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PubMed1980

[Effect on peripheral blood cells in children with acute leukemia during remission by intramuscular injected of arabinosyl-cytosine (author's transl)].

Investigation was done in children with acute leukemia in complete hematological remission. It was tested whether out-patient treatment by intramuscular administration of arabinosylcytosine (Ara-C) may obviate the continuous intravenous Ara-C regimen. Every therapy cycle lasted 5 days. Changes in peripheral blood cell counts after 5 days of continuous intravenous. Ara-C infusion against 5 days of intramuscular Ara-C application given every 12 h were tested. 129 cycles of intramuscular application and 11 cycles of intravenous application were evaluated. Evaluation was done by the Friedmanntest. There was a significant decrease in blood cell counts after intramuscular Ara-C treatment. When administered intramuscularly during the first cycle Ara-C was effective for at least 3 weeks, whereas after repeated cycles the decline of blood cells was only demonstrable for 14 days. A comparison of the effect between intravenous and intramuscular routes revealed similar results. No local side effects were noted when Ara-C was given intramuscularly.

Blood Cell Count↗
PubMed2004

Hollow-fiber assay for ligand-mediated cell adhesion.

BACKGROUND: The investigation of receptor-ligand interactions in the cellular context presents significant technical challenges, first, to immobilize the ligand in a manner that preserves functional properties and, second, to relate ligand properties to cell adhesion and other cellular processes. METHODS: Ligand-mediated cell adhesion was characterized by the development of a cellulose hollow-fiber adhesion assay in which ligand (protein A) was immobilized onto the cellulose membrane as a recombinant fusion protein containing a cellulose-binding domain affinity tag. Modules containing single cellulose hollow fibers were connected to a micro-flow system for cell deposition and detachment with fluid shear stress. The cell adhesion process that occurred inside a segment of hollow fiber was observed in real time by using an inverted microscope equipped with a CCD camera and digital frame grabber. Image analysis software was developed to count cells and record digital images. RESULTS: Cell adhesion strength was characterized by counting the number of cells that were detached by application of fluid shear stress with values that ranged from 2.3 to 185 dyne/cm2. The median shear stress of detachment of KG1a cells was directly related to the duration of membrane contact and the amount of immobilized monoclonal antibody (anti-CD34). CONCLUSIONS: The hollow-fiber assay provides a general method to determine functional properties of molecular domains that interact with cell surface receptors and markers.

Antibodies, Monoclonal↗
PubMed2006

Quantifying the amount of missing information in genetic association studies.

Many genetic analyses are done with incomplete information; for example, unknown phase in haplotype-based association studies. Measures of the amount of available information can be used for efficient planning of studies and/or analyses. In particular, the linkage disequilibrium (LD) between two sets of markers can be interpreted as the amount of information one set of markers contains for testing allele frequency differences in the second set, and measuring LD can be viewed as quantifying information in a missing data problem. We introduce a framework for measuring the association between two sets of variables; for example, genotype data for two distinct groups of markers, or haplotype and genotype data for a given set of polymorphisms. The goal is to quantify how much information is in one data set, e.g. genotype data for a set of SNPs, for estimating parameters that are functions of frequencies in the second data set, e.g. haplotype frequencies, relative to the ideal case of actually observing the complete data, e.g. haplotypes. In the case of genotype data on two mutually exclusive sets of markers, the measure determines the amount of multi-locus LD, and is equal to the classical measure r(2), if the sets consist each of one bi-allelic marker. In general, the measures are interpreted as the asymptotic ratio of sample sizes necessary to achieve the same power in case-control testing. The focus of this paper is on case-control allele/haplotype tests, but the framework can be extended easily to other settings like regressing quantitative traits on allele/haplotype counts, or tests on genotypes or diplotypes. We highlight applications of the approach, including tools for navigating the HapMap database [The International HapMap Consortium, 2003], and genotyping strategies for positional cloning studies.

