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Neutron flow exposure as a test for survival of Artemia salina spores.

Live and heat-inactivated Artemia salina spores (samples with the same mass and filling density) were exposed to a flow of thermal neutrons from a (252)Cf radioactive source at an equivalent dose power of about 1 microSv/h. Irradiation led to a 4-fold acceleration of nauplius development and to modification of the element profiles of live spores. The difference between absorption/diffusion of thermal neutrons by live and dead spores was revealed.

Animals↗

Influence of the administered mass of tellurium on plasma clearance in rabbits.

The combination of analytical techniques such as PNA and SIMS with a compartmental approach enables the study of the metabolism and biokinetics in humans of several elements by using stable isotopes as tracers. The techniques developed for Te require the administration of greater masses than those used for similar studies performed with radioactive tracers, therefore a test was carried out in rabbits in order to assess the possible influence of the administered amounts on the determination of the biokinetic parameters. The behaviour of the tracers was found to be similar for Te administration of up to 70 micrograms/kg of body weight. An inter-individual variability in the size of the transfer compartment was observed and has to be taken into account.

Animals↗

Aquatic mosses as a monitoring tool for 137Cs contamination in streams and rivers-a field study from central southern Norway.

Mosses are frequently used as biomonitors for trace element pollution in the aquatic environment. The purpose of this study was to assess their usefulness as a tool in monitoring and in regional surveys of radioactive contamination. Specimens of the aquatic mosses, Fontinalis antipyretica and F. dalecarlica, were transplanted from non-contaminated areas to streams and rivers in the Norwegian Jotunheimen Mountains and neighbouring lowland areas that had received radioactive fallout after the Chernobyl accident. Equilibrium concentrations of 137Cs in the exposed mosses were reached after a few weeks. Two series from 20 streams in 1994 and 1996 show linear correlations between activities in water and moss samples and biomagnification ratios of 10(4) - 10(5). We conclude that mosses are better suited for monitoring purposes than water samples, because they provide values integrated over weeks while the radioactivity in surface waters can be subject to rapid variations according to hydrological events. The activity concentrations in aquatic mosses can be easily measured with good precision even when aqueous concentrations are below the limit of detection. Use of aquatic mosses also reduces the logistic problems of transporting large volumes of water, especially in areas inaccessible by road.

Bryopsida↗

Vital staining of fungal elements in deep-seated mycotic lesions during experimental murine mycoses using the parenterally applied optical brightener Blankophor.

Optical brighteners of the diaminostilbene type are fluorescent dyes which are popular diagnostic tools in the mycology laboratory. While these dyes are conventionally used for the in vitro diagnosis of mycoses, their low toxicity and chemical reactivity have led us to investigate their potential use for in vivo staining of fungal elements in mycotic tissue. In mice we have established deep-seated candidiasis, cryptococcosis, aspergillosis and zygomycosis, as well as coccidioidomycosis, histoplasmosis and blastomycosis. After establishment of infection, which mostly required immunosuppression, a single dose of 100 microl of an aqueous solution (2.2 x 10(-4) M) of the optical brightener Blankophor P fluessig (4,4'-Bis [(4-anilino-6-substituted-1,3,5-triazine-2-yl) amino] stilbene-2,2'-disulfonic acid) was injected by the tail vein and the animals were sacrificed 1 h later. Sections of freshly prepared target organs were directly subjected to epifluorescence microscopy using an appropriate filter kit. In most cases, fluorescent fungal elements could be detected in the murine tissue. There was little evidence for uptake of the dye by non-infected tissues. It is suggested that radioactive labeling may render parenteral Blankophor suitable for radiographic localization of deep-seated mycotic foci in the host.

Animals↗

Morphological and biochemical studies of B cells of fetal rat endocrine pancreas in organ culture. Evidence for (pro) insulin biosynthesis.

