A strong, silk protein-inspired tissue adhesive with an enhanced drug release mechanism for transdermal drug delivery.

In transdermal drug delivery system (TDDS) patches, achieving prolonged adhesion, high drug loading, and rapid drug release simultaneously presented a significant challenge. In this study, a PHT-SP-Cu2+ adhesive was synthesized using polyethylene glycol (PEG), hexamethylene diisocyanate (HDI), trimethylolpropane (TMP), and silk protein (SP) as functional monomers which were combined with Cu2+ to improve the adhesion, drug loading, and drug release of the patch. The structure of the adhesion chains and the formation of Cu2+-p-π conjugated network in PHT-SP-Cu2+ were characterized and elucidated using different characterization methods including FT-IR, 13C NMR, XPS, SEM imaging and thermodynamic evaluation. The formulation of pressure-sensitive adhesive (PSA) was optimized through comprehensive research on adhesion, mechanics, rheology, and surface energy. The formulation of 3 wt.% SP and 3 wt.% Cu2+ provided superior adhesion properties compared to commercial standards. Subsequently, the peel strength of PHT-SP-Cu2+ was 7.6 times higher than that of the commercially available adhesive DURO-TAK® 87-4098 in the porcine skin peel test. The adhesion test on human skin confirmed that PHT-SP-Cu2+ could adhere to the human body for more than six days. Moreover, the drug loading, in vitro release test and skin permeation test were investigated using ketoprofen as a model drug, and the results showed that PHT-SP-Cu2+ had the efficacy of improving drug compatibility, promoting drug release and enhancing skin permeation as a TDDS. Among them, the drug loading of PHT-SP-Cu2+ was increased by 6.25-fold compared with PHT, and in the in vivo pharmacokinetic analysis, the AUC was similarly increased by 19.22-fold. The mechanism of α-helix facilitated drug release was demonstrated by Flori-Hawkins interaction parameters, molecular dynamics simulations and FT-IR. Biosafety evaluations highlighted the superior skin cytocompatibility and safety of PHT-SP-Cu2+ for transdermal applications. These results would contribute to the development of TDDS patch adhesives with outstanding adhesion, drug loading and release efficiency. STATEMENT OF SIGNIFICANCE: A new adhesive, PHT-SP-Cu2+, was created for transdermal drug delivery patches. Polyethylene glycol, hexamethylene diisocyanate, trimethylolpropane, silk protein, and Cu2+ were used in synthesis. Characterization techniques confirmed the structure and Cu2+-p-π conjugated networks. Optimal formulation included 3 wt.% SP and 3 wt.% Cu2+, exhibiting superior adhesion. PHT-SP-Cu2+ showed 7.6 times higher peel strength than DURO-TAK® 87-4098 on porcine skin and adhered to human skin for over six days. It demonstrated a 6.25-fold increase in drug loading compared to PHT, with 19.22-fold higher AUC in vivo studies. α-helix facilitated drug release, proven by various analyses. PHT-SP-Cu2+ showed excellent cytocompatibility and safety for transdermal applications. This study contributes to developing efficient TDDS patches.

Antileishmanial effects and drugability characteristics of a heterocyclic copper complex: An in silico, in vitro and molecular study.

Leishmaniasis caused by the protozoan Leishmania presents a severe illness, principally in tropical and subtropical areas. Antileishmanial metal complexes, like Glucantime®ï¸ with proven activity, are routinely studied to probe their potency. We investigated the effects of a Cu (II) homoleptic complex coordinated by two dimethyl-bipyridine ligands against Leishmania major stages in silico and in vitro. The affinity of this heterocyclic Cu (II) complex (CuDMBP) towards a parasitic metacaspase was studied by molecular docking. Key pharmacokinetic and pharmacodynamic properties of the complex were predicted using three web-based tools. CuDMBP was tested for in vitro antileishmanial activities using MTT assay, model murine macrophages, flow cytometry, and quantitative real-time polymerase chain reaction (qPCR). Molecular docking confirmed the tendency between the target macromolecule and the complex. ADMET evaluations highlighted CuDMBP's key pharmacological features, including P-glycoprotein-associated GI absorption and lack of trans-BBB permeability. MTT showed significant inhibitory effects against promastigotes. CuDMBP significantly increased the level of cellular IL-12 expression (p < 0.05), while the upregulation observed in the expression of iNOS was considered not significant (p > 0.05). It decreased the expression of IL-10 significantly (p < 0.05). Findings demonstrated that CuDMBP deserves to be introduced as a leishmanicidal candidate provided further studies are carried out.