Alleles↗
PubMedDate not supplied

Efficacy of an antibiotic coated indwelling catheter: a preliminary report.

This study was conducted to examine if an aminoglycoside (dibekacin sulfate, DKB) incorporated into a coating layer on outer and inner surface of indwelling catheters would be released into urine both in vitro and in clinical cases on a sustained basis and if the released DKB would have any efficacy on delaying lower urinary tract infections. Released amounts of DKB from the catheter silicone rubber (SR) catheter were periodically measured both in vitro and in clinical applications. During the clinical applications, the catheters were indwelled in 14 patients. Organism counts in the patients' urine were determined and organisms were isolated from bacteriuria (defined as greater than or equal to 10(4) CFU/mL) and MICs to DKB were measured. Observations on several combined symptoms frequently associated with indwelling catheterization were performed. Results showed that sustained release of DKB continued for more than 25 days and 13 days in vitro and in clinical cases, respectively. Clinical studies suggested that 8 days of sterile urine after catheterization might be expected in patients without systemic administration of antibiotics and more than 2 weeks if combined with it. No particular problems in its use and associated symptoms were recognized.

Catheters, Indwelling↗
PubMed1995

CCD microscopy and image analysis of cells and chromosomes stained by fluorescence in situ hybridization.

This paper reviews methods and applications of CCD microscopy for analysing cells and chromosomes subjected to fluorescence in situ hybridization (FISH). The current status of indirect and direct FISH staining methods with respect to probe labelling, detection sensitivity, multiplicity and DNA resolution is summarized. Microscope hardware, including special multi-band pass filters and CCD cameras required for FISH analysis, is described. Then follows a detailed discussion of current and emerging applications such as the automated enumeration of chromosomal abnormalities (counting of dots in interphase cells), comparative genomic hybridization, automated evaluation of radiation-induced chromosomal translocations, and high-resolution DNA mapping on highly extended chromatin. Finally, the limitations of the present methodology and future prospects are discussed.

Chromosome Mapping↗
PubMed1976

Colony growth of mouse bone marrow cells in agar contained in glass capillaries.

Mouse bone marrow cells were grown in semi-solid agar contained in glass capillary tubes. Several parameters affecting colony formation in the capillaries were studied. 10(4) cells in 100 mul incubation medium within one capillary produced 22 to 30 colonies of granulocytes and macrophages. Compared with the common petri dishes glass capillaries offer several advantages under the conditions used: 1. A twofold higher plating efficiency. 2. Applicability to optical scanning by light scattering and electronic counting, allowing automation and greatly improving sensitivity, statistical accuracy and reproducibility. Kinetics of colony growth can also be monitored. 3. Diminished risk of bacterial and fungal contamination. 4. A more than tenfold lower need for materials on similar statistical errors. Substituting methylcellulose for agar resulted in colonies of fibroblast-like cells adherent to glass surface. Glass non-adherent cells showed a threefold higher plating efficiency in agar.

Agar↗
PubMedDate not supplied

Elevated polymorphonuclear phagocytic function in thalassemia patients by chemiluminescence.

Twenty five patients with beta thalassemia major, with no evidence of infection were evaluated for their polymorphonuclear cell (PMN) metabolic function and serum opsonic activity by chemiluminescence assay. These were divided into Group I of normal adults (n = 21), Group II thalassemia major < 5 years (n = 9) and Group III thalassemia major > 5 years (n = 16). The ability of the chemiluminescence assay (CL) to reflect opsonic and phagocytic dysfunction suggested its potential application in the evaluation of phagocytic function. The peak count of Group I was (1.07 +/- 0.24 x 10(-5)), Group II (1.60 +/- 0.83 x 10(-5)) and Group III was (2.71 +/- 0.98 x 10(-5)) respectively in the presence of autologous sera. The peak count compared between Group I and III was found to be statistically significant (p < 0.05). The peak count of Group I and II when compared showed a trend in the increase activity not statistically significant. The polymorph function of all the groups were compared with autologous serum as well as normal serum. There was no increase in polymorph function of Group III in the presence of thalassemia serum, nor any decrease in the polymorph function of thalassemia patients of Group II and III. This concluded that polymorphs of thalassemia patients are active in the presence of autologous as well as normal serum. The increased activity of thalassemia polymorphs may be due to antigenic stimulation which may be due to multiple transfusion and not due to circulating iron load.

