Main Sites Of Accumulation Research Articles

Root exudate–assisted phytoremediation of copper and lead contamination using Rumex acetosa L. and Rumex K-1

Heavy metal pollutants can be effectively removed from soil through phytoremediation using root exudates. Herein, experiments were conducted to assess the phytoremediation capabilities of Rumex acetosa L. and Rumex K-1 root exudates for copper (Cu) and lead (Pb) contamination. Results indicated that these root exudates effectively adsorbed Cu and Pb. Furthermore, the optimal adsorption conditions of Cu by the root exudates of both plants were as follows: light duration of 36 h, light intensity of 8000 Lx, temperature of 25 °C and CO(NH2)2 concentration of 0 %. Moreover, the optimal adsorption conditions of Pb by Rumex acetosa L. and Rumex K-1 root exudates were light duration of 48 h and 24 h, respectively, light intensity of 8000 Lx, temperature of 25 °C and CO(NH2)2 concentration of 0 %. In addition, the root exudates from both plants enhanced the enrichment and transport of Cu and Pb. Moreover, the root was found to be the main accumulation site of Pb, while the stems and leaves were the main accumulation sites of Cu. With the application of root exudates, plant growth increased, with growth indices in Rumex acetosa L. and Rumex K-1 groups treated with exudates being 1.08–1.81-fold and 1.06–1.9-fold higher, respectively, compared with the untreated ones; physiological indexes showed 1.14–2.62-fold and 1.14–2.71-fold improvements, respectively. Remediation efficiency indexes showed 1.05–1.62-fold and 1.10–1.89-fold improvements, respectively. Rumex acetosa L. and Rumex K-1 exhibited promising potential for the phytoremediation of Cu and Pb, with root exudates playing a critical role in metal adsorption and stabilisation, suggesting their potential for enhancing remediation capabilities. This study sheds light on the mechanisms of root exudate–assisted phytoremediation and provides insights into alleviating heavy metal pollution.

Formation mechanism of glandular trichomes involved in the synthesis and storage of terpenoids in lavender

BackgroundLavender (genus Lavandula, family Lamiaceae) is an aromatic plant widely grown as an ornamental plant. The chemical composition of lavender is characterized by monoterpenoids, sesquiterpenoids, and other compounds, which are primarily synthesized and stored in epidermal secretory structures called glandular trichomes (GTs). Volatile organic compounds (VOCs) are responsible for the aroma characteristics of plant oil that drive consumer preference. Aroma is usually regarded as a characteristic trait for the classification of aromatic plants. Interestingly, VOCs are synthesized and stored in GTs. Lamiaceae species such as purple perilla, peppermint, basil, thyme, and oregano usually possess two types of GTs: peltate glandular trichomes (PGTs) and capitate glandular trichomes (CGTs). But the development process of PGTs in lavender has been reported in only a few studies to date.ResultsIn this study, we identified and quantified the VOCs in four lavender cultivars by headspace-solid phase micro extraction-gas chromatography mass spectrometry (HS–SPME–GC–MS). A total of 66 VOCs were identified in these four cultivars, the most prominent of which were linalyl acetate and linalool, and flowers were the main site of accumulation of these VOCs. Here, we examined the developmental process of PGTs, including the formation of their base, body, and apex. The apex cells contained secretory cavities, which produced VOCs. Based on the reference genome sequence of the lavender cultivar ‘Jingxun 2’, several R2R3-MYB subfamily genes related to GT formation were identified. These results will guide the engineering of GTs and molecular breeding of lavender for improving the VOC content.ConclusionsIn this study, we identified the VOCs in four lavender cultivars. We analyzed the formation of GTs, and compared the number and diameter size of PGTs among four lavender cultivars. Additionally, we identified four candidate genes belonging to the R2R3-MYB family.

