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The high prevalence of breast cancer is a global health concern, but there are no safe or effective treatments for it at its advanced stages. These facts urge the development of novel treatment strategies. Annexin A5 (ANXA5) is a natural human protein that binds with high specificity to phosphatidylserine, a phospholipid tightly maintained in the inner leaflet of the cell membrane on most healthy cells but externalized in tumor cells and the tumor vasculature. Here, we have developed a targeted photosensitizer for photothermal therapy (PTT) of solid tumors through the functionalization of single walled carbon nanotubes (SWCNTs) to ANXA5—the SWCNT-ANXA5 conjugate. The ablation of tumors through the SWCNT-ANXA5-mediated PTT synergizes with checkpoint inhibition, creating a systemic anti-cancer immune response. In vitro ablation of cells incubated with the conjugate promoted cell death in a dose-dependent and targeted manner. This treatment strategy was tested in vivo with the orthotopic EMT6 breast tumor model in female balb/cJ mice. Enhanced therapeutic effects were achieved by using intratumoral injection of the conjugate and treating tumors at a lower PTT temperature (45oC). When combined with checkpoint inhibition of anti-PD-1, SWCNT-ANXA5-mediated PTT increased survival and 80% of the mice survived for 100 days. Evidence of immune system activation by flow cytometry of splenic cells strengthens the hypothesis of an abscopal effect as a mechanism of prolonged survival. more...

This file contains includes 9 figures of supplementary data (S1-S9) and a description of live cell microscopy and the assessment of synergism for the combination therapies.

Mutant cystathionine gamma-lyase was targeted to phosphatidylserine exposed on tumor vasculature through fusion with Annexin A1 or Annexin A5. Cystathionine gamma-lyase E58N, R118L, and E338N mutations impart nonnative methionine gamma-lyase activity, resulting in tumor-localized generation of highly toxic methylselenol upon systemic administration of nontoxic selenomethionine. The described therapeutic system circumvents systemic toxicity issues using a novel drug delivery/generation approach and avoids the administration of nonnative proteins and/or DNA required with other enzyme prodrug systems. The enzyme fusion exhibits strong and stable in vitro binding with dissociation constants in the nanomolar range for both human and mouse breast cancer cells and in a cell model of tumor vascular endothelium. Daily administration of the therapy suppressed growth of highly aggressive triple-negative murine 4T1 mammary tumors in immunocompetent BALB/cJ mice and MDA-MB-231 tumors in SCID mice. Treatment did not result in the occurrence of negative side effects or the elicitation of neutralizing antibodies. On the basis of the vasculature-targeted nature of the therapy, combinations with rapamycin and cyclophosphamide were evaluated. Rapamycin, an mTOR inhibitor, reduces the prosurvival signaling of cells in a hypoxic environment potentially exacerbated by a vasculature-targeted therapy. IHC revealed, unsurprisingly, a significant hypoxic response (increase in hypoxia-inducible factor 1 α subunit, HIF1A) in the enzyme prodrug–treated tumors and a dramatic reduction of HIF1A upon rapamycin treatment. Cyclophosphamide, an immunomodulator at low doses, was combined with the enzyme prodrug therapy and rapamycin; this combination synergistically reduced tumor volumes, inhibited metastatic progression, and enhanced survival. Mol Cancer Ther; 16(9); 1855–65. ©2017 AACR. more...

Copper(II) is known to bind in the influenza virus His37 cluster in the homotetrameric M2 proton channel and block the proton current needed for uncoating. Copper complexes based on iminodiacetate also block the M2 proton channel and show reduced cytotoxicity and zebrafish-embryo toxicity. In voltage-clamp oocyte studies using the ubiquitous amantadine-insensitive M2 S31N variant, the current block showed fast and slow phases, in contrast to the single phase found for amantadine block of wild-type M2. Here, we evaluate the mechanism of block by copper adamantyl iminodiacitate and copper cyclooctyl iminodiacitate complexes and address whether the complexes can coordinate with one or more of the His37 imidazoles. The current traces were fitted to parametrized master equations. The energetics of binding and the rate constants suggest that the first step is copper complex binding within the channel, and the slow step in the current block is the formation of a Cu-histidine coordination complex. Solution-phase isothermal titration calorimetry and density functional theory (DFT) calculations indicate that imidazole binds to the copper complexes. Structural optimization using DFT reveals that the complexes fit inside the channel and project the Cu(II) toward the His37 cluster, allowing one imidazole to form a coordination complex with Cu(II). Electrophysiology and DFT studies also show that the complexes block the G34E amantadine-resistant mutant despite some crowding in the binding site by the glutamates. more...