Adult↗
PubMed1999

Is loss-free counting under statistical control?

A new formula for the statistical uncertainty of "loss-free counting" (LFC) is presented. Its validity is demonstrated by comparing with experimental data obtained with a HPGe gamma-ray spectrometer. Also, computer simulation data of nuclear counting with different types of count loss (pileup rejection, extending and nonextending dead time) are in agreement with the predicted counting uncertainty. The proposed formula for LFC uncertainty is applicable to spectrometers with a classical semi-Gaussian pulse-shaping amplifier as well as with a gated-integrator amplifier. Hence, achieving statistical control seems to be a feasible goal.

Computer Simulation↗
PubMed1999

Cystoid macular edema following immune recovery and treatment with cidofovir for cytomegalovirus retinitis.

BACKGROUND: Cytomegalovirus (CMV) retinitis is the most common opportunistic ocular infection in AIDS patients. Cidofovir has proved to be highly effective in treatment of CMV retinitis. Iritis and bulbar hypotony are known as the major complications after intravenous and intravitreal use of this antiviral drug. Cystoid macular edema (CME) after intravenous application of cidofovir has not been reported. METHODS: We analyzed retrospectively the incidence of CME after intravitreal or intravenous application of cidofovir and its correlation with CD4 cell counts of the patients. RESULTS: Two (22.2%) of 9 eyes in the intravenous and 3 (18.8%) of 16 eyes in the intravitreal injection group developed CME. It occurred between 3 and 48 weeks after cidofovir administration. In all eyes CMV retinitis was inactive. All patients received highly active antiretroviral treatment (HAART). CME was correlated with a rapid and sustained improvement in CD4 cell counts. CONCLUSION: We interpret the occurrence of CME as an immune recovery phenomenon for the following reasons. All CMEs were seen in eyes with inactive CMV retinitis and the unaffected contralateral side never developed CME. The time range of appearance between 3 and 48 weeks after cidofovir administration makes direct toxicity of cidofovir unlikely. All patients had a sustained improvement of CD4 cell counts due to HAART. No CME was reported during the use of cidofovir before the HAART era.

AIDS-Related Opportunistic Infections↗
PubMed1986

Motor, sympathetic and sensory innervation of rat skeletal muscles.

This study reports on the location, number and size of motor, sympathetic and sensory neurons innervating the following muscles of rat: quadriceps femoris (QF), tibialis anterior (TA), extensor digitorum longus (EDL), peroneus longus (PL), gastrocnemius medius (GM) and soleus (SOL). Cells were labelled by application of horseradish peroxidase (HRP) to transected muscle nerves. Counts of neurons were compared with counts of myelinated (MF) and unmyelinated (UMF) fibers in normal, deafferented and chemically sympathectomized nerves. The topographical arrangement of spinal motor nuclei resembled that reported previously in other mammals and birds. Sensory somata were aggregated without precise somatotopic organization, preferentially in one of the lumbar dorsal root ganglia at a segmental level corresponding to that of the motor innervation. Because lumbar sympathetic ganglia were often poorly circumscribed, the segmental position of sympathetic ganglion cells could not be localized with certainty. Sensory and sympathetic somata demonstrated a unimodal size-frequency distribution, while QF, TA and PL motoneurons could be subdivided according to size in alpha and gamma cells. For all muscles except unsuccessfully deafferented QF, counts of motor fibers after deafferentation correlated closely with counts of labelled motoneurons. Similarly, estimates of sympathetic axons, averaging 30,7% of the UMF, in most instances exceeded only marginally the ganglion cell population. In contrast, the number of peripheral afferent fibers outnumbered markedly that of sensory cell bodies, with an average of 2.8 axons per ganglion cell.

