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PubMed2004

Novel nanosensors for rapid analysis of telomerase activity.

Elevated telomerase levels are found in many malignancies, offering an attractive target for therapeutic intervention and diagnostic or prognostic purposes. Here we describe the use of a novel nanosensor developed for rapid screens of telomerase activity in biological samples. The technique utilizes magnetic nanoparticles that, on annealing with telomerase synthesized TTAGGG repeats, switch their magnet state, a phenomenon readily detectable by magnetic readers. We tested the efficacy of different telomerase inhibitors in crude human and murine samples and show that phosphorylation of telomerase regulates its activity. High-throughput adaptation of the technique by magnetic resonance imaging allowed processing of hundreds of samples within tens of minutes at ultrahigh sensitivities. Together, these studies establish and validate a novel and powerful tool for rapidly sensing telomerase activity and provide the rationale for developing analogous magnetic nanoparticles for in vivo sensing.

Biosensing Techniques↗
PubMed1992

A PC program for performing multigroup longitudinal comparisons using the Potthoff-Roy analysis and orthogonal polynomials.

A PC-program performing the Potthoff-Roy (PR) multigroup (G-sample) analysis of longtidinal data is described and illustrated. This program and the underlying statistical model are useful in the comparison of several longitudinal samples. Applications include the study of growth, development, adaptation, aging, and treatment effects (in short, any phenomenon in which the passage of time is important) for which serial data are available. Specifically, this method fits polynomials to the average growth curves in the samples, and tests hypotheses concerning the curves themselves and the individual coefficients of the polynomials. The program features the utilization of orthogonal polynomial regression coefficients (OPRCs) and is written in GAUSS, a relatively inexpensive yet comprehensive matrix programming language. It is documented that using OPRCs to comprise the within-individual or time design matrix has several advantages over the more usual choice of the successive-powers-of-t form of this matrix and an example of one important such advantage is provided. GAUSS was employed to make the program readily-accessible (i.e., executable code) to biomedical investigators. The GAUSS compiler is not required to run this program. Information regarding the availability of the program is provided in the Appendix.

Child↗
PubMed2003

Total protein determinations by particle beam/hollow cathode optical emission spectroscopy.

A novel method for quantitative total protein determinations is presented. Total protein content is determined by particle beam/hollow cathode optical emission spectroscopy (PB/HC-OES) through monitoring of carbon atomic emission. The PB/HC-OES offers such advantages as ease of operation, exclusion of labor-intensive sample pretreatment processes, rapid analysis, high sensitivity, and low detection limit. The method could also be adapted to be integrated to current microfluidics devices. Parametric optimization for sample introduction, nebulization, desolvation, and hollow cathode source conditions is performed for the analysis of aqueous bovine serum albumin solutions. Response curves of C (I) 193.0-nm emission were obtained under the optimized conditions with both 10% HCl and 100 micromL KCl added to the sample matrix as potential carriers. The detection limit for triplicate injections of bovine serum albumin standards was found to be on the single-nanogram level with 200-microL injections. The addition of KCl significantly improved the sensitivity, supporting the proposed "carrier effect" of chloride salts in the particle transport process. Results obtained here suggest a range of applications for the use of the PB/HC-OES source for total protein determinations; emphasis here is future use in assessing protein quantification in microfluidic systems.

Electrodes↗
PubMed2002

The relation between general cognitive level and adaptive behavior domains in individuals with autism with and without co-morbid mental retardation.

This study examined the association between adaptive behavior and general cognitive level in individuals with autism or PDD-NOS with and without comorbid mental retardation. Data from the screening version of the Vineland Adaptive Scales and the Wechsler Intelligence Scales were analysed in a sample of 67 subjects. While in the higher functioning individuals (IQ > 70, n = 34) IQ and adaptive behavior level differed significantly, performances were fairly comparable in subjects showing lower cognitive functioning (IQ < 70, n = 33). Regression models revealed a higher correlation between IQ and single adaptive behavior domains in the non-mentally retarded participants, with the domain Communication reaching the highest predictive power of the single adaptive behavior areas. Findings indicate, the relationship between adaptive and cognitive function in autistic disorders is mediated by the presence of a qualitative reduction of intelligence. Methodological limitations of the study are discussed.

Adaptation, Psychological↗
PubMed2006

Digitally reconstructed radiograph generation by an adaptive Monte Carlo method.

