The Chernobyl accident in 1986 resulted in the widespread identification of the post-accident presence of radioactive (or ‘hot’) particles across large areas of Eastern and Central Europe. Such particles arise from direct deposition and also from condensation and interactions on particle surfaces during and following the deposition of soluble fallout. Identification of the presence and nature of hot particles is necessary in order to determine the long-term ecological impact of radioactive fallout. This paper describes several techniques for the identification and characterization of hot particles in soil samples from Belarus. In addition to new results from the use of gamma spectrometry, we include two novel instrumentation approaches that have been developed and applied to Chernobyl fallout-contaminated soils. The first, ‘differential’ autoradiography, utilizes a photographic film sandwich to characterize the nature of the ionizing radiation emitted from samples. In this paper we show that differential autoradiography can not only identify hot particle presence in soil, but can also determine the dominant radionuclide in that particle. The second approach, sector field ICP-MS (ICP-SFMS), can provide rapid, high-precision determination of the actinides, including the transuranic actinides, that characteristically occur in hot particles originating from weapons fallout or fuel matrices. Here, ICP-SFMS is shown to yield sufficiently low detection limits for plutonium isotopes (with the exception of 238Pu) to enable us to confirm negligible presence of plutonium in areas outside the Chernobyl exclusion zone, but with high levels of fission-product contamination.

The effects of photoperiod and temperature on the translocation of triclopyr {[1(3,5,6-trichloro-2-pyridinyl)oxy] acetic acid}, picloram (4-amino-3,5,6-trichloropicolinic acid) and 2,4,5-T [(2,4,5-trichlorophenoxy)acetic acid] were studied on tanoak [Lithocarpus densiflorus (Hook. & Arn.) Rehd.], snowbush ceanothus (Ceanothus velutinus Dougl.), bigleaf maple (Acer macrophyllum Pursh), bean (Phaseolus vulgaris L. var. ‘Red Kidney’) and barley (Hordeum vulgare L. var. CM-67). Isolation of 14C and analysis for the radioactive herbicides revealed little metabolism of the herbicides. Regardless of herbicide or plant species herbicide movement was greatest under warm temperature and long day conditions. Among the herbicides tested, 14C associated with triclopyr was the most mobile in each species. Each herbicide moved readily in the symplast but root applications of each herbicide revealed limited apoplastic mobility.

Herbicide absorption and translocation patterns were determined following a wet blade method of application to dogfennel stems. Treatment consisted of severing stems with a pruning shear blade containing 5 μl of [14C]-triclopyr or [14C]-clopyralid solution. More than 73 or 82% of applied [14C]-triclopyr or [14C]-clopyralid was absorbed immediately, and uptake increased to more than 90%, 60 min after treatment. Both radiolabeled compounds appeared in the roots as soon as 12 h after treatment. By 48 h, 5 to 7% of the [14C] label was present in the roots, and 7 to 10% was exuded from the roots.

The metabolism of the herbicide 14C-glufosinate (GA) was studied in excised shoots and leaves of 20 weed and nonweed species; GA was fed through the xylem. After 24 or 48 h of incubation, the plant material was examined for phytotoxic symptoms, analyzed by autoradiography, and extracted. The extract was cleaned up and analyzed by high-performance liquid chromatography. GA and its metabolites were identified by cochromatography with authentic 14C-labelled reference compounds. 14C-GA was rapidly absorbed by the excised plant parts. Most of the radioactivity (91.3 to 99.7%) in the shoots and leaves was extractable. The main metabolite observed with all species was 3-(hydroxymethylphosphinyl)propionic acid (MPP); lower amounts of 2-hydroxy-4-(hydroxymethylphosphinyl)butanoic acid (MHB) were also found in 14 species. Metabolic rates of GA varied considerably ranging between 13.1 and 2,836.5 ng GA h−1 mg−1 protein. The species with the highest rates of metabolism of GA were Galium verum (2,836.5 ng GA h−1 mg−1 protein), Lythrum hyssopifolia (846.0 ng GA h−1 mg−1 protein), and Digitalis purpurea (494.8 ng GA h−1 mg−1 protein). The mean value across all species was 275.9 ng GA h−1 mg−1 protein.

