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PubMed1991

Relaxation distribution function of intracellular dielectric zones as an indicator of tumorous transition of living cells.

The response decay data of living cells subject to electric polarization is associated with their relaxation distribution function (RDF) and can be determined using the inverse Laplace transform method. A new polynomial, involving a series of associated Laguerre polynomials, has been used as the approximating function for evaluating the RDF, with the advantage of avoiding the usual arbitrary trial values of a particular parameter in the numerical computations. Some numerical examples are given, followed by an application to cervical tissue. It is found that the average relaxation time and the peak amplitude of the RDF exhibit higher values for tumorous cells than normal cells and might be used as parameters to differentiate them and their associated tissues.

Animals↗
PubMed2026

Next-Generation Sequencing Methods for Sensitive Hepatitis B Viral Genome Analysis: A European Study.

This multicentre study investigated the utility of next-generation sequencing (NGS) to detect and generate hepatitis B virus (HBV) genomes in samples of low viral load (from 0.2 to 6207 IU/mL). 23 HBV DNA-positive plasma samples of genotypes A-E and one HBV-negative control sample were assayed blindly via 9 established NGS methods from 6 European laboratories. Methods included untargeted metagenomics, pre-enrichment by probe-capture followed by Illumina sequencing, and HBV-specific PCR pre-amplification followed by sequencing with Nanopore or Illumina. Full HBV genomes were obtained only from samples with viral loads > 1000 IU/mL using probe-capture methods, > 200 IU/mL using PCR-Illumina methods, > 10 IU/mL using PCR-Nanopore methods, and in no samples using metagenomic methods. Contamination was observed in the negative control and samples with very low viral loads in PCR-based methods. Probe-capture and metagenomic methods detected additional viruses not routinely screened in blood donations, including polyomaviruses and herpesviruses; positive results were confirmed by PCR. In conclusion, NGS may delineate whole-genome sequences at low viral loads if supported by a PCR pre-amplification step. Probe-capture methods also reliably detect HBV without pre-amplification but show limited genome coverage for samples with low viral loads; they may additionally detect a wide range of blood-borne viruses.

Humans↗
PubMed1999

The ratio of X- and Y-bearing sperm in ejaculates of men with three or more children of the same sex.

PURPOSE: The present study evaluated the proportions of X-bearing and Y-bearing sperm within the semen of donors who were the declared fathers of three or more sons or daughters. METHODS: The proportions of sperm were determined using dual-color fluorescence in situ hybridization to identify the X and Y chromosomes. RESULTS: The only difference observed was in semen volume. There was no increase in the proportion of Y-bearing sperm for men with only sons (49.7 +/- 1.3%) or of X-bearing sperm for men with only daughters (44.8 +/- 2.6%). CONCLUSIONS: A preponderance of either sons or daughters in a family cannot be explained simply by an altered ratio of X-bearing and Y-bearing sperm in the father's semen.

Humans↗
PubMed1989

Effects of iron loading on free radical scavenging enzymes and lipid peroxidation in rat liver.

A comparison of the antioxidant protective system and presence of lipid peroxidation was made between rats iron-loaded by two different mechanisms. Superoxide dismutase activity, glutathione peroxidase activity, and reduced glutathione concentrations, together with malondialdehyde production, were measured in the livers of rats chronically iron-overloaded by (a) parenteral iron (primarily Kupffer cell iron deposition) and (b) dietary carbonyl iron (mainly parenchymal iron deposition). In carbonyl iron-treated rats, hepatic superoxide dismutase activity was significantly decreased, whereas hepatocyte lipid peroxidation, as measured by malondialdehyde levels, was significantly increased when compared with control rats at or above iron concentrations of 100 and 185 mumol/g dry wt, respectively. However, no significant decrease in superoxide dismutase activity or significant increase in malondialdehyde levels was observed in iron dextran-treated rats. Glutathione peroxidase activities and reduced glutathione concentrations in rats, iron-loaded by either method, were not significantly different from those of control animals. These results suggest that the deposition of iron in the reticuloendothelial cells of the liver does not lead to lipid peroxidation; however, iron deposited in the parenchymal cells of the liver may lead to an altered free radical antioxidant protective system, resulting in lipid peroxidation in these cells at a similar level of iron loading. We conclude that the cellular site of iron deposition as well as the hepatic iron concentration is important in determining iron-induced liver injury.

