The mesoporous nature of the prepared nanocomposites was observed from electron microscopy and surface area measurements. The electrochemical measurements show the bifunctional nature of mesoporous α-Fe2O3@g-C3N4-NCs in electrolysis of water for oxygen evolution and oxygen reduction reactions (OER/ORR) using 0.5 M KOH. Higher current density of mesoporous α-Fe2O3@g-C3N4-NCs reveals the enhanced electrochemical performance compared to pure Fe2O3 nanoparticles (NPs). The onset potential, over-potential and Tafel slopes of mesoporous α-Fe2O3@g-C3N4-NCs were found lower than that of pure α-Fe2O3-NPs. Rotating disc electrode experiments followed by the K-L equation were used to investigate 4e− redox system. Therefore, the mesoporous α-Fe2O3@g-C3N4-NCs bifunctional electro-catalysts can be considered as potential future low-cost alternatives for Pt/C catalysts, which are currently used in fuel cells.Mesoporous α-iron oxide@graphitized-carbon nitride nanocomposites (α-Fe Mesoporous carbon nitride has shown excellent photocatalytic hydrogen generation reactions due to its low band gap and high surface area (1.9 eV)18. The mesoporous g-C3N4 hetero-structured materials are reported as photo-catalysts in water splitting6 and dye degradation reactions19. The mesoporous hetero-structures of g-C3N4@FeNi3 were also used as an adsorbent in crude oil recovery20. Mesoporous nanostructured materials have been synthesized by various methods like solvothermal10, co-condensation14, hydrothermal20, microemulsion21 and microfluidic synthesis22. In this paper, we focus on facile synthesis of mesoporous nanostructured materials and their application in electrolysis of water (OER/ORR).The cost effective, template free, and environmental friendly synthesis of mesoporous nanostructured materials with controlled size and shape are of great interest till date. The variety of mesoporous nanostructured materials have been studied for various applications including sensing24. Noble metal based electro-catalysts are considered as the most capable catalysts for OER and ORR27. These electro-catalysts are very costly. Therefore, they are unwanted for the commercialization. The researchers are devoted to generate the cost-effective and durable electro-catalysts as alternative to replace the expensive noble metal electro-catalysts. Hematite phase of iron oxide (α-Fe2O3) is the most stable phase among other oxides of iron. This material is naturally abundant and inexpensive, which show great attention in several potential applications including supercapacitors28, batteries29, sensors30, adsorbent31 etc. Hematite suffers from the cycling stability during the electrochemical performance. Graphitized carbon nitride (g-C3N4) exhibits excellent chemical stability for ORR32. Therefore, we have designed mesoporous α-Fe2O3@g-C3N4 nanocomposites (NCs) electro-catalysts for electrolysis of water. The α-Fe2O3@g-C3N4-NCs were reported in photo-catalysis35, supercapacitors36 and photo-electrochemical reactions37. Porous core@shell Fe3C@NSC electro-catalyst was reported for efficient ORR in alkaline medium38. Suding Yan et al. have reported the g-C3N4/Fe2O3 composite as the photo-catalysts for the oxidation of bisphenol39. Other transition metal based nanostructured materials with controlled morphology and composition were also reported for electro-catalysis42 and supercapacitors43. g-C3N4 decorated FeNi3 and C-decorated iron oxide hybrid nanostructured materials were reported for energy storage45.Electrolysis of water has great interest in renewable energy resources for future development. The hydrogen evolution, oxygen evolution, and oxygen reduction reactions are three main processes in electrolysis of water for renewable energy conversion devices2O3@g-C3N4 nanostructured materials were synthesized by co-calcination47, ultrasonic35, hydrothermal48 methods and used for waste water treatment and photochemical reactions. Herein, we report the facile synthesis of mesoporous α-Fe2O3@g-C3N4-NCs using urea-formaldehyde (UF) resins at 400 °C/2 h. The structural and morphological characterizations of synthesized nanomaterials were carried out by powder X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), and electron microscopic techniques. 