Animals↗
PubMed1993

Localization of sympathetic postganglionic neurons innervating the femoral-saphenous vein in cats.

Physiological and histochemical studies have suggested that the limb veins are innervated by sympathetic adrenergic fibers. In the present experiment, horseradish peroxidase (HRP) was used as a retrograde tracer to identify and localize the sympathetic postganglionic neurons that innervate the femoral-saphenous vein in cats. In anesthetized cats, HRP was applied perivascularly on a femoral and a saphenous vein segment (4-8 mm in length for each segment) to allow uptake into the nerve endings. The sympathetic chains on both sides were dissected after the animal was sacrificed and fixed 60 h following the HRP application. Histological examination on serial section was done to count the HRP-labeled neurons in each sympathetic ganglion from L1 to S1. In 10 cats, the total number of HRP neurons amounted to 8569. Most neurons arose from L3 (47%) and then L4 (31%). The number of neurons became progressively decreasing towards both ends of the sympathetic chain. Few neurons (less than 2% of the total) were discovered in the contralateral sympathetic ganglia. In each ganglion, the distribution of HRP neurons appeared to be scattering. Our findings provide anatomical evidence to support that the femoral-saphenous vein of the cat was innervated by the sympathetic efferent fibers. The main origins of these neurons are the third and fourth lumbar sympathetic ganglia.

Animals↗
PubMed1991

Angiogenesis: modulation with opioids.

1. The effect of beta-endorphin (beta-EP) and morphine sulfate (MS), in presence and absence of naloxone (NX), on chicken chorioallantoic membrane was studied as a function of blood vessel proliferation. 2. A 50% reduction in blood vessel proliferation occurred by 10 micrograms of beta-EP or by 5 micrograms of MS per egg compared to controls. 3. An individual dose, i.e. 5 micrograms of beta-EP, did not significantly inhibit blood vessel counts after initial 24 hr period of the drug application when given alone compared to inhibition occurring with combined use of NX. 4. NX (1 microgram) did not significantly reverse the angiostatic effects of MS (10 micrograms) or of beta-EP (5 micrograms). 5. The observed modulation of angiogenesis by opioids suggests involvement of beta-EP and MS in the proliferation of vascular endothelial cells. 6. This may be due to an effect of beta-EP and MS on cell-mediated immunity factors such as interferons, interleukins and prostaglandin E2.

Animals↗
PubMed2004

Long-term parvovirus B19 viraemia associated with pure red cell aplasia after allogeneic bone marrow transplantation.

BACKGROUND: Parvovirus B19 infection is associated with a variety of symptoms like erythema infectiosum, anaemia and arthritis. In immunocompetent persons, viraemia is usually cleared a few weeks after infection. OBJECTIVE: An immunocompromised adult female patient was persistently infected with B19 after allogenic bone marrow transplantation (BMT) and developed chronic anaemia. STUDY DESIGN: B19-specific antibodies were determined by ELISA and viral load was assessed using a quantitative real time B19 PCR. The patient was evaluated clinically. RESULTS: Two years after successful BMT, the patient received intensified immunosuppressive treatment, erythropoetin and erythrocyte concentrates due to chronic graft-versus-host disease with renal failure. Despite of this treatment, the aplastic anaemia worsened. PCR revealed B19 viraemia with 10(12) geq/ml serum. After 7 months of repeated applications of immunoglobulins and reduction of immunosuppressive treatment, reticulocyte counts and haemoglobin levels normalized and the viral load finally dropped to 10(3) geq/ml serum. One of the back-up samples of the erythrocyte concentrates tested positive, the respective transfusion had been applied 2 months after the beginning of viraemia. CONCLUSIONS: The source of the primary infection remained unclear, but at least re-infection by blood transfusion is likely. Treatment did not result in virus elimination from peripheral blood but in resolvement of symptoms.