Prominent antibacterial effect of sub 5 nm Cu nanoparticles/MoS2composite under visible light.

Achieving an efficient and inexpensive bactericidal effect is a key point for the design of antibacterial agent. Recent advances have proved molybdenum disulfide (MoS2) as a promising platform for antimicrobial applications, while the combination of metal nanoparticle would promote the antibacterial efficiency. Nevertheless, the dispersivity, cheapness and safety of metal nanoparticle loaded on MoS2raised some concerns. In this paper, we successfully realized a uniform decoration of copper nanoparticles (CuNPs) on surface of MoS2nanosheets, and the size of CuNPs could be controlled below 5 nm. Under 5 min irradiation of 660 nm visible light, the synthesized CuNPs/MoS2composite demonstrated superior antibacterial performances (almost 100% bacterial killed) towards both Gram-negativeE. coliand Gram-positiveS. aureusover the single component (Cu or MoS2), while the bactericidal effect could last for at least 6 h. The synergism of photodynamic generated hydroxyl radical (·OH), oxidative stress without reactive oxygen species production and the release of Cu ions was considered as the mechanism for the antibacterial properties of CuNPs/MoS2. Our findings provided new insights into the development of two-dimensional antibacterial nanomaterials of high cost performance.

Liposomal formulations of anticancer copper(II) thiosemicarbazone complexes.

α-N-Heterocyclic thiosemicarbazones such as triapine and COTI-2 are currently investigated as anticancer therapeutics in clinical trials. However, triapine was widely inactive against solid tumor types. A likely explanation is the short plasma half-life time and fast metabolism. One promising approach to overcome these drawbacks is the encapsulation of the drug into nanoparticles (passive drug-targeting). In a previous work we showed that it was not possible to stably encapsulate free triapine into liposomes. Hence, in this manuscript we present the successful preparation of liposomal formulations of the copper(II) complexes of triapine and COTI-2. To this end, various drug-loading strategies were examined and the resulting liposomes were physico-chemically characterized. Especially for liposomal Cu-triapine, a decent encapsulation efficacy and a slow drug release behavior could be observed. In contrast, for COTI-2 and its copper(II) complex no stable loading could be achieved. Subsequent in vitro studies in different cell lines with liposomal Cu-triapine showed the expected strongly reduced cytotoxicity and DNA damage induction. Also in vivo distinctly higher copper plasma levels and a continuous release could be observed for the liposomal formulation compared to free Cu-triapine. Taken together, the here presented nanoformulation of Cu-triapine is an important step further to increase the plasma half-life time and tumor targeting properties of anticancer thiosemicarbazones.

Surface Decoration via Physical Interaction of Cupric Diethyldithiocarbamate Nanocrystals and Its Impact on Biodistribution and Tumor Targeting.

The vascular wall is the first physiologic barrier that circulating nanoparticles (NPs) encounter, which also is a key biological barrier to cancer drug delivery. NPs can continually scavenge the endothelium for biomarkers of cancer, and the chance of NPs' extravasation into the tumors can be enhanced. Here, we envision P-selectin as a target for specific delivery of drug nanocrystals to tumors. The cupric diethyldithiocarbamate nanocrystals (CuET NCs) were first prepared by an antisolvent method, and then nanocrystals were coated with fucoidan via physical interaction. The fucoidan-coated CuET nanocrystals (CuET@Fuc) possess high drug loading and have the ability to interact with human umbilical vein endothelial cells expressing P-selectin, which transiently enhances the endothelial permeability and facilitates CuET@Fuc extravasation from the peritumoral vascular to achieve higher tumor accumulation of drugs than bare CuET NCs. The CuET NC shows poorer anticancer efficacy than CuET@Fuc at the same dose of CuET. Upon repeated dosing of CuET@Fuc for 2 weeks, no mortality was observed in treated melanoma-bearing mice, while the mortality in the control group and excipient-treated groups reached 23%. The growth rate of melanoma in the CuET@Fuc-treated group was significantly lower than those in other groups. Furthermore, an acute toxicity study revealed that CuET@Fuc is a safe formulation for cancer treatment.

