" among Hominoids, is essentially correct. This study also concludes, however, that although the fossil is projected closer to Pan and Homo than to Pongo, the distances are considerably greater than found between behaviorally similar modern forms. Consequently, aside from assigning the fossil on morphometric grounds to the Hominoidea, little else can be said about its possible locomotor habits or its ancestry.

Animals↗

Pubic symphysis age distributions.

A further discussion of age assessment and palaeodemography requires detailed reviews of methods, especially pubic symphysis techniques. Before reanalysis of changes in symphyseal form, the initial steps in distributing ages must be examined. Use of the mean values for age scores gives age distributions that are not real, but subject to systematic distortions, and cumulative percentages of skeletal samples can be shown to reflect the mean ages. Distributing skeletal ages using 95% probability distributions provides a more accurate estimation of true ages for palaeodemography and a better basis for discussions of pubic symphysis aging techniques.

Adolescent↗

On aspects of skull form in African apes and orangutans, with implications for hominoid evolution.

The study of hominoid phylogeny is currently in a state of controversy and debate due to the discovery of new fossil material and reanalysis of the morphology of extant apes. An important key to the resolution of these debates lies in attaining a fuller understanding of the morphological differences in skull form between the African and Asian great apes. In this paper I have analyzed aspects of facial morphology and internal cranial anatomy in the great apes. Results from this study and previous ones suggest that Pongo is characterized by a marked dorsal deflection of the face relative to the basicranium. Many aspects of circumorbital, midfacial, palatal, and mandibular morphology in Pongo may be related to this airorynchous condition. This hypothesis is supported by Enlow's work on form and pattern in the primate and mammalian skull. The position of the face in known Sivapithecus appears to be similar to that seen in Pongo. Although Pongo may be specialized in its marked degree of airorynchy, it seems likely that an important derived feature linking African apes and hominids is a ventral rotation of the splanchnocranium on the neurocranium. The appearance of marked supraorbital tori and ethmofrontal sinuses are probably correlated developments. Additional implications of this work for debates about hominoid phylogeny are discussed.

Animals↗

Generic level relationships of the Papionini (Cercopithecoidea).

Phylogenetic hypotheses for the Old World monkey tribe Papionini based on molecular data are incongruent with those inferred from previous morphological analyses. Morphologists have often inferred a close relationship between Mandrillus and Papio based on their overall similarity. Theropithecus has been variously proposed to be either quite distantly related to these two genera, their sister taxon, or anywhere in between. Molecular and chromosomal analyses on the other hand unambiguously group Theropithecus and Papio together to the exclusion of Mandrillus. Additionally, molecular and chromosomal analyses reveal that mangabeys (Cerocebus) are paraphyletic. Morphologists have acknowledged this possibility resurrecting the genus name Lophocebus for one group of mangabeys. A review and reanalysis of the morphological characters put forth by various researchers find little to contradict the consensus phylogeny derived from analysis of chromosomal banding, nuclear RNA restriction mapping, alpha and beta hemoglobin sequences, albumin and transferrin microcomplement fixation, DNA-DNA hybridization, repetitive DNA patterns, immunodiffusion, hemoglobin and adenylate kinase isozymes, and mitochondrial cytochrome oxidase subunit II DNA sequences.

Animals↗

Analysis of dental attrition and mortality in the Medieval village of Tirup, Denmark.

New directions and new questions raised in the study of health in the past justify this reanalysis of the pattern of dental attrition in the Medieval Danish population of Tirup. Dental attrition was scored on all permanent molars from the Tirup skeletal sample. Scores were analyzed by means of logistic regression of the probability of having entered a given stage of wear for a given tooth in a way that is very similar to transition analysis. The primary determinant of dental attrition was age at death. In addition to age, the effects of sex, side, and dating were analyzed. In order to assess the homogeneity of the process of wearing teeth down, a third-order polynomial in age-at-death was also fitted to the transition probabilities. It was found that age is the single most important determinant of dental attrition, and that sex or side did not differentiate the rate of attrition. In several transitions, there was evidence of heterogeneity, indicating both random and systematic interpersonal differences in the rate of attrition and an association between the rate of attrition and age-at-death. It was found that attrition proceeded more quickly after AD 1300 than prior to that date. It is suggested that this was due to a possible general deterioration of living conditions in Northern Europe and an increased reliance on grain for food during the first half of the 14th century. The temporal effect on attrition rate accounts for some but not all the observed heterogeneity wear.

Age Factors↗

Reduction of joint pain in patients with knee osteoarthritis who have received monthly telephone calls from lay personnel and whose medical treatment regimens have remained stable.