Integrated mRNA and miRNA omics reveal the regulatory role of UV-B radiation in active ingredient biosynthesis of Chrysanthemum morifolium Ramat

Ultraviolet-B (UV-B) is an effective elicitor that enhances bioactive ingredient biosynthesis in different plants. Chrysanthemum morifolium Ramat (C. morifolium) is a well-known herbal medicine and beverage material, which is cultivated in many Asian countries and regions. The C. morifolium inflorescences are the main accumulation sites for active ingredients, which include flavonoids and chlorogenic acid. However, the molecular mechanisms underlying UV-B-mediated bioactive ingredient biosynthesis in C. morifolium inflorescences remain largely unknown. High-throughput sequencing technologies have been applied to identify the messenger RNAs (mRNAs) and microRNAs (miRNAs) responsive to UV-B radiation in C. morifolium inflorescences. Many UV-B responsive genes are enriched in flavonoid, unsaturated fatty acid, phenylpropanoid, and carotenoid biosynthesis pathways. In this study, we identified 169 miRNAs belonging to 38 typical families were detected in our constructed libraries. In total, 1954 unigenes were predicted to be targets of 118 miRNAs. Six miRNAs, PC-5p-7136_1319, PC-3p-78379_156, ath-MIR414-p5_2ss2CT21AC, PC-3p-154888_69, nta-miR171a_2ss8GT18GA and htu-miR403a were involved in the regulation of active ingredient biosynthesis by targeting 4CL-like 5/9, CYP81E8, UGT91D1 and UGT72B1 genes. Five miRNAs, PC-5p-79_56613, ath-miR858a_L-1, ath-MIR414-p5_2ss15AC18AT, ath-MIR414-p5_2ss2CT21AC, and aly-miR393a-5p were involved in regulation of flavonoid biosynthesis by targeting MYB-bHLH complex-related transcription factor genes. Our data provides a platform for identifying mRNAs and miRNAs involved in the UV-B-induced accumulation of bioactive ingredients in C. morifolium inflorescences.

Polymeric Nanomicelles Loaded with Anandamide and Their Renal Effects as a Therapeutic Alternative for Hypertension Treatment by Passive Targeting.

We have previously demonstrated significant in vitro natriuretic effects of anandamide (AEA) nanoformulation in polymeric nanoparticles, whose size prevents their accumulation in organs, such as the kidneys. Therefore, it is of particular interest to design and test nanostructures that can pharmacologically accumulate in these organs. In this regard, we prepared and characterized polymeric nanomicelles (~14 and 40 nm). Likewise, their biodistribution was determined. Spontaneously hypertensive rats (SHR) and normotensive rats (WKY), n = 3 per group, were divided into five treatment conditions: control, sham, free AEA freshly dispersed in aqueous solution or 24 h after its dispersion, and AEA encapsulated in nanomicelles. The kidneys were the main site of accumulation of the nanoformulation after 24 h. Freshly dispersed free AEA showed its classical triphasic response in SHR, which was absent from all other treatments. Nanoformulated AEA produced a sustained antihypertensive effect over 2 h, accompanied by a significant increase in fractional sodium excretion (FSE %). These effects were not observed in WKY, sham, or free AEA-treated rats after 24 h of its aqueous dispersion. Without precedent, we demonstrate in vivo natriuretic, diuretic, and hypotensive effects of AEA nanoformulation in polymeric nanomicelles, suggesting its possible use as a new antihypertensive agent with intravenous administration and passive renal accumulation.