Due to their non-toxicity and their ability to be functionalized, magnetite (Fe3O4) nanoparticles (NPs) are good candidates for a variety of biomedical applications. To better implement their applications, it is crucial to well understand the basic structural and magnetic properties of the NPs in correlation with their synthesis method. Here, we show interesting properties of Fe3O4 NPs of various sizes ranging from 5 to 100 nm and the dependence of these properties on particle size and preparation method. One synthetic method based on heating Fe(acac)3 with oleic acid consistently gives 5 ± 1 nm NPs. A second method using the thermal decomposition of Fe(oleate)3 in oleic acid led to larger NPs, greater than 8 nm in size. Increasing the amount of oleic acid caused the average NP size to slightly increase from 8 to 10 nm. Increasing both the reaction temperature and the reaction time caused the NP size to drastically increase from 10 to 100 nm. Powder X-ray diffraction and electron-microscopy imaging show a pure single crystalline Fe3O4 phase for all NPs smaller than 50 nm and spherical in shape. When the NPs get larger than 50 nm, they notably tend to form faceted, FeO core–Fe3O4 shell structures. Magnetometry data collected in various field-cooling conditions show a pure superparamagnetic (SPM) behavior for all NPs smaller than 20 nm. The observed blocking temperature, $T_{B}$ , gradually increases with NP size from about 25–150 K. In addition, the Verwey transition is observed with the emergence of a strong narrow peak at 125 K in the magnetization curves when larger NPs are present. Our data confirm the vanishing of the Verwey transition in smaller NPs. Magnetization loops indicate that the saturating field drastically decreases with NP size. While larger NPs show some coercivity ( $H_{c}$ ) up to 30 mT at 400 K, NPs smaller than 20 nm show no coercivity ( $H_{c} = 0$ ), confirming their pure SPM behavior at high temperature. Upon cooling below $T_{B}$ , some of the SPM NPs gradually show some coercivity, with $H_{c}$ reaching 45 mT at 5 K for the 10 nm NPs, indicating emergent interparticle couplings in the blocked state. more...

Resorcinarene-based deep cavitands including glutamic acid methyl resorcinarene (GMA) and glutamic acid undecyl resorcinarene (GUA) have been synthesised and their amine binding properties analysed. The cavitands are designed with hydrogen bonding groups on their upper rim (glutamic acids) and hydrophobic groups, on their lower rims. Along with the aromatic groups of the resorcinarene, these cavitands have many sites for guest binding. The cavitands form kite-like structures in dimethylsulfoxide, acetone, and water. The binding of GMA with amines, including guanidine containing drug compounds, was studied in DMSO by UV-vis titration and compared to that of phthalyl glutamic acid. The obtained binding constants for GMA to amines (K values) were in the range of 12–136 × 103 M−1 for benzyl amines and 4.3–153 × 103 M−1 for guanidine compounds. GMA shows a binding preference for more basic and less sterically encumbered amines and slightly stronger binding to S-enantiomers. more...

Identifying a universal biomarker for cancer treatment remains a major challenge in cancer therapy. Extracellular exposure of phosphatidylserine (PS) is tightly regulated and is an "eat me" signal for phagocytosis in healthy cells. Although cancer cells and vasculature express high levels of externalized PS, they do not undergo apoptosis, making them a promising biomarker for cancer treatment. Annexin A5 (ANXA5) is the native binding partner of PS and can actively target and deliver chemotherapies to the tumor microenvironment (TME) via PS expression. ANXA5 acts as a bridge between the innate and adaptive immune systems and contributes to an immunostimulatory profile in the TME. ANXA5-enzyme prodrug therapies allow for systemic delivery of prodrugs and targeted killing at the tumor site. ANXA5-carbon nanotube conjugates have been used to physically ablate tumors via photothermal therapy. This review aims to explore the expression of PS in cancer cells and how ANXA5 has been used as a chemotherapeutic and targeting agent for cancer. more...

The magnetic properties of ${\mathrm{Fe}}_{3}{\mathrm{O}}_{4}$ nanoparticle assemblies have been investigated in detail through a combination of vibrating sample magnetometry (VSM) and muon spin relaxation ($\ensuremath{\mu}\mathrm{S}\mathrm{R}$) techniques. Two samples with average particle sizes of 5 and 20 nm, respectively, were studied. For both samples, the VSM and $\ensuremath{\mu}\mathrm{S}\mathrm{R}$ results exhibit clear signatures of superparamagnetism at high temperature and magnetic blocking at low temperature. The $\ensuremath{\mu}\mathrm{S}\mathrm{R}$ data demonstrate that the transition from the superparamagnetic to the blocked state occurs gradually throughout the sample volume over an extended temperature range due to the finite particle size distribution of each nanoparticle batch. The transition occurs between approximately 3 and 45 K for the 5-nm nanoparticles and 150 and 300 K for the 20-nm nanoparticles. The VSM and $\ensuremath{\mu}\mathrm{S}\mathrm{R}$ data are further analyzed to yield estimates of microscopic magnetic parameters including the nanoparticle spin-flip activation energy ${E}_{A}$, magnetic anisotropy $K$, and intrinsic nanoparticle spin reversal attempt time ${\ensuremath{\tau}}_{0}$. These results highlight the complementary information about magnetic nanoparticles that can be obtained by bulk magnetic probes such as magnetometry and local magnetic probes such as $\ensuremath{\mu}\mathrm{S}\mathrm{R}$. more...