OBJECTIVE: We previously reported that monthly telephone contact by lay personnel, to promote self-care for patients with osteoarthritis (OA), was associated with improved joint pain and physical function after 1 year of followup. The present study was a secondary analysis to determine whether improvement was contingent on intensified medical treatment. METHODS: We reanalyzed control/treatment group differences in all 40 subjects with radiographically confirmed knee OA who had had no changes in antirheumatic drug therapy or institution of physical therapy during the period of observation. RESULTS: Group differences in measured pain remained significant (effect size [ES] = 0.65 SD, P less than 0.01). The same trend was observed for physical function (ES = 0.53 SD, P not significant). CONCLUSION: The findings in this reanalysis suggest that periodic telephone support interventions are effective enough to be regarded as an adjunctive treatment for OA.

Aged↗

Simplification of the Wertheimer-Leeper wire code.

Wire codes, introduced by Wertheimer and Leeper as a surrogate for residential magnetic fields, divide imputed exposure into several categories based on the configurations of electrical wiring within 40 m of homes. Using the data set gathered by Savitz et al. in the Denver, Colorado, area of the United States, we determined which of the wiring attributes that contribute to Wertheimer-Leeper coding are actually associated with low-power magnetic fields measured in bedrooms of subjects. The results led us to propose a considerably simplified three-category form of the Wertheimer-Leeper code that 1) drops the distinctions between thick and thin primary wires and between first-span and other secondary lines, 2) adds a new distinction between open (i.e., conductors not in physical contact) and spun secondaries, and 3) explains as much of the between-home variability in log-transformed bedroom fields as does the five categories of the original Wertheimer-Leeper code. The data necessary to classify residences using the modified code are considerably simpler to obtain and should lead to more reliable results. A separate reanalysis of the Denver data set of Savitz et al, shows that the modified code yields risk estimates that are both precise and markedly elevated for the highest exposure category, suggesting that this code may be useful in other studies.

Colorado↗

A validated chiral HPLC method for the determination of mebeverine HCl enantiomers in pharmaceutical dosage forms and spiked rat plasma.

A new, precise, simple and accurate HPLC method was developed for the first time to separate and determine mebeverine enantiomers. Enantiomeric resolution was achieved on a cellulose Tris (3,5-dimethylphenyl carbamate) column known as Chiralcel OD, with UV detection at 263 nm. The mobile phase consisted of n-hexane, isopropyl alcohol and triethylamine (90:9.9:0.1 v/v/v). Sample run time was 18 min. On using the chromatographic conditions described, mebeverine enantiomers were well resolved with mean retention times of about 11 and 14 min. A linear response (r>0.999) was observed over the concentration range 0.5-20 microg/mL racemic mebeverine. Precision, accuracy and stability were studied according to ICH guidelines. The limit of detection was found to be 0.05 microg/mL for each enantiomer of mebeverine. The proposed method was applied for analysis of mebeverine in commercially available tablets dosage formulations. Examples of application to biological samples are also given. Reanalysis of samples several weeks after the initial analysis showed no degradation of mebeverine.

Animals↗

Screening techniques for the detection of inborn errors of bile acid metabolism by direct injection and micro-high performance liquid chromatography-continuous flow/fast atom bombardment mass spectrometry.

Two methods, direct injection-continuous flow fast atom bombardment (DI-CF/FAB) mass spectrometry and micro-high-performance liquid chromatography (mu HPLC)-CF/FAB) mass spectrometry were developed for the clinical analysis of urinary bile acids and their conjugates, specifically for the diagnosis of disorders of bile acid metabolism. The method based upon DI of urine extracts into the CF/FAB flow stream was developed for the rapid screening of large numbers of patient samples. It allows injections to be made at 4 min intervals for high sample throughput. Analyses of standard mixtures of bile salts demonstrate that linear response curves are obtained over more than a hundred-fold concentration range with a minimum detectable amount in the picogram range. Oxo bile salts are identified by reduction with sodium borodeuteride followed by reanalysis for detection of any reduction products. This methodology has been used for the analysis of over 1000 specimens. A reverse-phase mu HPLC-CF/FAB mass spectrometric method was also developed for use in the confirmation or further investigation of screening results. With no additional sample preparation this method permits the separation, identification and quantitation of urinary bile salt isomers within a 45 min analysis time.

Bile Acids and Salts↗

Improved survival after acute myocardial infarction in patients with advanced Killip class.