Uptake and distribution of difenoconazole in rice plants under different culture patterns

The effects of spraying and root irrigation on the uptake and transport of the fungicide difenoconazole under hydroponic and soil cultivation were investigated. Rice was used as the crop for a short-term exposure experiment. A modified QuEChERS pre-treatment combined with ultra-high-performance liquid chromatography–tandem mass spectrometry was used to extract and detect difenoconazole from rice plants, water and soil. The recoveries of difenoconazole were in the range of 72.8–110.5%, with a relative standard deviation of 2.4–19.5% for all the samples when spiked with 0.01, 0.1 and 1 mg kg−1 of difenoconazole, respectively. The limit of quantitation (LOQ) of this method was 0.01 mg kg−1. The exposure results showed that difenoconazole could be absorbed by rice plants and transmitted to different parts of rice plants in all the treatments. In the hydroponic experiment, difenoconazole was mainly distributed in the roots of rice regardless of whether irrigation or spraying was used. For rice cultivated in soil, difenoconazole mainly accumulated in leaves after the root irrigation treatment, whereas after the spraying treatment, the rice roots were the main site of accumulation of difenoconazole. This experiment extends our knowledge of the influence of the cultivation system and application mode on the translocation of difenoconazole in rice plants.

Management of Urdhwag Amlapitta with Jaladhauti and lifestyle modification: A case study

Amlapitta is a gastrointestinal disorder. It comes under lifestyle disorder. Improper diet and habits, stress, and spicy food are some causative factors that vitiate Pitta and Kapha by Ama. The objective of this study was to access the role of lifestyle modification and Ayurvedic management in Urdhwag Amlapitta. A 35-year-old female patient with a known case of Urdhwag Amlapitta presented with complaints of sour belching, burning sensation in the chest, headache, exertion without work, nausea, indigestion, and thirst for 1 year. Appropriate modifications were made in diet and lifestyle. A proper diet chart was provided. Three settings of Jaladhauti Shuddhikriya (~voluntarily induced vomiting) then Hingwashtak Churna, Avipattikar Churna, and Dadimavaleh were advised for 1 month. Follow-up was taken on the 7th day and 15th day. Subjective symptoms reduced remarkably and no recurrence of Amlapitta was seen. Lifestyle modification plays an important role in Nidanparivarjan (avoid intake of risk factors) to control recurrence. Jaladhauti shuddhikriya corrects the pathology by eliminating disease causative factor Kapha/Ama from its main site of accumulation. Jatharagni Deepan and Pachan of residual Aam are done by Hingwashtak Churna. Avipattikar Churna works as Pittavirechak. By this study, it can be concluded that lifestyle modification helps in controlling recurrence and Jaladhauti Shuddhikriya is a safe and effective treatment for Urdhwag Amlapitta.

Review on Lagrangian Particle Tracking Modeling for Floating Objects around the Korean Peninsula

Marine debris is becoming an increasingly serious problem. Using a Lagrangian particle model one can calculate the trajectories of various floating objects including marine debris. We reviewed Lagrangian particle tracking of floating objects in the seas around Korea from three recent studies. The first one was on the fate of marine debris from lands through the rivers discharging into the seas around the Korean Peninsula. The main path of the debris depended on their geographical locations. The debris from the rivers on the east coast of China moved southward along the Chinese coast, to the Korea Strait, or to the North Pacific Ocean via the Kuroshio Current (KC). That from the rivers located on the west coast of the Korean Peninsula moved southward to reach the KC, and then to the North Pacific Ocean. The main pathway of the debris from the Nakdong Rivers was toward the East Sea (ES). The particles released from the Tumen River were limited to the ES. In the second example, the pathways of the debris released through the Tumen River located between the North Korea and Russia during the flooding induced by the typhoon Lionrock in August 2016 were investigated. The debris was transported southward along the east coast of the Korean Peninsula by the southward North Korea Cold Current while affected by the wind. Three potential hotspots of debris accumulation along the coast were also found. In the third example, the most probable route of a wave buoy found at Cape Mendocino, California, USA was deduced. The buoy moved from Jeju Island toward Taiwan, and then to the North Pacific via the KC. These model results would help us to establish efficient cleaning and management plans of marine debris by showing us the main accumulation sites, pathways and source areas.