A tall waterhemp population from Missisippi was suspected to be resistant to glyphosate. Glyphosate dose response experiments resulted in GR50 (dose required to reduce plant growth by 50%) values of 1.28 and 0.28 kg ae ha−1 glyphosate for the glyphosate-resistant (GR) and -susceptible (GS) populations, respectively, indicating a five-fold resistance. The absorption pattern of 14C-glyphosate between the GR and GS populations was similar up to 24 h after treatment (HAT). Thereafter, the susceptible population absorbed more glyphosate (55 and 49% of applied) compared to the resistant population (41 and 40% of applied) by 48 and 72 HAT, respectively. Treatment of a single leaf in individual plants with glyphosate at 0.84 kg ha−1, in the form of 10 1-µl droplets, provided greater control (85 vs. 29%) and shoot fresh weight reduction (73 vs. 34% of nontreated control) of the GS plants compared to the GR plants, possibly indicating a reduced movement of glyphosate in the GR plants. The amount of 14C-glyphosate that translocated out of the treated leaves of GR plants (20% of absorbed at 24 HAT and 23% of absorbed at 48 HAT) was significantly lower than the GS plants (31% of absorbed at 24 HAT and 32% of absorbed at 48 HAT). A potential difference in shikimate accumulation between GR and GS populations at different concentrations of glyphosate was also studied in vitro. The IC50 (glyphosate concentration required to cause shikimate accumulation at 50% of peak levels measured) values for the GR and GS populations were 480 and 140 µM of glyphosate, respectively, resulting in more shikimate accumulation in the GS than the GR population. Sequence analysis of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), the target site of glyphosate, from GR and GS plants identified a consistent single nucleotide polymorphism (T/C, thymine/cytosine) between GR/GS plants, resulting in a proline to serine amino acid substitution at position 106 in the GR population. The GR and GS plants contained equal genomic copy number of EPSPS, which was positively correlated with EPSPS gene expression. Thus, glyphosate resistance in the tall waterhemp population from Mississippi is due to both altered target site and nontarget site mechanisms. This is the first report of an altered EPSPS-based resistance in a dicot weed species that has evolved resistance to glyphosate.

The effect of formulation and adjuvants on absorption and translocation of 14C-clethodim was determined at 1, 4, 12, 24, 48, and 72 h after treatment (HAT) in bermudagrass under greenhouse conditions. Absorption of 14C-clethodim with the 0.12 kg L−1 (15 to 85%) formulation was higher than with the 0.24 kg L−1 (5 to 40%) formulation, regardless of presence or absence of adjuvant. There was considerable variation in the effect of adjuvant on 14C-clethodim absorption. When either ammonium sulfate (AMS) or AMS plus crop oil concentrate (COC) was added to the 0.12 kg L−1 formulation, 14C-clethodim absorption increased significantly at all harvest times except at 12 HAT compared with 0.12 kg L−1 formulation alone, whereas, 14C-clethodim absorption after addition of COC to the 0.12 kg L−1 formulation was similar to the 0.12 kg L−1 formulation alone up to 24 HAT. Conversely, COC enhanced 14C-absorption at all harvest times when added to 0.24 kg L−1 formulation. Most of 14C-clethodim (79 to 100% of absorbed) remained in the treated leaf, independent of formulation or adjuvant. Formulation did not have an impact on distribution of absorbed 14C-clethodim; however, presence of an adjuvant increased movement of 14C-clethodim out of treated leaf. Of the absorbed 14C-label, most remained in the treated leaf. 14C-clethodim that translocated out of the treated leaf remained in the shoot, and negligible amount of 14C-clethodim translocated to roots. These results demonstrated improved absorption of clethodim with formulations containing half the active ingredient (0.12 kg L−1) and inclusion of both AMS and COC.

The use of 75Selenomethionine labelled cercariae of Schistosoma mansoni for ultrastructural localization of parasites in host tissues has been evaluated. Both gamma counting of tissue, and autoradiography of resin-embedded tissue were successful. The autoradiographic technique was more sensitive and schistosomula were readily located in pulmonary tissue up to 24 days post-infection.

The effect of exogenous steroid hormones on the egg output of Heligmosomoides polygyrus (Nematoda: Trichostrongylidae) was examined in vitro. Using worms raised in female mice, it was found that estradiol, testosterone, and cortisone each significantly decreased egg output. Although similar trends were found using H. polygyrus raised in male mice, none of the decreases found was significant. No significant differences were found with ecdysone or progesterone treatments using worms from female or male mice. Treatment of worms with cortisone did not significantly affect retention of eggs within the uterus of H. polygyrus. Titration of the effect of cortisone on egg output indicated that levels of reduction were significant for concentrations of 5·6 × 10−6 M to 5·6 × 10−3 M in worms from female mice and for concentrations of 5·6 × 10−8, × 10−7, × 10−5 and × 10−3 in worms from male mice. Radioisotope labelling experiments showed incorporation of 3H-corticosterone in the nucleus of intestinal cells of H. polygyrus suggesting that its effect on egg production may be via a modulatory effect on the intestinal cells.