The greatest contributors to cancer mortality are metastasis and the consequences of its treatment. Here, we present a novel treatment of metastatic breast cancer that combines photothermal therapy with targeted single-walled carbon nanotubes (SWCNTs) and immunostimulation with a checkpoint inhibitor. We find that the selective near-infrared photothermal ablation of primary orthotopic EMT6 breast tumors in syngeneic BALB/cJ mice using an annexin A5 (ANXA5) functionalized SWCNT bioconjugate synergistically enhances an anti-cytotoxic T-lymphocyte-associated protein 4 (anti-CTLA-4)-dependent abscopal response, resulting in an increased survival (55%) at 100 days after tumor inoculation. In comparison, there was no survival at 100 days for either photothermal therapy by itself or immunostimulation by itself. Prior to photothermal therapy, the SWCNT-ANXA5 bioconjugate was administered systemically at a relatively low dose of 1.2 mg/kg, where it then accumulated in tumor vasculature via ANXA5-dependent binding. During photothermal therapy, the average maximum temperature in the tumor reached 54 °C (duration 175 s). The mechanism of prolonged survival resulting from combinatorial photothermal ablation and immune stimulation was evaluated by flow cytometric quantification of splenic antitumoral immune effector cells and serum cytokine quantification. more...

Copper(II) is known to bind in the influenza virus His37 cluster in the homotetrameric M2 proton channel and block the proton current needed for uncoating. Copper complexes based on iminodiacetate also block the M2 proton channel and show reduced cytotoxicity and zebrafish-embryo toxicity. In voltage-clamp oocyte studies using the ubiquitous amantadine-insensitive M2 S31N variant, the current block showed fast and slow phases in contrast to the single phase found for amantadine block of WT M2. Here we evaluate the mechanism of block by copper adamantyl iminodiacitate (Cu(AMT-IDA)) and copper cyclooctyl iminodiacitate (Cu(CO-IDA)) complexes and address whether the complexes can covalently bind to one or more of the His37 imidazoles. The current traces were fitted to parametrized master equations. The energetics of binding and the rate constants suggest that the first step is copper-complex binding within the channel and the slow step in the current block is the covalent bond formation between copper complex and histidine. Isothermal titration calorimetry (ITC) indicates that a single imidazole binds strongly to the copper complexes. Structural optimization using density functional theory (DFT) reveals that the complexes fit inside the channel and project the Cu(II) towards the His37 cluster allowing one imidazole to form a covalent bond with the Cu(II). Electrophysiology and DFT studies also show that the complexes block the G34E amantadine-resistant mutant in spite of some crowding in the binding site by the glutamates. more...

Technetium poses an environmental hazard because of its radioactivity and long half-life. It exists in the form of pertechnetate in the environment and can be modeled by the nonradioactive ion perrhenate, since pertechnetate and perrhenate have the same geometry and similar chemical properties. In this research, a new zinc cyclen resorcinarene cavitand (ZCR) column was used in ion chromatography (IC) to efficiently separate perrhenate. Ion chromatography has the advantage of requiring almost no sample preparation for water samples. The ZCR column demonstrated the ability to separate anions: fluoride, chloride, nitrate, sulfate, phosphate, perchlorate, and perrhenate by gradient 2–60 mM NaOH. Unlike other columns, the new column material was selective in retaining perrhenate. The ZCR column also gave a linear range from 2.0 to 1000 mg L−1 for perrhenate with R2 > 0.997. There was a logarithmic relationship between the concentration of perrhenate and its retention time. Excellent perrhenate recovery was achieved on the ZCR column when river water was spiked with perrhenate and perrhenate was preconcentrated. The efficient separations of perrhenate by the ZCR column will potentially assist in pertechnetate separations. more...

This study validates bacterial anionic phospholipids (APs) as a putative molecular target in a novel antibiotic treatment against the Gram-positive bacterium Listeria monocytogenes and the Gram-negative bacterium Escherichia coli. Bacterial AP expression was targeted with its associated protein-ligand partner, annexin A5 (ANXA5). This protein was functionalised with the covalent addition of the antibiotic ampicillin (AMP) and separately with the antibiotic moxifloxacin (MOX). Functionalised ANXA5 serves as a delivery vehicle, directing the antibiotic to bacterial AP expression. The results presented here suggest that this ANXA5-AMP bioconjugate participates in a positive feedback loop where APs, the target of the delivery vehicle ANXA5, are upregulated by the chemotherapeutic payload of the bioconjugate. Importantly, the ANXA5 delivery vehicle is non-toxic to bacterial cells by itself and neither is the ANXA5–antibiotic bioconjugate toxic to human vascular endothelial cells. As measured by the IC50, conjugation to ANXA5 resulted in increasing the antibiotic activity of AMP against L. monocytogenes and E. coli by more than 4 and 3 orders of magnitude, respectively, compared with free AMP, whilst the activity of MOX against L. monocytogenes is increased by 4 orders of magnitude. Given the conservation of AP expression in pathologies such as oncogenesis and other bacterial/viral/parasitic infections, we hypothesise that a therapeutic modality targeting AP expression may be a viable chemotherapeutic strategy in many infectious diseases. more...