Pharmacodynamic and Pharmacokinetic Properties of Full Phosphorothioate Small Interfering RNAs for Gene Silencing In Vivo

State-of-the-art small interfering RNA (siRNA) therapeutics such as givosiran and fitusiran are constructed from three variable components: a fully-modified RNA core that conveys metabolic stability, a targeting moiety that mediates target-cell uptake, and a linker. This structural complexity poses challenges for metabolite characterization and risk assessment after long-term patient exposure. In this study, we show that basic phosphorothioate modification of a siRNA targeting the oncoprotein Lin28B provides a useful increase in metabolic stability, without greatly compromising potency. We found that its stability in vitro matched that of nanoparticle-free patisiran in serum and surpassed it in liver tritosome extracts, although it did not reach the stability of the fitusiran siRNA core structure. Liver and kidney were the main sites of accumulation after its subcutaneous administration in mice. Despite the lack of a delivery agent-free antitumor effect, we anticipate our study to be a starting point to develop alternative siRNA scaffolds that can be degraded into naturally-occurring metabolites and help alleviate the aforementioned challenges. Furthermore, Lin28B is a promising target for cancers, and the development of such simplified siRNA analogs, possibly together with novel targeting units, holds potential.

Spatial distribution of structural elements in leaves of Ilex paraguariensis: physiological and ecological implications

Calcium accumulation in crystals (mainly in the midrib) and silicon on the external surface of the upper epidermis suggests physiological and/or ecological roles. Concentration, form, and location of elements in leaf tissue of Ilex paraguariensis A. St.-Hil. can influence remobilization patterns and release of substances during its infusion for making yerba mate beverages. Even though, spatial distribution of elements and characterization of crystal formation in I. paraguariensis has not been elucidated. For filling this gap, this study used microanalytical techniques to evaluate accumulation and spatial distribution of elements in yerba mate leaves, which was done using chemical analyses and scanning electron microscopy combined with energy-dispersive spectroscopy (SEM–EDS). Chemical analyses confirmed high concentrations of potassium (K) and manganese (Mn) in leaves. Cell walls were predominantly composed of carbon (C) and oxygen (O). Compartmentalization of silicon (Si) occurred on the external surface of the upper epidermis between cell walls and the leaf cuticle. Precipitation of calcium (Ca) was higher in the midrib than in the leaf blade with the mesophyll and palisade mesophyll being the main accumulation sites. Microscopy–spectroscopy analysis revealed different patterns of elemental accumulation in leaves, which suggests physiological and/or ecological roles in yerba mate plants. Additionally, the abundance of Ca oxalates could explain the low Ca solubility during the beverage infusion process.

Porous Silicon as a Platform for Radiation Theranostics Together with a Novel RIB-Based Radiolanthanoid.

Mesoporous silicon (PSi) is biocompatible and tailorable material with high potential in drug delivery applications. Here, we report of an evaluation of PSi as a carrier platform for theranostics by delivering a radioactive ion beam- (RIB-) based radioactive lanthanoid into tumors in a mouse model of prostate carcinoma. Thermally hydrocarbonized porous silicon (THCPSi) wafers were implanted with 159Dy at the facility for radioactive ion beams ISOLDE located at CERN, and the resulting [159Dy]THCPSi was postprocessed into particles. The particles were intratumorally injected into mice bearing prostate cancer xenografts. The stability of the particles was studied in vivo, followed by ex vivo biodistribution and autoradiographic studies. We showed that the process of producing radionuclide-implanted PSi particles is feasible and that the [159Dy]THCPSi particles stay stable and local inside the tumor over seven days. Upon release of 159Dy from the particles, the main site of accumulation is in the skeleton, which is in agreement with previous studies on the biodistribution of dysprosium. We conclude that THCPSi particles are a suitable platform together with RIB-based radiolanthanoids for theranostic purposes as they are retained after administration inside the tumor and the radiolanthanoid remains embedded in the THCPSi.

Accumulation and tolerance characteristics of lead in Althaea rosea Cav. and Malva crispa L.

