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PubMed1995

Psychological adaptation among residents following restart of Three Mile Island.

Psychological adaptation is examined in a sample of residents who remained in the vicinity of Three Mile Island following the restart of the nuclear generating facility which had been shut down since the 1979 accident. Findings indicate a lowering of psychological symptoms between 1985 and 1989 in spite of increased lack of control, less faith in experts and increased fear of developing cancer. The suggestion is made that reduced stress might have been related to a process of adaptation whereby a cognition of emergency preparedness was integrated by some of these residents as a modulating cognitive element. Findings also indicate that "loss of faith in experts" is a persistently salient cognition consistent with the "shattered assumptions" theory of victimization.

Adaptation, Psychological↗

PubMed1980

The reactions of l nm particles of plutonium-238 dioxide and curium-244 dioxide with lung fluid.

The reactions of 1 nm particles of plutonium-238 dioxide and curium-244 dioxide with rat lung fluid have been studied both in vitro and in vivo. The plutonium-238 particles are positively charged and combine by electrostatic attraction with the negative pulmonary surfactant which mediates the transfer of radioactivity to the blood. In contrast the curium-244 particles are negative and are assumed to diffuse passively through the alveolar walls. The results emphasise that electrostatic charge is an important factor governing the reaction of 1 nm actinide oxide particles with macromolecules.

Actinoid Series Elements↗

PubMed1998

Pharmacologic importance of the reversible fatty acid conjugation of budesonide studied in a rat cell line In vitro.

Functional implications of the recently described fatty acid conjugation of budesonide (BUD) (Tunek, A., K. Sjödin, and G. Hallström, Drug Metabol. Dispos. 1997;25:1311-1317; Miller-Larson, A., E. Hjertberg, H. Mattsson, M. Dahlbäck, A. Tunek, and R. Brattsand, Am. J. Respir. Crit. Care Med. 1997;155:A353 [Abstr.]) were studied in a rat cell line, Rat1, transfected with the activation protein-1 (AP-1)-controlled regulatory element (TRE) driving the reporter gene beta-galactosidase. TRE is downregulated by glucocorticosteroids (GCS) through interaction with the AP-1 complex. BUD was compared to fluticasone propionate (FP), a potent glucocorticosteroid that does not form fatty acid conjugates. The kinetics and metabolism of the GCS were studied after incubation of either 3H-BUD or 3H-FP with transfected Rat1 cells. Up to 20% of added BUD was taken up into the cells over 24 h. The great majority of the intracellular radioactivity (80-90%) consisted of lipophilic BUD conjugates. After removing extracellular 3H-GCS, the outflow of radioactivity was studied. Only free BUD and not fatty acid conjugates was detected extracellularly, suggesting that hydrolysis of the conjugates was required to release BUD from the cell. During 165 min, less BUD (about 65% of totally incorporated) was released than FP (more than 90%). In the functional studies, FP was about six times more potent than BUD in downregulating TRE after 24 h continuous exposure. However, after a 6-h pulse of GCS, the effect of BUD persisted unchanged 18 h later, whereas FP had almost lost its efficacy (P < 0.05 between the drugs). In addition, the reversible conjugation process of BUD resulted in transferable GCS effects. Medium containing released BUD from previously loaded cells mediated nearly the same downregulatory effect after addition to naive cells as did continuous treatment. No such transferable effect was seen for FP. In conclusion, the reversible fatty acid conjugation of BUD resulted in prolonged cellular retention and anti-inflammatory activity after pulse exposure in this in vitro system. This fatty acid conjugation mechanism appears to add to the beneficial pharmacologic profile of BUD.

Androstadienes↗

PubMed1984

Immunity repressor of bacteriophage P2. Identification and DNA-binding activity.

The product of gene C of the temperate bacteriophage P2, the immunity repressor, can be detected as a unique band eluting from phosphocellulose columns at 0.12 M-potassium phosphate when differentially labelled with a radioactive amino acid: the band is absent when phages that either have lost gene C through deletion or carry a suppressor-sensitive mutation in the gene are used. The repressor in its monomeric form is about 11,000 in molecular weight. At near physiological salt concentrations, the form predominantly recovered is the dimer. In filter-binding assays, the partially purified repressor binds wild-type P2 DNA strongly. It does not bind DNA of P2 vir94, a deletion that removes all the genetic elements involved in the regulation of lysogeny; it also does not bind, or binds inefficiently, DNA of P2 vir3, a mutation in the operator that controls the early replicative functions of P2. At the concentrations employed, the dimer is the active form in binding. The P2 repressor clearly differs in several features from the well-studied immunity repressor of bacteriophage lambda.

