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PubMed2006

Sox9 mRNA expression in the developing palate and craniofacial muscles and skeletons.

BACKGROUND: SOX9 is a critical transcription factor for chondrogenesis and sex determination. Haploinsufficiency mutations of Sox9 in humans lead to campomelic dysplasia. Inactivation of Sox9 in the craniofacial region of mice results in an absence of endochondral bones and in malformation of other structures. This suggests that Sox9 plays multiple roles in craniofacial development and these remain to be elucidated. In order to study the functions of Sox9 in craniofacial development, a preliminary expression examination was performed. MATERIAL AND METHODS: To detect the expression of Sox9 mRNA, antisense riboprobe was synthesized by in vitro transcription. Radioactive in situ hybridization was performed on sagittal and coronal sections of mice head from organogenesis to the early postnatal stage. RESULTS: It was found that Sox9 was expressed in multiple stages and distinct processes. Besides the expression in cartilage, it was seen in the fusing stage of palatogenesis. Sox9 was also present during differentiation and maturation of craniofacial muscles. In addition, it was observed in intramembranous skeletal elements at restricted sites and stage. CONCLUSIONS: The expression pattern suggests that Sox9 serves broad roles in craniofacial development.

Animals↗

PubMed1986

Human IgA as a heterovalent ligand: switching from the asialoglycoprotein receptor to secretory component during transport across the rat hepatocyte.

Asialoglycoproteins are taken up by the rat liver for degradation; rat polymeric IgA is taken up via a separate receptor, secretory component (SC), for quantitative delivery to bile. There is negligible uptake of these ligands by the converse receptor, and only a low level of missorting of ligands to opposite destinations. The two pathways are not cross-inhibitable and operate independently (Schiff, J.M., M. M. Fisher, and B. J. Underdown, 1984, J. Cell Biol., 98:79-89). We report here that when human IgA is presented as a ligand in the rat, it is processed using elements of both pathways. To study this in detail, different IgA fractions were prepared using two radiolabeling methods that provide separate probes for degradation or re-secretion. Behavior of intravenously injected human polymeric IgA in the rat depended on its binding properties. If deprived of SC binding activity by affinity adsorption or by reduction and alkylation, greater than 80% of human IgA was degraded in hepatic lysosomes; radioactive catabolites were released into bile by a leupeptin-inhibitable process. If prevented from binding to the asialoglycoprotein receptor by competition or by treatment with galactose oxidase, human IgA was cleared and transported to bile directly via SC, but its uptake was about fivefold slower than rat IgA. Untreated human IgA was taken up rapidly by the asialoglycoprotein receptor, but depended on SC binding to get to bile: the proportion secreted correlated 1:1 with SC binding activity determined in vitro, and the IgA was released into bile with SC still attached. These results demonstrate that human IgA is normally heterovalent: it is first captured from blood by the asialoglycoprotein receptor, but escapes the usual fate of asialoglycoproteins by switching to SC during transport. Since the biliary transit times of native human and rat IgA are the same, it is probable that the receptor switching event occurs en route. This implies that the two receptors briefly share a common intracellular compartment.

Animals↗

PubMed1983

Repeated nucleotide sequence arrays in Balbiani ring 1 of Chironomus tentans contain internally nonrepeating and subrepeating elements.

Balbiani rings in Chironomus are large puffs on salivary gland polytene chromosomes that contain functionally related, but nonidentical genes that code for tissue-specific secretory polypeptides. In situ hybridization was used to select a recombinant plasmid (pCtBR1-1) that contained an insert of Chironomus tentans genomic DNA that originated from Balbiani ring 1. Mapping with restriction endonucleases demonstrated that the insert was 385 bp (base pair) and it contained duplicate clusters of certain cleavage sites about 250 bp apart. This repeat was shown to be part of tandem sequence arrays in the genome by hybridization of radioactive pCtBR1-1 to nitrocellulose blots containing limit and partial restriction endonuclease digests of nuclear DNA. Subsequent sequence analysis of the cloned DNA confirmed the presence of one complete copy of a 246-bp repeat comprised of a 114-bp internally nonrepeating segment and a 132-bp segment containing four 33-bp subrepeats. The subrepeats apparently evolved from a simple 9-bp sequence encoding a consensus tripeptide (Lys-Pro-Ser) in which the first two codons (AAA-CCA) were highly conserved at the nucleotide level. Comparisons between intragenic and interspecific (BRb in Chironomus thummi) copies of corresponding sequences revealed that, during the evolution of these tandemly repeated protein-coding sequences, internally nonrepeated segments were highly conserved and most likely became interspersed by variable segments containing subrepeats that arose from reduplication and divergence of 9-bp repeats.