Two ornamental plants of Althaea rosea Cav. and Malva crispa L. were exposed to various concentrations of lead (Pb) (0, 50, 100, 200 and 500 mg·kg−1) for 70 days to evaluate the accumulating potential and the tolerance characteristics. The results showed that both plant species grown normally under Pb stress, and A. rosea had a higher tolerance than M. crispa, while M. crispa had a higher ability in Pb accumulation than A. rosea. Besides, lower Pb concentration (50 mg·kg−1) stimulated the shoot biomass in both plant species. Pb accumulation in plants was consistent with the increase of Pb levels, and the main accumulation sites were the roots and the older leaves. In addition, the photosynthetic pigments content and chlorophyll fluorescence parameters were influenced by Pb stress. In such case, both of the plants could improve the activities of antioxidant enzymes of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX), and the contents of the total soluble sugar and soluble protein, which reached the highest value at Pb 100 mg·kg−1, as well as the accumulation of the total thiols (T-SH) and non-protein thiols (NP-SH) to adapt to Pb stress. Thus, it provides the theoretical basis and possibility for ornamental plants of A. rosea and M. crispa in phytoremediation of Pb contaminated areas.

English

Among antibiotics, tetracyclines are the most commonly used and detected in the environment. In this study, the amount of tetracycline taken up from soil by pea seedlings was analyzed, identified its main site of accumulation in plants and determined also changes in the protein profile of pea. The study demonstrates that pea seedlings take up tetracycline from soil and transport the drug via roots to over-ground parts and then accumulate it in the youngest parts, such as upper stem and leaves. After the taken up of drug, the activity of guaiacol peroxidase is modified and changes in the profile of proteins, as determined by two-dimensional gel electrophoresis occur. The majority of proteins (∼40%) visualized possessed molecular weight between 25 and 37 kDa. Only 8% of the proteins had molecular weight lower than 20 kDa, and 2% greater than 75 kDa. The number of spots in the control samples was 194, which is less by 49 than at the concentration of 150 mg kg of soil. Isoflavone reductase was present only in seedlings growing with tetracycline. Tetracycline uptake from soil modify mainly the changes in biochemical processes connected with protein. © 2016 Friends Science Publishers

Determination of phenolic compounds in flowers of Hypericum species native to South Brazil and Peruvian Páramos

The flowers constitute one of the main sites of accumulation of phenolic compounds in plants of the Hypericum genus. In addition to their important pharmacological activities, some metabolites found in species from the section Brathys and Trigynobrathys appear to have chemotaxonomic significance according to the literature. HPLC analyses were carried out to assess the pattern and accumulation of the dimeric phloroglucinols, benzophenones, benzopyrans, flavonoids and a phenolic acid in flowers of Hypericum species native to southern Brazil and Peruvian Paramos. Qualitative and quantitative differences are reported. Uliginosin B and hyperoside were the main components detected in all species and with maximum concentrations up to 0.188 % in H. caprifoliatum and 5.987 % in H. andinum, respectively. The content of japonicin A varied from 0.003 to 0.087 % in H. myrianthum, while the yield of hyperbrasilol B ranged from 0.006 % in H. laricifolium to 0.011 % in H. caprifoliatum. The major compounds in H. polyanthemum and H. carinatum were the benzopyrans 6-isobutyryl-5,7-dimethoxy-2,2-dimethyl-benzopyran (HP1 = 0.200 %), 7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethyl-benzopyran (HP2 = 0.225 %) and 5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethyl-benzopyran (HP3 = 0.327 %) and benzophenones cariphenone A (0.309 %) and cariphenone B (0.062 %), respectively. Maximum amounts of chlorogenic acid, isoquercitrin, quercitrin and guaijaverin were observed, respectively, in H. campestre (1.458 %), H. andinum (1.161 %), H. carinatum (0.231 %) and H. laricifolium (0.404 %). The results obtained support the taxonomic evidence of the dimeric phloroglucinol derivatives at the section level.