Fetal rat pancreases explanted on the 18th day of gestation and maintained in organ culture for 1-10 days were utilized for this series of studies. Ultrastructurally, at the time of explantation, the majority of fetal B cells was sparsely granulated and characterized by numerous free ribosomes and undeveloped rough endoplasmic reticulum (RER) and Golgi complexes. During the culture period, extensive development of the RER and Golgi complexes preceded an increasing accumulation of beta-granules. This later increase in the number of beta-granules and in the concentration of immunoreactive insulin was paralleled by a reduction of RER and Golgi complex activity. High resolution radioautographic studies of pulse-chase experiment over a 1 hr period demonstrated the shift of silver grains from the elements of the RER, through the Golgi region, and finally to the beta-granules. Incubation with (14)C-labeled leucine demonstrated the incorporation of radioactivity into molecules possessing the immunological and electrophoretic properties of insulin. These studies indicate that de novo synthesis of (pro)insulin occurs also during culture of fetal rat pancreas explanted relatively late in gestation.

Animals↗

Development of a laser safety hazard evaluation procedure for the research university setting.

Laser safety often receives less attention and fewer resources than other institutional safety and health program elements. This diminished support occurs despite the fact that the potential for laser-related occupational injuries is much greater than the potential for injury from radioactive materials used in biomedical research. Therefore, a small emphasis on laser safety can potentially produce a much greater return on investment than more conventional health physics programs. Developing and implementing a formal laser hazard evaluation procedure provides all efficient mechanism for identifying potential laser safety hazards and offers a great professional development opportunity in this important area of health physics expertise. This paper outlines a process for developing such a university laser hazard evaluation program.

Forms and Records Control↗

Power-law kinetics of tracer washout from physiological systems.

Recent studies suggest that the tail of the washout of tracer-labeled substances from physiological systems can exhibit power-law behavior. In this work we develop a theoretical interpretation of the power-law behavior of the flow-limited washout of tracer-labeled water from the myocardium. Using minimal assumptions concerning the complicated structure of the coronary network we show that the washout from a heterogeneous flow system is given by h(t) approximately equal to A x p1 (V/t)(-beta), where beta is close to 3, p1 is the probability density of flows through the system, V is a constant volume associated with each pathway, and A is a constant. This prediction fits observed power-law washout behavior of tracer water in the heart. This theory is general enough to lead us to speculate that close examination of transport in other heterogeneity-perfused systems is likely to reveal similar power-law behavior.

Animals↗

Isolation of an endocytic compartment from A431 cells using a density modification procedure employing a receptor-specific monoclonal antibody complexed with colloidal gold.

Our objective was to isolate a prelysosomal compartment involved in receptor-mediated endocytosis in human epidermoid carcinoma (A431) cells. The isolation protocol involves density modification of endosome elements in A431 cells, caused by the receptor-dependent binding and internalization at 20 degrees C of colloidal gold-transferrin receptor antibody (B3/25) particles. The use of 125I-labelled gold-B3/25 provides a radioactive marker for the endosome compartment, the major peak being recovered at the bottom of a continuous sucrose gradient at a density of 1.23g ml-1. Enzyme markers characteristic of other cytoplasmic compartments are present only in negligible amounts in this fraction and L-[35S]methionine-labelling of the cells indicates approximately a 200-fold enrichment of 125I-labelled gold-B3/25 versus protein. Electron microscopy of the endosome-rich fraction reveals that we have isolated a highly purified population of small gold-containing vesicles and tubules from which the transferrin receptor can be immunoprecipitated using the B3/25 antibody. Gel electrophoresis and fluorography of L-[35S]-methionine-labelled cells suggests that these elements contain a characteristic profile of approximately 10 major proteins of which three appear to be specifically enriched. In cells incubated with [125I]transferrin, 12% of the ligand sediments with the gold-labelled elements. We conclude, therefore, that the components we have isolated play a role in the intracellular processing of the transferrin-transferrin receptor complexes.

Antibodies, Monoclonal↗

A fat body derived protein is selectively sulfated in the stick insect ovary by transcytosis through the follicular epithelium.