Bacteriophages↗

PubMed1966

The in vivo protein synthetic activities of free versus membrane-bound ribonucleoprotein in a plasma-cell tumor of the mouse.

Cytoplasmic extracts of the transplantable RPC-20 plasma-cell tumor were fractionated by sucrose density gradient centrifugation. Four major fractions were distinguished: (a) microsomes and mitochondria; (b) membrane-free polyribosomes; (c) free monomeric ribosomes; and (d) soluble fraction. The fractions were analyzed for RNA and lipid phosphorus, and their particulate components were characterized by electron microscopy. Particular attention was paid to the problem of membrane contamination of the free polyribosome fraction. It was shown that this contamination was small in relation with the total content of ribosomes in the fraction, and that it consisted primarily of smooth-surfaced membranes which were not physically associated with the polyribosomes themselves. In vivo incorporation studies were carried out by injecting tumor-bearing animals intravenously with leucine-C(14), removing the tumors at various times thereafter, and determining the distribution of protein radioactivity among the gradient-separated cytoplasmic fractions. The free polyribosome and the microsome-mitochondria fractions constituted active centers for protein synthesis. It was shown that nascent protein of the free polyribosome fractions was not associated significantly with the contaminating membranes. The kinetics of labeling during incorporation times up to 11 min suggested that protein synthesized on the free polyribosomes was rapidly transferred in vivo to the soluble fraction of the cell, while protein synthesized by the microsomes and mitochondria remained localized within these elements. It was estimated that the free polyribosome fraction and the microsome-mitochondria fraction accounted for approximately equal proportions of the total cytoplasmic protein synthesis in vivo.

Animals↗

PubMed1984

Spatial distribution of DNA loop attachment and replicational sites in the nuclear matrix.

Biochemical fractionation was combined with high resolution electron microscopic autoradiography to study the localization in rat liver nuclear matrix of attached DNA fragments, in vivo replicated DNA, and in vitro synthesized DNA. In particular, we determined the distribution of these DNA components with the peripheral nuclear lamina versus more internally localized structural elements of isolated nuclear matrix. Autoradiography demonstrated that the bulk of in vivo newly replicated DNA associated with the nuclear matrix (71%) was found within internal matrix regions. A similar interior localization was observed in isolated nuclei and in situ in whole liver tissue. Likewise, isolated nuclear lamina contained only a small amount (12%) of the total matrix-bound, newly replicated DNA. The structural localization of matrix-bound DNA fragments was examined following long-term in vivo labeling of the DNA. The radioactive DNA fragments were found predominantly within interior regions of the matrix structure (77%), and isolated nuclear lamina contained less than 15% of the total nuclear matrix-associated DNA. Most of the endogenous DNA template sites for the replicative enzyme DNA polymerase alpha (approximately 70%) were also sequestered within interior regions of the matrix. In contrast, a majority of the endogenous DNA template sites for DNA polymerase beta (a presumptive repair enzyme) were closely associated with the peripheral nuclear lamina. A similar spatial distribution for both polymerase activities was measured in isolated nuclei before matrix fractionation. Furthermore, isolated nuclear lamina contained only a small proportion of total matrix-bound DNA polymerase alpha endogenous and exogenous template activities (3-12%), but a considerable amount of the corresponding beta polymerase activities (47-52%). Our results support the hypothesis that DNA loops are both anchored and replicated at nuclear matrix-bound sites that are predominantly but not exclusively associated with interior components of the matrix structure. Our results also suggest that the sites of nuclear DNA polymerase beta-driven DNA synthesis are uniquely sequestered within the characteristic peripheral heterochromatin shell and associated nuclear envelope structure, where they may potentially participate in DNA repair and/or replicative functions.

Animals↗

PubMedDate not supplied

GFAP gene expression is altered in young rats following developmental low level lead exposure.

The astroglia cytoskeletal element, glial fibrillary acid protein GFAP, is a generally accepted sensitive indicator for neurotoxic effects in the mature brain. We used GFAP RNA as a marker for molecular biological changes in rat brain related to chronic low level lead exposure during the developmental period. Brains in the third week of postnatal life, following a continuous prenatal and postnatal exposure to lead of low dose were investigated. Sections were processed for in situ hybridization with a 35S labeled riboprobe, which is specific for mouse GFAP RNA. Radioactive signals which resulted from the RNA-RNA-hybridization process, were achieved by exposure of the sections to X-ray film. The gray values on the X-ray films were investigated by a computer assisted image analyzer system. Our data show that low level lead exposure affects the GFAP RNA-amount. Interestingly these effects were spatially diverse and different in the brain regions investigated. The gray values were significantly enhanced in the CA1 and CA2 regions of the hippocampus, in the striatum and in the cerebellum. The GFAP gene expression seems to reflect the reaction of the astroglia in accordance to the electrophysiological functions of the NMDA receptor in the diverse brain regions and their alterations following low level lead exposure.