Digitally reconstructed radiograph (DRR) generation is an important step in several medical imaging applications such as 2D-3D image registration, where the generation of DRR is a rate-limiting step. We present a novel DRR generation technique, called the adaptive Monte Carlo volume rendering (AMCVR) algorithm. It is based on the conventional Monte Carlo volume rendering (MCVR) technique that is very efficient for rendering large medical datasets. In contrast to the MCVR, the AMCVR does not produce sample points by sampling directly in the entire volume domain. Instead, it adaptively divides the entire volume domain into sub-domains using importance separation and then performs sampling in these sub-domains. As a result, the AMCVR produces almost the same image quality as that obtained with the MCVR while only using half samples, and increases projection speed by a factor of 2. Moreover, the AMCVR is suitable for fast memory addressing, which further improves processing speed. Independent of the size of medical datasets, the AMCVR allows for achieving a frame rate of about 15 Hz on a 2.8 GHz Pentium 4 PC while generating reasonably good quality DRR.

Algorithms↗
PubMed2006

Possible involvement of CCT5, RGS3, and YKT6 genes up-regulated in p53-mutated tumors in resistance to docetaxel in human breast cancers.

BACKGROUND: Present study was aimed to investigate the relationship of p53 mutation status with response to docetaxel in breast cancers. In addition, attempts were made to identify the genes differentially expressed between p53-wild and p53-mutated breast tumors and to study their relationship with response to docetaxel. METHODS: Mutational analysis of p53 was done in 50 breast tumor samples obtained from primary breast cancer patients (n = 33) and locally recurrent breast cancer patients (n = 17) before docetaxel therapy. Response to docetaxel was evaluated clinically. Gene expression profiling (n = 2,412) was conducted by adapter-tagged competitive-PCR in 186 tumor samples, which were also analyzed in their p53 mutational status in order to identify the differentially expressed genes according to p53 mutation status and their relationship with response to docetaxel. RESULTS: Response rate of p53-mutated tumors (44%) was lower than that of p53-wild tumors (62%) though there was no statistical significance (P = 0.23). Of 2412 genes, mRNA expression of 13 genes was significantly different between p53-wild and p53-mutated tumors. Of these 13 genes, mRNA expression of CCT5, RGS3, and YKT6 was significantly up-regulated in p53-mutated tumors and associated with a low response rate to docetaxel. Treatment of MCF-7 cells with siRNA specific for CCT5, RGS3, or YKT6 resulted in a significant enhancement of docetaxel-induced apoptosis. CONCLUSIONS: CCT5, RGS3, and YKT6 mRNA expressions, which are up-regulated in p53-mutated breast tumors, might be implicated in resistance to docetaxel and clinically useful in identifying the subset of breast cancer patients who may or may not benefit from docetaxel treatment.

Antineoplastic Agents, Phytogenic↗
PubMedDate not supplied

Routine monitoring of cyclosporine in whole blood and in kidney tissue using high performance liquid chromatography.

A simple isocratic reversed-phase high pressure liquid chromatographic procedure for the specific estimation of cyclosporine in 1 mL of whole blood using cyclosproin D as internal standard is described. The chromatographic conditions chosen afford a high degree of resolution and sensitivity, a prolonged analytical column life (four months), and complete analysis of ether extracts within 8 min. The limit of detection is 10 micrograms/L. Thirty-five patient samples can be analyzed in a single batch with a turnaround time of 7 hr. This method has been in use in the authors' therapeutic drug monitoring laboratory for nearly two years. It can be simply adapted for the analysis of cyclosporine in kidney samples as a possible aid in the diagnosis of therapeutic failure or toxicity in renal allograft recipients. Examples are given to illustrate the usefulness of frequent monitoring of cyclosporine in stabilizing graft function after organ transplantation.

Adult↗
PubMed1984

Airway receptor activity in the developing opossum.

Characteristics of airway receptor discharge were evaluated in pentobarbital-anesthetized, gallamine-paralyzed, artificially ventilated, open-chest suckling and weanling opossums (Didelphis marsupialis). Animals were tested with single-unit vagal recordings at 20, 30, 55, and 90 days of age; results were compared with previous data from adults. Rapidly adapting airway receptors ( RARs ) comprised a smaller percentage of the sampled population at 20 and 30 days of age than in older groups. The static firing properties of slowly adapting airway receptors (SARs) were examined at 0-20 cmH2O transpulmonary pressure (Ptp). At the higher Ptp levels, average receptor discharge rates increased with increasing age; these results are similar to findings in placental mammals. At 5 cmH2O, however, only the 20-day-old animals exhibited reduced rates of SAR discharge. The dynamic response of SARs in 20- and 30-day-old animals was reduced compared with adult values, as measured by the adaptation of their discharge frequencies; however, the reduction was proportional to the lower receptor firing rates. About 80% of tested SARs were inhibited by CO2 at 20 and 55 days; similar results were obtained in adults. Other results have shown that morphologic development of the vagi in opossums at 50 days of age compares with placental mammals similarly studied at birth. In contrast, the static SAR discharge in the 50-day-old opossum at low Ptp (i.e., 5 cmH2O) is similar to that observed in adults, whereas the newborn placental mammal has a greatly reduced SAR firing rate. This suggests that early utilization of the lungs can contribute to functional maturation of airway receptors.