Amino Acid Sequence↗

PubMed1991

Cs-134/137 contamination and root uptake of different forest trees before and after the Chernobyl accident.

The Cs-134/137 activities were measured from different tree organs of spruce, larch and sycamore maple. Two locations in South Bavaria were monitored during a period of 2.5 years following the Chernobyl accident. Samples taken in 1985 allow to determine the Cs-137 contamination before the accident. Increasing Cs-137 activities from older to younger needle years of Picea abies caused by root-uptake of the global weapons' fallout are due to the high phloem mobility of this element and the remaining of the needles at the tree for about 6-7 years. In contrast, the Cs-137 activity was much smaller in leaves of larch and sycamore maple. After the Chernobyl accident, the higher contamination of spruce greater than larch greater than sycamore maple is dependent on the roughness of bark, absolute bark surface and the existence of leaves during the deposition of Chernobyl-derived radioactivity. The Cs-134/137 activity (Bq/kg d.w.) was about 25-times higher in bark compared to wood of Picea abies and 1.5-4.7 times higher in directly contaminated twig-axes than in leaves. Till the end of the investigation the major contamination of the shoots was due to direct deposition of cesium on the trees. A maximum of 5-15% of the total activity of the directly contaminated branches of the plants was calculated to be part of root-uptake, depending on the amount of initial retention. 20% of the translocated cesium into new leaves of larch and about 50% into sycamore maple resulted from root-uptake 2.5 years after the accident.

Accidents↗

PubMed2005

Basic considerations for Monte Carlo calculations in soil.

Monte Carlo codes are extensively used for probabilistic simulations of various physical systems. These codes are widely used in calculations of neutron and gamma ray transport in soil for radiation shielding, soil activation by neutrons, well logging industry, and in simulations of complex nuclear gauges for in soil measurements. However, these calculations are complicated by the diversity of soils in which the proportions of solid, liquid and gas vary considerably together with extensive variations in soil elemental composition, morphology, and density. Nevertheless use of these codes requires knowledge of the elemental composition and density of the soil and its physical characteristics as input information for performing these calculations. It is shown that not always all of the soil parameters are critical but depend on the objectives of the calculations. An approach for identifying soil elemental composition and some simplifying assumptions for implementing the transport codes are presented.

Algorithms↗

PubMed2000

Transcriptional activation analysis using bioluminescent reporter assays.

Analysis of promoter and enhancer DNA sequences provides the researcher with valuable information regarding the expression patterns of genes. Insertion of small DNA fragments containing the regulatory sequence of interest into vectors carrying reporter genes allows for the accurate quantitative analysis of the gene's expression patterns and responses to various stimuli. The use of bioluminescent reporter genes provides a simple, rapid, and inexpensive system that generates virtually no toxic or radioactive waste products. In addition, bioluminescent reporter vectors are more sensitive than previous methods such as the chloramphenicol acetyl transferase (CAT) systems, that require the use of hazardous chemicals and isotopically labeled reagents.

Animals↗

PubMedDate not supplied

Aluminum effect on dissolution and precipitation under hyperalkaline conditions: I. Liquid phase transformations.