BACKGROUND: The continuing applicability of the Killip classification system to the effective stratification of long-term and short-term outcome in patients with acute myocardial infarction (MI) and its influence on treatment strategy calls for reanalysis in the setting of today's primary reperfusion treatments. HYPOTHESIS: Our study sought to test the hypothesis that Killip classification, established on admission in patients with acute MI, is an effective tool for early prediction of in-hospital mortality and long-term survival. METHODS: A series of 909 consecutive Olmsted County patients admitted with acute MI to St. Marys Hospital, Mayo Clinic, between January 1988 and March 1998 was analyzed. Killip classification was the primary variable. Endpoints were in-hospital death, major in-hospital complications, and post-hospital death. RESULTS: Patients analyzed included 714 classified as Killip I, 170 classified as Killip II/III, and 25 classified as Killip IV. Increases in in-hospital mortality and prevalence of in-hospital complications correspond significantly with advanced Killip class (p < 0.01), with in-hospital mortality 7% in class I, 17.6% in classes II/III, and 36% in class IV patients (p < 0.001). Killip classification was strongly associated with mode of therapy administered within 24 h of admission (p < 0.01). Killip IV patients underwent primary angioplasty most commonly and were less likely to receive medical therapy. CONCLUSIONS: Killip classification remains a strong independent predictor of in-hospital mortality and complications, and of long-term survival. Early primary angioplasty has contributed to a decrease in mortality in Killip IV patients, but effective adjunctive medical therapy is underutilized.

Aged↗

A multigene test for the risk of sporadic breast carcinoma.

BACKGROUND: Although the identification of the BRCA1 and BRCA2 genes have been of great interest, these genes account for less than 5% of all breast carcinoma cases. The remaining cases are sporadic. Reanalysis of a large twin study suggested that genetic factors may play a significant role in sporadic breast and other carcinomas. Sporadic breast carcinoma is polygenically inherited. Multiple genes are likely to have an additive effect, each gene accounting for a fraction of the variance. One factor that may have an impact on the development of hormonally responsive breast tumors is the duration of exposure of the breast to estrogen. Therefore, one of the demographic risk factors for breast carcinoma is an early age of onset of menarche. The current study was based on the hypothesis that genes that play a role in demographic risk factors may be breast carcinoma risk genes in their own right. The authors hypothesized that six genes relevant to the timing of the onset of menarche and related risk factors might be candidate genes for breast carcinoma. These were the leptin gene (LEP), the leptin receptor gene (LEPR), the catechol-0-methyltransferase gene (COMT), the dopamine D(2) receptor gene (DRD2), the estrogen 1 receptor gene (ESR1), and the androgen receptor gene (AR). METHODS: The authors examined 67 women with postmenopausal sporadic breast carcinoma and 145 gender and race-matched controls. RESULTS: Five of these genes accounted for a significant percent of the variance (r(2)) of breast carcinoma. The following r(2) and P values were calculated: LEP: 0.073, P < or = 0.0001; LEPR: 0.064, P < or = 0.0002; COMT: 0.073, P < or = 0.0001; AR: 0.040, P < or = 0.0035; and DRD2: 0.018, P < or = 0.05. When evaluated in a multivariate regression analysis, they accounted collectively for 24% of the variance of breast carcinoma (P < or = 0.0001). These genes accounted for 40% of the variance (P < or = 0.00001) in a subset of age-matched cases. Individual gene scores were added to form a breast carcinoma risk score (BCRS) that ranged from 0 to 17. When the BCRS was evaluated in a receiver operator characteristic plot, the area under the curve was 0.80 for the full set and 0.869 for the age-matched set. The relative breast carcinoma risk for the different BCRS scores ranged from 0.10 to 11.9. CONCLUSIONS: These results demonstrate a potentially powerful method of evaluating the additive effect of multiple breast carcinoma risk genes to form a potentially clinically useful assessment of women's risk for sporadic breast carcinoma.

Adult↗

Three-color immunofluorescence analysis of Leu antigens on human peripheral blood using two lasers on a fluorescence-activated cell sorter.

A fluorescence-activated cell sorter was modified to quantify simultaneously three immunofluorescence stains on a population of cells. A dye laser containing rhodamine 6G was used to obtain 600 nm light to excite Texas Red coupled avidin. An argon ion laser operating at 488 nm excited both fluorescein and phycoerythrin directly conjugated antibodies. The emission from these fluorophores could be independently quantified as demonstrated by the histograms generated by samples labeled separately with each of the three stains. This three-color detection system was used to analyze human peripheral blood mononuclear cells for the expression of three antigens: Leu 2, Leu 7, and Leu 11. Upon reanalysis of the list mode data, several discrete subpopulations of lymphocytes could be identified based on the quantitative expression of these three antigens. Of particular interest in the normal sample studied was a population of dimly labeled Leu 2+ cells which were predominantly Leu 11+, a phenotype which is seen infrequently in normal individuals. This technique expands the combinations of antigens that can be studied at any one time and will facilitate the detection and functional analysis of cells in heterogeneous populations.