A new sensor for Hg2+ and cysteine detection has been synthesized and tested. The sensor based on cinamaldehyde and pyrimidine absorbs visible light when exposed to Hg2+ and becomes colored. The sensor only changes color due to Hg2+ and does not change when exposed to some other metal ions. Calculations show the change in color is due to lowering of the HOMO-LUMO transition. Titrations show the sensor-Hg2+ complex is composed of two sensor molecules per Hg2+. The limit of detection for Hg2+ is 0.39 μM. The sensor-Hg2+ complex selectively responds to cysteine (Cys), but does not show a color change due to other amino acids. The limit of detection of Cys by the sensor-Hg2+ complex is 0.10 μM. more...

Hypochlorite is used as a water disinfectant and it is also produced by biological organisms. Its detection and quantification is important and could lead to its mechanism of reactivity in cells. We have synthesized a new fluorescence sensor for hypochlorite based on bithiophene and furan-carbohydrazide. The sensor shows increased fluorescence as a function of hypochlorite and is selective for hypochlorite. Fluorescence enhancement due to hypochlorite is observed when the sensor is used in aqueous solutions at neutral pH values. Using the sensor, the detection limit for hypochlorite is 4.2 µM, making the sensor practical to determine hypochlorite in water. Applying the sensor to aide in the detection of hypochlorite in zebrafish, showed localization of ClO−/HClO in the air bladders and eyes of zebrafish. more...

New M2 blockers effective against the ubiquitous amantadine-resistant S31N M2 mutation in influenza A are needed. Six copper complexes, 2, 4, 6, 8, 9, and 10, were synthesized and found to block both wild type and S31N M2. Free Cu2+ also blocks M2 S31N but not S31N/H37A. The copper complexes do not block M2 H37A (either S31 or S31N). The complexes were effective against three influenza A strains in cell-culture assays, but less toxic to cells than CuCl2. For example 4, Cu(cyclooctylamineiminodiacetate), which was stable at pH > 4 in the buffers used, had an EC50 against A/Calif/07/2009 H1N1 of 0.7 ± 0.1 μM with a CC50 of 147 μM (therapeutic index, averaged over three strains, 67.8). In contrast, CuCl2 had an EC50 of 3.8 ± 0.9 μM and CC50 of 19 μM. Because M2 H37 is highly conserved, these complexes show promise for further testing as drugs against all strains of influenza A. more...

Formation of crystalline hexagonal ZnO prisms from a sol–gel method is presented. The method requires zinc acetate, water, and diethanolamine to create a zinc hydroxide/zinc hydroxide acetate gel, which in the presence of water and upon heating forms hexagonal prisms. Ethanolamines were found to be critical for gel and prism formation, while other bases such NaOH and triethylamine produce nanoparticles. Scanning electron microscope images showed hexagonal prisms with uniform size (approx. 0.5 × 2 µm) and very distinct edges. Transmission electron microscope and electron diffraction images showed that the prisms were highly crystalline with rough surfaces. Transmission electron microscope also showed a change from randomly oriented particles in the gel to ordered crystals after heating. Using an X-ray diffraction high-temperature chamber to characterize the gel, it was determined that it formed ZnO at 100 °C. Along with heating, acetate and water were found to be necessary for prism formation. In a potential application, the prisms efficiently acted as a photocatalyst in the degradation of organic dye. more...

Since their discovery in the 1990s, carbon nanotubes have been intensively studied in several different fields, including recently for the treatment of cancer. A property of carbon nanotubes that has been used extensively in cancer treatment studies is their strong absorbance of light in the near-infrared range (700–1400 nm). This strong absorbance of light leads to heating of the nanotubes and destruction of cancer cells that have taken up or are in the vicinity of nanotubes. Photothermal therapy studies for cancer treatment both in vitro and in vivo will be discussed, including the functionalization of the nanotubes for targeting and also the in vivo coadministration of other agents such as antibodies to stimulate the immune system and thereby increase the therapeutic response. For the work in vivo, research will be discussed about the biodistribution of carbon nanotubes after being injected and their elimination and/or degradation, including toxicology evaluations. more...