Tight Junction Proteins and Oxidative Stress in Heavy Metals-Induced Nephrotoxicity

Kidney is a target organ for heavy metals. They accumulate in several segments of the nephron and cause profound alterations in morphology and function. Acute intoxication frequently causes acute renal failure. The effects of chronic exposure have not been fully disclosed. In recent years increasing awareness of the consequences of their presence in the kidney has evolved. In this review we focus on the alterations induced by heavy metals on the intercellular junctions of the kidney. We describe that in addition to the proximal tubule, which has been recognized as the main site of accumulation and injury, other segments of the nephron, such as glomeruli, vessels, and distal nephron, show also deleterious effects. We also emphasize the participation of oxidative stress as a relevant component of the renal damage induced by heavy metals and the beneficial effect that some antioxidant drugs, such as vitamin A (all-trans-retinoic acid) and vitamin E (α-tocopherol), depict on the morphological and functional alterations induced by heavy metals.

Evaluation of a 99mTc-Labeled AnnexinA5 Variant for Non-invasive SPECT Imaging of Cell Death in Liver, Spleen and Prostate

We investigate radio-labeling and pharmacokinetics of a new AnnexinA5 variant (HYNIC-cys-AnxA5) and then assess its utility for the non-invasive detection of cell death in liver, spleen and prostate. AnnexinA5 binds to phosphatidylserine expressed on the surface of apoptotic and necrotic cells. Contrary to other AnnexinA5 variants, the new cys-AnxA5 allows for site-specific conjugation of a hydrazinonicotinamide-maleimide moiety and subsequent radio-labeling with (99m)Tc at a position not involved in the AnxA5-phosphatidylserine interaction. Distribution of (99m)Tc-HYNIC-cys-AnxA5 was studied in rats, both invasively and via SPECT/CT. Cycloheximide was used to induce cell death in liver and spleen, whereas apoptosis in the prostate was induced by castration. HYNIC-cys-AnxA5 was efficiently and reproducibly labeled with (99m)Tc. Blood clearance of radioactivity after iv-injection was adequately described by a two-compartment model, the renal cortex representing the main site of accumulation. Cycloheximide treatment resulted in increased accumulation of intravenous-injected (99m)Tc-HYNIC-cys-AnxA5 in liver and spleen over controls, which correlated well with TUNEL staining for cell death in corresponding tissue sections. However, the increase in TUNEL-positive prostate epithelial cells observed following castration was not paralleled by greater (99m)Tc-HYNIC-cys-AnxA5 accumulation. (99m)Tc-HYNIC-cys-AnxA5 appears a suitable tracer for assessment of cell death in liver and spleen, but not prostate.

Methods applied in studies of benthic marine debris

The ocean floor is one of the main accumulation sites of marine debris. The study of this kind of debris still lags behind that of shorelines. It is necessary to identify the methods used to evaluate this debris and how the results are presented and interpreted. From the available literature on benthic marine debris (26 studies), six sampling methods were registered: bottom trawl net, sonar, submersible, snorkeling, scuba diving and manta tow. The most frequent method used was bottom trawl net, followed by the three methods of diving. The majority of the debris was classified according to their former use and the results usually expressed as items per unity of area. To facilitate comparisons of the contamination levels among sites and regions some standardization requirements are suggested.

The R2R3‐MYB transcription factor HAG1/MYB28 is a regulator of methionine‐derived glucosinolate biosynthesis in Arabidopsis thaliana