With the onset of vitellogenesis, the follicular epithelium overlying the oocyte in stick insect ovarioles becomes highly polarized and patent by formation of wide intercellular spaces. The aim of the present study was to provide experimental support to the notion that the follicular epithelium in this insect species may be involved in transcytosis. Data demonstrate that the follicular epithelium carries out sulfo-conjugation of a 85 kDa fat body derived protein by allowing it ot transit from one cell pole to another. Along the basal end, follicle cells branch into a number of cytoplasmic finger-like projections. At the opposite end facing the oocyte they taper off into lance-head shapes. Different vesicular elements are evident at both these extremities. In vivo exposure to horseradish peroxidase shows that the vesicular elements present along the apical end provide an endocytic entry. In contrast, those present along the basal end are labeled with sodium [35S]-sulfate, suggesting that they may be exocytic vesicles containing a sulfo-conjugated secretory product. In vivo exposure to sodium [35S]-sulfate caused radioactivity to appear over the Golgi apparatus and some nearby vesicles of the follicle cell cytoplasm, including the exocytic vesicles. The intracellular pathway of the follicle cells was also examined by immunogold labeling using a monoclonal antibody raised against a 85 kDa fat body derived protein. Under these conditions, gold particles were consistently detected over the Golgi apparatus and the vesicular elements lying along both poles of the follicle cell membrane. Based on this evidence, it is concluded that follicular cells in stick insect ovarioles are endowed with the ability to undergo transcytosis by providing an endocytic entry along the apical end and by releasing exocytically a sulfo-conjugated 85 kDa protein along the baso-lateral domain of the follicle cell membrane.

Animals↗

Simultaneous detection of neuropeptides and messenger RNA in the magnocellular hypothalamo-neurohypophysial system by a combination of non-radioactive in situ hybridization histochemistry and immunohistochemistry.

A protocol was developed combining non-radioactive in situ hybridization histochemistry with enzyme based immunohistochemistry, detect the expression of mRNA in phenotypically defined neurons. Free-floating brain sections were hybridized with the oligonucleotide probes which have been 3'-end labelled with biotin-11-dUTP. The hybridized probe was visualized by a combined avidin-biotin bridge method, anti-avidin immunohistochemistry, and horseradish peroxidase detection using diaminobenzidine as a substrate. The in situ hybridization step yielded a very stable reaction product enabling subsequent immunohistochemical reactions using horseradish peroxidase and benzidine dihydrochloride as a chromogen. Magnocellular neurons of the hypothalamo-neurophypophysial system synthesize either vasopressin or oxytocin; water deprivation and chronic saline ingestion are potent stimuli for the expression of both of the genes encoding these neuropeptides. A number of other neuropeptides with putative transmitter action are synthesized in magnocellular neurons during such stimulation. Experiments were performed to explore whether neuropeptide Y immunoreactivity is present within magnocellular vasopressin mRNA-expressing neurons of the hypothalamo-neurophypophysial system. The results clearly demonstrated that neuropeptide Y-immunoreactive elements were present within a number of magnocellular vasopressin mRNA-containing cells. In addition, immunohistochemical detection of the neuropeptides ocytocin and cholecystokinin was carried out on sections hybridized non-radioactively for vasopressin; as expected vasopressin mRNA did not co-exist with cholecystokinin, whereas a few oxytocin immunoreactive neurons in osmotically stimulated animals also contained vasopressin mRNA. The developed method makes possible the immunohistochemical detection of intracellular antigens with concomitant detection of intracellular mRNA.

Animals↗

[Radioecological condition of cleansing structures on the territory contaminated with radionuclides as a result of the accident on the Chernobyl Nuclear Power Station and the problems handing with the forming precipitates of sewage in republic Belarus].

This article deals with the problems, connected with the formation of radioactive deposits in sewage (D.S.). It's shown that the main places of the accumulation of D.S. are the fields of filtration, silty sites and biological pools. In order to reduce the extent of the risk of the secondary contamination of environment we could offer the modernization of the elements of the cleansing structures, the development of refining technologies and reduction of mobility of radiocaesium.

Humans↗

Electron microscope autoradiographic evidence for specific transneuronal transport in the mouse accessory olfactory bulb.