Anemia, Aplastic↗
PubMed1998

Time-of-flight PET.

Time-of-Flight (TOF) positron emission tomographs (PET) were developed during the 1980s and early 1990s. Initial investigations into the potential improvements in signal-to-noise ratios if TOF information was utilized were published in 1980. By 1982, two groups (Washington University and Commissariat á l'Energie Atomique-Laboratorie d'Electronique et de L'Informatique [CEA-LETI]) were designing and building the first TOF tomographs. A third group at the University of Texas also designed and built a TOF system. These systems were optimized for high count-rate imaging of short-lived radiotracers for applications such as cardiac blood flow. The first system put into operation for patient scans was the Super PETT I built at Washington University by Michel Ter-Pogossian and his colleagues. The Washington University group went on to design two additional versions of TOF systems and the CEA-LETI group developed two basic tomograph designs. As Bismuth Germanate (BGO)-based scanners were refined, it became clear that the TOF systems could not provide the same high spatial resolution as offered by the newer systems. The use of the fast scintillators required for TOF systems also resulted in lower intrinsic sensitivity that was only partially compensated for by the effective gain in sensitivity offered by TOF image reconstruction techniques. Further development of TOF systems was suspended in the early 1990s. With the development of new scintillators that provide more light output and are denser than those available in the 1980s and considerably faster than BGO, there is new interest in the application of TOF techniques for future tomograph designs.

Humans↗
PubMed1999

Fast scanning and efficient photodetection in a simple two-photon microscope.

Two-photon laser scan microscopy carries many advantages for work on brain slices and bulk tissue. However, it has very low signal levels compared to conventional fluorescence microscopy. This is disadvantageous in fast imaging applications when photon shot noise is limiting. Working on brain slices with excitation powers of 8-10 mW at the specimen plane, the resting signal from cerebellar Purkinje cell somas loaded with 10 microM Oregon Green 488 BAPTA-1 averaged 4 detected photons/micros; axons of interneurons loaded with 200 microM of this indicator yielded about 1 photon/micros. To obtain satisfactory images at high time resolution, long pixel dwell times are required and data collection should be restricted to as few pixels as necessary. Furthermore, a large proportion of total measurement time (duty cycle) should be available for data collection. We therefore developed a method for scanning small regions of interest with line repetition rates two to four times higher than conventional ones and a duty cycle of 70%. We also compared the performance of several photodetectors and found the optimum choice to depend strongly on the photon flux during a given application. For fluxes smaller than 5 photons/micros, the photon counting avalanche photodiode shows the best signal to noise ratio. At larger fluxes, photomultipliers or intensified photodiodes are superior.

Animals↗
PubMed1997

Response to chemotherapy has predictive value for further survival of patients with advanced non-small cell lung cancer: 10 years experience of the European Lung Cancer Working Party.