Adaptation, Physiological↗
PubMed1988

Aids and adaptations for the elderly at home: underprovided, underused, and undermaintained.

A random sample of 140 elderly people aged over 75 was selected from the age-sex register of an urban general practice to assess the provision and use of aids and adaptations in their homes. Many of the aids that the elderly had were faulty, including half of the walking aids and 15% of hearing aids, reading spectacles, and dentures, and up to half of the aids were not used. Yet despite this underuse there were many disabled elderly people who required aids for the bath and toilet. When screening of elderly people is carried out in general practice assessment of aids and adaptations should be included to see that they are provided where needed, are used, and are adequately maintained.

Aged↗
PubMed1997

A computerized adaptive testing system for speech discrimination measurement: the Speech Sound Pattern Discrimination Test.

A computerized, adaptive test-delivery system for the measurement of speech discrimination, the Speech Sound Pattern Discrimination Test, is described and evaluated. Using a modified discrimination task, the testing system draws on a pool of 130 items spanning a broad range of difficulty to estimate an examinee's location along an underlying continuum of speech processing ability, yet does not require the examinee to possess a high level of English language proficiency. The system is driven by a mathematical measurement model which selects only test items which are appropriate in difficulty level for a given examinee, thereby individualizing the testing experience. Test items were administered to a sample of young deaf adults, and the adaptive testing system evaluated in terms of respondents' sensory and perceptual capabilities, acoustic and phonetic dimensions of speech, and theories of speech perception. Data obtained in this study support the validity, reliability, and efficiency of this test as a measure of speech processing ability.

Adolescent↗
PubMed1984

Biokinetic analyses of adaptation and succession: microbial activity in composting municipal sewage sludge.

The interactions between temperature and the microbial communities in composting municipal sewage sludge were studied to determine the optimal temperature range for efficient decomposition (stabilization) of the sludge. Information concerning thermophilic successions in such communities was also obtained. Samples were taken from several different temperature areas in a production-scale composting pile throughout the 19-day processing run. Optimum temperatures for microbial activity, determined as the rate of [14C]acetate incorporation into microbial lipids, were determined for each sample. Biomass was determined from the lipid phosphate content of the sample. Maximal activities were generally found in samples coming from lower-temperature areas (25 to 45 degrees C), whereas samples from high temperatures (55 to 74 degrees C) usually had relatively little activity. The temperature giving the optimum activity in samples incubated at a variety of temperatures during the assay tended to increase as the composting time progressed, but never exceeded about 50 degrees C. Many of these temperature response curves were similar in nature to curves reported for purified enzyme systems and pure cultures of bacteria. Comparisons of the apparent energies of activation calculated for different temperature ranges over time also indicated that the overall community was better adapted to higher temperatures during the latter part of the composting run. It was also found that the relationship between the apparent energies of activation and the apparent energies of inactivation (apparent heats of denaturation) consistently changed with sample temperature throughout the composting run, suggesting that the microbial communities from hotter samples were better adapted to high temperatures than those from cooler samples, and vice versa.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗
PubMed1997

Adaptation and the color statistics of natural images.

Color perception depends profoundly on adaptation processes that adjust sensitivity in response to the prevailing pattern of stimulation. We examined how color sensitivity and appearance might be influenced by adaptation to the color distributions characteristic of natural images. Color distributions were measured for natural scenes by sampling an array of locations within each scene with a spectroradiometer, or by recording each scene with a digital camera successively through 31 interference filters. The images were used to reconstruct the L, M and S cone excitation at each spatial location, and the contrasts along three post-receptoral axes [L + M, L - M or S - (L + M)]. Individual scenes varied substantially in their mean chromaticity and luminance, in the principal color-luminance axes of their distributions, and in the range of contrasts in their distributions. Chromatic contrasts were biased along a relatively narrow range of bluish to yellowish-green angles, lying roughly between the S - (L + M) axis (which was more characteristic of scenes with lush vegetation and little sky) and a unique blue-yellow axis (which was more typical of arid scenes). For many scenes L - M and S - (L + M) signals were highly correlated, with weaker correlations between luminance and chromaticity. We use a two-stage model (von Kries scaling followed by decorrelation) to show how the appearance of colors may be altered by light adaptation to the mean of the distributions and by contrast adaptation to the contrast range and principal axes of the distributions; and we show that such adjustments are qualitatively consistent with empirical measurements of asymmetric color matches obtained after adaptation to successive random samples drawn from natural distributions of chromaticities and lightnesses. Such adaptation effects define the natural range of operating states of the visual system.