The distribution of radioactive material was examined autoradiographically 8 h after application of [3H] proline to the vomeronasal organ in mice. Labelled material was transported along the axons of the vomeronasal nerves to their terminals in the glomerular layer of the accessory olfactory bulb (AOB). A lesser but consistent amount of radioactivity was found in the external plexiform layer (EPL) of the AOB. Electron microscopic autoradiography was used to determine which of the components of the EPL contained this labelled material. The method of proportional grain counts showed that the highest concentration of silver grains lay over the mitral cell dendrites, which are the elements immediately postsynaptic to the vomeronasal nerve axons. However, a fairly high proportion of grains also lay over the peripheral processes of granule cells. By application of a method of 'crossfire analysis' (which is explained in detail) it was possible to show that the observed grain distribution is best explained by the assumption that the radioactive material is confined to mitral cells, and the labelling over granule cell processes is due to crossfire from these sources. Im one animal at 5 days after [3H]proline administration label was found to have extended from mitral cells to granule cells, suggesting that the transsynaptically transported radioactive material, which was confined to the mitral cells at 8 h, may have become further redistributed at longer survivals. In a control experiment, [3H]proline was applied directly to the surface of the AOB. This gave rise to a completely different distribution of radioactivity in the EPL: radioactive material was present in all tissue components.

Animals↗

A colorimetric assay for measuring activation of Gs- and Gq-coupled signaling pathways.

Current assays for functional activation of Gs-coupled receptors usually involve quantitation of adenylyl cyclase or measurement of cAMP concentration by radioimmunoassay. The activation of Gq-coupled receptors is commonly assayed by measurement of the production of inositol triphosphate or diacylglycerol from phosphatidylinositol 4,5-bisphosphate or of changes in intracellular calcium. These assays generally require large numbers of cells (10(5)-10(6)) and/or the use of radioactive materials. We have developed a rapid nonradioactive colorimetric assay that utilizes a beta-galactosidase (lacZ) gene fused to five copies of the cyclic AMP response element (CRE) to detect the activation of CRE-binding protein that results from an increase in intracellular cAMP or calcium. This assay can be performed using as few as 30,000 cells in a 96-well format with the end products measured simultaneously in a microplate reader. Consequently, a single individual can readily assay 1000 samples a day. Using this assay, the fold increase in beta-galactosidase activity was similar in magnitude to increases in cAMP or adenylyl cyclase activity and was approximately linear from 0.01 to 0.27 fmol/cell of intracellular cAMP. Furthermore, pharmacological characterization of one of the melanocortin receptors, mMC5-R, using this assay resulted in a similar order of potency for several melanocortin peptides to that obtained with a commonly used adenylyl cyclase enzyme assay. This assay is also useful for the characterization of Gq-coupled receptors as is demonstrated here using cells transfected with the mouse bombesin receptor. The large-scale capacity of this assay makes it an excellent method for screening molecules of interest acting on Gs- and Gq-coupled receptors.

Base Sequence↗

Hormonal control of manganese transport in the mouse thyroid.

The present study deals with a possible mechanism controlling the transport of manganese (Mn), an essential trace element, from the circulation to the thyroid. Mice were pretreated with propylthiouracil (PTU) or triiodothyronine (T3), and a measurement of the thyroid:serum concentration ratio (T/S) of radioactive manganese (54Mn) was carried out. The T/S of 54Mn was greatly enhanced by PTU, but reduced by T3. Several methods were used to demonstrate that the T/S of 54Mn depends upon the level of thyroid-stimulating hormone (TSH) in the serum. First, bovine TSH was injected into mice; an increase in the T/S resulted. Secondly, serum thyroxine and T3 levels measured by radioimmunoassay (RIA) suggested that PTU produced an increase in serum TSH and T3 a decrease. However, direct measurement of mouse TSH by RIA for rat TSH failed to produce proof of any changes in TSH level, owing to poor cross-reactivity. Taking all the information into account, it is concluded that Mn-transport into the thyroid is controlled by the thyroid state.

Animals↗

Molecular characterization of Mycobacterium tuberculosis isolates presenting various drug susceptibility from Greece using three DNA typing methods.

