SEARCH · Search PubMed
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Explore the source record for details and available documents.
We have made a quantitative and concurrent analysis of B cell frequencies and VH gene family expression to study the influence of tissue type and age on the development and establishment of the primary B cell repertoire. Using LPS-mediated limiting dilution analysis and a panel of antigens we show that the newly generated B cell specificities from bone marrow get distributed without a bias to peripheral tissues such as spleen and Peyer's patches throughout the lifetime of the animal. Comparison of the B cell frequencies in animals of four different age groups (2-4 days old, 3, 12, and 18 months old) reveals that while the neonatal repertoire is comparable to that of adults, there was a selective twofold increase in the generation and distribution of B cells reactive with autologous mouse red blood cells in older mice compared to young ones. By means of a novel technique that employs fluorescent in situ hybridization and flow cytometry, we have also compared the VH gene family usage in large numbers of single B cells from these mice. Analysis of the same cell population (surface Ig+, functional B lineage cells) for expression of 7183, J558, and S107 VH families shows a preferential twofold increase in the use of VH 7183 in neonates compared to adults, while all three families show no significant difference in levels of expression during adult life or between primary and secondary lymphoid tissues. Taken together, our data indicate that specific and selective changes occur in both VH gene usage and antibody frequencies during murine ontogeny.
PURPOSE: To quantify optical coherence tomography (OCT) images of the central retina in patients with blue-cone monochromatism (BCM) and achromatopsia (ACH) compared with healthy control individuals. METHODS: The study included 15 patients with ACH, 6 with BCM, and 20 control subjects. Diagnosis of BCM and ACH was established by visual acuity testing, morphologic examination, color vision testing, and Ganzfeld ERG recording. OCT images were acquired with the Stratus OCT 3 (Carl Zeiss Meditec AG, Oberkochen, Germany). Foveal OCT images were analyzed by calculating longitudinal reflectivity profiles (LRPs) from scan lines. Profiles were analyzed quantitatively to determine foveal thickness and distances between reflectivity layers. RESULTS: Patients with ACH and BCM had a mean visual acuity of 20/200 and 20/60, respectively. Color vision testing results were characteristic of the diseases. The LRPs of control subjects yielded four peaks (P1-P4), presumably representing the RPE (P1), the ovoid region of the photoreceptors (P2), the external limiting membrane (ELM) (P3), and the internal limiting membrane (P4). In patients with ACH, P2 was absent, but foveal thickness (P1-P4) did not differ significantly from that in the control subjects (187 +/- 20 vs. 192 +/- 14 microm, respectively). The distance from P1 to P3 did not differ significantly (78 +/- 10 vs. 82 +/- 5 microm) between ACH and controls subjects. In patients with BCM, P3 was lacking, and P2 advanced toward P1 compared with the control subjects (32 +/- 6 vs. 48 +/- 4 microm). Foveal thickness (153 +/- 16 microm) was significantly reduced compared with that in control subjects and patients with ACH. CONCLUSIONS: Quantitative OCT image analysis reveals distinct patterns for controls subjects and patients with ACH and BCM, respectively. Quantitative analysis of OCT imaging can be useful in differentiating retinal diseases affecting photoreceptors. Foveal thickness is similar in both normal subjects and patients with ACH but is decreased in patients with BCM.
Secretory otitis media (SOM) is a common childhood disease without a completely clarified etiology. A chronic inflammatory condition in the nasopharynx, presumably caused by an increased bacterial load, is one factor of probable etiological importance. In the present study a flow cytometric method for analysis of adenoid lymphoid cell populations was developed to facilitate quantitative comparisons between children with SOM and children without ear disease. Adenoids removed from 18 children due to adenoid hyperplasia and obstructive symptoms were studied. Results of the flow cytometric analysis correlated well with the findings from immunohistological studies of five of the adenoids. PCA-1 and CD25 were found to be good markers of increased cellular activity after non-specific stimulation in cell culture. It is concluded that the flow cytometric method is suitable for further quantitative analysis of adenoid tissue.
Automated quantitative image analysis (QIAF) was used to measure and compare the adrenergic nerve plexuses of 4 blood vessels from the guinea pig, demonstrated by glyoxylic acid fluorescence (GAF). The results showed considerable quantitative variation of plexus density, size of bundles, and numbers of varicosities. A range of alternative procedural and anatomical sources of variability were investigated and assessed. The carotid artery was found to have a dense plexus with more nerves than that of the mesenteric artery; the mesenteric vein and abdominal aorta had sparse plexuses. The carotid artery plexus, despite the density of its nerves, possessed only half the number of varicosities of the mesenteric artery plexus. This sparse varicosity population was shown to have a similar density to the varicosities demonstrated by QIAF in the scattered nerves of the mesenteric vein and abdominal aorta. QIAF confirmed visual estimates of adrenergic plexus density, and was able to demonstrate less obvious differences of nerve density and size, and varicosity populations, between the different plexuses studied. The method is applicable to stretch preparations and transverse sections of many adrenergically innervated tissues.