Substantial amounts of self-boiling, Al-rich, hyperalkaline, and saline high-level waste fluids (HLWF) were deposited to the vadose zone at the Hanford Site, in Washington State. The objective of this study was to investigate the effects of similar fluids on the extent of dissolution and precipitation in the sediments. Metal- and glass-free systems were used to conduct batch experiments at 323 K under CO2 and O2 free conditions. Base-induced dissolution of the soil minerals was rapid in the first 48 h as indicated by immediate releases of Si and Fe into the soil solution. Potassium release lagged behind and dissolution of K-bearing minerals (mica and K-feldspar) proceeded faster only after 2 to 3 d of the experiment. Silicon and Fe release exhibited high dependence on aqueous [Al] (rate orders <-1), because Al decreased free OH concentration in the contact solution and probably inhibited soil mineral dissolution. Initial K release exhibited low dependence on [Al] (fractional rate orders). Initial dissolution rates calculated based on Si release varied with aqueous [Al] from 29.47 to 4.35 x 10(-12) mol m(-2) s(-1). Aluminum participated in the formation of the secondary phases (precipitation rates of 10(-8) mol s(-1)) but the overall precipitation rate of alumino-silicate secondary phases was probably controlled by aqueous [Si] (rates of 10(-9), and rate constants between 0.0054 and 0.0084 h(-1)). The changes in the soil solution chemistry (release of K, Si, Fe, and other elements) may play a significant role in the fate of radionuclides and contaminants like Cs, Sr, Cr, and U in the Hanford sediments.

Accidents↗

PubMed1983

An examination of the environmental half-time for radionuclides deposited on vegetation.

After radionuclides are deposited on vegetation, environmental removal processes combine with radioactive decay to reduce the quantity of initial contamination. The time in which one-half of the radioactivity is removed from vegetation by environmental processes alone is referred to as the environmental half-time, Tw. For long-lived radionuclides, the dose to man via ingestion of contaminated terrestrial foods may be directly influenced by values of Tw, provided that environmental removal processes dominate root uptake and time is sufficiently long between initial exposure of vegetation and harvest. Values of Tw reported in the literature for various radionuclides and methods of deposition are examined. Factors affecting the variability of Tw are related in part to the physicochemical form of the depositing substance, vegetation type and growth form, climate, season, and experimental procedure. For growing vegetation, values of Tw are generally lower than those reported for dormant vegetation. Values of Tw for iodine vapor and iodine particulate are less than values of Tw reported for particulates of other elements. Values of Tw determined on a per unit vegetation mass basis are also less than values of Tw determined on a per unit ground area basis, the differences being attributable to the effects of growth dilution. Although an assessor has a range from which to select a value of Tw for the purpose of estimating contamination by long-lived radionuclides, the variability in Tw is small in comparison to many other parameters used in environmental radiological assessments.

Half-Life↗

PubMed2005

Neutron activation of patients following boron neutron capture therapy of brain tumors at the high flux reactor (HFR) Petten (EORTC Trials 11961 and 11011).

BACKGROUND AND PURPOSE: At the High Flux Reactor (HFR), Petten, The Netherlands, EORTC clinical trials of Boron Neutron Capture Therapy (BNCT) have been in progress since 1997. BNCT involves the irradiation of cancer patients by a beam of neutrons, with an energy range of predominantly 1 eV to 10 keV. The patient is infused with a tumor-seeking, (10)B-loaded compound prior to irradiation. Neutron capture in the (10)B atoms results in a high local radiation dose to the tumor cells, whilst sparing the healthy tissue. Neutron capture, however, also occurs in other atoms naturally present in tissue, sometimes resulting in radionuclides that will be present after treatment. The patient is therefore, following BNCT, radioactive. The importance of this induced activity with respect to the absorbed dose in the patient as well as to the radiation exposure of the staff has been investigated. MATERIAL AND METHODS: As a standard radiation protection procedure, the ambient dose equivalent rate was measured on all patients following BNCT using a dose ratemeter. Furthermore, some of the patients underwent measurements using a gamma-ray spectrometer to identify which elements and confirm which isotopes are activated. RESULTS: Peak levels, i.e., at contact and directly after irradiation, are of the order of 40-60 muSv/h, falling to < 10 muSv/h 30-50 min after treatment. The average ambient dose equivalent in the first 2 h at a distance of 2 m from the patient is in the order of 2.5 muSv. The ambient dose equivalent rate in 2 m distance from the patient's head at the earliest time of leaving the reactor center (20 min after the end of treatment) is far less than 1 muSv/h. The main radioisotopes were identified as (38)Cl, (49)Ca, and (24)Na. Furthermore, in two patients, the isotopes (198)Au and (116m)In were also present. The initial activity is predominantly due to (49)Ca, whilst the remaining activity is predominantly due to (24)Na. CONCLUSION: The absorbed dose resulting from the activated isotopes in the irradiated volume is in the order of < 1% of the prescribed dose and therefore does not add a significant contribution to the absorbed dose in the target volume. In other parts of the patient's body, the absorbed dose by induced activity is magnitudes smaller and can be neglected. The levels of radiation received by staff members and non-radiation workers (i.e., accompanying persons) are well below the recommended limits.