The aim of this study was the assessment of the predictive value for survival of an antitumoral response to three courses of chemotherapy in association with various pretreatment characteristics in patients with non-resectable non-small cell lung cancer treated by cisplatin- (or carboplatin)-based combination regimens. Patients considered for this study were eligible patients with advanced non-small cell lung cancer registered in one of the seven trials conducted by the European Lung Cancer Working Party from December 1980 to August 1991. All these trials tested chemotherapy regimens with platinum derivatives (cisplatin and/or carboplatin). In this population of 1052 eligible patients, 752 were assessed in this analysis. Data were prospectively collected on 23 pretherapeutic variables and objective response after three chemotherapy cycles. The predictive value of response to chemotherapy on survival (measured from the time of response assessment i.e. 12 weeks after registration in the trial) was studied by univariate analysis as well as by multivariate methods (adjustment of the impact of several covariates simultaneously on the dependent variable) with adjustment for the pretreatment prognostic variables. After three cycles of chemotherapy, the global estimated median survival time was 24 weeks with a 95% confidence interval of 22-25 weeks. By univariate analysis, we identified an objective response to chemotherapy as a highly significant discriminant marker (P < 0.0001) for further survival with estimated median survival times of 41 weeks (95% CI: 38-46) and 19 weeks (95% CI: 17-20), respectively, for the responding and non-responding patients. In a Cox regression model fitted to the data using a forward stepwise procedure, this variable was the first selected explanatory variable. Its effect was adjusted by the introduction in the model of initial disease extent, Karnofsky performance status, serum calcium level and white blood cell count. These results were consistent with those obtained by application of recursive partitioning and amalgamation algorithms (RECPAM) which led to a classification of the patients into three homogeneous subgroups. Our results, using a classical Cox regression model consistent with those highlighted by application of a RECPAM analysis, found an objective response to chemotherapy to be a predominant predictive factor for further survival, although it did not allow any conclusion about a causal relationship. The RECPAM results led to a classification of the patients into three subgroups which needs to be validated in other series.

Adult↗
PubMed2004

Receptor model comparisons and wind direction analyses of volatile organic compounds and submicrometer particles in an arid, binational, urban air shed.

The relationship between continuous measurements of volatile organic compounds sources and particle number was evaluated at a Photochemical Assessment Monitoring Station Network (PAMS) site located near the U.S.-Mexico Border in central El Paso, TX. Sources of volatile organic compounds (VOCs) were investigated using the multivariate receptor model UNMIX and the effective variance least squares receptor model known as Chemical Mass Balance (CMB, Version 8.0). As expected from PAMS measurements, overall findings from data screening as well as both receptor models confirmed that mobile sources were the major source of VOCs. Comparison of hourly source contribution estimates (SCEs) from the two receptor models revealed significant differences in motor vehicle exhaust and evaporative gasoline contributions. However, the motor vehicle exhaust contributions were highly correlated with each other. Motor vehicle exhaust was also correlated with the ultrafine and accumulation mode particle count, which suggests that motor vehicle exhaust is a source of these particles at the measurement site. Wind sector analyses were performed using the SCE and pollutant data to assess source location of VOCs, particle count, and criteria pollutants. Results from this study have application to source apportionment studies and mobile source emission control strategies that are ongoing in this air shed.

Air Pollutants↗
PubMed2002

Effectiveness of chlorine washing disinfection and effects on the appearance of artichoke and borage.

AIM: Optimal conditions for chlorine application to obtain a reasonable decrease in the microbial counts without damaging the appearance of artichoke and borage have been established. METHODS AND RESULTS: The influence of chlorine concentration (0-200 mg l(-1)), pH, addition of organic acids, contact time and presence of protective structures on the microflora and vegetal appearance were studied. When pH was not controlled the effect of chlorine depended on its concentration until the pH increase caused by addition of chlorine reached 8.8. Any further increase in chlorine concentration was nullified by the pH increase. When pH was adjusted to 4.5 with acetic acid, the effectiveness increased with concentration. However, the use of citric acid to control pH caused a sharp decrease in effectiveness at concentration about 250 mg l(-1). The higher effectiveness of chlorine on homogenized plant extracts compared with the whole plant showed the impact of the vegetal structures on the resistance of the microorganisms. For artichoke, a relationship between the effectiveness of chlorine disinfection and its structures was also found. Extended washing times did not affect the total counts. However, in both vegetables, the appearance was affected by the extended contact times. CONCLUSIONS: The solutions rendering the highest microbial reduction with minimum damages were: 50 mg l(-1) free chlorine without pH control for artichoke and 100 mg l(-1) free chlorine at pH 7.0 for borage. SIGNIFICANCE AND IMPACT OF THE STUDY: Specific conditions for chlorine disinfection of artichoke and borage were determined to reduce the microorganisms in minimally processed artichoke and borage without damaging their appearance.

Acetic Acid↗