"Organization and Strategy, Roger G. Harrison Analytical Considerations in the Development of Protein Purification Processes, Vincent Anicetti and William S. Hancock Cell Breakage, Cady R. Engler Crossflow Membrane Filtration, Zhi-Guo Su and Clark K. Cotton Liquid-Liquid Extraction, Dennis J. Kubek Differential Precipitation of Proteins: Science and Technology, Fred Rothstein Conventional Chromatography, John M. Simpson Biospecific Affinity Chromatography, Nikos K. Harakas Freeze Drying: A Practical Overview, Larry A. Gatlin and Steven L. Nail " more...

Following our call to join in the discussion over the suitability of implementing a reporting checklist for bio–nano papers, the community responds.

Objectives The bleak prognosis associated with pancreatic cancer (PDAC) drives the need for the development of novel treatment methodologies. Here, we evaluate the applicability of 3 enzyme prodrug therapies for PDAC, which are simultaneously targeted to the tumor, tumor vasculature, and metastases via annexin V. In these therapies, annexin V is fused to an enzyme, creating a fusion protein that converts nontoxic drug precursors, prodrugs, into anticancer compounds while bound to the tumor, therefore mitigating the risk of side effects. Methods The binding strength of fusion proteins to the human PDAC cell lines Panc-1 and Capan-1 was measured via streptavidin-horseradish peroxidase binding to biotinylated fusion proteins. Cytotoxic efficacy was evaluated by treatment with saturating concentrations of fusion protein followed by varying concentrations of the corresponding prodrug plus docetaxel. Results All fusion proteins exhibited strong binding to PDAC cells, with dissociation constants between 0.02 and 1.15 nM. Cytotoxic efficacy was determined to be very good for 2 of the systems, both of which achieved complete cell death on at least 1 cell line at physiologically attainable prodrug concentrations. Conclusions Strong binding of fusion proteins to PDAC cells and effective cytotoxicity demonstrate the potential applicability of enzyme prodrug therapy to the treatment of PDAC. more...

Monitoring Zn2+ levels in biological environments with fluorescent sensors is important. This paper gives the synthesis and properties of a new Zn2+ sensor based on quinoline and pyridylaminophenol. The sensor is selective for Zn2+ and remains fluorescent when bound to Zn2 even in the presence of other metal ions. Along with fluorescing when bound to Zn2+, the sensor becomes colored when Cu2+ or Co2+ is added to it. These two metal ions result in the sensor becoming yellow. The crystal structure of the Cu–sensor complex shows that all of the sensor's nitrogens are bound to the metal ion. In studies with living cells, the fluorescence intensity of the sensor correlates to the concentration of Zn2+. more...

The extent of reduction of unsupported iron-based high-temperature water-gas shift catalysts with small (

The synthesis, Zn 2+ binding, crystal structure, and cell imaging studies of a new pyrazole amine quinoline receptor with a flexible binding pocket are described. Upon coordination to Zn 2+ , the absorption of the receptor increases at 364 nm and it fluoresces at 500 nm. The fluorescence response to Zn 2+ is selective for Zn 2+ and does not occur with other metal ions, not even Cd 2+ . In solution, the receptor forms 1:1 complexes with Zn 2+ , but in the solid-state two Zn 2+ ions coordinate to the receptor. The aqueous solubility of the receptor allows for imaging of Zn 2+ in living cells. Cells exposed to receptor and Zn 2+ fluoresce when excited with visible light. more...

Molecular conjugates are important to link light sensitized materials to electron acceptors. We have synthesized oligothiophenes and oligothiophene–ruthenium complexes and bound them to CdSe nanoparticles. The absorption and fluorescence properties of the oligothiophenes bound to CdSe were measured. Steady-state luminescence and time correlated single photon counting were used to observe the effects on fluorescence and fluorescence lifetimes before and after binding. It was found that fluorescence of CdSe nanoparticles was quenched when they were bound to the oligothiophenes, and that the fluorescence of the oligothiophenes was also quenched. The fluorescence lifetimes of the quenched species were shortened and suggest electron transfer from oligothiophene to nanoparticle is on the order of one nanosecond. Orbital energy calculations predict that the Ru bound oligothiophenes have HOMO–LUMO energies of correct energy to allow electron and hole transfer. These experiments show that the oligothiophenes efficiently transfer optical energy between CdSe nanoparticles and could potentially be used as charge transfer junctions. more...

People with chronic kidney disease suffer from uremic toxins which accumulate in their bodies. Detection and quantification of uremic toxins help diagnose kidney problems and start patient care. The aim of this research was to seek a new method to assist this diagnosis by trace level detection and separation of guanidine containing uremic toxins in water and urine. To detect and quantify the uremic toxins, new stationary phases for ion chromatography (IC) columns based on glutamic acid functionalized resorcinarenes bound to divinylbenzene macroporous resin were prepared. The new column packing material afforded separation of the five compounds: guanidinoacetic acid, guanidine, methylguanidine, creatinine, and guanidinobenzoic acid in 30min. Peak resolutions ranged from 7.6 to 1.3. Gradient elutions at ambient temperature with methanesulfonic acid (MSA) solution as eluent resulted in detection levels in water from 10 to 47ppb and in synthetic urine from 28 to 180ppb. Limits of quantification for the analytes using pulsed amperometric detection were 30-160ppb in water and 93-590ppb in urine. Trace levels of creatinine (1ppm) were detected in the urine of a healthy individual using the columns. more...

