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Minichromosomal repetitive DNA in Trypanosoma cruzi: its use in a high-sensitivity parasite detection assay.

We have isolated genomic clones containing members of a tandemly repeated DNA family from Trypanosoma cruzi. This family, which contains a 195-base pair (bp) repeating unit, is the most abundant repetitive DNA in this organism. DNA sequencing analysis of three adjacent tandem repeats as well as two independent nonadjacent repeats showed relatively little sequence heterogeneity. Surprisingly, the three tandem elements contained a 585-bp open reading frame. However, blot hybridization of RNA from epimastigotes as well as blood-form trypomastigotes failed to show evidence for transcription of these sequences. Fractionation of whole T. cruzi DNA in sucrose gradients or in agarose gels followed by hybridization with appropriate radioactive probes showed that the size distribution of DNA bearing the 195-bp repetitive element is distinct from that of kinetoplast DNA as well as from that of DNA bearing tubulin genes. Hybridization of the 195-bp element probe with DNA from six different T. cruzi strains was positive; hybridization with DNA of other protozoa was negative with the single exception of Leptomonas collosoma , which displayed a weak cross-hybridization signal. Clones bearing this repetitive element are shown to be useful as probes for identification and counting of T. cruzi cells by dot-blot hybridization. The sensitivity of this assay permits detection of the DNA of 30 parasites in blood samples.

Base Sequence↗

Recent developments in medical applications of SIMS microscopy.

The purpose of this review is to present the recent developments in the medical applications of SIMS microscopy. This technique is one of the microanalytical mass spectrometry methods which allow in theory the detection of all the elements of the Mendeleiev table as well as the separation of stable and radioactive isotopes. It is based on a phenomenon whereby a biological sample surface is sputtered by bombardment with an energetic 'primary ion' beam. Part of the sputtered matter is ionized and the resulting 'secondary ions' are characteristic of the atomic composition of the analyzed area. These secondary positive or negative ions are collected and separated in a mass spectrometer at low or high mass resolution, which is dependent on both the element studied and its concentration. An analytic image which conserves the tissue distribution of the selected element is displayed on a fluorescent screen linked to an image processing system. Local elemental concentration can also be measured. Results are highly dependent on the techniques used for sample preparation which should preserve both the chemical and the structural integrity of the tissue. Further, the ionic images must be correlated with corresponding images of the same areas of the serial sections observed in a photonic microscope. With our SIMS microscope (lateral resolution approximately 0.5 microns, and mass resolution 300 to 12,000) we have demonstrated that this microscopic imaging technique is suitable for physiopathological studies. We revisited thyroid iodine metabolism by mapping chemical elements such as 32S and 127I, characteristic of hormonal physiology. Newly organified iodine (radioiodine) can be evaluated in relation to previously stored iodine (127I) in a given follicle, thus allowing an appraisal of glandular adaptation to aging and iodine overload. Another area in which SIMS can be used in medicine, is for the localization of drug markers in tumor tissue (e.g. fluorine-5-fluouracil, iodine in iododeoxyrubicin). This could facilitate the evaluation of the intratumor drug concentration at the onset of the treatment. Likewise, SIMS can be used to localize radiopharmaceuticals used in diagnosis (e.g. technetium) and therapy (131I of metaiodobenzylguanidine). This would permit a better evaluation of the radiation dose delivered to tissue. Further prospects are within reach with the imminent advent of higher lateral resolution (0.05 microns) SIMS microscopes.

3-Iodobenzylguanidine↗

Localized vascular regression during limb morphogenesis in the chicken embryo. I. Spatial and temporal changes in the vascular pattern.

Morphogenesis of the vertebrate limb bud depends upon reciprocal interactions between epithelial and mesenchymal tissues. A characteristic limb vascular pattern is essential for normal limb outgrowth. The vascular pattern in the distal portion of the wing bud was examined by ink injection and compared to the sites of cartilage differentiation, as indicated by [35S]-incorporation. During development, avascular areas arose in three distinct locations within the vascularized mesoderm. These areas corresponded to the distal skeletal elements, referred to as digits 2, 3, and 4. Incorporation of radioactive sulfate was high in the avascular areas and low in the adjacent vascular tissue. Examination of autoradiographs of ink-injected limbs suggested that the appearance of avascular regions preceded the accumulation of sulfated cartilage matrix. These results indicate that remodeling of the limb vasculature is related to the formation of the skeletal pattern.

Animals↗

The lesson of the Chernobyl disaster.