Body Burden↗

PubMed2002

Food chains and biogeochemical pathways: contributions of fallout and other radiotracers.

This paper reviews examples of how measurements of global fallout in the environment and related tracer radionuclides have been used to enhance our basic knowledge of biogeochemical processes and food-chain pathways. Because it is these fundamental, natural processes that control the transport and accumulation of such trace substances in the environment, direct measurements of trace substances over time and space reveal strong insights into these processes. The necessity to monitor global fallout transport, although largely motivated by human health concerns, gave rise to a plethora of new information about plants, animals, and natural and agricultural ecosystems and how they function. This review provides a small selection of examples in the areas of plant and animal physiology, productivity and energy transfer in food chains, biogeochemical cycles of certain elements and their analogues, feeding relationships and movements of organisms, and the agriculture-based human food chain. It is concluded that if society is to cope successfully with continued growth of the human population and resource consumption, more knowledge is still required about these fundamental processes. The use of radiotracers can contribute greatly to this need, but current funding priorities, societal attitudes, and onerous regulations on the use of radioactivity may continue to limit such applications.

Animals↗

PubMed1985

Association of 239Pu with lysosomes in rat, Syrian hamster, and Chinese hamster liver as studied by carrier-free electrophoresis and electron microscopic autoradiography with 241Pu.

The binding of injected monomeric plutonium in the liver of rats, Syrian hamsters, and Chinese hamsters (species which show profound differences in their ability to eliminate 239Pu from the liver) was investigated by carrier-free electrophoresis using 239Pu and electron microscopic autoradiography with 241Pu. These studies are part of a program designed to obtain a better understanding of the mechanisms of the clearance of transuranium elements from liver of different mammals and man. Between 4 and 9 days after nuclide injection, a clear correlation between the majority of the 239Pu and lysosomal enzymes was observed when the mitochondrial-lysosomal (ML) fraction of the livers was analyzed by carrier-free electrophoresis. In the two hamster species, a second 239Pu peak exists from the beginning and increases with time to comprise 50% of the total radioactivity at later times. During electron microscopic examination 4 days after 241Pu injection, beta tracks were frequently observed over globular structures resembling dense bodies in Chinese hamster liver. They were also observed frequently over chromatin-rich portions of the cell nuclei. These results, together with those from previous density gradient studies, show that lysosomes are the primary deposition site for 239Pu in the liver cytoplasm of these three rodent species. The hypothesis of a morphologic transformation of these lysosomes with time in hamster liver and of rapid bulk exocytosis of the lysosomes in rats are still possible explanations for the extreme differences in the elimination among the three species.

Animals↗

PubMed1979

[Biosynthesis of rat hemoglobin fractions under normal and increased erythropoiesis].