There is an ongoing need for sensitive and specific markers of bacterial infection. In this prospective study, standard clinical laboratory data (neutrophil count, serum C reactive protein level, erythrocyte sedimentation rate) and quantitative flow cytometric analysis of neutrophil complement receptors, CR1 and CR3, were obtained from 289 hospitalized febrile patients. After microbiological confirmation or clinical diagnosis, 135 patients were found to have either bacterial (n = 89) or viral (n = 46) infection. The patient data was compared to 60 healthy controls. In bacterial infections, all measured variables were significantly increased, particularly the average amounts of CR1 and CR3 on neutrophils were over three-fold and two-fold higher, respectively, compared to viral infections and controls. We described a novel marker of local and systemic bacterial infections designated 'clinical infection score (CIS) point', which incorporates quantitative analysis of complement receptors on neutrophils and standard clinical laboratory data. CIS point varied between 0 and 8, and displayed 98% sensitivity and 97% specificity in distinguishing between bacterial and viral infections [average (S.D.); CIS points: 6.2 (1.7) vs. 0.6 (1.0); p < 0.001]. These findings suggest that the proposed CIS-based diagnostic test could potentially assist physicians in deciding whether antibiotic treatment is necessary.
Isolated cold stress testing (ICST) has been used to assess cold stress performance or digital thermoregulation, but statistical analysis of the results has been limited to visual comparisons of data. In this prospective study, 11 patients who underwent complete digital replantation were followed serially with ICST at 6 weeks, 3 months, 6 months, 1 year, and 2 years postoperatively, and the results were analyzed quantitatively. For that analysis, we devised a mathematical method that provided a cooling and a warming coefficient to fit the data. Differences in these cooling and warming coefficients were then regressed against time after replantation, patient age, number of digits replanted, hand dominance, and clinical evidence of sensory recovery. There was a significant correlation between response to isolated cold stress testing (thermoregulation) and degree of sensory recovery (p less than or equal to 0.02). This method for quantitative analysis of isolated cold stress testing data allows objective evaluation of digital blood flow patterns based on temperature, thereby providing a reliable and objective assessment of the recovery of thermoregulation in the replanted human digit.
In the present study, quantitative electroencephalographic (EEG) analysis was performed at rest and during photic stimulation (5, 10, and 15 Hz) in nine patients with presenile dementia of the Alzheimer type (AD; mean age at onset, 55 years) and nine sex- and age-matched control subjects. Compared with the normal controls, the AD patients had a significantly lower alpha-2 and beta band power in the resting EEG as well as a significant increase in delta and theta band power. EEG analysis during the photic stimulation demonstrated that the AD patients had a significantly lower EEG power during photic stimulation for the alpha (9.8-10.2 Hz) and beta bands (14.8-15.2 Hz) corresponding to photic stimulation at 10 Hz and 15 Hz, respectively. In addition, when we examined EEG changes from rest to the stimulus condition, the AD patients were found to show significantly smaller changes in EEG power mainly over the posterior regions, irrespective of the stimulus frequency. These findings provide evidence that AD patients have EEG abnormalities in both non-stimulus and stimulus conditions, and suggest diminished EEG reactivity to photic stimulation.