Methionine-derived glucosinolates belong to a class of plant secondary metabolites that serve as chemoprotective compounds in plant biotic defense reactions and also exhibit strong anticancerogenic properties beneficial to human health. In a screen for the trans-activation potential of various transcription factors toward glucosinolate biosynthetic genes, we could identify the HAG1 (HIGH ALIPHATIC GLUCOSINOLATE 1, also referred to as MYB28) gene as a positive regulator of aliphatic methionine-derived glucosinolates. The content of aliphatic glucosinolates as well as transcript levels of aliphatic glucosinolate biosynthetic genes were elevated in gain-of-function mutants and decreased in HAG1 RNAi knock-down mutants. Pro(HAG1):GUS expression analysis revealed strong HAG1 promoter activity in generative organs and mature leaves of A. thaliana plants, the main sites of accumulation of aliphatic glucosinolates. Mechanical stimuli such as touch or wounding transiently induced HAG1/MYB28 expression in inflorescences of flowering plants, and HAG1/MYB28 over-expression reduced insect performance as revealed by weight gain assays with the generalist lepidopteran herbivore Spodoptera exigua. Expression of HAG1/MYB28 was significantly induced by glucose, indicating a novel transcriptional regulatory mechanism for the integration of carbohydrate availability upon biotic challenge. We hypothesize that HAG1/MYB28 is a novel regulator of aliphatic glucosinolate biosynthesis that controls the response to biotic challenges.

Avoidance responses to aluminium in the freshwater bivalve Anodonta cygnea

This study examined the effect of aluminium (Al) on the filtering behaviour (shell opening or gape) of the freshwater bivalve Anodonta cygnea L in neutral fresh water. Parallel measurements of Al concentration in the soft tissues were made to examine the relationship between changes in behaviour and accumulation of Al. The number of lysosomal granules in the gill, kidney and digestive gland were counted, as lysosomes are known to be involved in the excretion and detoxification of trace metals. The bivalves were exposed to two environmentally relevant concentrations of added Al i.e. 250 and 500 μg l −1 (9.25 and 18.5 μM l −1) at neutral pH for 15 days and shell movement monitored continuously. Aluminium affected the mussels’ filtering activity, producing an avoidance reaction whose magnitude was concentration-dependent; 250 μg l −1 added Al produced no detectable change, while 500 μg l −1 Al reduced mean duration of shell opening by 50%. This effect was irreversible over a 15 day recovery period. Tissue levels of Al after 15 days exposure were an order of magnitude higher in animals exposed to 250 μg l −1 added Al than in those exposed to 500 μg l −1. This was consistent with the inhibition of filtering activity due to valve closure at the higher concentration, which may have prevented uptake of Al. In addition, probable different chemical speciation of Al in the water column (soluble for 250 and colloidal for 500 μg l −1) may lead to marked differences in tissue uptake. The kidney and digestive gland were the main sites of accumulation of Al and concentrations remained significantly elevated 15 days after transfer of animals to clean water. It is suggested that mucus plays a role in the exclusion of Al as elevated concentrations were measured in the pseudofaeces of animals during and after exposure. Lysosomal granules may be involved in the intracellular handling and detoxification of Al as numbers increased significantly in all organs during exposure and continued to increase after the animals were transferred to clean water. The present study provides evidence for the bioavailability and toxicity of Al to mussels at neutral pH and at concentrations which are known to enter neutral freshwaters when mobilised by natural or anthropogenic acidity. The changes in behaviour and uptake of Al in the mussel observed in this investigation are, therefore, likely to be reflected in the natural environment and the degree to which Al affects the ‘fitness’ of the mussel populations and the transfer of Al through the food chain merit investigation.

Extracellular complexation of Cd in the Hartig net and cytosolic Zn sequestration in the fungal mantle of Picea abies – Hebeloma crustuliniforme ectomycorrhizas