Animals↗

PubMed1993

PCR product quantification by non-radioactive hybridization procedures using an oligonucleotide covalently bound to microwells.

Oligonucleotides derived from IS6110, an insertion sequence from Mycobacterium tuberculosis, have been covalently immobilized on polystyrene Covalink NH microwells to develop a sandwich and a competitive non-radioactive hybridization assay for the quantitative determination of the DNA fragments obtained by polymerase chain reaction (PCR). Using the appropriate standard DNA, the method can be employed for the quantitative analysis of PCR fragments. The sandwich assay can detect as little as 3 fmol of target DNA per well and the standard curve may be used with quantities ranging from 3 to 300 fmol per well. The competitive hybridization assay is less sensitive since it is quantitative between 100 and 8000 fmol per well. We show here that both kinds of assays can be used to identify M. tuberculosis strains isolated from clinical samples. The non-radioactive hybridization procedures using an oligonucleotide covalently bound to microwells involve few and simple operations, and are thus suitable for routine diagnosis. Moreover, when stored at 5 degrees C, precoated strips can still be used for hybridization up to at least 10 months.

Base Sequence↗

PubMed2003

Transport of trace elements in lenses of normal and hereditary cataract UPL rats.

The multitracer technique was applied to the determination of the uptake of trace elements in the lenses of normal and hereditary cataract UPL rats to investigate the transport mechanisms of trace elements during cataract development. Be, Na, Sc, V, Cr, Mn, Fe, Co, Zn, As, Se, Rb, Sr, Y, Zr, Tc, Ru and Rh accumulate in normal and UPL cataract rat lenses. The rates of uptake of trace elements differ among species and also differ between normal and UPL rat lenses. The uptakes of V and Sr are greater in normal rat lenses, while the uptakes of Mn and Co are greater in UPL rat lenses. High concentrations of Zn are transported into normal rat lenses in comparison with other elements. However, the uptake of Se was highest in the lenses of UPL cataract rats. In addition, the difference in Se uptake between the normal and UPL rat lenses was greatest among the tested trace elements. The present study suggests that the transport characteristics of trace elements are different in the lenses of normal and UPL cataract rats. The different transport characteristics of trace elements in the lenses of normal and UPL cataract rats, especially the higher accumulation of Se in UPL rat lenses, may be implicated in cataract development.

Animals↗

PubMed1994

Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification.

Simple sequence repeats (SSR), or microsatellites, are ubiquitous in eukaryotic genomes. Here we demonstrate the utility of microsatellite-directed DNA fingerprinting by polymerase chain reaction (PCR) amplification of the interrepeat region. No sequencing is required to design the oligonucleotide primers. We tested primers anchored at 3' or 5' termini of the (CA)n repeats, extended into the flanking sequence by 2 to 4 nucleotide residues [3'-anchored primers: (CA)8RG, (CA)8RY, and (CA)7RTCY; and 5'-anchored primers: BDB(CA)7C, DBDA(CA)7, VHVG(TG)7 and HVH(TG)7T]. Radioactively labeled amplification products were analyzed by electrophoresis, revealing information on multiple genomic loci in a single gel lane. Complex, species-specific patterns were obtained from a variety of eukaryotic taxa. Intraspecies polymorphisms were also observed and shown to segregate as Mendelian markers. Inter-SSR PCR provides a novel fingerprinting approach applicable for taxonomic and phylogenetic comparisons and as a mapping tool in a wide range of organisms. This application of (CA)n repeats may be extended to different microsatellites and other common dispersed elements.

Animals↗

PubMed2002

Assessing the transfer of pentachlorophenol through soil columns using 13[C]isotope.

The transfer of organic pollutants was studied through soil columns using 13[C]-labelled pentachlorophenol (PCP) as a model compound. The organic carbon content and the 13[C]/12[C] ratio were measured in two soil sections, 0-3 cm and 3-6 cm, and in percolated water using an Elemental Analyser coupled with a Magnetic Mass Sector. The mass balance of carbon was evaluated and the amount of PCP was calculated in each compartment of the soil-water systems. The results show that more than 80% of the PCP-derived 13[C] remained in the upper layer of the soil column. Approximately 20% was transferred to the lower soil layer, and less than 1% was found in the water leachates. The 13[C]-labelled tracers may thus be used as an alternative to radioactive compounds to follow the fate of organic pollutants in soil and water under field conditions.

