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PubMed1997

Prognostic role of cyclin D1 in non small cell lung cancer: an immunohistochemical analysis.

Lung cancer is a worldwide problem and in many countires it is the most lethal malignancy. Because relapse is frequent after resection of non small cell lung cancer, an urgent need exists to define prognostic factors which could help in choosing the best therapeutic approach. We performed immunohistochemistry on 60 formalin-fixed paraffin-embedded non small cell lung cancer specimens in order to evaluate the frequency of cyclin D1 overexpression, and to relate it to the degree of malignancy of these tumors and to the overall survival time of the patients. All specimens were positive for cyclin D1 immunostaining. We found cyclin D1 overexpression in 30 (50%) of our specimens, with no significant difference among the different histological types. Cyclin D1 overexpression correlates in a statistical manner with short-term patient survival. Mantel-Cox analysis of these data generated a significant P value = 0.003. The mean survival time and the five-year survival rate also differed statistically. We did not find any statistically significant correlation between cyclin D1 overexpression and histological grading, tumor stage or TNM status. We concluded that cyclin D1 overexpression in 30 patients is a frequent event in non small cell lung cancer pathogenesis and may have prognostic relevance.

Biomarkers, Tumor↗

PubMed2006

Assessment of airways with three-dimensional quantitative thin-section CT: in vitro and in vivo validation.

PURPOSE: To prospectively validate the ability of customized three-dimensional (3D) software to enable bronchial tree skeletonization, orthogonal reconstruction of the main bronchial axis, and measurement of cross-sectional wall area (WA) and lumen area (LA) of any visible bronchus on thin-section computed tomographic (CT) images. MATERIALS AND METHODS: Institutional review board approval and patient agreement and informed consent were obtained. Software was validated in a phantom that consisted of seven tubes and an excised human lung obtained and used according to institutional guidelines. In vivo validation was performed with multi-detector row CT in six healthy subjects (mean age, 47 years; range, 20-55 years). Intra- and interobserver agreement and reproducibility over time for bronchial tree skeletonization were evaluated with Bland-Altman analysis. Concordance in identifying bronchial generation was assessed with the kappa statistic. WA and LA obtained with the manual method were compared with WA and LA obtained with validated software by means of the Wilcoxon test and Bland-Altman analysis. RESULTS: WA and LA measurements in the phantom were reproducible over multiple sessions (P > .90) and were not significantly different from WA and LA assessed with the manual method (P > .62). WA and LA measurements in the excised lung and the subjects were not different from measurements obtained with the manual method (intraclass correlation coefficient > 0.99). All lobar bronchi and 80.8% of third generation bronchi, 72.5% of fourth generation bronchi, and 37.7% of fifth generation bronchi were identified in vivo. Intra- and interobserver agreement and reproducibility over time for airway skeletonization and concordance in identifying bronchial generation were good to excellent (intraclass correlation coefficient > 0.98, kappa > 0.54, respectively). CONCLUSION: This method enables accurate and reproducible measurement of WA and LA on reformatted CT sections perpendicular to the main axis of bronchi visible on thin-section CT scans.

Adult↗

PubMed2000

Sequence-based analysis of mutagenized mice.

Treating mice with ethylnitrosourea (ENU) is an efficient means for mutagenizing spermatogonial cells, and this treatment has been proven effective in a variety of screens for both dominant and recessive mutations. However, a significant problem for this technology is that the efficiency of mutagenesis is assessed most often by the empiric determination of a per-locus mutation frequency by using the specific locus test, which is expensive, time-consuming, and logistically difficult. To approach this question more directly and more efficiently, one can utilize methods of PCR-based mutation detection for the characterization of progeny of mutagenized mice. Since this analysis can be done after a single generation of breeding, it is useful as a rapid means for the assessment of the efficiency of mutagen treatment. Furthermore, it is readily imaginable that this strategy can be applied for the general determination of gene function in a systematic manner. Theoretical considerations and empirical analysis suggest that the per-base mutation frequency for a fractionated-dose treatment protocol is on the order of 1 sequence change per 10(5) bp.