The present study was performed as part of a systematic examination of glycine's coexistence with other classical transmitters and neuropeptides in neuronal populations of the larval tiger salamander retina. Substance P immunocytochemistry was combined with either glycine immunocytochemistry or autoradiography of glycine high-affinity uptake to examine whether tiger salamander substance P-amacrine cells express these glycine markers. Double-label analyses revealed two populations of substance P-amacrine cells that express glycine immunoreactivity and glycine high-affinity uptake. The large majority of double-labeled cells were situated in the innermost cell row of the inner nuclear layer, while a smaller number were located in the inner nuclear layer in the second cell row distal to the inner plexiform layer. Double-label immunocytochemistry revealed that these double-labeled cells accounted for 91.7% of substance P-immunoreactive amacrine cells. A slightly lower percentage (90.1%) of substance P-amacrine cells were found to exhibit a glycine high-affinity uptake mechanism. Substance P-amacrine cells that did not co-label for markers of glycine activity were situated in the innermost cell row of the inner nuclear layer. Substance P-immunoreactive displaced amacrine cells were not observed to co-label for either glycine immunoreactivity or glycine high-affinity uptake.

The present study reveals that the large majority of substance P-amacrine cells in the larval tiger salamander retina co-express markers of glycine activity. This finding suggests a functional diversity in the population of tiger salamander substance P-amacrine cells relative to their coexisting relationship with a major inhibitory neurotransmitter.

Two physiologically distinct, HRP-marked turtle retinal ganglion cells were examined for their morphology, GABAergic, glycinergic, and bipolar cell synaptic inputs, using electron-microscopic autoradiography and postembedding immunocytochemistry. One cell was a color-opponent, transient ON/OFF ganglion cell. Its center response to red was a sustained hyperpolarization, and its center response to green was a depolarization with increased spiking at onset. The HRP-injected cell most resembled G6, from previous Golgi-impregnation studies (Kolb, 1982; Kolb et al., 1988). It was a narrow-field bistratified cell, whose two broad dendritic strata peaked at approximately levels L20–25 (sublamina a) and L60 (sublamina b) of the inner plexiform layer. Bipolar cell synapses onto G6 were found evenly distributed between its distal and proximal dendritic strata, spanning L20–75. These inputs probably originated from several different bipolar cells, reflecting the complexity of the center response. GABAergic inputs were found onto both the distal and proximal strata, from near L20–L85. Only a few glycinergic inputs, confined to dendrites at L50–70, were observed.

A second ganglion cell type that we physiologically characterized and HRP-injected had sustained ON-center, sustained OFF-surround responses. Two examples were studied; both were bistratified in sublamina b, near L60–70 and L85–100, with branches up to near L40. They resembled G10, from previous Golgi-impregnation studies (Kolb, 1982; Kolb et al., 1988). One cell was partially reconstructed to look at the distributions of GABAergic and glycinergic amacrine cell, and bipolar cell inputs. Although synapses from bipolar cells were equally divided between the two major dendritic strata of G10, the inputs to the distal stratum were close to the soma, and the inputs to the more proximal stratum were on the peripheral dendrites. This arrangement may reflect input from two distinct types of ON-bipolar cell. GABAergic and glycinergic inputs to G10 costratified to both strata and to the distal branches; but where glycinergic inputs were found distributed throughout the arbor, GABAergic inputs appeared to be confined to peripheral dendrites. We hypothesize on the neural elements involved and the circuitry that may underlie the physiologically recorded receptive fields of these two very different ganglion cell types in the turtle retina.

The projection of the thalamic intralaminar nuclei (ILN) upon the visual cortex in the squirrel monkey was studied using anterograde, autoradiographic techniques. In area 17, the ILN afferents terminate in the inner and outer portions of lamina V, whereas in areas 18 and 19 the fibers terminate more diffusely along the laminae V–VI boundary. Widespread labeling of layer I is seen throughout the occipital cortex.

Immunocytochemical and autoradiographic methods were used to identify neurons in the pure cone retina of the lizard (Anolis carolinensis) that are likely to employ glutamate (GLU) or aspartate (ASP) as a neurotransmitter.

GLU immunocytochemistry demonstrated high levels of endogenous GLU in all cone types and numerous bipolar cells. Moderate GLU levels were found in horizontal and ganglion cells. Müller cells and most amacrine cells had very low GLU levels. GLU immunoreactivity (GLU-IR) in the cones was present from the inner segment to the synaptic pedicle. A large spherical cell type with moderate GLU-IR was identified in the proximal inner plexiform layer (IPL). These cells also contain ASP and have been tentatively identified as amacrine cells. Uptake of [3H]-L-GLU labeled all retinal layers. All cone types and Müller cells sequestered [3H]-D-ASP, a substrate specific for the GLU transporter.