OBJECTIVES: The purpose of the present study was the molecular characterization of Mycobacterium tuberculosis clinical isolates using three DNA typing methods. METHODS: One hundred nineteen independent (77 susceptible to all antituberculous drugs, 17 rifampin-resistant and 25 isoniazid-resistant), and nine related Mycobacterium tuberculosis isolates obtained over a 3-years period (1997-1999) from Greece were typed with restriction fragment length polymorphism (RFLP), using the non-radioactive IS6110 probe (IS6110-RFLP), and two PCR-based molecular methods: random amplification of polymorphic domains (RAPD) using four different primers and double repetitive element-PCR (DRE-PCR). RESULTS: IS6110-RFLP and RAPD-PCR using IRIS primer were proved to be the most discriminatory methods, while DRE-PCR gave satisfactory results and RAPD-PCR methods using the other three primers (A1245, B1245 and Leg2) were not so effective. The related strains, isolated from affected members of four families, gave similar PCR and RFLP patterns, while the independent strains presented a high degree of polymorphism. In terms of cost effectiveness and technical simplicity, the PCR-based methods were found to be superior. CONCLUSIONS: So, they may serve as screening methods to classify a large number of isolates into clusters for further subtyping and recognition of well-defined genotype families.

Adult↗

Isotopic ratio of 129I/127I in seaweed measured by neutron activation analysis with gamma-gamma coincidence.

129I is a long-lived (1.6 x 10(7) y) radionuclide that is produced in nature as the result of spontaneous fission of heavy elements and reaction of xenon with cosmic rays. Recently, however, artificial sources from nuclear power plants and nuclear test explosions have become a significant component of environmental radioactive iodine. Coincidence gamma-ray detection using Ge detectors makes it possible to simultaneously resolve the numerous gamma-rays produced by neutron activation. In this study, the coincidence gamma-ray detection technique was combined with neutron activation analysis to determine the radioactive iodine composition of seaweed. The ratio of 129I/127I in this common Japanese food item collected from the Ibaraki prefecture has been derived without the need for radiochemical purification. The isotopic ratio of 129I/127I in Kajime algae is 3.5(5) x 10(-10).

Gamma Rays↗

Dietary intakes of seven elements of importance in radiological protection by asian population: comparison with ICRP data.

Within the framework of a Coordinated Research Project (CRP) organized by the International Atomic Energy Agency, Vienna, the daily dietary intakes of seven elements by adult populations living in nine Asian countries were estimated. The countries that participated in the study were Bangladesh, China, India, Indonesia, Japan, Pakistan, Philippines, South Korea (Republic of Korea, ROK), and Vietnam and together they represented more than half of the world population. The seven elements studied were calcium, cesium, iodine, potassium, strontium, thorium, and uranium. These elements have chemical and biological similarity to some of the radionuclides abundantly encountered during nuclear power production and therefore data on these elements could provide important information on their biokinetic behavior. Analyses of diet samples for these seven elements were carried out using highly sensitive and reliable analytical techniques. One thousand one hundred and sixty analytical determinations were made on two hundred and twenty samples of typical diets consumed in these countries to estimate the daily intakes of these elements by the adult Asian population. The median daily dietary intakes for the adult Asian population were found to be 0.45 g calcium, 7 microg cesium, 90 microg iodine, 1.75 g potassium, 1.65 mg strontium, 1 microg thorium, and 1 microg uranium. When compared with the intakes proposed for ICRP Reference Man by International Commission for Radiological Protection, these intakes were lower by factors of 0.41 for calcium, 0.7 for cesium, 0.45 for iodine, 0.53 for potassium, 0.87 for strontium, 0.33 for thorium, and 0.52 for uranium. The lower daily intakes of calcium, cesium, and iodine by Asian population could be due to significantly lower consumption of milk and milk products, which are rich in these elements. The significantly lower intake of calcium in most of the Asian countries may lead to higher uptake of fission nuclide 90Sr and could result in perhaps higher internal radiation dose. The use of highly sensitive and reliable analytical methods resulted in accurate and lower intake values obtained for thorium and uranium, which suggest that radiation dose from their ingestion at natural background levels is likely to be lower than what may be concluded from ICRP data.

Asia↗