Objectives Use of electronic cigarettes has dramatically increased in the United States since 2010, with a forecasted growth of 37% between 2014 and 2019. There is little research on e-liquid nicotine concentration from domestic manufacturers. However, limited research outside of the United States found wide inconsistencies between the labeled concentration of nicotine in e-liquids and the actual nicotine concentration. Methods The 7 most popular online manufacturers or distributors were identified. E-liquid samples of the 5 most popular flavors from each manufacturer were purchased in nicotine concentrations of 0 and 18 mg/mL. Of the samples purchased (n = 70), all were labeled as produced in the United States of America. The researchers anonymized the samples before sending them to an independent university laboratory for testing. Results The 35 e-liquid samples labeled 18 mg/mL nicotine measured between 11.6 and 27.4 mg/mL (M = 18.7, SD = 3.3) nicotine. The labeled 18 mg/mL samples measured as little as 35% less nicotine and as much as 52% greater nicotine. In the 35 samples labeled 0 mg/mL, nicotine was detected (>0.01 mg/mL) in 91.4% of the samples (range 0-23.9 mg/mL; M = 2.9, SD = 7.2). Six samples from 2 manufacturers labeled as 0 mg/mL were found to contain nicotine in amounts ranging from 5.7 to 23.9 mg/mL. Conclusion This study demonstrates the nicotine labeling inaccuracies present in current e-liquid solutions produced in the United States. Incorrect labeling poses a significant risk to consumers and supports the recent regulation changes enacted by the US Food and Drug Administration. Additional routine testing of nicotine concentrations should be conducted to evaluate the effectiveness of the regulations on future e-liquid production. more...

Approximately 75% of ovarian cancer is diagnosed once metastasis to the peritoneal cavity has occurred. A large proportion of patients eventually develop platinum-resistive tumors, which are considered terminal. In order to provide an alternative a novel fusion protein, mCTH-ANXA5, has been developed for the treatment of recurrent, metastatic ovarian cancer. The fusion protein combines annexin V (ANXA5), an ovarian tumor and tumor vasculature targeting protein, with mutated cystathionine gamma-lyase (mCTH), an enzyme that converts selenomethionine (SeMet) into toxic methylselenol, which generates reactive oxygen species and eventual tumor cell death. In order to further enhance the therapeutic efficacy, anti-CD73 and anti-OX40 immunostimulants were combined with mCTH-ANXA5, resulting in an increase of survival by 100% from 12 to 24 days post-therapy and decrease tumor burden in mice with orthotopic metastatic ovarian cancer. Further evaluation of the combination therapy revealed a strong antibody-mediated immune response, and an increased infiltration of cytotoxic T-cells along with a decrease in tumor promoting immune cells. This study demonstrates the efficacy of a synergistic, multi-drug system by attacking the tumor as well as enlisting the body's own defense system to treat the patient. more...

A series of novel chloro-complexes of Mn2+, Co2+, Ni2+ and Fe3+ with 2,4,6-tris-(2-pyridyl)-1,3,5-triazine (L1) and thiourea (L2) with the formulae, [Mn(L1)Cl2(EtOH)]L2 (1), [Co(L1)Cl2(H2O)]L2.1/2H2O (2), [Ni(L1)Cl2(H2O)]L2.1/2H2O (3) and [Fe(L1)Cl3]L2 (4) were investigated utilizing convenient methods. Conversely, similar reaction resulted in complex with structure [Mn(L1)(OAc)(H2O)Cl]·H2O (5) in presence of sodium acetate. Pentagonal-bipyramidal geometry around Mn2+ ion is suggested. The solid state ESR spectra of Mn2+, Co2+ and Ni2+ complexes parameters show g|| > g┴ > 2.0023 indicating that the ground state is dx2-y2. Electronic spectra of two Mn2+ and the other complexes and the values of magnetic moments suggest octahedral geometry for complexes while pentagonal-pyramidal for [Mn(L1)Cl2(EtOH)]L2. Biological activity of complexes was tested versus alpathogenic bacteria (E. Coli, S. Aureus and C. Albicans). Further, the cytotoxic activity was screened for in vitro against (HeLa), (MCF-7), (HePG2) and (HeP-2). The role of ionic radii of the metal ion on the biological activity was investigated. more...

Every discussion of space power in the United States these days begins with the so-called “three C's,” an abbreviation for congested, competitive, and contested space. Reducing complex strategic pr... more...