Antibodies, Monoclonal↗

Single-cell analysis of the relationship among transferrin receptors, proliferation, and cell cycle phase in K562 cells.

Multiparameter single-cell analysis by flow cytometry was used to distinguish between size-related changes in K562 cell transferrin receptor (TfR) expression and changes in membrane receptor density throughout the cell cycle and over time in culture. Light-scatter pulse-width time-of-flight, a direct and readily calibrated measure of cell diameter, was used to calculate receptor density as the average number of receptors per unit cell surface area. Cell surface TfRs were unimodally distributed over the cell population and were present throughout the cell cycle. The number of receptors increased as cells progressed through the cell cycle, but cell cycle phase was also correlated with cell volume. However, when size heterogeneity was factored out by reanalysis of listmode data, there was a clear cell-cycle effect: among cells of the same size, both the number of receptors per cell and the receptor density increased from G1 to S to G2/M. TfR expression was also followed over time in culture after dilution into fresh medium. A decrease in growth rate after four days was preceded by one to two days by a decrease in both number of TfRs per cell and mean receptor density, indicating that decreased TfR expression represented true "down-regulation" and not just decreased cell size or an increase in the proportion of smaller G1 cells. This type of analysis is generally applicable for resolving the effects of cell size heterogeneity and cell cycle on membrane protein distribution and for other studies of ligand-receptor interaction.

Cell Cycle↗

Reproducibility of FCM DNA content from replicate paraffin block samples.

Fifty-five paraffin blocks of morphologically and clinically normal colon specimens were divided in half and the two halves treated as separate aliquots for determination of DNA content using propidium iodide staining and FCM analysis. Forty-five of the 55 samples showed a peak channel difference between the two aliquots from the same block of greater than 1 channel. Seven of the 55 specimens showed peak channel difference greater than or equal to 10 channels. Similar results were seen if the mean channel of the G0G1 peak was used in place of the peak channel, with 5 of the 55 specimens showing mean channel differences of greater than or equal to 10 channels. The seven samples with peak channel difference greater than or equal to 10 channels were reprocessed and five of the seven showed reduced peak channel difference on reanalysis. The mean peak channel difference of the 55 samples was 4.3 +/- 10.5 (mean +/- 2 S.D.) using the first analysis. These data suggest that caution should be used in interpreting ploidy of nuclei extracted from paraffin block material if the interpretation is based on comparing two aliquots from the same block, even if they are processed and analyzed in the same batch.

Cell Nucleus↗

Regenerative and predictive medicine of cardiovascular disease: the 9th Leipziger Workshop and the 2nd International Workshop on slide based cytometry.

Slide-based cytometry (SBC) and related techniques offer unique tools to perform complex immunophenotyping, thereby enabling diagnostic procedures at very early disease stages. Multicolor or polychromatic analysis of cells by SBC is of special importance, not only as a cytomics technology platform but also for patients with low blood volume such as neonates. The exact knowledge of the location of each cell on the slide allows restaining and subsequent reanalysis of the specimen. These separate measurements of the same specimen can be fused to one data file (merging), thus increasing the information obtained per cell. Relocalization and optical evaluation of the cells, a feature typical of SBC, can be of integral importance for cytometric analysis. Due to this feature, artifacts can be excluded and morphology of measured cells can be documented. Predictive medicine aims at the detection of changes in patient's state before the manifestation of the disease or its complications. Such instances concern multiorgan failure in sepsis or noninfectious posttraumatic shock in intensive care patients or the pretherapeutic identification of high-risk patients undergoing cancer cytostatic therapy. Early anti-infectious or antishock therapy and curative chemotherapy in combination with stem cell transplantation may provide better chances of patients' survival at concomitant cost containment. Predictive medicine that guides early individualized decrease or cessation of therapy may lower or abrogate potential therapeutic side effects (individualized medicine). Regenerative medicine concerns patients who have diseased and injured organs and may be treated with transplanted organs. However, there is a severe shortage of donor organs that is worsening yearly given the aging population. Regenerative medicine and tissue engineering apply the principles of cell transplantation, material science, and bioengineering to construct biological substitutes that will restore and maintain normal function in diseased and injured tissues. Neovascularization is promoted by bone marrow-derived endothelial progenitor cells that lead to the formation of entirely new vessels into ischemic tissue. With this knowledge, many therapeutical borders can be skipped. Diseases formerly uncontrolled can be corrected with stem cells to provide causal healing with regeneration processes. The 9th Leipziger Workshop combined with the 2nd International Workshop on SBC aimed to offer new methods in image cytometry and SBC for solutions in clinical research. It moved toward practical applications in clinics and the clinical laboratory. This development will be continued in 2005 at the upcoming Leipziger Workshop and the 3rd International Workshop on SBC.