On April 26, 1986 a major nuclear disaster took place at 1 h 24 min local time, destroying the fourth reactor of the Chernobyl plant. Five years later the consequences of the disaster are still not fully known. Nevertheless the long term future of nuclear energy in the world is uncertain. Questions need to be answered by observing hard facts if emotional attitudes are not to prevail over reality. The reactor and its core were destroyed by an explosion, causing two radioactive jet emissions of iodine 131, followed by caesium 137. Both elements are mainly incorporated in the body via food. The Chernobyl disaster was a consequence of inadequate safety regulations and human error. Enforcement of strict regulations are likely to be highly effective in preventing a further catastrophe. However, governments should consider another possibility. What would be the consequences for public health if a terroristic act deliberately destroyed a nuclear power station?

Accidents↗

Concentration levels of 210Pb and 210Po in dry tobacco leaves in Greece.

Tobacco leaves are large and have sticky exudates that retain the radon decay products once they deposit on the leaves. The study of 210Po in tobacco is required, because of the cumulative alpha-radiation dose delivered to humans from inhaled 210Po in cigarette smoke. 210Pb is the other element of interest since it is the 210Po precursor in the radioactive decay chain. In the present study, the concentrations of these two radionuclides were determined in tobacco samples from seven regions in Greece. 210Po was determined by alpha spectrometry using a surface barrier detector after radiochemical separation and spontaneous deposition of polonium on a nickel disk. The 210Pb activity in the samples was determined via the 210Po resulting from the decay of 210Pb. The results of the present study indicate that 210Po concentrations ranged from 3.6 to 17.0 mBqg(-1) (average 13.1 mBqg(-1)) of dry tobacco, while 210Pb concentrations ranged from 7.3 to 18.0 mBqg(-1) (average 13.4 mBqg(-1)). The mean value of the annual committed effective dose for smokers (20 cigarettes per day) of Greek tobacco was estimated to be 287 microSv (124 microSv from 210Po and 163 microSv from 210Pb). The inhalation dose for smokers is on average about 12 times higher than for non-smokers living in the mid-latitudes of the northern hemisphere.

Greece↗

Bone uptake of vanadium in mice: simultaneous tracing of V, Se, Sr, Y, Zr, Ru and Rh using a radioactive multitracer.

The radioactive multitracer technique, which enables simultaneous tracing of a number of elements in a variety of bio-systems and an accurate comparison of their biobehavior, was employed in a study on uptake behavior of trace elements in various mice tissues, focusing on uptake of V by bone. We found that the bone uptake behavior of trace V closely resembles that of trace Sr and Y. It is suggested that V is a bone-seeking element which probably plays an important role in bone formation. The multitracer technique was thus shown to be useful for comparative studies of behavior of trace elements in living organisms.

Animals↗

Geophysical natural gamma-ray well logging and spectrometric signatures of south AL-Abter phosphatic deposits in Syria.

A combination of several exploration techniques was carried out in AL-Abter region, aiming at outlining the main characteristics of the phosphatic layers in this region. The techniques used in this research are natural 7-ray well logging, gamma and alpha-spectrometry. It is shown that uranium concentrations in the samples taken from the wells studied vary between 42.8 and 112.5 ppm with a standard deviation of 15.2 ppm. The P2O5 content varies between 20.5% and 28.31% with a standard deviation of 3.11% and the radioactive intensities vary between 150 and 275 counts per second (cps) with a standard deviation of 31 cps. The ratio of 234U/238U in the analyzed samples indicates that the study region is in radioactive equilibrium. The affinity of uranium and P2O5 to some trace elements, such as V, Sr, Cu and Ni has been verified through studying the correlation matrix of these elements.

Journal Article↗

[A rapid method for the measurement of major and minor elements in vanadium titanium magnetite sample].

A high resolution (203 eV/Mn Kalpha) portable X-ray fluorescence spectrometer has been developed and the measurement of major and minor elements in vanadium titanium magnetite sample was carried out. An 241Am radioactive source for X-ray excitation, a Si-PIN detector and a 2048 channel analyzer for the measurement of vanadium titanium magnetite direct powder sample were used. The correction methods for interferences, calibration and drift are presented. The method is simply, accurate and rapid. The detection limit is an order of magnitude lower compared with a sealed proportional counter system.

English Abstract↗

Ontogenesis of the hypothalamic catecholaminergic system in rats: synthesis, uptake and release of catecholamines.