Carbon Radioisotopes↗

PubMed2005

Concrete waterproofing in nuclear industry.

One of the main points of aggregate safety during the transportation and storage of radioactive materials is to supply waterproofing for all constructions having direct contact with radiating substances and providing strength, seismic shielding etc. This is the problem with all waterside structures in nuclear industry and concrete installations in the treatment and storage of radioactive materials. In this connection, the problem of developing efficient techniques both for the repair of operating constructions and the waterproofing of new objects of the specified assignment is genuine. Various techniques of concrete waterproofing are widely applied in the world today. However, in conditions of radiation many of these techniques can bring not a profit but irreparable damage of durability and reliability of a concrete construction; for instance, when waterproofing materials contain organic constituents, polymers etc. Application of new technology or materials in basic construction elements requires in-depth analysis and thorough testing. The price of an error might be very large. A comparative analysis shows that one of the most promising types of waterproofing materials for radiation loaded concrete constructions is "integral capillary systems" (ICS). The tests on radiation, thermal and strength stability of ICS and ICS-treated concrete samples were initiated and fulfilled in RFNC-VNIITF. The main result is--ICS applying is increasing of waterproofing and strength properties of concrete in conditions of readiation The paper is devoted to describing the research strategy, the tests and their results and also to planning of new tests.

Construction Materials↗

PubMed1988

Comparative immunochemical and structural similarities of five stress proteins from various tissue types.

1. Incorporation of radioactive phosphate or leucine by stress proteins from rodent lymphoma, submaxillary and liver nuclei were observed in two-dimensional gels following chemical and environmental stress. 2. The stress proteins were isolated from second-dimension gels and their similarities compared. Mol. wt determinations, immunochemical blotting and protease V8 peptide mapping confirmed the identical nature of the stress proteins possessing identical Mr, but from diverse tissue types. 3. These data imply that highly similar stress proteins exist in diverse tissues, are conserved during evolution, and possess some elemental and essential function.

Animals↗

PubMed1985

Transport of elements from soil to human diet: an alternative approach to pathway analysis.

Transport from soil to human dietary intake is conventionally treated by analyzing the various pathways through uptake by plant roots and transfer from animal feed into milk and meat. An alternative approach is proposed requiring only the average daily human ingestion of each element and its abundance in soil, both readily available for most elements. The two methods are compared quantitatively and give results consistent within a mean factor of four, which is roughly the standard deviation in estimates from the pathway analysis. Neither method gives consistently larger or smaller transfer rates. Each method has important advantages in some applications.

Animal Feed↗

PubMed1991

Molecular cloning and characterization of form I antigen genes of Shigella sonnei.

Sau3AI-generated DNA fragments of the Shigella sonnei large plasmid encoding the form I antigen were cloned into Escherichia coli with cosmid vector pHSG262. One resulting plasmid, designated pJK1137, was studied further. Restriction endonuclease mapping and analysis of transposon Tn3 insertion mutants demonstrated that the form I antigen genes were located within a region of about 12.6 kb consisting of the two contiguous HindIII fragments of 1.26 kb and 12.4 kb. The results of complementation studies between Tn3 insertion mutants of pJK1137 and recombinant plasmids carrying different parts of the form I antigen genes indicated that the 12.6 kb DNA sequence contained at least four gene clusters, regions A, B, C and D. Analysis of radioactively labelled proteins in minicells demonstrated that the DNA sequence of about 12.6 kb coded for at least four specific proteins of 42, 23, 48 and 39 kDa. The former two were coded by region A, the latter two by region D.

Agglutination Tests↗

PubMed1996

Patellar tendon and anterior cruciate ligament have different mitogenic responses to platelet-derived growth factor and transforming growth factor beta.