Acute Toxicity of Cu-MOF Nanoparticles (nanoHKUST-1) towards Embryos and Adult Zebrafish.

Metal-organic frameworks (MOFs) demonstrate unique properties, which are prospective for drug delivery, catalysis, and gas separation, but their biomedical applications might be limited due to their obscure interactions with the environment and humans. It is important to understand their toxic effect on nature before their wide practical application. In this study, HKUST-1 nanoparticles (Cu-nanoMOF, Cu3(btc)2, btc = benzene-1,3,5-tricarboxylate) were synthesized by the microwave (MW)-assisted ionothermal method and characterized by X-ray powder diffraction (XRD) and transmission electron microscopy (TEM) techniques. The embryotoxicity and acute toxicity of HKUST-1 towards embryos and adult zebrafish were investigated. To gain a better understanding of the effects of Cu-MOF particles towards Danio rerio (D. rerio) embryos were exposed to HKUST-1 nanoparticles (NPs) and Cu2+ ions (CuSO4). Cu2+ ions showed a higher toxic effect towards fish compared with Cu-MOF NPs for D. rerio. Both forms of fish were sensitive to the presence of HKUST-1 NPs. Estimated LC50 values were 2.132 mg/L and 1.500 mg/L for zebrafish embryos and adults, respectively. During 96 h of exposure, the release of copper ions in a stock solution and accumulation of copper after 96 h were measured in the internal organs of adult fishes. Uptake examination of the major internal organs did not show any concentration dependency. An increase in the number of copper ions in the test medium was found on the first day of exposure. Toxicity was largely restricted to copper release from HKUST-1 nanomaterials structure into solution.

Hyaluronic acid/polyethyleneimine nanoparticles loaded with copper ion and disulfiram for esophageal cancer.

In the clinical treatment of cancer, improving the effectiveness and targeting of drugs has always been a bottleneck problem that needs to be solved. In this contribution, inspired by the targeted inhibition on cancer from combination application of disulfiram and divalent copper ion (Cu2+), we optimized the concentration of disulfiram and Cu2+ ion for inhibiting esophageal cancer cells, and loaded them in hyaluronic acid (HA)/polyethyleneimine (PEI) nanoparticles with specific scales, in order to improve the effectiveness and targeting of drugs. The in vitro cell experiments demonstrated that more drug loaded HA/PEI nanoparticles accumulated to the esophageal squamous cell carcinoma (Eca109) and promoted higher apoptosis ratio of Eca109. Both in vitro and in vivo biological assessment verified that the disulfiram/Cu2+ loaded HA/PEI nanoparticles promoted the apoptosis of cancer cells and inhibited the tumor proliferation, but had no toxicity on other normal organs.

Influence of culinary treatment on the concentration and on the bioavailability of cadmium, chromium, copper, and lead in seafood.