The applications of resorcinarene derivatives in modern separation techniques including high performance liquid chromatography (HPLC), gas chromatography (GC), capillary electrophoresis (CE), ion chromatrography (IC), and liquid membranes are reviewed in this paper. Resorcinarenes are macrocyclic compounds which can be modified with various substituents on upper and lower rims to provide specific functionality and selectivity. The derivatives can be adsorbed or covalently bound to the stationary phase of HPLC, GC, and IC, or used as pseudo-stationary phase in CE to separate organic or inorganic species. They have been tested as carriers to selectively transfer species from source phase to receiving phase in bulk liquid membranes, supported liquid membranes, and polymer inclusion membranes. These broad applications of resorcinarene-based macrocyclic ligands indicate that they are a promising class of compounds that are worthwhile to be explored in both synthesis and separation applications. more...

Cu-doped ZnO nanoparticles were synthesized by a simple chemical method at low temperature with Cu:Zn atomic ratio from 0 to 5 %. The synthesis process was based on the hydrolysis of zinc acetate dehydrate and copper acetate tetrahydrate heated under reflux to 65 °C using methanol as a solvent. X-ray diffraction (XRD) analysis reveals that the Cu-doped ZnO crystallize in a wurtzite structure with a change of crystal size from 12 nm for undoped ZnO to 5 nm for Cu-doped ZnO. These nano size crystallites of Cu doped ZnO self-organized into microspheres. The XRD patterns, Scanning electron microscopy and transmission electron microscopy micrographs of doping of Cu in ZnO confirmed the formation of microspheres and indicated that the Cu2+ is successfully substituted into the ZnO host structure of the Zn2+ site. Cu doping shifts the absorption onset to blue from 373 to 350 nm, indicating an increase in the band gap from 3.33 to 3.55 eV. A relative increase in the intensity of the deep trap emission of Cu-doped ZnO is observed when increasing the concentration of Cu. Magnetic measurements indicate that Cu-doped ZnO samples are ferromagnetic at room temperature except pure ZnO. more...

Zinc sensor molecules containing quinoline have been synthesized, which show fluorescence in the presence of Zn2+. The nitrogen in quinoline is critical to fluorescence and fluorescence enhancement is promoted by deprotonating the sensor’s amide. One of the sensors is highly selective for Zn2+ over Cd2+ and other cations such as Hg2+, Fe2+, Mn2+ and Ca2+. This selectivity can be attributed to the increased absorption of the sensor in the presence of Zn2+ and the strong binding of Zn2+. Structural studies, including X-ray and NMR, show the ability of dipicolylamine (DPA) to bind in facial and meridional manners to Zn2+. Crystal structures of different compounds show Zn2+ coordinating to three, four, and five nitrogens from the compounds. They also indicate that the selectivity of DPA containing compounds towards Zn2+ may originate from Zn2+ being stable in different coordination environments. more...

A zinc sensor based on quinoline and morpholine has been synthesized. The sensor selectively fluoresces in the presence of Zn2+, while not for other metal ions. Absorbance changes in the 350nm region are observed when Zn2+ binds, which binds in a 1:1 ratio. The sensor fluoresces due to Zn2+ above pH values of 6.0 and in the biological important region. The Zn2+-sensor complex has the unique ability to detect both Hg2+ and HS-. The fluorescence of the Zn2+-sensor complex is quenched when it is exposed to aqueous solutions of Hg2+ with sub-micromolar detection levels for Hg2+. The fluorescence of the Zn2+-sensor complex is also quenched by aqueous solutions of hydrosulfide. The sensor was used to detect Zn2+ and Hg2+ in living cells. more...

Mutant cystathionine gamma-lyase was targeted to phosphatidylserine exposed on tumor vasculature through fusion with Annexin A1 or Annexin A5. Cystathionine gamma-lyase E58N, R118L, and E338N mutations impart nonnative methionine gamma-lyase activity, resulting in tumor-localized generation of highly toxic methylselenol upon systemic administration of nontoxic selenomethionine. The described therapeutic system circumvents systemic toxicity issues using a novel drug delivery/generation approach and avoids the administration of nonnative proteins and/or DNA required with other enzyme prodrug systems. The enzyme fusion exhibits strong and stable in vitro binding with dissociation constants in the nanomolar range for both human and mouse breast cancer cells and in a cell model of tumor vascular endothelium. Daily administration of the therapy suppressed growth of highly aggressive triple-negative murine 4T1 mammary tumors in immunocompetent BALB/cJ mice and MDA-MB-231 tumors in SCID mice. Treatment did not result in the occurrence of negative side effects or the elicitation of neutralizing antibodies. On the basis of the vasculature-targeted nature of the therapy, combinations with rapamycin and cyclophosphamide were evaluated. Rapamycin, an mTOR inhibitor, reduces the prosurvival signaling of cells in a hypoxic environment potentially exacerbated by a vasculature-targeted therapy. IHC revealed, unsurprisingly, a significant hypoxic response (increase in hypoxia-inducible factor 1 α subunit, HIF1A) in the enzyme prodrug–treated tumors and a dramatic reduction of HIF1A upon rapamycin treatment. Cyclophosphamide, an immunomodulator at low doses, was combined with the enzyme prodrug therapy and rapamycin; this combination synergistically reduced tumor volumes, inhibited metastatic progression, and enhanced survival. Mol Cancer Ther; 16(9); 1855–65. ©2017 AACR. more...