4-e transferred process in alkaline medium is more desirable for ORR. Therefore, bifunctional electro-catalytic performances of α-Fe2O3@g-C3N4 nanostructured materials were investigated in detail using alkaline medium.Recently, α-Fe2O3@g-C3N4-NCs and α-Fe2O3-NPs were shown in Fig. 2O3. The entire xrd patterns match with the JCPDS number of 86-0550. A small peak was also detected in XRD at ~26.50, which reveals the presence of graphitized (g) carbon nitride (C3N4) in the nanocomposites at 400 °C  or Fe (0) in XPS. The peaks between 397–400 eV correspond to the graphitized nitrogen  as also reported24. Moreover, FTIR studies clearly show the difference between mesoporous α-Fe2O3@g-C3N4-NCs and α-Fe2O3-NPs in the spectra as shown in Fig. −1 represent to the vibrations of C=C, C-N and C-H in the mesoporous α-Fe2O3@g-C3N4-NCs  correspond to Fe-O49. Figure 3N4, mesoporous α-Fe2O3@g-C3N4-NCs and α-Fe2O3-NPs. The characteristic Raman bands of α-Fe2O3-NPs were identified at ~221, ~294, ~410, ~498, and ~607 cm−1, which resemble to the A1g, E1g, E1g, A1g, and E1g Raman modes, respectively. Raman peaks were also observed at ~220, ~480, ~708, ~760, and ~980 cm−1 for g-C3N4. The most intense peak of g-C3N4 at ~708 cm−1 represented the s-triazine ring50 and also consistent with FTIR results.Figure NCs Fig.  while noted Fig. . A small3N4 Fig. . The ban2O3@g-C3N4-NCs and α-Fe2O3-NPs. TEM and high magnification TEM studies of α-Fe2O3@g-C3N4-NCs clearly show the porous nature of the materials and also reveal that the α-Fe2O3-NPs are embedded to the g-C3N4 matrix at 400 °C/2 h  of 0.05–0.35. Figure 2O3-NPs and α-Fe2O3@g-C3N4-NCs. The resulting surface area of α-Fe2O3-NPs and α-Fe2O3@g-C3N4-NCs were found to be ~26 and ~115 m2g−1 respectively. N2 adsorption-desorption analysis of α-Fe2O3-NPs and α-Fe2O3@g-C3N4-NCs exhibit the type IV isotherms followed by hysteresis-3 type as shown in Fig. 2O3-NPs and α-Fe2O3@g-C3N4-NCs were found to be 15 and 65 Å respectively  has been loaded on the working electrode (0.07 cm2), which is used to calculate the current density. The cathodic and anodic sweeps in cyclic voltammetry (CV) show the bifunctional redox behavior (OER/ORR) within the potential window of −1 to +1 V at 50 mV/s  to confirm the water oxidation reaction. Figure 2O3@g-C3N4-NCs than that of pure α-Fe2O3-NPs at 0.5 V for 600 seconds. Figure 2O3@g-C3N4-NCs and α-Fe2O3-NPs for ORR (cathodic sweep) at 50 mV/s vs Ag/AgCl. Low onset potential and high current density of α-Fe2O3@g-C3N4-NCs were observed for OER and ORR as compared to Fe2O3-NPs. Inset of Fig. 2O3@g-C3N4-NCs (~280 mV/dec) and α-Fe2O3-NPs (~320 mV/dec) for OER while the Tafel values of α-Fe2O3@g-C3N4-NCs and α-Fe2O3-NPs were found to be ~90 and ~215 mV/dec for ORR. The over-potentials of the α-Fe2O3@g-C3N4-NCs and α-Fe2O3-NPs for OER were found to be ~425 and ~550 mV, respectively, for OER at the current density (η10) of 10 mA/cm2. However, while the over-potentials of the α-Fe2O3@g-C3N4-NCs and α-Fe2O3-NPs for ORR were found to be ~350 and ~530 mV, respectively, at 10 mA/cm2. The Nernst equation42 has been used to convert the potential from Ag/AgCl to reversible hydrogen electrode in order to understand the over-potential of water oxidation reactions. The following reactions could be summarized on the basis of OER and ORR from the electrolysis of water in alkaline medium (0.1MKOH) i.e. 4OH− → O2(g) + 2H2O(l) + 4e− (for OER) and O2 + 2H2O(l) + 4e− → 4OH− (for ORR). The energy conversion tools include electro-catalytic ORR and OER at cathode and anode of an electrolytic cell consisting of two half-cells reactions. The resulting onset potentials, over-potentials, and Tafel slopes are inversely proportional to the electro-catalytic activity, while the resulting current density is directly proportional to the electro-catalytic activity of the materials.Electrochemical measurements of α-FeV/s Fig. . Figure 2O3@g-C3N4-NCs were carried out for ORR by rotating disk electrode using 0.5 M KOH vs Ag/AgCl at the scan rate of 25 mV/s  in ORR. The following K-L equation has been used for the calculation of transferred electrons during electrolysis of water:Moreover, LSV measurements of mesoporous α-FeV/s Fig. . The rot−2); JK: Kinetic current density (A.cm−2); JL: Diffusion-limiting current densities (A.cm−2); F: Faraday’s constant (C.mol−1); Do: Diffusion coefficient of O2 in 0.5 M KOH; ν: Kinematic viscosity of the electrolyte (cm2s−1), Co: Saturation concentration of O2 in 0.5 M KOH at 1 atm O2 pressure (mol.cm−3); ω: rotation rate (rad.s−1).J: Current density (A.cm−1cm2) and ω−1/2 (rpm−1/2) reveals that the K-L plots follow the 1st order reaction kinetics with around four electrons ORR process in 0.5 M KOH. Our results are in good agreement with the reported values of transferred electrons during the water redox reactions52. Based on the present studies, mesoporous α-Fe2O3@g-C3N4-NCs show excellent electro-catalytic behavior for OER/ORR compared to pure α-Fe2O3-NPs in alkaline medium and other reported works. The synergic effect arises with the interaction of iron oxide and g-C3N4, which is also important for better ORR performance of the nanocomposite. The mesoporous nature of the prepared materials provides high surface area as well as more active sites for electro-chemical reactions to enhance the ORR performance. Table 2O3@g-C3N4-NCs could be used as potential low-cost alternatives Pt/C electrode materials in fuel cells.Linear fitting curves of 1/current density  as compared to pure α-Fe2O3-NPs using 0.5 M KOH electrolyte. Our results show low energy loss with α-Fe2O3@g-C3N4-NCs (~90 mV/dec) as compared to pure α-Fe2O3-NPs (~215 mV/dec) during electrolysis of water for ORR. Low onset potential, low over-potential, low Tafel slope, high current density and excellent stability of mesoporous α-Fe2O3@g-C3N4-NCs have been observed, which makes it an alternative electro-catalyst over expensive noble metal based electro-catalysts. Therefore, mesoporous low cost α-Fe2O3@g-C3N4-NCs can be considered as potential candidate in electrochemical water splitting reactions for renewable energy conversion devices in near future.Successfully, we have prepared the low cost mesoporous α-Fe2O3@g-C3N4 nanocomposites have been synthesized using urea-formaldehyde UF resins at 400 °C/2 h. A typical UF resin was obtained from aqueous urea  and formaldehyde (0.2 mole) at pH of 10. Thereafter, 0.01 mole of FeCl3 solution was made with 100 mL of DI water. These two systems were mixed together on magnetic stir at 80 °C for 30 minutes. The resulting brown colored precipitates were appeared. The precipitates were filtered and then dried at 80 °C/24 h. These precipitates were used as the single source precursor for the synthesis of mesoporous α-Fe2O3@g-C3N4-NCs at 400 °C/2 h. The Fe2O3 nanoparticles (NPs) were also synthesized using the same precursor at 500 °C/24 h. Figure Mesoporous α-Fe−10 Torr. FTIR data was measured on Bruker TENSOR-27 spectrometer. The powder samples were pelletized with KBR and run the FTIR measurements under transmittance mode in the range from 400 to 4000 cm−1. Raman measurements were carried out on Bruker Sentera Raman microscope. High