Animals↗

PubMed1999

Analysis of flanking sequences from dissociation insertion lines: a database for reverse genetics in Arabidopsis.

We have generated Dissociation (Ds) element insertions throughout the Arabidopsis genome as a means of random mutagenesis. Here, we present the molecular analysis of genomic sequences that flank the Ds insertions of 931 independent transposant lines. Flanking sequences from 511 lines proved to be identical or homologous to DNA or protein sequences in public databases, and disruptions within known or putative genes were indicated for 354 lines. Because a significant portion (45%) of the insertions occurred within sequences defined by GenBank BAC and P1 clones, we were able to assess the distribution of Ds insertions throughout the genome. We discovered a significant preference for Ds transposition to the regions adjacent to nucleolus organizer regions on chromosomes 2 and 4. Otherwise, the mapped insertions appeared to be evenly dispersed throughout the genome. For any given gene, insertions preferentially occurred at the 5' end, although disruption was clearly possible at any intragenic position. The insertion sites of >500 lines that could be characterized by reference to public databases are presented in a tabular format at http://www.plantcell. org/cgi/content/full/11/12/2263/DC1. This database should be of value to researchers using reverse genetics approaches to determine gene function.

Arabidopsis↗

PubMed1996

Three-dimensional metabolic and hemodynamic imaging of the normal and ischemic rat brain.

Unique insights into the topography of local metabolism/blood flow interrelationships in focal cerebral ischemia have afforded by the recent development of powerful image-processing techniques permitting three-dimensional (3D) autoradiographic image-averaging and analysis of replicate studies by a novel method termed "disparity analysis". This method, based upon a linear affine transformation model, directly estimates scaling, translation and rotation parameters simultaneously. The method was validated in awake Wister rats studied for local cerebral glucose metabolism (lCMRgl) with 14C-2-deoxyglucose. Brains were subserially sectioned, aligned by disparity analysis, and mapped into a common template so as to generate aggregate 3D data sets of the mean and standard deviation of the entire series (n = 9). Internal anatomic architecture was faithfully represented in the average image, and Fourier analysis revealed satisfactory retention of low-frequency information. The method was then applied to study metabolism/blood flow relationships in the acute focal ischemic penumbra of Sprague-Dawley rats subjected to distal photothrombotic middle cerebral artery (MCA) occlusion, coupled with permanent ipsilateral and 1 h contralateral common carotid artery occlusions. Matched series were studied for lCBF at 1.5 h and for lCMRgl at 1.25-2 h post-occlusion. The averaged lCBF image revealed the ischemic penumbra (defined as lCBF 20-40% of control) to form a "shell" around the cortical ischemic core and a confluent aggregate at the anterior and posterior poles of the core-zone. lCMRgl in the penumbra was heterogeneous, ranging from near-normal to markedly increased. An average lCMRgl/lCBF ratio data set revealed marked metabolism-flow uncoupling in penumbral pixels, averaging nearly five-fold above control ratio values. Sustained deflections of the DC potential were recorded in the penumbra, the site of marked uncoupling. This analysis defined for the first time the 3D topography of the ischemic penumbra and substantiated marked metabolism/flow dissociation, which is believed to be a metabolic consequence of the energy demand imposed by repeated peri-infarct depolarizations.

Animals↗

PubMed2003

Noncontacting lateral transportation using gas squeeze film generated by flexural traveling waves--numerical analysis.

This paper presents the theory describing the dynamical behavior of a noncontacting lateral transportation of planer objects by means of a gas squeeze film created by traveling flexural waves of a driving surface. An oscillating motion in the normal direction between two surfaces can generate a gas film with an average pressure higher than the surrounding. This load-carrying phenomenon arises from the fact that a viscous flow cannot be instantaneously squeezed; therefore, fast vibrations give rise to a cushioning effect. Equilibrium is established through a balance between viscous flow forces and compressibility forces. When the oscillatory motion between two surfaces creates traveling waves, lateral viscous forces are generated in addition to the normal levitation forces. These forces are produced as a result of nonuniform pressure gradients in the lateral direction between the surfaces. The combination of normal and lateral forces could be used for transporting objects without any direct contact with the driving surface. The numerical algorithm in this work couples the squeeze film phenomenon, which is represented by means of finite difference equations, to model a variant of the Reynolds equation, together with the equations describing the dynamics of the floating object. Numerical simulations are presented and investigated to highlight noteworthy topics.