BACKGROUND: Seafood present important advantages for human nutrition, but it can also accumulate high levels of toxic and potentially toxic elements. Culinary treatments could influence seafood chemical element content and element bioavailability. In this study, the influence of culinary treatments on the total concentration and on the bioavailability of Cd, Cr, Cu and Pb in shark, shrimp, squid, oyster, and scallop was assessed. METHODS: Boiling, frying, and sautéing with or without seasonings (salt, lemon juice and garlic) were evaluated. Total concentration and bioavailability of Cd, Cr, Cu and Pb in seafood after all these culinary treatments were compared with those in uncooked samples. Analytes were determined by triple-quadrupole inductively coupled plasma mass spectrometry (ICP-MS/MS). An alternative to express the results avoiding underestimated or overestimated values was proposed. RESULTS: The analytes concentration in seafood without culinary treatment varied from 0.0030 µg g-1 (shrimp) to 0.338 µg g-1 (oyster) for Cd; 0.010 µg g-1 (squid) to 0.036 µg g-1 (oyster) for Cr; 0.088 µg g-1 (scallop) to 8.63 µg g-1 (oyster) for Cu, and < 0.005 µg g-1 (shrimp, squid and oyster) to 0.020 µg g-1 (shark) for Pb. Only Cd (in scallop) was influenced by culinary treatments (reduction from 37 to 53 % after boiling, frying, and sautéing). Bioavailability percentage varied from 11% (oyster) for Cd; 18% (oyster) to 41% (shark) for Cr; 6% (shark) for Cu, and 8% (oyster) for Pb. Bioavailability percentage was not influenced by culinary treatments. CONCLUSION: Cadmium concentration was reduced in scallop after some culinary treatments (reduction o 37-53% after boiling, frying, and sautéing), but bioavailability percentage was not influenced. The employed analytical method was adequate for the purpose, presenting import results for food safety assessment about the influence of culinary treatments on metals concentration and bioavailability in seafood.

Effects of three biochars on copper immobilization and soil microbial communities in a metal-contaminated soil using a metallophyte and two agricultural plants.

Biochar (BC) is a porous, carbonaceous material produced by slow pyrolysis of biomass under oxygen-limited conditions. BC production has been attracting research interest because it modifies soil physicochemical characteristics and improves the growth of plants in problem soils. These benefits may be best actualized for soils contaminated by metals, where remediation is hampered by metal toxicity to both plants and soil microbial communities. The objectives of this study were to evaluate the impact of the addition of chicken manure biochar (CMB), oat hull biochar (OHB), or pine bark biochar (PBB) on copper (Cu) bioavailability in a Cu-contaminated soil, the effectiveness of these BCs promoting plant growth, and its effects on soil microbial communities supporting these plants. A sandy soil (338 mg Cu kg-1) was amended with CMB, OHB, and PBB, and the metallophyte Oenothera picensis or the agricultural species Solanum lycopersicum and Lolium perenne were grown for 3 months. The BCs produced an increase in soil pH, reduced the exchangeable Cu, and increased Cu bound to organic matter and residual fractions. All BCs enhanced the quality of contaminated soil and increased the plant biomass production, notably for S. lycopersicum, which grew until 12 times more than plants in non-amended soil. While BC addition reduced the concentration of Cu in soil pore water, the amendment did not reduce the concentrations of Cu in shoot tissues. BC additions also stimulated soil microorganisms, increasing basal respiration and DHA activity and modifying microbial communities, especially in soils supporting L. perenne. These results indicate that BCs represent an effective tool to remediate Cu-contaminated sandy soils.

Assessing the toxicity and accumulation of bulk- and nano-CuO in Hordeum sativum L.

The effects of bulk- and nano-CuO were monitored on barley (Hordeum sativum L.) in hydroponic conditions. The anatomical and cyto-/morphometric parameters of plants, exposed to both types of CuO in different doses (300 and 2000 mg/L) were recorded. The germination rate, root and shoot lengths decreased in a dose-dependent manner. Exposure to nano-CuO significantly increased Cu content in the H. sativum roots; however, the translocation rates of dissolved Cu were low and showed less accumulation in above-ground tissues. The differences between nano- and bulk-CuO treated plants were sufficiently evident, but at lower concentrations, these differences were non-significant. The relative seed germination inhibition was noted up to 11% and 22% under the high dose of bulk- and nano-CuO, respectively; however, at low dose, it was non-significant. The relative root length was reduced 3.6 fold by bulk- and 1.5 fold by nano-CuO, and shoot lengths decreased 1.6 fold by bulk- and 1.4 fold by nano-CuO under the high dose after growth of 30 days. It indicated more morphological effects on H. sativum caused by bulk- than the nano-CuO. The cytomorphometric analysis indicated the average cortex cell, total cortex, and total central cylinder areas of root cells and the average areas of chlorenchyma leaf cells were increased as compared to control in both bulk- and nano-CuO treated plants. It showed destructive effects of nano- and bulk-CuO on cellular organizations of H. sativum anatomy. Thus, at the low dose, the minimal effects of nano-CuO were observed than the bulk. Therefore, the finding could be interest for the safe application of nano-CuO.