ABSTRACTCompartmentation of heavy metals on or within mycorrhizal fungi may serve as a protective function for the roots of forest trees growing in soils containing elevated concentrations of metals such as Cd and Zn. In this paper we present the first quantitative measurements by X‐ray microanalysis of heavy metals in high‐pressure frozen and cryosectioned ectomycorrhizal fungal hyphae. We used this technique to analyse the main sites of Cd and Zn in fungal cells of mantle and Hartig net hyphae and in cortical root cells of symbiotic Picea abies – Hebeloma crustuliniforme associations to gain new insights into the mechanisms of detoxification of these two metals in Norway spruce seedlings. The mycorrhizal seedlings were exposed in growth pouches to either 1 mM Cd or 2 mM Zn for 5 weeks. The microanalytical data revealed that two distinct Cd‐ and Zn‐binding mechanisms are involved in cellular compartmentation of Cd and Zn in the mycobiont. Whereas extracellular complexation of Cd occurred predominantly in the Hartig net hyphae, both extracellular complexation and cytosolic sequestration of Zn occurred in the fungal tissue. The vacuoles were presumed not to be a significant pool for Cd and Zn storage. Cadmium was almost exclusively localized in the cell walls of the Hartig net (up to 161 mmol kg−1 DW) compared with significantly lower concentrations in the cell walls of mantle hyphae (22 mmol kg−1 DW) and in the cell walls of cortical cells (15 mmol kg−1 DW). This suggests that the apoplast of the Hartig net is a primary accumulation site for Cd. Zinc accumulated mainly in the cell walls of the mantle hyphae (111 mmol kg−1 DW), the Hartig net hyphae (130 mmol kg−1 DW) and the cortical cells (152 mmol kg−1 DW). In addition, Zn occurred in high concentrations in the cytoplasm of the fungal mantle hyphae (up to 164 mmol kg−1 DW) suggesting that both the cell walls and the cytoplasm of fungal tissue are the main accumulation sites for Zn in P. abies resulting in decreased Zn transfer from the fungus to the root.

Pharmacokinetics and biodistribution of a nucleotide-based thrombin inhibitor in rats.

To characterize the pharmacokinetic and tissue distribution profiles of a nucleotide-based thrombin inhibitor (GS522, phosphodiester oligonucleotide, GGTTGGTGTGGTTGG) following intravenous administration to rats. Pharmacokinetic study: 10 mg/kg, 20 mg/kg, 30 mg/kg (6 animals/dose) were administered to rats by rapid injection into the femoral vein. Blood samples were collected over a 45 minute period. Plasma concentrations of GS522 were determined using capillary gel electrophoresis with laser-induced fluorescence detection. Biodistribution Study: 10 mg/kg (400 microl, 31.46 microCi/ml) of 3H-GS522 was administered to rats by rapid injection into the femoral vein. The animals were sacrificed by decapitation at 1, 5, 10, 30, 60, 360 minutes post-dose (3 rats/point). Brain, blood, duodenum, eyes, heart, kidney, liver, lungs, muscle, pancreas, skin, spleen and vein samples were collected, processed and quantitated using liquid scintillation counting. The pharmacokinetic profile declines in multiexponential manner, exhibiting extremely fast distribution and elimination (t1/2 = 7.6-9.0 min, Cl = 22.0-28.0 ml/min, V = 83.9-132.4 ml/kg). GS522 follows linear pharmacokinetics, with the area under the curve being proportional to the dose (Rsq = 0.9744). Highest radioactivity levels were detected in kidney, liver and blood (39.7, 15.7 and 15.3% dose/ respective organ). Less than 1% of the dose was detected in the heart, spleen and lungs, and >0.3% of the dose was found in the brain and eyes. The oligonucleotide associated radioactivity was uniformly distributed between the brain regions (left and right lobe and cerebellum). Six hours following the dose administration a statistically significant increase (p < 0.05) in radioactivity levels was observed in the brain, eyes, skin, liver, pancreas and vein. The pharmacokinetic and biodistribution profiles of GS522 following intravenous administration to rats at three doses were characterized. The oligonucleotide associated radioactivity was widely distributed in tissues. The amount of radioactivity sharply decreased with time in most tissues. Kidney, liver and muscle were the main sites of accumulation. The oligonucleotide associated radioactivity did not cross the blood brain barrier to an appreciable extent. In addition, a statistically significant increase (p < 0.05) in the radioactivity levels observed in select tissues suggested a re-uptake mechanism for intact oligonucleotide or its degradation products.