Animals↗
PubMed2004

Effect of retrobulbar injection of lidocaine on saccadic velocities.

PURPOSE: To determine whether exposing the extraocular muscles (EOMs) to lidocaine via retrobulbar injection for cataract surgery has a demonstrable negative effect on subsequent function of the muscle. SETTING: York Finch Eye Associates, Humber River Regional Hospital, and Toronto Western Hospital Research Institute, Toronto, Ontario, Canada. METHODS: This study comprised 37 eyes that had phacoemulsification and posterior chamber intraocular lens implantation; 13 eyes had retrobulbar lidocaine with hyaluronidase and 24 eyes, topical anesthesia. The postoperative saccadic velocities were compared with the preoperative velocities using a sensitive recording device. The results were compared within and between the retrobulbar lidocaine and topical anesthesia groups. RESULTS: No detectable decrement in postoperative saccadic velocities was detected in any patient, and no difference was found between the groups. CONCLUSIONS: Exposing EOMs to lidocaine for cataract surgery had no detectable negative effect on saccadic velocities 1 week after surgery.

Aged↗
PubMed2005

Monte Carlo simulation study of droplet nucleation.

A new rigorous Monte Carlo simulation approach is employed to study nucleation barriers for droplets in Lennard-Jones fluid. Using the gauge cell method we generate the excess isotherm of critical clusters in the size range from two to six molecular diameters. The ghost field method is employed to compute the cluster free energy and the nucleation barrier with desired precision of (1-2)kT. Based on quantitative results obtained by Monte Carlo simulations, we access the limits of applicability of the capillarity approximation of the classical nucleation theory and the Tolman equation. We show that the capillarity approximation corrected for vapor nonideality and liquid compressibility provides a reasonable assessment for the size of critical clusters in Lennard-Jones fluid; however, its accuracy is not sufficient to predict the nucleation barriers for making practical estimates of the rate of nucleation. The established dependence of the droplet surface tension on the droplet size cannot be approximated by the Tolman equation for small droplets of radius less than four molecular diameters. We confirm the conclusion of ten Wolde and Frenkel [J. Chem. Phys. 109, 9901 (1998)] that integration of the normal component of the Irving-Kirkwood pressure tensor severely underestimates the nucleation barriers for small clusters.

Journal Article↗
PubMed1999

Analysis of CD34 populations in mobilised peripheral blood stem cell harvests and in bone marrow by fluorescent in situ hybridisation for the bcr/abl gene fusion in patients with chronic granulocytic leukaemia.

For those patients ineligible for allogeneic bone marrow transplant and who are non-responsive to interferon, autotransplant with peripheral blood stem cells (PBSC) mobilised after intensive chemotherapy, may provide a novel approach to improve prognosis in patients with chronic granulocytic leukaemia. PBSC harvests are assessed for CD34-positive cell numbers, which serve as an indicator of engraftment potential, and are also analysed cytogenetically to ascertain tumour cell contamination. However, a more accurate assessment of PBSC harvest contamination requires investigation of the Philadelphia (Ph) status of the CD34pos population, in which the cells that provide long-term engraftment are contained. In this study, we have analysed these levels in mobilised PBSC and also in bone marrow (BM) harvests, taken several weeks prior to mobilising chemotherapy. Using fluorescent in situ hybridisation for the bcr/abl gene fusion, we have shown that the median number of Ph negative cells in CD34pos isolated populations was 14.95% in BM compared to 79.05% in PBSC harvests and that in all PBSC samples tested, Ph positivity in CD34pos populations was always detectable either by FISH or one round PCR methods. In paired assessments of both PBSC and BM harvests, higher levels of Ph negative CD34pos cells (> or = 14%) isolated from BM harvests, taken prior to intensive chemotherapy, correlated with higher levels of Ph negative CD34pos cells (> or = 78.5%) in PBSC harvests. These data may aid in the selection of patients for whom PBSC harvesting, after mobilisation, is more likely to achieve an autograft product containing predominantly Ph negative CD34pos cells and may exclude those patients for whom the risk, morbidity and expense of stem cell harvesting may have no apparent benefit over a chronic phase BM harvest.

Adult↗
PubMed2002

Quantitative evaluation of nonablative laser technology.