The development of the hypothalamic catecholaminergic system during ontogenesis in rats has been studied with glyoxylic acid histofluorescent method in vivo and with isotopic biochemical technique in vitro. It has been demonstrated that at the 15th fetal day the catecholaminergic system was functionally inactive at least in its ability for the uptake and K(+)-stimulated release of catecholamines. Since the 16th fetal day, hypothalamic neuronal elements gained an ability for synthesis of catecholamines, their specific uptake and K(+)-evoked release. Over the subsequent two days, the intensity of the fluorescent intraneuronal product rose considerably showing the increase of either synthesis or accumulation of catecholamines. Simultaneously, the values of the uptake and K(+)-stimulated release of the exogenous radioactively-labelled dopamine increased significantly. The intensity of the fluorescence of the hypothalamic neuronal elements dropped from 20th fetal until the ninth postnatal day, whereas the specific uptake doubled over the same period reaching its adult level. By the 21st postnatal day the reaccumulation of the fluorescent product occurred.

Animals↗

Bile acid absorption kinetics in Crohn's disease on elemental diet after oral administration of a stable-isotope tracer with chenodeoxycholic-11, 12-d2 acid.

A non-radioactive tracer, deuterium, incorporated in chenodeoxycholic acid, was used to study bile acid absorption kinetics employing gas chromatography-mass fragmentography for its analysis. Eight patients with Crohn's disease and four control volunteers were administered chenodeoxycholic-11, 12-d2 acid. In order to obtain constant dietary conditions, they were fed by an elemental diet during the study. There were significant reductions of the biological half-life of chenodeoxycholic-11, 12-d2 acid, the pool size of chenodeoxycholic acid, and the total bile acid pool size in patients with Crohn's disease as compared with those in normal subjects. There was a marked increase in the relative concentration of ursodeoxycholic acid with a concomitant decrease of deoxycholic acid in patients with Crohn's disease, although such differences did not reach statistical significance. The present study confirms that there are some impairments of enterohepatic circulation of bile acids in patients with Crohn's disease.

Adolescent↗

Evaluation of trace elements in human lung tissue. II. Recovery and analysis of inhaled particulates.

Inhaled particulate material, deposited in the lungs of eight Belgian individuals, is differentiated from the tissue with a digestion procedure using the enzyme ficin. This method is simple and efficient and yields a small amount of insoluble dust. The total element concentration of the tissue samples was determined by neutron activation analysis. The radioactive residues obtained after the digestion of the tissue were counted and are considered to represent the insoluble dust fraction. A major part of the Co, Cr, Sb and Sc was recovered in the residues, but only small fractions of the Cs, Rb and Zn. These findings confirm the assumption that Co, Cr, Sb and Sc are enriched in the lungs as a result of the accumulation of atmospheric contaminants. Further proof can be derived from the analogy between the elemental composition of the residue and that of atmospheric particulate matter collected in Belgium.

Adolescent↗

Radiotracers for the determination of the accuracy of trace element analyses.

Many analytical techniques for the determination of trace elements in environmental samples are plagued by ill-defined systematic errors. Investigations of analytical techniques with radioactively labelled analytes can identify systematic errors, lead to their elimination, and can improve the accuracy of the results. Radiotracers are directly detected with high sensitivity and great rapidity in most cases by gamma-ray counters. Examples of the use of radiotracer techniques for the investigation of the following steps of analytical procedures are presented: presampling events, sampling and sample handling, mineralization of samples, separation and preconcentration, and determination. Most of the examples discussed in the text were taken from the analytical chemistry of mercury, sulfur, lead, sodium, zinc and selenium. Other examples are summarized in tables. The results of these investigations clearly show the superiority of the radiotracer technique over other techniques. A more frequent use of the radiotracer technique in investigations aimed at improving the accuracy of analytical procedures is certainly desirable.

Evaluation Studies as Topic↗

Evaluation of the potential of laser-induced breakdown spectroscopy for detection of trace element in liquid.

The analytical figure of merit of the potential of laser-induced breakdown spectroscopy (LIBS) has been evaluated for detection of trace element in liquid. LIBS data of Mg, Cr, Mn, and Re were studied. Various optical geometries, which produce the laser spark in and at the liquid sample, were tested. The calibration curves for Mg, Cr, Mn, and Re were obtained at the optimized experimental conditions with bulk liquid and in liquid jet. It was found that measurements using a liquid jet provide better detection limits than bulk liquid measurements. The limits of detection (LOD) of Mg, Cr, Mn, and Re in the present liquid jet measurement are found to be 0.1, 0.4, 0.7, and 8 ppm, respectively. The LOD of Mg using Mg 279.55 nm was compared with the values found in other liquid work.