Journal Article↗

PubMed1999

Procedural competency in emergency medicine: the current range of resident experience.

OBJECTIVES: To evaluate the recorded range of procedures tracked by emergency medicine (EM) programs, and to determine whether differences in procedural experience occur in various types of residency or hospital settings. METHODS: The program directors of 112 approved EM programs were asked to send actual procedure logs. The requested information included the average total number of a given procedure per graduating resident, for all procedures that were tracked. Data were categorized by program format, hospital type, and ED volume. To assess the global procedural experience among programs, a set of 22 "index procedures" were identified; all procedures the EM residency review committee (RRC-EM) required to be tracked were included in this set. The means per graduating resident for each index procedure were added together to generate a "mean index procedure sum" (MIPS) per graduating resident for each residency program. These MIPSs for a residency were then compared by program format, hospital type, and ED volume. A similar analysis was performed for all resuscitations, and a "mean index resuscitation sum" (MIRS) per graduating resident was generated. RESULTS: An overall response rate of 82% was achieved; a number of programs had not graduated a residency class and were not included. Sixty-five of 85 eligible programs (76%) provided procedural data. The average number of a given procedure per graduating resident (95% CI in parentheses) for selected procedures is as follows: oral intubation 65 (46 to 85), intubation unspecified 75 (62 to 87), nasal intubation 6 (4 to 9), cricothyroidotomy 2 (1 to 2), subclavian catheter 23 (16 to 30), chest tubes 17 (14 to 20), intraosseous line 2 (1 to 3), thoracotomy 3 (2 to 5), and vaginal deliveries 17 (13 to 21). The only statistically significant differences in subgroup comparisons were in diagnostic peritoneal lavage, trauma resuscitations, and pediatric medical resuscitations when compared by postgraduate year format, and intubation-unspecified and cricothyroidotomy when compared by hospital type. There was no statistically significant difference when MIPSs were compared by format, hospital type, or ED volume. CONCLUSIONS: To the authors' knowledge, this is the first study of the range of EM resident procedure experience across the spectrum of EM residency types and settings. Overall, there are few statistically significant differences in procedure experience among different program formats. Similar experiences are recorded in a variety of different hospital types or ED volumes. However, some programs report very limited EM resident experience with selected critical procedures. There is a large variation in the types and numbers of procedures recorded by EM programs.

Analysis of Variance↗

PubMed1977

Adaptive distortions in the generator potential of semicircular canal sensory afferents.

The generator potential in sensory afferents of frog crista ampullaris was extracellularly recorded from the cut end of the posterior ampullary nerve by means of suction electrodes. A servocontrolled turntable allowed suitable rotatary stimulations. The analysis of the recorded generator potential revealed a different time course from that predicted on the basis of the pendulum model. Adaptation and undershoots in the responses to velocity ramps, steps and sinusoids, were mainly responsible for the deviations, which became very evident only when fairly high acceleration rates were applied. Both adaptation and undershoots were produced presumably by the activation of an electrogenic pump, probably located in nerv terminals contacting the hair cells. In fact, the time course of the generator potential became much more consistent with the predictions from the pendulum model under treatments capable of hindering the ion pump activity.

Acceleration↗

PubMed2004

Development of AFLP-derived STS markers for the selection of 5-methyltryptophan-resistant rice mutants.