A new method for treating facial rhytides and acne scars with nonablative laser and light source techniques has recently been introduced. Given the inherent limitations of photographic and clinical evaluation to assess subtle changes in rhytides and surface topography, new noninvasive, objective instruments were used to accurately assess the outcome of these procedures. This study measures and objectively quantifies facial skin by using 2 novel, noninvasive measuring systems after 3 to 5 treatment sessions with the 1,064 nm QS Nd:YAG laser in patients with photodamage and acne scarring. One system measures the skin surface topography and the other characterizes the biomechanical properties of the skin. Patients undergoing facial rejuvenation procedures were analyzed before and after therapy with a 32 x 32 mm in vivo three-dimensional microtopography imaging system (PRIMOS, GFM, Teltow, Germany). The imaging system projects light on to a specific surface of the skin with a Digital Micromirror Device (DMD; Texas Instruments, Irving, TX) and records the image with a CCD camera. Skin Surface microtopography is reconstructed using temporal phase shift algorithms to generate three-dimensional images. Measurements were taken at baseline, at various times during the treatment protocol, and then at 3 and 6 month follow-up visits. Silicone skin replicas (FLEXICO, Herts, England) were also made before and after the laser treatment protocol for comparison to in vivo acquisition. Skin stiffness and compliance were measured with the BTC-2000 (BTC 2000, S.R.L.I. Inc. Nashville, TN). This instrument measures the response of the skin during dynamic stress created by suction. An infrared targeting laser measures the vertical displacement of the skin during the cycle. Pressure and deformation are graphically displayed on the monitor and embedded biomechanical algorithms determine the aforementioned parameters based on the average of 3 repetitive cycles. Four sites on the face were analyzed and compared including the right and left cheek and the right and left forehead. Measurements were taken at baseline and then at 1-, 3- and 6-month follow-up visits. Skin roughness decreased 11% from baseline in the patient with photodamaged skin, while the patients with acne scarring showed a 33% improvement from baseline after 3 treatment sessions. Six-months after the fifth treatment session, a 26% improvement in skin smoothness was documented in the patients with photodamaged skin, while a 61% improvement was recorded in the subject with acne scarring. Biomechanical analysis of the skin showed a 23% decrease in skin stiffness and a 30% increase in skin compliance at the 6-month follow-up in the patients with photodamage. Alternatively, the patient with acne scarring showed a 50% increase in skin stiffness, and a 30% decrease in skin compliance at the 6-month follow-up. Three-dimensional in vivo optical skin imaging provided a rapid and quantitative assessment of surface topography and facial fine lines after multiple treatment sessions with a 1064-nm QS Nd:YAG laser, correlating with clinical and subjective responses. Biomechanical analysis provided technical understanding of structural changes in photodamaged skin and acne scarring following nonablative laser treatment. Future applications of these devices may include comparison of nonablative laser technology, optimization of treatment regimens, and objective evaluation of other aesthetic procedures performed by dermatologists.

Cicatrix↗
PubMed1997

Successful management of external duodenal fistula in a specialized unit.

BACKGROUND: High-output external duodenal fistula remains a challenging condition to treat. Despite advances in metabolic and nutritional care, mortality rates remain high. METHOD: Retrospective casenote review of patients managed on a specialised unit. RESULTS: Thirteen of 388 admissions (3.4%, 11 males, median age 51 years) had an external duodenal fistula. Management was by eradication of abdominal and systemic sepsis and maintenance of nutritional status by the administration of total parenteral nutrition. The majority of fistulas followed surgery for peptic ulcer disease. Eight of 13 fistulas closed spontaneously, and of those requiring operative closure, surgery was successful in 4 of 5 cases. There were 2 mortalities. CONCLUSION: High-output external duodenal fistula may be successfully managed in a specialised unit.

Adult↗
PubMed2002

High-frequency probe EUS-assisted endoscopic mucosal resection: a therapeutic strategy for submucosal tumors of the GI tract.