resolution transmission electron microscopic (HRTEM) images were captured on JEOL (JSM-2100F) using C-coated Cu TEM grid to identify the morphology and size of the particles. Nitrogen adsorption–desorption isotherm data were collected on the Micromeritics ASAP-2020 physisorption at 77 K. The nanoparticles were degassed at 150 °C for 10 h to remove the moisture, contaminants and adsorbed gases on the surface of the materials. The BET surface area was calculated from the adsorption data obtained at the relative pressure ranging from 0 to 1. Pore size distribution plots were figured using the desorption isotherms followed by the Barrett, Joyner, and Halenda (BJH) method.Powder X-ray diffraction (XRD) data was recorded on Rigaku MiniFlex using Ni-filtered-CuKα radiation (λ = 1.54056 Å). The data was collected in two theta range from 10 to 60° with step size and step time of 0.05° and 1 s respectively. X-ray photoelectron spectroscopy (XPS) was used to identify the oxidation states of the elements. XPS data was recorded in an ultra-high vacuum chamber (Kratos Axis Ultra-DLD electron-spectrometer) with the pressure of 5 × 102) was used to calculate the current density of the electrodes. 0.5 M KOH solution was used as an electrolyte in the electrochemical studies of the electrode materials. The electro-catalysts (5.0 mg of) and isopropanol (1.0 ml) were mixed with nafion (0.2 ml) and sonicated for 10 minutes. One drop of the suspension was put on to the glassy carbon (GC) surface and then dried. The loaded amount of electro-catalysts was of ~0.21 mg/cm2 on the working electrode. Cyclic voltammetry (CV), linear sweep voltammetry (LSV) and Tafel studies were carried out for electrolysis of water to OER and ORR in 0.5 M KOH vs Ag/AgCl at at 50 mV/s. In order to investigate the number of involved electrons in electrolysis of water, we have conducted rotating disc electrode (RDE) experiments with the rotation of 400–2000 rpm in ORR followed by Koutecky–Levich (K–L) equation54. Note that all the electrochemical measurements were conducted at room temperature and repeated three times to check the reproducibility of the results.The electrolysis of water was investigated on potentiostat/galvanostat electrochemical work station (CHI 660E). The reference, counter and working electrodes were Ag/AgCl, Pt-wire, and glassy carbon respectively. The area of working electrode (0.07 cm


Torotrogla paenae n. sp. (Acariformes: Syringophilidae) parasitising the Kalahari scrub-robin Cercotrichas paena (Smith) (Passeriformes: Muscicapidae) in Namibia is described based on the external morphology and DNA barcode data . Females of T. paenae n. sp. morphologically differ from the most similar species T. lusciniae Skoracki, 2004 by the total body length (780–830 vs 645–715 µm in T. lusciniae) and the presence of hysteronotal shields (vs absence), apunctate propodonotal and pygidial shields (vs punctate), apunctate coxal fields (vs punctate), the fan-like setae p’ and p” of legs III–IV provided with c.10 tines (vs 14–15) and the length of setae si (140–180 vs 190–210 µm) and se (160–185 vs 210–225 µm). The male of T. paenae n. sp. morphologically differs from T. lusciniae by the lateral branch of peritremes composed of 4 chambers (vs 7–8 chambers) and lengths of setae ve (45 vs 70–75 µm) and se (120 vs 165 µm).A new quill mite species  Torotrogla Kethley, 1970 is widespread globally with proven occurrence in the Palaearctic, Nearctic, Neotropical and Saharo-Arabian zoogeographical regions   are tiny, permanent bird ectoparasites inhabiting quills of feathers. Syringopilids spend there all their life-cycle and also feed by piercing quill walls with dagger-like chelicerae and suck liquid components of surrounding soft tissue Casto, . The famTorotrogla paenae n. sp. parasitising the