Influence of mycorrhiza and fly ash on the survival, growth and heavy metal accumulation in three Acacia species grown in Cu-Ni mine soil.

Acacia albida, Acacia luederitzii, and Acacia tortilis are dominant acacia species in Botswana and have the potential to rehabilitate the heavy metal degraded environment. To establish this claim, experiments to assess the influence of mycorrhizal inoculation and fly ash amendments on the survival, growth and heavy metal accumulation of these species in mine tailings were conducted. A two-factor (AM inoculation × fly ash) in CRD was done on each of the three Acacia species consisting of four treatments: control (no mycorrhizal, no fly ash coded as - AM/- FA), with mycorrhizal but no fly ash (+ AM/- FA), no mycorrhizal but with fly ash (- AM/+ FA), and with mycorrhizal and with fly ash (+ AM/+ FA). After 24 weeks, results showed that the survival and dry matter yield of all Acacia species were enhanced by 10% with fly ash amendments. However, mycorrhiza inoculation alone improved the survival of A. albida and A. luederitzii but reduced that of the A. tortilis in mine tailings. Fly ash amendments increased the pH of the mine tailings, reduced the availability of Cu, Ni, Pb, Mn and Zn and consequently reduced the concentration of these metals in shoots. On the other hand, it increased the availability of As in the mine tailings. In addition, mycorrhizal inoculation reduced the concentration of these metals in shoots regardless of fly ash amendments. Overall, combined mycorrhizal inoculation and fly ash amendment enhanced the establishment of A. luederitzii in heavy metal-contaminated soils by reducing the heavy metal availability and metal uptake, thus increasing the survival and dry matter yield of plants.

Nickel and copper accumulation strategies in Odontarrhena obovata growing on copper smelter-influenced and non-influenced serpentine soils: a comparative field study.

The present investigation is the first in situ comparative study for the identification of Ni and Cu accumulation strategies involved in Odontarrhena obovata (syn. Alyssum obovatum (C.A. Mey.) Turcz.) growing in Cu-rich smelter-influenced (CSI) and non-Cu-influenced (NCI) sites. The total and Na2EDTA (disodium ethylenediaminetetraacetic acid)-extractable metal concentration in soils and plant tissues (roots, stem, leaves and flowers) were determined for CSI and NCI sites. High concentrations of total Ni, Cr, Co and Mg in the soil suggest serpentine nature of both the sites. In spite of high total and extractable Cu concentrations in CSI soil, majority of its accumulation was restricted to O. obovata roots showing its excluder response. Since the translocation and bioconcentration factors of Ni > 1 and the foliar Ni concentration > 1000 µg g-1, it can be assumed that O. obovata has Ni hyperaccumulation potential for both the sites. No significant differences in chlorophyll content in O. obovata leaves were observed between studied sites, suggesting higher tolerance of this species under prolonged heavy metal stress. Furthermore, this species from CSI site demonstrated rather high viability under extreme technogenic conditions due to active formation of antioxidants such as ascorbate, free proline and protein thiols. The presence of Cu in higher concentration in serpentine soil does not exert detrimental effect on O. obovata and its Ni hyperaccumulation ability. Thus, O. obovata could act as a putative plant species for the remediation of Cu-rich/influenced serpentine soils without compromising its Ni content and vitality.

A comparative study on the effects of three different metals (Cu, Zn and Cd) at similar toxicity levels in common carp, Cyprinus carpio.

To improve our understanding of underlying toxic mechanisms, it is important to evaluate differences in effects that a variety of metals exert at concentrations representing the same toxic level to the organism. Therefore, the main goal of the present study was to compare the effects of waterborne copper (Cu(II)), zinc (Zn(II)) and cadmium (Cd (II)) on a freshwater fish, the common carp (Cyprinus carpio), at concentrations being 0%, 25%, 50% and 100% of the 96 h LC50 (the concentration which is lethal to 50% of the population in 96 h). All the exposures were performed for a period of 1 week at 20°C. Our results show a rapid increase in the amount of copper and cadmium accumulated in the gills, while zinc only started to increase by the end of the experiment. All three metal ions increased metallothionein gene expression in both gills and liver. However, clear adverse effects were mainly observed for the Cu exposed group. Cu caused a decrease in Na level in gill tissue; it altered the expression of genes involved in ionoregulation such as Na+ /K+ -ATPase and H+ -ATPase as well as the expression of oxidative stress-related genes, such as catalase, glutathione reductase and glutathione S-transferase. Zinc and cadmium exposure did not alter the ion levels in the gills. In addition, no obvious effect of oxidative stress was observed, except for a transient increase in glutathione reductase at the highest cadmium concentration.