PURPOSE: To retrospectively compare quantitative and qualitative methods of assessing magnetic resonance (MR) imaging contrast enhancement as the basis for diagnosing renal malignancy. MATERIALS AND METHODS: MR imaging was performed by using a gadolinium-enhanced breath-hold fat-suppressed three-dimensional T1-weighted gradient-echo sequence in 71 patients (48 men and 23 women; mean age, 62 years; age range, 26-87 years) with 93 renal lesions for which pathologic correlation was available. For quantitative measurements of enhancement, the relative increase in signal intensity values was measured by one investigator with manually defined regions of interest, and the threshold of an increase of 15% or greater was used to distinguish malignant from benign masses. For qualitative assessment, two investigators independently reviewed the subtracted images of all lesions and subjectively determined whether enhancement was present or absent. The sensitivity, specificity, and positive and negative predictive values for each method were calculated and compared. Mean (+/- standard deviation) and median values of relative enhancement were also calculated for benign and malignant lesions. RESULTS: At pathologic analysis, 74 (80%) of the 93 lesions were malignant, and 19 (20%)-including seven oncocytomas-were benign. For diagnosing malignancy based on enhancement alone, sensitivity and specificity, respectively, were 95% (70 of 74 lesions) and 53% (10 of 19 lesions) at quantitative analysis and 99% (73 of 74 lesions) and 58% (11 of 19 lesions) at qualitative analysis. All seven oncocytomas were considered to be malignant with both methods. When the oncocytomas were excluded, specificities increased to 83% (10 of 12 lesions) and 92% (11 of 12 lesions) for the quantitative and qualitative evaluations, respectively. Three of the four malignant lesions incorrectly characterized as benign at quantitative assessment were hyperintense on unenhanced MR images; all were diagnosed correctly at qualitative evaluation. CONCLUSION: Image subtraction enables accurate assessment of renal tumor enhancement, particularly in the setting of masses that are hyperintense on unenhanced MR images.
A computer-aided quantitative method for a complex analysis of gel electrophoretograms is presented. The analysis consists of several steps: (i) determination of the background image by methods of mathematical morphology and its subtraction from the gel image, (ii) selection of an appropriate part of the gel lane including curved lanes and lanes with a nonuniform width, (iii) computation of the lane densitogram by averaging several lane-parallel scans, (iv) decomposition of the lane densitogram into component bands using a data selecting algorithm and Marquardt's minimizer. Several different functions for component bands are utilized. It is shown that the densitogram can be decomposed into component bands with reasonable accuracy only if an appropriate model function is chosen. The algorithms are tested on several different gel electrophoretograms which show typical features as a nonuniform background, curved lanes, an asymmetrical band shape and a superposition of small bands on the shoulders of big ones. It is shown that overlapped bands are best approximated by an asymmetrical Gausian curve and an asymmetrical Gauss-Cauchy function. Linear response to the serial dilution of the protein sample is tested.
BACKGROUND: The identification of the function of all genes that contribute to specific biological processes and complex traits is one of the major challenges in the postgenomic era. One approach is to employ forward genetic screens in genetically tractable model organisms. In Drosophila melanogaster, P element-mediated insertional mutagenesis is a versatile tool for the dissection of molecular pathways, and there is an ongoing effort to tag every gene with a P element insertion. However, the vast majority of P element insertion lines are viable and fertile as homozygotes and do not exhibit obvious phenotypic defects, perhaps because of the tendency for P elements to insert 5' of transcription units. Quantitative genetic analysis of subtle effects of P element mutations that have been induced in an isogenic background may be a highly efficient method for functional genome annotation. RESULTS: Here, we have tested the efficacy of this strategy by assessing the extent to which screening for quantitative effects of P elements on sensory bristle number can identify genes affecting neural development. We find that such quantitative screens uncover an unusually large number of genes that are known to function in neural development, as well as genes with yet uncharacterized effects on neural development, and novel loci. CONCLUSIONS: Our findings establish the use of quantitative trait analysis for functional genome annotation through forward genetics. Similar analyses of quantitative effects of P element insertions will facilitate our understanding of the genes affecting many other complex traits in Drosophila.
The continuous monitoring of spontaneous locomotor activity has emerged as one of the most widely used metrics in rodent circadian research. This behavioral measure is also extremely useful for the description of the effects of aging on circadian rhythms. The present study describes the successful use of a log-survivorship approach to identify discrete bouts of hamster wheel-running activity and provides a detailed description of the age-related fragmentation in the 24-h profile of this behavioral variable. In addition, stepwise discriminant analysis identified the most important quantitative measures for distinguishing between the individual patterns of wheel-running activity of young (3 mo) and old (17-18 mo) golden hamsters. The results suggest that this method of bout analysis can be a valuable tool for the study of genetic, developmental, neurochemical, physiological, and environmental factors involved in the temporal control of rodent locomotor behavior.