Kalahari scrub-robin Cercotrichas paena (Smith) (Passeriformes: Muscicapidae) in Namibia is described based on morphological and DNA barcode data. Cercotrichas paena is a new host species for the family Syringophilidae and the first record of the genus Torotrogla in the Afrotropical realm.In this paper, a new quill mite species Animal material and morphological analysisMite material used in the study was acquired from the collection of feathers deposited in the Smithsonian Institution, National Museum of Natural History, Department of Vertebrate Zoology, Division of Birds, Washington, DC, USA (USNM) (September 2014). Bird specimen was collected in Namibia, 2009. Drawings were made with an Olympus BH2 microscope with differential interference contrast (DIC) optics and a camera lucida. All measurements are in micrometres. The idiosomal setation follows Grandjean  with modcox1) and MG952940 (D1 region of the 28S rRNA gene).Material depositories and abbreviations: AMU, Adam Mickiewicz University, Poznań, Poland; USNM, Smithsonian Institution, National Museum of Natural History, Washington, DC, USA. The voucher slides and corresponding DNA samples are deposited in the collection of the AMU under the identification numbers as indicated below. The sequences were deposited in the GenBank database under the accession numbers MG948551  gene and the D1 region of the nuclear 28S rRNA gene. cox1 was amplified by PCR with degenerate primers Aseq01F (5′-GGA ACR ATA TAY TTT ATT TTT AGA-3′) and Aseq03R (5′-GGA TCT CCW CCT CCW GAT GGA TT-3′)  as described by Dabert et al. . We usedFamily Syringophilidae Lavoipierre, 1953Subfamily Syringophilinae Lavoipierre, 1953GenusTorotroglaKethley, 1970Torotrogla paenaen. sp.Type-host: Cercotrichas paena (Smith) (Passeriformes: Muscicapidae), Kalahari scrub-robin.Type-locality: Omaheke, Aminuis , Namibia.Type-material: Holotype female, 2 female and 1 male paratypes (USNM 642341) are deposited in the USNM, 2 female paratypes in the AMU; coll. E. Glowska (10.viii.2009).Representative sequences: GenBank accession numbers for molecular voucher code KR003: MG948551 (cox1) and MG952940 (D1 of the 28S rRNA gene).ZooBank registration: To comply with the regulations set out in article 8.5 of the amended 2012 version of the International Code of Zoological Nomenclature ; Figs. vi, ve, si and c1. Length ratio of setae vi:ve:si 1:1.6–1.7:1.7–2.6. Setae se situated anterior to level of setae c1. Hysteronotal shields represented by a pair of ovate plates bearing bases of setae d1. Length ratio of setae d2:d1:e2 1:1.1–1.4:1–1.5. Pygidial shield well sclerotised and apunctate. Agenital series represented by 5–7 setae on each body side. Legs. All coxal fields apunctate. Setae tc’ and tc” of legs III–IV subequal in length. Fan-like tarsal setae p’ and p” of legs III–IV with c.10 tines ; ve 130 (105–120); si 140 (160–180); se 185 (160–185); c2 195 (195–215); c1 230 (215–245); d1 170 (155–185); d2 140 (130–150); e2 195 (155–200); f1 55 (75–90); f2 325 (305–370); h1 65 (65–70); h2 405 (345–375); ps1 and ps2 35 (35–40); g1 and g2 40 (45–55); 3b 30 (50–60); 3c 80 (70–90); 4b 45 (30–60); 4c 70 (60–105); tc’III–IV 70 (65–70); tc”III–IV 60 (65–75); l’RIII 40 (55–65); l’RIV 45 (35–45).Male [Based on 1 paratype; Figs. vi, ve, si and c1. Length ratio of setae vi:ve:si 1:1.1:3. Setae se situated anterior to level of setae c1. Hysteronotal shield bearing bases of setae d1, e2, f2 and h2. Length ratio of setae d2:d1:e2 1.5:1:1.5. Setae h2 5.8 times longer than f2. Agenital series represented by 3 setae on each body side. Setae g1,2 and ps1,2 subequal in length. Legs. All coxal fields apunctate. Setae tc’ and tc” of legs III–IV subequal in length. Fan–like setae p’ and p” of legs III–IV with 8 tines. Lengths of setae: vi 40; ve 45; si 120; se 120; c2 120; c1 120; d1 