To increase the specific free amino acid content in the japonica rice (Oryza sativa L.) cultivar Donganbyeo, mutant cell lines resistant to growth inhibition by 5-methyltryptophan (5MT) were selected from embryo-cultured callus irradiated with 50 Gy gamma-rays. Four 5MT-resistant homozygous M4 lines, MRI-40, MRI-116, MRII-8, and MRII-12, were obtained. The mean content of nine free essential amino acids were 70.1, 72.5, 31.7, and 35.4% greater than the original variety in these four mutant lines, respectively. For AFLP analysis, 8 EcoRI (+2) and 8 MseI (+3) primers used in 45 primer combinations generated a total of 3,684 bands with a mean of 82 bands, of which 361 (9.8%) were clearly polymorphic with the control cultivar, the four 5MT-resistant mutants, and five sensitive lines. The lines were grouped into three clusters through cluster analysis using unweighted pair grouping method of averages. The 36 polymorphic PCR products present only in the four homozygous 5MT-resistant lines were cloned and sequenced, and 10 of these sequenced products were converted into sequence tagged site (STS) markers. These STS primer sets were designated OSMR1-OSMR10. Six STS primer sets (OSMR1, OSMR2, OSMR3, OSMR4, OSMR5, and OSMR6) generated a single monomorphic PCR product identical in size to the original AFLP fragments. The broad applicability of these STS markers for the screening of 5MT resistance was evaluated with seven putative 5MT-resistant M2 plants (PM-1 to PM-7). Four STS markers (OSMR1, OSMR2, OSMR4, and OSMR5) out of six STS primer sets were revealed as polymorphic products between the control cultivar and the seven M2 plants. These markers can be utilized for the fine selection of 5MT resistance in rice, and this PCR-screening technique is less time-consuming, less labor-intensive, and more accurate and reliable than selection based solely on phenotypic evaluation involving soaking in 5MT solutions.

Amino Acids↗

PubMed2006

CHILD syndrome in 3 generations: the importance of mild or minimal skin lesions.

BACKGROUND: CHILD syndrome (congenital hemidysplasia with ichthyosiform nevus and limb defects, Online Mendelian Inheritance in Man 308050) is an X-linked dominant trait with lethality for male embryos. The disorder is caused by mutations in NSDHL (Online Mendelian Inheritance in Man 300275), a gene playing an important role in the cholesterol biosynthetic pathway. Most reports deal with sporadic cases, and only 5 cases of mother-to-daughter transmission have been documented. We present here a family with mild features of CHILD syndrome in 3 generations. Molecular analysis was used to confirm the diagnosis. OBSERVATIONS: We studied 14 members of a family with CHILD syndrome. The 23-year-old proposita, her mother, 2 aunts, and her grandmother presented with mild or minimal skin lesions that had been present since infancy. Analysis of the NSDHL gene showed missense mutation c.370G-->A in these 5 patients. This mutation was absent in the 9 clinically unaffected family members tested. CONCLUSIONS: In this family, we recognized CHILD syndrome with mild or minimal features in 3 generations because we were able to verify our clinical diagnosis by means of molecular analysis. We assume that many cases that so far have been considered sporadic may in fact be familial when a meticulous physical examination of female family members is combined with molecular testing.

3-Hydroxysteroid Dehydrogenases↗

PubMed1991

Analysis of the human CD36 leucocyte differentiation antigen by means of the monoclonal antibody NL07.

The murine monoclonal antibody (MoAb) NL07 was generated by immunization with human platelet extracts. NL07 MoAb recognized a molecule expressed by human platelets, monocytes, and endothelial cells, as well as by the myelomonocytic line U937 and by some melanoma cells or lines. Normal endothelial cells and the melanoma cells express the NL07 epitope only while adhering to a substrate. SDS-polyacrylamide gel electrophoresis and two-dimensional gel analysis indicate that the molecule recognized by NL07 MoAb on platelets is a single chain structure featuring a molecular weight of 85 kDa under reducing conditions, with an acidic isoelectric point ranging from 5.2 to 5.5. The specific phenotype distribution and the biochemical structure indicate that NL07 MoAb recognizes the platelet GPIV (CD36) molecule, a surface glycoprotein with a functional role of thrombospondin receptor. The results of competition tests with OKM5 MoAb (specific for the CD36 molecule) confirm the molecular specificity and epitope coincidence. Furthermore, upon binding to the platelets, NL07 MoAb is able to transmit via CD36 an activation signal which is followed by a potent aggregation. On the contrary, there is lack of evidence concerning the ability of the CD36 molecule of transmitting signal(s) on the U937 cells.