Structural characterization and pharmacological assessment in vitro/in vivo of a new copper(II)-based derivative of enrofloxacin.

Enrofloxacin (EFX) was selected as the medicinal ligand to afford a new copper(ii)-based complex, EFX-Cu, which was structurally characterized by spectroscopic analyses including X-ray single crystal diffraction. It was also stable and could retain the coordination state in aqueous solution. The in vitro antibacterial activity of EFX-Cu against a panel of pathogenic bacteria was about the same as that of EFX, except that it was twice as active against E. coli. The in vivo test on mice gave a LD50 value of 8148 mg kg-1 for EFX-Cu, which was much lower than those for EFX (LD50, 5312 mg kg-1) and its clinically used sodium salt, EFX-Na (LD50, 1421 mg kg-1). In addition, no obvious lesions in the organs of the dead mice were found by histopathological examination. Pharmacokinetic studies on rats suggested similar pharmacokinetics between EFX-Cu and EFX. On the other hand, EFX-Cu showed higher acute toxicity than EFX-Na in zebrafish, which was inconsistent with that in mice. The ROS-related inflammation and anti-inflammatory assay of EFX-Cu, respectively, in normal cells and zebrafish could be ascribed to its ROS-related redox property. Unfortunately, the final in vivo therapeutic assay in the E. coli-infected mouse model indicated that the therapeutic effect of EFX-Cu, mainly in terms of mortality in mice, was found to be lower than that of EFX-Na at the same dosage (800 mg kg-1, continuous gavage), although the contradictory factors between toxicity and antibacterial activity could not be excluded in this trial.

Graphene Oxide/Copper Nanoderivatives-Modified Chitosan/Hyaluronic Acid Dressings for Facilitating Wound Healing in Infected Full-Thickness Skin Defects.

PURPOSE: Wound healing, especially of infected wounds, remains a clinical challenge in plastic surgery. This study aimed to manufacture a novel and multifunctional wound dressing by combining graphene oxide/copper nanocomposites (GO/Cu) with chitosan/hyaluronic acid, providing significant opportunities for the therapy of wound repair in wounds with a high risk of bacterial infection. METHODS: In this study, GO/Cu-decorated chitosan/hyaluronic acid dressings (C/H/GO/Cu) were prepared using sodium trimetaphosphate (STMP) crosslinking and the vacuum freeze-drying method, and chitosan/hyaluronic acid dressings (C/H) and GO-incorporated chitosan/hyaluronic acid dressings (C/H/GO) served as controls. The surface characterization, in vitro degradation under various pH values, antimicrobial potential, cytocompatibility and in vivo therapeutic efficacy in a bacteria-infected full-thickness skin defect model were systematically evaluated. RESULTS: Our experimental results indicated that the acidic environment facilitated the release of copper (CuNPs and Cu2+) from the dressings, and prepared C/H/GO/Cu dressings exhibited significant in vitro antimicrobial activities against the two tested bacterial strains (ATCC35984 and ATCC25923). All three dressings showed satisfactory cytocompatibility with mouse fibroblasts (NIH/3T3-L1). Moreover, remarkably accelerated wound healing was found in the C/H/GO/Cu group, with controlled inflammatory infiltration and improved angiogenesis in granulation tissues. In addition, no pathological damage was noted in the tissue structures of the tested organs (heart, lung, liver and kidney) in any of the four groups. CONCLUSION: Collectively, GO/Cu-incorporated chitosan/hyaluronic acid dressings suggested a synergistic antimicrobial efficacy and acceptable biocompatibility both in vitro and in vivo, as well as a significantly accelerated healing process of bacteria-infected wounds. Thus, the multifunctional C/H/GO/Cu composite is expected to be a potential alternative for wound dressings, especially for the management of intractable wounds caused by bacterial infection.