INTRODUCTION AND OBJECTIVES: Use of ablative therapy has the potential to treat bladder cancer resistant to BCG and chemotherapy. However, early efforts with photodynamic therapy and intravenous application of the photosensitizer resulted in significant complications. We have combined Annexin V targeting of phosphatidylserine on tumor surface with single-walled carbon nanotubes (SWNTs) to target bladder tumor cells and treat with near infrared (NIR) light for thermal ablation of tumor in the preclinical setting. METHODS: In vitro studies were conducted on mouse bladder cancer (MB49) and human bladder cancer (J82) cell lines. The dissociation constant for AV binding strength was determined. The specific binding results were confirmed with fluorescence microscopy. Quantitative analysis of the number of cell surface bound AVs was conducted for each cell line. Alamar blue preand post-cell viability assays were conducted to determine the cytotoxic effects of NIR and SWNT-AVs alone as well as a combined approach on both bladder cancer lines. In vivo studies were conducted to determine the biodistribution of intravesically delivered SWNT-AVs in MB49 orthotropic models. FT-Raman analysis was conducted to determine SWNT-AV accumulation in harvested organs. An in vivo NIR tolerance test followed by H&E staining was conducted with a 360 radiating fiber to confirm minimized non-specific tissue damage. A therapeutic efficacy study combining SWNT-AVs and NIR was performed. RESULTS: In vitro binding studies confirmed a strong binding affinity of AV to MB49 (Kd 1⁄4 4.14 1.28 nM) and J82 (Kd 1⁄4 0.38 0.20 nM) cells. Subtoxic levels of docetaxel increased the number of bound AVs per MB49 cell but had no effect on the J82 cells. Inducing SWNT-AV heating due to NIR confirmed significant cancer cell death as compared to untreated controls for both cell lines. The in vitro tests provided statistically significant validation for the potential of this targeted ablation therapy. In vivo testing on C57BL-6 mice was conducted to confirm the efficacy of this treatment even further. A biodistribution study followed by FT-Raman analysis verified no non-specific accumulation of SWNT-AVs in various organs. NIR power tolerance tests confirmed that no healthy tissue damage occurred at 50 J/cm2. In vivo data will also be presented for the effect of SWNT-AV and NIR combination therapy on MB49 mouse bladder cancer. CONCLUSIONS: SWNT-AVs have proven to preferentially target bladder cancer cells and in conjunction with NIR cause significant cytotoxicity in vitro. The results of this study show promise for NIR thermally heated SWNT-AVs as a viable therapeutic option for recurrent bladder cancers. more...

UV-vis and electron paramagnetic resonance (EPR) spectroscopic studies have been carried out on the multinuclear copper(II) complex Cu4BpaRes. The copper atoms are in a tetragonal distorted geometry with nitrogens from bispicolylamine (Bpa) coordinating in the equatorial plane and water molecules or anions completing four coordination sites. The interaction of anions with this polynuclear copper complex in aqueous solution supports the formation of different complex species, which depend on the type and the concentration of the anions. In the presence of excess anions, frozen solution EPR parameters show the formation of species in which the in-plane coordination is characterised by the presence of three nitrogen atoms coming from the ligand and a donor atom from the specific anion. For the bidentate anion ligands and especially for malonate, UV-vis titrations indicate the formation of a 1:4 (Cu4BpaRes:anion) species. EPR experiments support the formation of such a species and indicate that the four copper centres are equivalent and reach penta-coordination via the coordination of both oxygens from the bidentate ligand. more...

Co-doped ZnO nanoparticles were synthesized by a simple chemical method at low temperature with Co:Zn atomic ratio from 0 to 7 %. The synthesis process is based on the hydrolysis of zinc acetate dihydrate and cobalt acetate tetrahydrate heated under reflux to 65 °C using methanol as a solvent. X-ray diffraction analysis reveals that the Co-doped ZnO crystallizes in a wurtzite structure with crystal size of 12–5 nm. These nanocrystals self-aggregated themselves in a highly spherical superstructure of broad size distribution. High-resolution transmission electron microscopy image also shows that each sphere is made up of numerous nanocrystals with average interfringe distance of ~0.28 nm. The X-ray diffraction patterns, energy dispersive X-ray, scanning electron microscopy and high-resolution transmission electron microscopy micrographs of doping of Co in ZnO confirmed the formation of spherical superstructure and indicated that the Co2+ is successfully substituted into the ZnO host structure of the Zn2+ site. The optical band gap of ZnO nanoparticles was remarkably from 3.32 to 4.12 eV with increase of Co doping levels from 0 to 7 %. Photoluminescence measurements confirm these results. more...