Antibodies, Monoclonal↗

PubMed1990

Automated conformational analysis and structure generation: algorithms for molecular perception.

Many methodologies for performing automated conformational analysis require some means of "perceiving" a molecule to determine features of interest. Algorithms for finding rings, bond orders, and stereocenters and detecting the presence of substructural fragments have been developed. These algorithms are described, emphasizing their importance in conformational analysis.

Artificial Intelligence↗

PubMed1989

[Spectrum analysis of blood pressure and cardiac rate in the conscious rat].

Intrafemoral pulsatile blood pressure of conscious rats was computed to generate evenly spaced signals (systolic, diastolic, mean blood pressure, heart rate) at 200 ms intervals. This equidistant sampling allowed a direct spectral analysis using a Fast Fourier Transform algorithm. Systolic blood pressure and heart rate exhibited low frequency oscillations (Mayer waves, 20-605 mHz) and a high frequency peak related to respiration (1,765 mHz). The diastolic blood pressure and the mean blood pressure only exhibited low frequency oscillations. This procedure could be useful to analyze the various components of blood pressure variability.

Analysis of Variance↗

PubMed1999

A novel method of extraction of TnC from skeletal muscle myofibrils.

Incubation of mechanically skinned barnacle myofibrillar bundles in 10 mM orthovanadate (pH 6.6) results in the loss of Ca2+-dependent force generation, which reduces to 0.98+/-0.006% (mean +/-SEM, n=25) of control levels. Analysis of myofibrillar bundles by gel electrophoresis showed that tension loss is primarily due to the extraction of troponin C (TnC) (65.4+/-5.04% mean +/-SEM, n=5). This is a novel finding, since treating cardiac fibres with orthovanadate results in the removal of both TnC and troponin I (TnI) (28). Ca2+ dependence was restored to the myofibrillar bundles following reconstitution with either native isoform of barnacle TnC (BTnC1: 78. 72+/-12.8%, n=9, BTnC2: 82.73+/-20.3%, n=3). The reversible loss of Ca2+-dependent tension generation following the removal and replacement of TnC indicates that the regulation of contraction in the barnacle is controlled by thin-filament regulatory proteins.

Animals↗

PubMed2001

Electrophysiological analysis of chronic atrial fibrillation associated with mitral valve disease by using spectral analysis.

Several authors have suggested that periodic activation is related to maintenance of atrial fibrillation (AF). The aim of this study was to examine periodic electrical activations in both atria that may lead to the generation and maintenance of AF associated with valvular diseases by means of fast Fourier transform (FFT) analysis. Atrial electrograms (AEGs) were analyzed in 15 persistent AF patients, who underwent pulmonary vein orifice (PVO) isolation with mitral valve surgery. Intraoperatively, AEGs of 4 seconds duration were acquired at 48 epicardial sites, 24 each from the right and left atrium (RA, LA). Dominant peaks (DPs) examined using FFT were present in 26% of all sampling points (380/1440). Prominent clustering of DPs was mostly observed on the LA. The mean AF cycle length (mAFCL) estimated from DP frequency was significantly shorter in LA than that in RA (178 +/- 32 msec vs 247 +/- 58, p = 0.0003). The shortest AF cycle length in each patient was mainly found in the LA. The PVO isolation procedures successfully eliminated AF in 87% of patients (13/15). In cases of recurred AF (2/15), the difference in mAFCL between bilateral atria was significantly smaller than in the case of successful AF elimination (17 +/- 7 msec vs 76 +/- 56, p = 0.042). FFT analysis of intraoperative mapping data clarified that periodic activity was present predominantly in LA. It may be possible to predict the efficacy of surgical procedures for eliminating AF using this technique.

Adult↗

PubMedDate not supplied

Predictors of hospital length of stay after heart transplantation.