Adaptation, Ocular↗
PubMed2001

Adaptations to pressure in the RBC metabolism of diving mammals.

Marine mammals are known to dive up to 2000 m and, therefore, tolerate as much as 200 atm. of hydrostatic pressure. To examine possible metabolic adaptations to these elevated pressures, fresh blood samples from marine and terrestrial mammals were incubated for 2 h at 37 degrees C under 136 atm (2000 psi) of hydrostatic pressure. The consumption of plasma glucose and the production of lactate over the 2-h period were used to assess glycolytic flux in the red cells. The results indicate that glycolytic flux as measured by lactate production under pressure can be significantly depressed in most terrestrial mammals and either not altered or accelerated in marine mammals. The data also suggest that there is a significant shift in the ratio of lactate produced to glucose consumed under pressure. Interestingly, human and dolphin blood do not react to pressure. These combined data imply a metabolic adaptation to pressure in marine mammal RBC that may not be necessary in human or dolphin cells due to their unique patterns of glucose metabolism.

Animals↗
PubMed2001

Downsizing proteolytic digestion and analysis using dispenser-aided sample handling and nanovial matrix-assisted laser/desorption ionization-target arrays.

An efficient technique for enzymatic digestion of proteins in nanovial arrays and identification by peptide mass fingerprinting using matrix-assisted laser desorption/ionization (MALDI-MS) is presented in this work. Through dispensing of a protein solution with simultaneous evaporation the protein (substrate) is concentrated up to 300 times in-vial. At higher substrate concentrations the catalytic turnover numbers increase according to the Michaelis-Menten kinetics. Therefore, the dispenser-aided nanodigestion is valuable for identification of low-level proteins (10 nM-500 nM) as well as for automatic high efficiency digestions performed in 0.2-10 min. As an example of low-level protein identification, a 10 nM solution of lysozyme C was unambiguously identified after 5 min of nanodigestion. Moreover, only 30 s nanodigestion was sufficient to identify hemoglobin (10 microM), exemplifying the fast catalysis of the nanodigestion technique. The developed silicon flow-through piezoelectric dispenser is adapted for low-volume and preconcentrated samples in the nL-microL range and provides fast, accurate and contact-free sample positioning into the nanovials. In this work, the properties of the nanodigestion concept regarding proteins of different characteristics are explored. Furthermore, the potential of automated protein identification using precoated proteolytic nanovial-arrays is demonstrated.

Animals↗
PubMed1997

A microassay for the detection of low levels of cytochrome P450 O-deethylation activities with alkoxyresorufin substrates.

A microfluorometric method for the detection of low levels of cytochrome P450 was developed to increase the sensitivity of the assay, since a low level of CYP450 associated enzymatic activities was expected in human placenta tissues and small samples of placenta (approximately 10 g) could be easily collected, stored and processed. The dual fluorescence assay of Kennedy et al. [1], which was developed to simultaneously quantitate microsomal proteins and ethoxyresorufin-O-deethylase (EROD) activity was adapted for 96 wells microtiter plates. Placental microsomes samples were analyzed. For samples obtained from non-smoking mothers from the general southern Quebec population, results ranged from less than 1-3.3 pmol/mg protein.min. Samples collected from smoking mothers showed activity levels ranging from 30-69 pmol/mg protein.min. These results showed the suitability of the microassay for measuring low level of CYP450 activity in tissues such as placenta.

Alkylation↗
PubMed2000

Capillarity, fibre types and fibre morphometry in different sampling sites across and along the tibialis anterior muscle of the rat.