Studies of erythroid elements of hematopoietic organs and peripheric blood and bone marrow cell populations fractionated by precipitation in an albumin density gradient and synchronized by actinomycin D revealed that the rate of synthesis of individual haemoglobin fractions in the course of erythropoiesis is changed. The proliferating cells are characterized by a predominant accumulation of haemoglobin in fractions with beta c-and in the maturing cells--with beta b-chains. The radioactivity in the whole population of hematopoietic organs under phenylhydrazine anemia is mainly increased in the beta c-chains, whereas under the effects of erythropoietic factors (erythropoietin, animal sera)--that of beta b-chains is increased. In early erythroblasts the erythropoietic factors activate the formation of beta c-chains and in late ones--that of beta b-chains. Similar time dependence was observed in case of synthesis and activation of alpha b- and alpha a-chains. The specific cell responses in the erythroid series of bone marrow, spleen and blood to the effects of erythropoietic factors and anemia were established. The molecular mechanisms involved in the switch-over of the synthesis of various haemoglobin chains are discussed.

Animals↗

PubMed2000

The social psychology of public response to warnings of a nuclear power plant accident.

This article reviews the process of public response to warnings of an impending nuclear power plant emergency. Significant evidence exists to suggest that people engage in protective action in response to warnings based upon the substance and course through which emergency warning information is disseminated. The three basic components of a warning system are defined, and the elements of public response to warnings are summarized. Popular myths about public response to warnings are outlined and dispelled based upon current research verification. The conclusion provides an overview and synthesis of the warning response process.

Communication↗

PubMed1980

Autoradiography of GABA in the rat hypothalamic median eminence.

Light microscopy autoradiographs of the rat hypothalamic median eminence were prepared after injection of high specific activity tritiated GABA and GABA structural analogs. Following intracardiac injection of labeled GABA with short (15 min) survival time, a dense accumulation of silver grains was observed over the external layer of the median eminence. The silver grains appeared much less numerous and randomly scattered over the internal layer. No conspicuously labeled cells could be detected in the median eminence. A similar pattern of labeling was observed after 10 min in vitro incubation of the median eminence with a low concentration (2.5 x 10(-7) M) of labeled GABA. Clusters of silver grains were also visible over the external layer following intraventricular injection of labeled GABA. In this latter case, however, other sites of labeling were revealed over the internal and ependymal layers. The dense labeling over the external layer with tritiated GABA was partially reduced by a simultaneous intracardiac injection of a 50-fold excess of non-radioactive cis-aminocyclohexane carboxylic acid--a reported preferential substrate for GABA neuronal uptake--but it was not displaced by a 2000-fold excess of non-radioactive beta-alanine--a reported specific substrate for GABA glial uptake. Intracardiac injection of triated beta-alanine led to a faint and even labeling over the entire median eminence with no preferential accumulation of silver grains over the various layers. Following intraventricular injection of labeled beta-alanine the tanycytes and their processes as well as numerous glial cells appeared heavily labeled. These results suggested that there exist cell elements in the external layer of the hypothalamic median eminence which are capable of accumulating exogenous GABA according to its neuronal uptake characteristics. Although the exact nature of these cells is not readily apparent at this stage of our investigations, these findings led us to speculate that there might be a subpopulation of GABAergic nerve endings in the vicinity of the primary plexus capillaries.

Alanine↗

PubMedDate not supplied

Carbamoylation of peptides and proteins in vitro by S-(N-methylcarbamoyl)glutathione and S-(N-methylcarbamoyl)cysteine, two electrophilic S-linked conjugates of methyl isocyanate.

The reactivity toward peptides and proteins of S-(N-methylcarbamoyl)glutathione (SMG), the glutathione conjugate of methyl isocyanate, and the corresponding cysteine adduct, S-(N-methylcarbamoyl)cysteine (SMC), was investigated with the aid of in vitro model systems. Incubation of SMC or a trideuteriomethyl analogue of SMC with either the reduced or oxidized forms of oxytocin afforded similar mixtures of mono-, bis- and tris-N-methylcarbamoylated peptides. Structure elucidation of the mono and bis adducts by fast atom bombardment tandem mass spectrometry indicated that carbamoylation of oxytocin occurred preferentially at Cys-6 and that Cys-1 and/or Tyr-2 were secondary sites of modification. Upon incubation of S-[N-([14C]methyl)carbamoyl]glutathione (14C-SMG) with native bovine serum albumin (BSA), radioactivity became bound covalently to the protein in a time- and concentration-dependent fashion. "Blocking" of the lone Cys-34 thiol group of BSA in the form of a disulfide prior to exposure of the protein to 14C-SMG failed to decrease significantly the extent or time course of this covalent binding. It is concluded that carbamate thioester conjugates of MIC are reactive, carbamoylating entities which can donate the elements of MIC to nucleophilic functionalities on peptides and proteins. Free thiols appear to be preferred sites for such carbamoylation processes, a phenomenon that may have important toxicological consequences in the pathology of tissue lesions induced by MIC and related isocyanates.