Calibration↗

[Principles of a canteen feeding organization of railway transport workers in repair-restorative work in regions polluted with radiation].

Dietary intake and actual food consumption of the railway transport workers are employed in repair in radioactive polluted regions. Disbalance of dining rations and deficiency in vegetable fat, some mineral elements and vitamins was established. For the correction of revealed defects the methodological recommendations for organization of a canteen feeding of the workers of railway transport was developed.

Dietary Services↗

Environmental biochemistry of current environmental levels of heavy metals: preparation of radiotracers with very high specific radioactivity for metallobiochemical experiments on laboratory animals.

Environmental toxicology research on dose-response relationships of heavy metals requires experiments on laboratory animals exposed to "low doses" of trace elements which should reflect "present or actual environmental levels" characteristic of polluted environments. Unfortunately no criteria exist to establish the "low doses" to which laboratory animals must be exposed, in practice the choice of the level used is made in an almost arbitrary manner. In order to define the "present environmental levels" of heavy metals which should be administered to laboratory animals an approach is suggested, based upon knowledge of the concentrations of trace elements in the diet, air and food as well as the fractions absorbed. Today daily intakes of trace elements by man are of the order of few micrograms or nanograms thus requiring the use of extremely sensitive analytical techniques to determine the very low amounts of heavy metals in tissues and cellular components. In these fields of research the use of radiotracers with very high specific radioactivity appears particularly advantageous but requires considerable care during their preparation and use. The first part of this paper deals with a definition of the ranges of concentrations of trace elements which should be used for metabolic studies on laboratory animals when they are exposed via different routes such as ingestion, inhalation in injection; the second part describes the production of radiotracers with very high specific radioactivity by proton activation in the cyclotron and by neutron irradiation in the nuclear reactor. Their use to label present levels of heavy metals under conditions adapted for biochemical purposes, as well as the preparation of different metal-labelled chemical species is also reported. Particular attention is directed to quality control of the radiotracer solutions which are administered to the animals including those of radioactivity concentrations, radioisotopic purity, radiochemical purity, carrier content and chemical impurities.

Animals↗

Non-destructive method of characterisation of radioactive waste containers using gamma spectroscopy and Monte Carlo techniques.

During the decommissioning of the SATURNE accelerator at CEA Saclay (France), a number of concrete containers with radioactive materials of low or very low activity had to be characterised before their final storage. In this paper, a non-destructive approach combining gamma ray spectroscopy and Monte Carlo simulations is used in order to characterise massive concrete blocks containing some radioactive waste. The limits and uncertainties of the proposed method are quantified for the source term activity estimates using 137Cs as a tracer element. A series of activity measurements with a few representative waste containers were performed before and after destruction. It has been found that neither was the distribution of radioactive materials homogeneous nor was its density unique, and this became the major source of systematic errors in this study. Nevertheless, we conclude that by combining gamma ray spectroscopy and full scale Monte Carlo simulations one can estimate the source term activity for some tracer elements such as 134Cs, 137Cs, 60Co, etc. The uncertainty of this estimation should not be bigger than a factor of 2-3.

Algorithms↗

Radiological states around the Kraton-4 underground nuclear explosion site in Sakha.

A radiological survey around the site of Kraton-4, an underground nuclear explosion (Yield of 20 kt, depth of 560 m, 1978) in Sakha was carried out in March 1998. Gamma survey and in-situ spectroscopy on the ground exhibited quite normal levels: a dose rate of 0.022 microSv/h and Cs-137 surface contamination of less than 1.1 kBq/m2 around the hypocenter. The results suggested no remarkable leakage of radioactivity from the epicenter to the ground surface at least not for non-rare gas elements as of 1998.

Dose-Response Relationship, Radiation↗

Nucleosynthesis in stars: recent developments.

The development of new observational, experimental, and computational technologies is changing our understanding of the origins of the elements by thermonuclear burning in stars. Gamma-ray lines from newly made radioactive nuclei have been identified using instruments onboard low-Earth orbiting satellites. Grains in meteorites have isotopic anomalies which suggest that the grains were put together in a stellar explosion such as a supernova. Computer simulations allow such anomalies to be used to probe how these events happen. The simulations are being independently tested by experiments with high-energy density lasers. These developments are beginning to provide a quantitative diagnostic of galactic evolution, and of the epoch of formation of the first stars and galaxies.

Astronomical Phenomena↗