Reactor produced [64Cu]CuCl2 as a PET radiopharmaceutical for cancer imaging: from radiochemistry laboratory to nuclear medicine clinic.

OBJECTIVE: Copper-64 is a useful theranostic radioisotope that is attracting renewed interest from the nuclear medicine community in the recent times. This study aims to demonstrate the utility of research reactors to produce clinical-grade 64Cu via 63Cu(n,γ)64Cu reaction and use it in the form of [64Cu]CuCl2 as a radiopharmaceutical for PET imaging of cancer in human patients. METHODS: Copper-64 was produced by irradiation of natural CuO target in a medium flux research reactor. The irradiated target was radiochemically processed and detailed quality control analyses were carried out. Sub-acute toxicity studies were carried out with different doses of Cu in Wistar rats. The biological efficacy of the radiopharmaceutical was established in preclinical setting by biodistribution studies in melanoma tumor bearing mice. After getting regulatory approvals, [64Cu]CuCl2 formulation was clinically used for PET imaging of prostate cancer and glioblastoma patients. RESULTS: Large-scale (~ 30 GBq) production of 64Cu could be achieved in a typical batch and it was adequate for formulation of clinical doses for multiple patients. The radiopharmaceutical met all the purity requirements for administration in human subjects. Studies carried out in animal model showed that the toxicity due to "cold" Cu in clinical dose of [64Cu]CuCl2 for PET scans would be negligible. Clinical PET scans showed satisfactory uptake of the radiopharmaceutical in the primary cancer and its metastatic sites. CONCLUSIONS: To the best of our knowledge, this is the first study on use of reactor produced [64Cu]CuCl2 for PET imaging of cancer in human patients. It is envisaged that this route of production of 64Cu would aid towards affordable availability of this radioisotope for widespread clinical use in countries with limited cyclotron facilities.

Enzymatic and non-enzymatic detoxification in Lycosa terrestris and Pardosa birmanica exposed to single and binary mixture of copper and lead.

Organisms employ various enzymatic and non-enzymatic detoxification mechanisms to minimize the harmful effects of metal pollution in the terrestrial environment. We examined the effects of copper (Cu), lead (Pb) and their mixture (Cu + Pb) on glutathione (GSH), metallothionein (MTs), cytochrome P450 (CYP 450), carboxylesterase (CarbE), acetylcholinesterase (AchE) and glutathione S-transferase (GST) in Lycosa terrestris and Pardosa birmanica via two exposure routes, i.e., soil and food for 10, 20 and 40 days. The present results revealed that the accumulation of Cu and Pb in both spiders' species increase with exposure duration and depend on the route of exposure and type of metal. The activities of CarbE, GST, and MTs significantly increased with increasing metal body burden for all experimental treatments. The CYP 450 activity exhibited a significant time-dependent decrease with increasing Cu concentration in both species. The AchE activity was significantly inhibited on Pb exposure via soil and Cu + Pb exposure via both routes. The decrease in the level of GSH was measured on Cu + Pb exposure via both routes. Thus, all these enzymatic and non-enzymatic responses are sensitive to the metals tested and could serve as early warning indicators for assessing the effects of metal pollution in these species.

Influence of calcium and EDTA on copper ion bioavailability in copper nanoparticle toxicity tests improves understanding of nano-specific effects.