BACKGROUND: Sensitive monitoring of minimal residual disease may improve the treatment of neuroblastoma in children. To detect and monitor neuroblastoma cells in blood and bone marrow, we developed a quantitative method for the analysis of tyrosine hydroxylase mRNA. METHODS: We used real-time reverse transcription-PCR. The calibrator was constructed from a segment of tyrosine hydroxylase mRNA that included the target. Blood and bone marrow samples from 24 children with neuroblastoma and 1 child with ganglioneuroma were analyzed. Controls were blood samples from the cords of 40 babies, from 58 children 6 months to 15 years of age, and from 34 healthy adults, as well as from 12 children with other diseases. RESULTS: The detection limit was approximately 70 transcripts/mL. All 144 blood controls were below this limit. At diagnosis, blood tyrosine hydroxylase mRNA was higher in children with widespread disease (stage 4/4S; n = 6; range, 203-46,000 transcripts/mL) than in patients with localized disease (stages 1-3; n = 6; </=83 transcripts/mL; P = 0.002). Bone marrow from all five children with localized disease had concentrations <72 transcripts/mL, whereas five of six stage 4 patients had increased concentrations (6,000-8,000,000 transcripts/mL; P <0.05). In nine children in whom tyrosine hydroxylase mRNA was measured repeatedly, the results corresponded to the clinical course. CONCLUSION: Quantitative analysis of tyrosine hydroxylase mRNA in blood and bone marrow is reliable and easy to perform and may be used for upfront staging, prognostic assessment, and treatment monitoring of neuroblastoma.
A GC/low-resolution MS method for the quantitative isotope dilution analysis of 39 mono- to heptabrominated diphenyl ethers was developed. The effects of two different ionization sources, electron impact (EI) and electron capture negative ionization (ECNI), and the effects of their parameters on production of high-mass fragment ions [M - xH - yBr](-) specific to PBDEs were investigated. Electron energy, emission current, source temperature, ECNI system pressure, and choice of ECNI reagent gases were optimized. Previously unidentified enhancement of PBDE high-mass fragment ion [M - xH - yBr](-) abundance was achieved. Electron energy had the largest impact on PBDE high-mass fragment ion abundance for both the ECNI and EI sources. By monitoring high-mass fragment ions of PBDEs under optimized ECNI source conditions, quantitative isotope dilution analysis of 39 PBDEs was conducted using nine (13)C(12) labeled PBDEs on a low-resolution MS with low picogram to femtogram instrument detection limits.
Quantitative parameters characterizing the shape and interlocation of the vertebrae were determined in roentgenograms of 60 healthy persons and of 61 patients suffering from chronic pain syndrome caused by cervical osteochondrosis. The calendar age of the persons examined has correlated mostly with a group of parameters characterizing the shape of vertebrae (the highest correlation was with the length of vertebrae). Using the multiple correlation method the authors have worked out highly reliable (p < 0.00005) regressive models of the calendar age of healthy persons and have confirmed the increased rate of age changes in a cervical segment of the spine in patients suffering from cervical pain syndrome.
A simple quantitative method of gait analysis in parkinsonism is presented. In this method, spatial and temporal parameters (stride length and duration, stance duration and double support duration) and cinematic parameters (velocity) were evaluated. A pilot trial performed in 6 patients highlighted the pathological aspect of the parkinsonian gait as compared to the gait of healthy elderly subjects. A significant degradation (reduced speed, diminished stride length, short duration of the double support stage) was noted, even when parkinsonians were compared with the oldest of the control subjects.
Quantitative information on the normal size range of the masticatory muscles needed for successful treatment of facial asymmetry is sparse. Our purpose in this study was to define morphological indicators for the volume of the masticatory muscles on CT. Cross-sectional areas and volumes of the masseter and medial pterygoid muscles were measured from CT scans of 65 patients. The muscles were scanned from origin to insertion and the maximum cross-sectional area (MCSA) and its location together with the total volume were determined. The mandibular foramen was used as a landmark to determine the location of MCSA. The location of MCSA of masseter muscle was centred 8 mm above the mandibular foramen, and that of medial pterygoid muscle at the foramen. The MCSA of both muscles was highly correlated with volume suggesting that this parameter might serve as an indicator of their volume. Knowledge of the location of the MCSA would facilitate its measurement and so reduce radiation exposure. There was a positive correlation between the two muscles for both volume and MCSA. This finding could be relevant for future studies of the relationship between the morphology, biomechanics and pathology of the masticatory muscles.
Quantitative data are presented on the composition of the major phospholipids in isolated giant barnacle muscle fibers. It is shown, using internal perfusion techniques, that the high specific activity of labeling of polyphosphoinositides in vivo is attained by the activities of specific kinases. Electrical stimulation causes a reduction in the specific activity of labeling of PtdInsP2. This phospholipid, which is the immediate precursor for the release of InsP3, is found at a significant concentration (40 nmol/g wet weight) in single barnacle muscle fibers, sufficient to support a role as precursors of second messengers. The rapid catabolization of PtdInsP2 in the absence of external Ca2+ suggests that E-C coupling in barnacle muscle may be associated with a voltage-dependent, Ca(2+)-independent, activation of the breakdown of polyphosphoinositides.