Capillarity, fibre types, fibre cross-sectional areas and perimeters were studied along and across the rat tibialis anterior muscle. The muscle was sectioned at three different levels (proximal, equatorial and distal) choosing five sampling fields for measurements at each level (from anterior to posterior and lateral to medial zones). Significant differences were found in the percentage of fibre types and capillarity between different fields of the same muscle section. Slow oxidative fibres were confined to the posterior muscle zone with a maximum of 3.7%. The posterior fields also had a greater percentage of fast oxidative glycolytic fibres at proximal (72.3%) and equatorial (61.3%) levels, but a lower value at the distal level (44.8%) and lower capillary density counts in total cross-section means (758 vs. 1,069 capillaries/mm(2) in equatorial and 1,035 capillaries/mm(2) at proximal levels). The uneven distribution of both fast fibre types and the different degrees of capillarisation along the muscle are statistically significant and may be due to different biomechanical performances along the rat tibialis anterior. Fibre size was significantly larger at the distal level, but no morphometric differences were found across the section of the same level. At the distal level, the mean total fibre area of fast glycolytic fibres (5,130 microm(2)) and fast oxidative glycolytic fibres (2,493 microm(2)) contrasted with values at the proximal (fast glycolytic: 4,070 microm(2), fast oxidative glycolytic: 1,970 microm(2)) and equatorial (fast glycolytic: 3,535 microm(2), fast oxidative glycolytic: 1,714 microm(2)) levels. The differences along and across the muscle show the need to design a standardised procedure for sample location when performing comparative studies of morphofunctional adaptive changes in skeletal muscle. A significant difference between individuals (animals) in all parameters was evident and should be taken into consideration when analysing the variability: the factor 'animal' should be considered in multiway ANOVAs, especially when low sample sizes are used.

Analysis of Variance↗
PubMed2002

Effects of repeated biopsying on muscle tissue in horses.

The collection of muscle samples in horses to estimate their performance potential, adaptations to training and diagnosis of exertional myopathies is of increasing interest. This study was designed to examine whether repeated biopsying of the gluteus medius muscle at 7 week intervals alters morphology and muscle fibre variables in the sampling area. The gluteus had been biopsied 3 times previously, each biopsy separated by a 7 week interval. Seven weeks after the last biopsy, this study was initiated. Percutaneous needle biopsies were taken from left and right muscles at 2 depths (2 and 6 cm) and at 2 sites for each depth (22 and 25 cm from the tuber coxae) of 7 mature Thoroughbreds. The 22 cm site was located close (2 cm caudal) to an area of the muscle that had been biopsied previously. The 25 cm site was 5 cm apart from this area. A total of 52 samples were available for histology, electron microscopy and immunohistochemistry. The latter was used to study percentages, sizes and capillaries of muscle fibre types. Most muscle samples collected (75%) had normal morphology, but the remaining 25% showed signs of muscle repair. In some circumstances, they showed regenerative signs of complete muscle repair, while in others ineffective muscle repair (scar formation) was evident. Nevertheless, sufficient normal tissue was always available for measuring routine muscle variables. Samples collected 3 cm laterally apart showed large differences with regard to muscle fibre type variables examined. These results show that repeated muscle samples in intervals of 7 weeks do not have effects that would impair evaluation of muscle fibre variables for diagnosis of effectivity of conditioning programmes. Furthermore, they emphasise that the only way of keeping variations through sample collection technique small is to standardise all biopsying techniques accurately.

Animals↗
PubMed1991

Carbohydrate Metabolism in Taproots of Medicago sativa L. during Winter Adaptation and Spring Regrowth.

Our objective was to identify amylases that may participate in starch degradation in alfalfa (Medicago sativa L.) taproots during winter hardening and subsequent spring regrowth. Taproots from field-grown plants were sampled at intervals throughout fall, winter, and early spring. In experiment 1, taproots were separated into bark and wood tissues. Concentrations of soluble sugars, starch, and buffer-soluble proteins and activities of endo- and exoamylase were determined. Starch concentrations declined in late fall, whereas concentrations of sucrose increased. Total amylolytic activity (primarily exoamylase) was not consistently associated with starch degradation but followed trends in soluble protein concentration of taproots. This was especially evident in spring when both declined as starch degradation increased and shoot growth resumed. Activity of endoamylase increased during periods of starch degradation, especially in bark tissues. In experiment 2, a low starch line had higher specific activity of taproot amylases. This line depleted its taproot starch by late winter, after which taproot sugar concentrations declined. As in experiment 1, total amylolytic activity declined in spring in both lines, whereas that of endoamylase increased in both lines even though little starch remained in taproots of the low starch line. Several isoforms of both amylases were distinguished using native polyacrylamide electrophoresis, with isoforms being similar in bark and wood tissues. The slowest migrating isoform of endoamylase was most prominent at each sampling. Activity of all endoamylase isoforms increased during winter adaptation and in spring when shoot growth resumed. Endoamylase activity consistently increased at times of starch utilization in alfalfa taproots (hardening, spring regrowth, after defoliation), indicating that it may serve an important role in starch degradation.

Journal Article↗