Metal-based nanoparticles (NPs) can release metal ions that are toxic to aquatic organisms; however, whether the toxicity is from metal ions rather than unique "nano-scale" effects of the NPs is unresolved. The present study aimed to compare the toxicity of Cu2+ and Cu-NPs in larval zebrafish (Danio rerio) to clarify whether toxic effects are attributable to release of Cu ions and to determine the effect of the chelating agent ethylenediaminetetraacetic acid (EDTA) and calcium hardness (as CaCO3) on the Cu toxicity. First, the acute toxicity (96-h lethality) of Cu-NPs was determined in comparison to aqueous Cu in larvae exposed to CuSO4, and subsequently, sublethal tests with Cu-NPs and CuSO4 were conducted with additions of EDTA or calcium ions to evaluate alterations in expression of metallothionein-2 (MT2) gene transcripts (quantitative real-time polymerase chain reaction). Acute toxicity of Cu in larvae exposed to CuSO4 was greater (LC50 = 226 µg Cu/L) than for larvae exposed to Cu-NPs (LC50 = 648 µg Cu/L). The expression of MT2 increased with Cu concentration (p < 0.05), and the slope of the linear regression was significantly greater in fish exposed to CuSO4 (slope = 0.090) compared to Cu-NPs (slope = 0.011). Cu2+ was 2.9-fold more toxic than Cu-NPs. The presence of 5 mg/L EDTA and 220 mg/L CaCO3 significantly reduced the expression of MT2 (1.8-fold for EDTA, 2.3-fold for CaCO3) in larvae exposed to CuSO4. For larvae exposed to Cu-NPs, the presence of EDTA reduced the expression of MT2 (1.7-fold) relative to Cu-NP concentration. While Cu-NPs induced MT2 expression, the differences in concentration relationships of MT2 expression between Cu-NPs and CuSO4 indicated that factors other than release of Cu ions from Cu-NPs influenced acute toxicity of Cu-NPs. The conclusion drawn from this ecotoxicological risk assessment was that EDTA and calcium significantly decreased Cu toxicity in freshwater fish.

Demonstrating the Reliability of bio-met for Determining Compliance with Environmental Quality Standards for Metals in Europe.

The importance of considering the bioavailability of metals in understanding and assessing their toxicity in freshwaters has been recognized for many years. Currently, biotic ligand models (BLMs) are being applied for the derivation and implementation of environmental quality standards (EQS) for metals under the Water Framework Directive in Europe. bio-met is a simplified tool that was developed for implementing bioavailability-based EQS for metals in European freshwaters. We demonstrate the reliability of the relationship between the full BLM predictions and the thresholds (hazardous concentration affecting 5% of species [HC5] values) predicted by bio-met in 3 stages, for the metals copper, nickel, and zinc. First, ecotoxicity data for specific species from laboratory tests in natural waters are compared with predictions by the individual species BLMs included in the full BLMs. Second, the site-specific HC5 values predicted by bio-met for the natural waters used for ecotoxicity testing are compared with those provided by the full BLMs. The reliability of both relationships is demonstrated for all 3 metals, with more than 80% of individual species BLM predictions being within a factor of 3 of the experimental results, and 99% of bio-met local HC5 predictions being within a factor of 2 of the full BLM result. Third, using a larger set of European natural waters in addition demonstrates the reliability of bio-met over a broad range of water chemistry conditions. bio-met is therefore an appropriate tool for performing compliance assessments against EQS values in Europe, due to the demonstrated consistency with the toxicity test data. Environ Toxicol Chem 2020;39:2361-2377. © 2020 SETAC.

Effects of the Ionic and Nanoparticle Forms of Cu and Ag on These Metals' Bioaccumulation in the Eggs and Fry of Rainbow Trout (Oncorhynchus mykiss W.).

Nanotechnology is a rapidly growing field of science, and an increasing number of nanoproducts, including nanometals, can be found on the market. Various nanometals and the products that are manufactured based on them can help to fight bacteria and fungi, but they can also penetrate organisms and accumulate in them. This study aimed to compare the effects of two metals, silver (Ag) and copper (Cu), with known antibacterial and fungicidal properties in their ionic (AgNO3, CuSO4·5H2O) and nanoparticle (AgNPs, CuNPs) forms on rainbow trout eggs and fry. Concentrations of metals ranging from 0 to 16 mg/L were used during egg swelling for 2 h. The swelling of eggs in Cu solutions resulted in an increase in Cu content in the eggs (just like in the case of Ag); however, the changes in fry were not significant in the case of both Ag and Cu. The concentrations of these metals in eggs was greatly affected by the applied form of Ag and Cu. Because CuNPs penetrated the embryo in fish eggs at lower concentrations compared to AgNPs, it would be worth considering them for antibacterial applications during egg incubation.