Factors that predict hospital length of stay after heart transplantation were identified from retrospective data of 65 patients (82% male, mean age, 43.3 years). Multiple regression analysis with a stepwise procedure was used to generate three predictive models for length of stay: (1) a model to be used before operation, (2) a model that combines preoperative and donor information, and (3) a model that takes preoperative, donor, and postoperative factors (complete model) into consideration. Hospital length of stay ranged from 15 to 45 days after heart transplantation (median length of stay, 22.5 days; mean length of stay, 24.4 +/- 6.4 days). In the preoperative model, diagnosis, duration of cardiac symptoms, severity of heart failure, and pulmonary vascular resistance were significantly related to length of stay and together accounted for 36% of the variance in length of stay. When donor information (for example, size and ischemic time) was added to preoperative information, the resultant model failed to account for appreciably more of the variance in length of stay. A model that considered preoperative, donor, and postoperative factors accounted for 71% of the variance in length of stay. Significant variables in the model were the month in which the patient had transplantation in the program, duration of cardiac symptoms before transplantation, preoperative severity of heart failure, pulmonary vascular resistance, and postoperative incidence of severe acute rejection. Patient age, sex, and postoperative infections were not related to length of stay. In conclusion, there are cardiopulmonary and immunologic factors that can predict length of stay. The model also suggests that a program's experience with heart transplantation affects length of stay.

Adolescent↗

PubMed1994

[Analysis of antihypertensive effectiveness by means of ambulatory blood pressure monitoring. Usefulness of the Discrete Fourier Transform Model].

In order to assess the usefulness of the Discrete Fourier Transform Model (DFT) to evaluate time-course drug effects on hypertensive patients studied with Ambulatory Blood Pressure Monitoring (ABPM) a number of experiments were carried out. A total of 10 mild to moderate hypertensive patients were evaluated under placebo and after 8 weeks of active treatment with Enalapril 20 mg per day using ABPM. Systolic and Diastolic blood pressure (SBP and DBP) were registered every 15 minutes during daytime and every 30 minutes at night. Pressure profiles of each patient were initially smoothed by hourly means. DFT was then applied to these profiles. The minimum number of harmonics necessary to generate a statistically significant fitting of the blood pressure profile, were obtained by residuals analysis (run test and analysis of variance of the mean sum of residual squares with each new harmonic incorporated to the model). A profile of the blood pressure differences (treatment-placebo) with the rough data of each patient was smoothed by hourly means. DFT was applied again on these substraction profiles. To estimate peak and trough drug effects for the blood pressure decrease function, maximum, minimum and inflexion points were calculated defining the following parameters: T peak: time from drug administration to maximum pressure decrease; T late response: time from drug administration to the inflexion point following the last minimum previous to the next dose; BP peak: the maximum blood pressure decrease amplitude; and the slope BP peak/T peak. The stability of the individual circadian rhythm was confirmed for both ABPM controls comparing times of maximum and minimum on the DFT smoothed profiles.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1977

Correlation analysis of delta activity generated in cerebral hypoxia.

1. Differences in spatial and temporal characteristics between polymorphous delta activity (PDA) and "standard slow complexes" (SSCS) resulting from grave cerebral hypoxia have been demonstrated by means of digital correlation analysis and on line recording of correlograms. 2. PDA observed in the human EEG and the ECoG of dogs is characterized by the absence of periodic organization (high extinction indices of auto- and crosscorrelation functions) and absence of spatial synchronization when recorded from widely separated areas of the hemispheres (low correlation coefficients, significant and variable time shifts), with bilateral symmetry of slow waves being preserved. 3. Delta waves recorded from nucleus amygdalae, nucleus caudatus, thalamus, hypothalamus and midbrain in the dog differ from cortical PDA by a more synchronous and rhythmically arranged process. 4. During the establishment of SSC activity correlograms show a synchronous process common to all cortical aand subcortical areas characteized by very high values of correlation coefficients. 5. It is suggested that the unorganized slow wave PDA process could arise under conditions of hypoxic dysfunction of distant cortico-cortical relationships, accompanied by weak subcortical pacemaker influences. The process reflects local coexcitation of relatively limited cortical areas. The particular statistical characteristics of SSCs generated during further increase in oxygen deficit can be accounted for by simultaneous generation of responses in some subcortical areas to single volleys of impulses from a hypothetical pacemaker, accompanied by a passive wave propagation towards a cortex completetly depressed by hypoxia

Adolescent↗