The healing responses of the anterior cruciate ligament and the patellar tendon differ markedly. The anterior cruciate ligament fails to heal, whereas the patellar tendon heals slowly. The basis of these differences is unknown. Since cellular proliferation is a critical element of healing, we investigated the response to explants of anterior cruciate ligament and patellar tendon from sheep knees to platelet-derived growth factor-AB and transforming growth factor beta 1 as a function of time and dose. Explants cultured for 48, 72, and 96 hours with transforming growth factor beta 1 (0-100 ng/ml) or platelet-derived growth factor-AB (0-200 ng/ml) were radiolabeled for the final 24 hours with [3H]thymidine, and DNA synthesis was quantified as trichloroacetic acid-precipitable radioactivity normalized to dry tissue weight. Statistical analyses (analysis of variance) showed that transforming growth factor beta 1 induced a significant proliferative response in the anterior cruciate ligament at 96 hours with equivalent responses at 10, 50, and 100 ng/ml, whereas the patellar tendon only responded to one condition, 10 ng/ml at 96 hours. Conversely, the patellar tendon had a significant dose-dependent response to platelet-derived growth factor-AB at 72 and 96 hours, whereas the anterior cruciate ligament showed no proliferative response to platelet-derived growth factor-AB. The minimal response of anterior cruciate ligament to platelet-derived growth factor-AB could explain, at least in part, the poor repair capacity of this tissue. The response of the anterior cruciate ligament to transforming growth factor beta suggests that exogenous transforming growth factor beta may promote initial healing. Although growth factors have the potential to modulate soft-tissue repair, tissue responses in tendons and ligaments may vary at different anatomic sites.

Animals↗

PubMed2005

Further evaluation of a new standard of efficacy for stored platelets.

BACKGROUND: The proposal to assess the viability capabilities of platelets (PLTs) collected, treated, or stored in a developmental system against "fresh" PLTs from the same subject poses several important methodologic issues pertaining to the timing and manner of the collecting and separating the fresh PLTs. This study extended the previous validation of this method of comparing fresh and stored PLTs, applying it to an assessment of apheresis PLTs stored for 7 days with a newly standardized radiolabeling protocol. STUDY DESIGN AND METHODS: Eighteen normal subjects donated 1 unit of leukoreduced PLTs, pheresed with a standard, approved system. They received an aliquot radiolabeled with 51Cr on Day 7 simultaneously with 111In-labeled fresh PLTs that had been separated by a manual method. Recovery and survival were compared to determine whether the stored PLTs were not inferior to the criterion of 67 percent of recovery and 50 percent of survival of fresh PLTs. Separate studies were undertaken to document the similarity of recovery and survival with 51Cr and 111In radiolabeling in PLTs stored to 8 days and to determine the importance of correcting the radioactivity in timed samples for the activity remaining in blood beyond the life span of the retransfused PLTs. RESULTS: PLTs stored for 7 days demonstrated 88.7 +/- 35.2 percent of the recovery and 89.9 +/- 21.2 percent of the survival of PLTs collected via a nonproprietary, manual system and thus met the comparative criterion. In a separate study (n = 12), labeling Day 8 PLTs with 51Cr or 111In resulted in recoveries and survivals that were not different. Radiolabel eluted from labeled PLTs in vitro was taken up by cellular blood elements in a reuptake incubation. CONCLUSION: Apheresis PLTs stored for 7 days met the criterion proposed for comparison with fresh PLTs. This analytic approach is feasible with PLTs collected and prepared via a manual method. A standardized protocol for radiolabeling PLTs with 51Cr and 111In and analyzing the results in a standardized fashion was employed successfully, with the two radioisotopes yielding similar results. The importance of correcting for residual activity after disappearance of injected cells was noted.

Blood Platelets↗

PubMed1994

Effects of pulmonary blood flow on the fractal nature of flow heterogeneity in sheep lungs.

The spatial heterogeneity of pulmonary blood flow can be described by the relative dispersion (RD) of weight-flow histograms (RD = SD/mean). Glenny and Robertson (J. Appl. Physiol. 69: 532-545, 1990) showed that RD of flow in the lung is fractal in nature, characterized by the fractal dimension (D) and RD for the smallest realizable volume element (RDref). We studied the effects of increasing total pulmonary blood flow on D and RDref. In eight in situ perfused sheep lung preparations, 15-microns radio-labeled microspheres were injected into the pulmonary artery at five different blood flows ranging, in random order, from 1.5 to 5.0 l/m. The lungs were in zone 2 at the lower flows and in zone 3 at the higher flows. The lungs were removed, dried, cut into 2 x 2 x 2-cm3 pieces, weighed, and then counted for microsphere radioactivity. Fractal plots of log(weight) vs. log(RD) were constructed by iteratively combining neighboring pieces and then calculating RD with the increasingly larger portion size. D, which is one minus the slope of the fit through this plot, was 1.14 +/- 0.09 and did not change as blood flow increased. However, RDref decreased significantly (P < 0.01) as total flow increased. We conclude that the fractal nature of pulmonary blood flow distribution is not altered by changes in overall flow.

Animals↗