BACKGROUND: The aim of this study was to evaluate the efficacy and safety of high-frequency probe EUS (HFPE)-assisted endoscopic mucosal resection in the management of submucosal tumors of the GI tract. METHODS: HFPE-assisted endoscopic mucosal resection was attempted in 28 patients with submucosal tumors less than 2 cm in diameter. HFPE was performed with a 20-MHz "through-the-scope" probe. Saline solution was injected into the submucosa. After confirming detachment of the lesion from the muscularis propria by repeat HFPE, endoscopic mucosal resection was performed with a lift-and-cut or endoscopic mucosal resection cap technique. Follow-up endoscopy was performed in all patients. RESULTS: Submucosal tumors from the following areas were included: esophagus 3, stomach 4, duodenum 3, and colon 18. The submucosal tumors were located in the upper third (n = 3), middle third (n = 18), and lower third (n = 7) of the submucosa. Twenty-one submucosal tumors were removed by the lift-and-cut technique and 6 by the cap method. One patient required surgical resection after unsuccessful endoscopic mucosal resection. The origin and depth of penetration of all lesions was accurately depicted by HFPE. Median tumor diameter was 9 mm (range 3-20 mm). Resection was successful and complete in 93% of the cases. There were no immediate postprocedure complications (exact 95% CI [0%, 12.3%]). During a median follow-up of 21.5 months (range 2-74 months) no recurrence was found. CONCLUSIONS: HFPE-assisted endoscopic mucosal resection is safe and effective for the management of selected submucosal tumors of the GI tract. A management algorithm based on endoscopic and HFPE findings is proposed.

Adult↗
PubMed1993

Evaluation of a novel fluorescence polarization immunoassay for teicoplanin.

A fluorescence polarization immunoassay (FPI) for teicoplanin that uses the TDx Instrument System (Abbott, Irving, Tex.) as an automated analyzer has been developed by Innotron of Oregon Inc. and was evaluated in patients with staphylococcal infections enrolled in a clinical trial of the antibiotic. The assay proved accurate in estimating concentrations of between 5 and 100 mg/liter. The intraassay coefficient of variation was < 7.3%, while the interassay variance was < 11.6% against three commercially prepared standards at known concentrations of approximately 5, 35, and 75 mg/liter. Against routinely prepared standards at 10 concentrations between 5 and 100 mg/liter analyzed in a single run, the coefficient of variance did not exceed 4.3%. Compared with bioassay, the FPI demonstrated good correlation in terms of reliability (r = 0.909) in samples containing teicoplanin only and specificity (r = 0.916) in samples containing both teicoplanin and gentamicin. With a turnaround time of 20 min and with only 50 microliters of serum needed for estimation of the amount of drug in a sample, the FPI described here should provide a useful method of teicoplanin measurement in routine diagnostic laboratories.

Biological Assay↗
PubMedDate not supplied

Stem cell regeneration of the nucleus pulposus.

Low back pain due to disc degeneration is one of the largest health problems faced in this nation when judged by lost work time and direct as well as indirect costs. Many experimental methods are being explored to treat or to reverse the effect of disc degeneration. This article reviews the strategy of a tissue engineering approach to disc regeneration. Mesenchymal stem cells (MSC) such as those obtained from marrow stroma when exposed to the appropriate microenvironment (hypoxia, growth factor, three dimensional culture) differentiate into nucleus pulposus-like cells. These then may be candidates for transplantation and nucleus repopulation. While the work is in its infancy, there is significant optimism that next steps will lead to organ culture models of disc degeneration and regeneration, and ultimately to in vivo rescue of degenerating discs with stem cell treatment.

Humans↗
PubMed1988

Evaluation of a portable measure of expired-air carbon monoxide.

There are a variety of independent methods of estimating smoking status; measurement of expired air carbon monoxide has proven to be reliable and convenient. This article describes a new, inexpensive, portable analyzer for measuring expired-air carbon monoxide --the Bedfont EC50. We have compared this analyzer with the standard instrument for measuring expired-air carbon monoxide in 138 normal subjects and found good correlation and agreement. The Bedfont was demonstrated to be better at identifying smoking status, and the relevance of this finding for classification of smokers and nonsmokers by fixed-threshold carbon monoxide levels is discussed. The features of the Bedfont EC50 are particularly appealing for field and clinical use.

Adult↗
PubMed2001

Panning and selection of proteins using ribosome display.

Eukaryotic ribosome complexes can be used as a means to display a library of proteins, and isolate specific binding reagents by screening against target molecules. Here we present, as an example, a method for the display of a library of immunoglobulin variable-like domains (VLDs) for the production of stable mRNA/ribosome/protein complexes. These complexes are produced by the addition of specific in vitro transcriptional promoter elements and translation control sequences to the template DNA. Furthermore, an appropriate spacer (anchor) domain is included for efficient folding of the nascent translated protein, which remains attached to the ribosome complex. Ribosome complexes are panned against hen egg lysozyme-conjugated magnetic beads and genes encoding specific, binding, V-like domains are recovered by RT-PCR and cloned into an Escherichia coli expression vector.