Anti-ASP labeling was observed in cones, horizontal cells, amacrine cells, and cells in the ganglion cell layer. ASP immunoreactivity (ASP-IR) in the cones was confined to the inner segment. One ASP-containing pyriform amacrine cell subtype ramifying in IPL sublamina b was identified.

Analysis of GLU-IR, ASP-IR, and GABA-IR on serial sections indicated that there were two distinct populations of horizontal cells in the Anolis retina: one containing GABA-IR, GLU-IR, and ASP-IR; and another type containing only GLU-IR and ASP-IR. Light GLU-IR was frequently found in GABA-containing amacrine cells but ASP-IR was not.

The distinct distributions of GLU and ASP may indicate distinctly different roles for these amino acids. GLU, not ASP, is probably the major neurotransmitter in the cone-biploar-ganglion cell pathway of the Anolis retina. Both GLU and ASP are present in horizontal cells and specific subpopulations of amacrine cells, but it is unclear if GLU or ASP have a neurotransmitter role in these cells.

Abstract An autoradiographic study was made of leucine and thymidine incorporation into the meristematic root primordia and hypocotyl tips of seeds of the recalcitrant mangrove species, Avicennia marina. The investigations show that although there is a temporary reduction of protein synthesis at shedding, root primordia and surrounding hypocotyl cells of the axis never wholly cease incorporation of [3H]leucine and regain pre-shedding levels of activity within a day. Precursor studies using methyl-[3H]thymidine show that, at shedding, there is a temporary cessation of incorporation into root meristem nuclei that lasts no longer than 48 h and, within a day, pre-shedding levels are regained in the meristem nuclei. Analysis of DNA fragmentation patterns in root tips at the time of shedding, and their ability to repair radiation-induced DNA damage, indicate that DNA repair processes are markedly compromised in these cells if water loss reaches 22%. Protein synthesis and DNA replication are reduced by more than half by a water loss of 18% and 16%, respectively. DNA replication does not fully recover on rehydration after only 8% water loss. DNA fragmentation to nucleosomes indicates a programme of cell death at a water loss of 10%. We suggest that the feature of continuous protein synthesis activity with only a temporary interruption in active cell cycling in A. marina root primordia helps to explain both the rapidity in seedling establishment and the extreme vulnerability to desiccation.

Anatomical findings are presented that identify cortical and subcortical sources of afferents to the nucleus reticularis tegmenti pontis (NRTP) and basal pontine nuclei. Projections from the middle temporal visual area (MT), medial superior temporal visual area (MST), lateral intraparietal area (LIP), and areas 7a and 7b to the basal pontine nuclei were studied using 3H-leucine autoradiography. The results complemented a parallel study of retrograde neuronal labeling attributable to injecting WGA-HRP into NRTP and neighboring pontine nuclei. Small 3H-leucine injections confined to MT, MST, LIP, area 7a, or area 7b, produced multiple patches of pontine terminal label distributed as follows: (1) An injection within MT produced terminal label limited to the dorsolateral and lateral pontine nuclei. (2) Injections restricted to MST or LIP showed patches of terminal label in the dorsal, dorsolateral, lateral, and peduncular pontine nuclei. (3) Area 7a targets the dorsal, dorsolateral, lateral, peduncular, and ventral pontine nuclei, whereas area 7b projects, additionally, to the dorsomedial and paramedian pontine nuclei. Notably, no projections were seen to NRTP from any of these cortical areas. In contrast, injections made by other investigators into cortical areas anterior to the central sulcus revealed cerebrocortical afferents to NRTP, in addition to nuclei of the basal pontine gray. With our pontine WGA-HRP injections, retrograde neuronal labeling was observed over a large extent of the frontal cortex continuing onto the medial surface which included the lining of the cingulate sulcus and cingulate gyrus. Significant subcortical sources for afferents to the NRTP and basal pontine nuclei were the zona incerta, ventral mesencephalic tegmentum, dorsomedial hypothalamic area, rostral interstitial nucleus of the medial longitudinal fasciculus, red nucleus, and subthalamic nucleus. The combined anterograde and retrograde labeling data indicated that visuo-motor cortico-pontine pathways arising from parietal cortices target only the basal pontine gray, whereas the NRTP, together with select pontine nuclei, is a recipient of afferents from frontal cortical areas. The present findings implicate the existence of parallel direct and indirect cortico-pontine pathways from frontal motor-related cortices to NRTP and neighboring pontine nuclei.