Cyanates↗

PubMedDate not supplied

Evaluation of 99mtechnetium-radiopharmaceutical binding to blood elements using different trichloroacetic acid concentrations.

Secure determination of the binding of 99mTc-radiopharmaceuticals to plasma (P) and blood cell (BC) constituents can help to understand the biodistribution of radiophamaceuticals. The reported precipitation studies of blood with radiopharmaceuticals have shown that the results can not be easily compared between studies. We decided to determine the "gold standard" concentration of trichloroacetic acid (TCA) to evaluate the binding to blood elements for several radiopharmaceuticals used in routine nuclear medicine. We have studied phytic (99mTc-PHY), diethylenetriaminepentaacetic (99mTc-DTPA), glucoheptonic (99mTc-GHA) and dimercaptosuccinic (99mTc-DMSA) acids. Blood was incubated with radiopharmaceuticals, centrifuged and P and BC separated. Samples of P and BC were also precipitated with TCA concentrations (20.0, 10.0, 5.0, 1.0, 0.5 and 0.1 percent) and soluble (SF) and insoluble fractions (IF) were isolated. The percent radioactivity (percent rad) in IF-P depends on TCA concentration. It varied from 36.4 to 65.0 (99mTc-PHY), from 17.9 to 32.0 (99mTc-DTPA), from 11.5 to 38.8 (99mTc-GHA) and from 52.8 to 66.2 (99mTc-DMSA). The results for the binding of 99mTc-PHY to IF-P show that there was no differences in the percent rad when TCA concentrations of 0.1 to 1.0 percent were used. For 99mTc-DTPA, 5.0 percent is the best TCA concentration. For 99mTc-GHA, low values of percent rad bound to IF-P is found with TCA concentrations of 0.1, 0.5 and 1.0. Interestingly, with 99mTc-DMSA, high values of bound radioactivity are not dependent on TCA concentrations (0.1 to 10.0). Radioactivity in IF-BC depends on TCA concentration and it varied for 99mTc-PHY (80.1 to 54.1) and for 99mTc-GHA (85.5 to 61.7). With 99mTc-DTPA and with 99mTc-DMSA the percent rad in IF-BC seems independent of TCA concentration. We suggest that the evaluation of the binding of the various 99mTc-radiopharmaceuticals to blood constituents, using only one TCA concentration, should be avoided.

Animals↗

PubMed1997

Use of stable isotopically labeled tracers for studies of metabolic kinetics: an overview.

Stable isotopically labeled tracers offer a reliable and safe alternative to the use of radioactive tracers for studies of metabolic kinetics. This overview examines some of the principles and technical issues regarding mass spectrometry instrumentation, and reviews some of the approaches used in the application of stable isotopically labeled tracers to studies of protein, lipid, and carbohydrate metabolic kinetics.

Elements↗

PubMed1994

Household humidifiers as a radiological dose pathway.

Household humidifiers can inadvertently release contaminants to the atmosphere with water. Four types of humidifiers were tested to estimate the release rate of elements from the water. Stable nickel, selenium, iodine, and uranium tracers were used as analogues of some important radionuclides in the assessment of the disposal of used nuclear fuel. Humidifiers that inject water droplets into the air were the most efficient at releasing the elements from the water. Calculations of potential radiological dose to humans indicate that doses are less than 0.2 mSv y-1 if water was contaminated up to the maximum regulatory levels for several radionuclides.

Household Articles↗