Cloning, Molecular↗
PubMed1999

Colorectal cancer screening in general practice. A survey of current practice and attitudes in Victoria.

OBJECTIVE: To determine the views and practices of Victorian general practitioners (GPs) on the early detection of colorectal cancer (CRC). METHODS: A 1995/1996 quantitative mail survey was used to determine GP knowledge and practice in relation to CRC screening and diagnosis in asymptomatic and symptomatic patients. This includes the amount of investigation, the type and process of diagnostic testing and attitudes to CRC screening. RESULTS: GPs are investigating symptomatic patients but few asymptomatic patients. In general, only patients in the increased risk groups are being investigated. Colonoscopy is the most common method of examination. Most GPs prefer faecal occult blood tests (FOBTs) to be interpreted via laboratory and endoscope procedures to be performed by a specialist. CONCLUSION: GPs were unlikely to screen asymptomatic patients unless there is a family or personal history of CRC or adenomatous polyps. Colonoscopy is the preferred practice for examining such patients. The use of FOBTs for screening standard risk asymptomatic patients was lower than recent research recommends. If population screening for CRC is endorsed, professional and public education in CRC tests, particularly for asymptomatic standard risk patients would be required. Patient initiated screening was highly favoured, supporting the need for public awareness programs.

Colorectal Neoplasms↗
PubMed2003

Optical metrology for analysis of lobster-eye x-ray optics.

A new method that uses optical microscopy to determine the physical structure of lobster-eye x-ray optics is described. This approach offers the ability to predict x-ray performance without having to take an x-ray measurement. An overlapping series of images of the entrance and exit faces of an optic are obtained and examined by purpose-built software. A 24-parameter description of each channel is obtained from which a quantitative analysis of all the major optic defects, except surface roughness, is performed. Results for a planar lobster-eye optic are used to illustrate this technique and discuss its abilities as well as directions for future enhancements.

Journal Article↗
PubMed2006

Non-competitive inhibitory activities of morphinan and morphine derivatives at the alpha 3 beta 4 Neuronal nicotinic acetylcholine receptor determined using nonlinear chromatography and chemometric techniques.

PURPOSE: A series of morphine and morphinan derivatives were chromatographed on a column containing immobilized cellular membranes from a cell line expressing the alpha 3 beta 4 neuronal nicotinic acetylcholine receptor (alpha 3 beta 4 nAChR). METHODS: The results were analyzed using chemometric and molecular modeling techniques in order to predict the noncompetitive inhibitory (NCI) activity of these compounds, the molecular basis for the predicted activity and the binding sites of the inhibitors. RESULTS: The data demonstrated that seven of seven morphinans were NCIs and bound in the central lumen of the nAChR while only 2 of 13 morphine derivatives had NCI activity and these compounds most likely bound at the quinacrine binding site on the nAChR. The predicted activities were confirmed using functional inhibition studies. CONCLUSIONS: The results indicate that this approach can be used to rapidly assess pharmacological activity and to guide new drug design.

Binding Sites↗
PubMed1985

Effect of infant age on aminopyrine breath test results.

UNLABELLED: The aminopyrine breath test has been used in adults as a measure of hepatic N-demethylase activity. In order to study maturational changes in enzyme function, 13C aminopyrine (2 mg/kg) was administered orally to infants (n = 16) between the ages of 1 and 38 wk. Breath samples were collected for 6 h after administration of the labeled aminopyrine for the measurement of 13CO2 enrichment. Using a number of different scoring methods to quantitate 13CO2 elimination of breath, demethylation of aminopyrine was found to be positively correlated to age. By 20 wk of age, some infants had rates of elimination similar to those measured in adults. Absorption was excluded as a limiting variable, because no improvement in oxidation rates was found when the aminopyrine was readministered as an intravenous bolus. Changes in nutritional status and route of feeding (enteral versus parenteral) did not prevent the effect of maturation on aminopyrine elimination. CONCLUSIONS: 1) maturational differences are seen in the metabolism of aminopyrine; 2) these differences may reflect immaturity of N-demethylase activity or diversion of the liberated formaldehyde into biosynthetic rather than oxidative pathways.

Age Factors↗