Cardiovascular Diseases↗

Systems biology and clinical cytomics: The 10th Leipziger Workshop and the 3rd International Workshop on Slide-Based Cytometry, Leipzig, Germany, April 2005.

Despite very significant technical and software improvements in flow cytometry (FCM) since the 1980's, the demand for a cytometric technology combining both quantitative cell analysis and morphological documentation in Cytomics became evident. Improvements in microtechnology and computing permit nowadays similar quantitative and stoichiometric single cell-based high-throughput analyses by microscopic instruments, like Slide-Based Cytometry (SBC). SBC and related techniques offer unique tools to perform complex immunophenotyping, thereby enabling diagnostic procedures during early disease stages. Multicolor or polychromatic analysis of cells by SBC is of special importance not only as a cytomics technology platform but also because of low quantities of required reagents and biological material. The exact knowledge of the location of each cell on the slide permits repetitive restaining and reanalysis of specimens. Various separate measurements of the same specimen can be ultimately fused to one database increasing the information obtained per cell. Relocation and optical evaluation of cells as typical SBC feature, can be of integral importance for cytometric analysis, since artifacts can be excluded and morphology of measured cells can be documented. Progress in cell analytic: In the SBC, new horizons can be opened by the new techniques of structural and functional analysis with the high resolution from intracellular and membrane (confocal microscopy, nanoscopy, total internal fluorescence microscopy (TIRFM), and tissue level (tissomics), to organ and organism level (in vivo cytometry, optical whole body imaging). Predictive medicine aims at the detection of changes in patient's state prior to the manifestation of the disease or the complication. Such instances concern immune consequences of surgeries or noninfectious posttraumatic shock in intensive care patients or the pretherapeutic identification of high risk patients in cancer cytostatic therapy. Preventive anti-infectious or anti-shock therapy as well as curative chemotherapy in combination with stem cell transplantation may provide better survival chances for patient at concomitant cost containment. Predictive medicine-guided optimization of therapy could lead to individualized medicine that gives significant therapeutic effect and may lower or abrogate potential therapeutic side effects. The 10th Leipziger Workshop combined with the 3rd International Workshop on SBC aimed to offer new methods in Image- and Slide-Based Cytometry for solutions in clinical research. It moved towards practical applications in the clinics and the clinical laboratory. This development will be continued in 2006 at the upcoming Leipziger Workshop and the International Workshop on Slide-Based Cytometry.

Cytophotometry↗

Hyperchromatic cytometry principles for cytomics using slide based cytometry.

BACKGROUND: Polychromatic analysis of biological specimens has become increasingly important because of the emerging new fields of high-content and high-throughput single cell analysis for systems biology and cytomics. Combining different technologies and staining methods, multicolor analysis can be pushed forward to measure anything stainable in a cell. We term this approach hyperchromatic cytometry and present different components suitable for achieving this task. For cell analysis, slide based cytometry (SBC) technologies are ideal as, unlike flow cytometry, they are non-consumptive, i.e. the analyzed sample is fixed on the slide and can be reanalyzed following restaining of the object. METHODS AND RESULTS: We demonstrate various approaches for hyperchromatic analysis on a SBC instrument, the Laser Scanning Cytometer. The different components demonstrated here include (1) polychromatic cytometry (staining of the specimen with eight or more different fluorochromes simultaneously), (2) iterative restaining (using the same fluorochrome for restaining and subsequent reanalysis), (3) differential photobleaching (differentiating fluorochromes by their different photostability), (4) photoactivation (activating fluorescent nanoparticles or photocaged dyes), and (5) photodestruction (destruction of FRET dyes). Based on the ability to relocate cells that are immobilized on a microscope slide with a precision of approximately 1 microm, identical cells can be reanalyzed on the single cell level after manipulation steps. CONCLUSION: With the intelligent combination of several different techniques, the hyperchromatic cytometry approach allows to quantify and analyze all components of relevance on the single cell level. The information gained per specimen is only limited by the number of available antibodies and sterical hindrance.

Animals↗