20; d2 30; e2 30; f2 35; h2 205; ps1,2 10; g1,2 10; 3b 30; 3c 60; 4b 45; 4c 45; tc’III–IV 45; tc”III–IV 45; l’RIII 30; l’RIV 20.DNA barcodescox1 sequence data were generated from three females of Torotrogla paenae n. sp. All specimens shared the same cox1 haplotype (GenBank: MG948551). For one specimen a 305-bp fragment coding for the region D1 of 28S rRNA gene was sequenced as DNA-barcode for nuclear DNA (GenBank: MG952940).The Torotrogla paenae n. sp. is morphologically most similar to T. lusciniae Skoracki, 2004 described from the common nightingale Luscinia megarhynchos Brehm (Muscicapidae) from Italy , the hysteronotal shields are present (vs absent), the propodonotal and pygidial shields are apunctate (vs punctate), all coxal fields are apunctate (vs punctate), the fan-like tarsal setae p’ and p” of legs III–IV are provided with c.10 tines (vs 14–15) and the length of setae si (140–180 vs 190–210 µm) and se (160–185 vs 210–225 µm). Males of T. paenae n. sp. differ from T. lusciniae in having the lateral branch of peritremes composed of 4 chambers (vs 7–8 chambers), and lengths of setae ve (45 vs 70–75 µm) and se (120 vs 165 µm).This new species


O-glucosylation was observed with two anomers of HT2 glucoside being present in contaminated oats. In contrast, only one anomer of T2 glucoside was found. The second aspect of this study addresses the stability of the glucosides during thermal food processing. Oat flour was artificially contaminated with T2 and HT2 glucosides individually and extruded at varying initial moisture content and temperature. All four glucosides appear to be more stable during food extrusion than the parent compounds with the glucosidic bond not being hydrolyzed.Plant-derived phase II metabolites of T-2 toxin (T2) and HT-2 toxin (HT2) were first described in 2011 and further characterized in the following years. Since then, some efforts have been made to understand their biosynthesis, occurrence, toxicity, toxicokinetics, and finally relevance for consumers. Thus, the probably most important question is whether and how these metabolites contribute to toxicity upon hydrolysis either during food processing or the gastrointestinal passage. To answer this question, firstly, knowledge on the correct stereochemistry of T2 and HT2 glucosides is important as this affects hydrolysis and chemical behavior. So far, contradictory results have been published concerning the number and anomericity of occurring glucosides. For this reason, we set up different strategies for the synthesis of mg-amounts of T2, HT2, and T2 triol glucosides in both α and ß configuration. All synthesized glucosides were fully characterized by NMR spectroscopy as well as mass spectrometry and used as references for the analysis of naturally contaminated food samples to validate or invalidate their natural occurrence. Generally, 3- Fusarium-toxins T-2 toxin -12,13-epoxy-trichothec-9-ene, T2, Fig. Fusarium sporotrichioides, F. poae, and F. langsethiae  of 100 ng/kg body weight/day that was established by the European Food Safety Authority’s Panel on Contaminants in the Food Chain EFSA  and receFusarium culture material  but also conjugates with minor organic acids such as ferulic acid and glutathione are described . Whole oat flour and maize grits were purchased from Herrnmühle  and white rice flour was obtained by Ziegler & Co. . Naturally contaminated oat kernels that were used as a reference of naturally occurring T2 and HT2 glucosides were provided by Jens C. Meyer .Chemicals in the highest purity available were purchased from either Sigma-Aldrich , Carl Roth , or Fisher Scientific  if not mentioned otherwise. ASTM type 1 water was used at all times unless its non-use is explicitly mentioned, produced with a Purelab Flex 2 system from Veolia Water Technologies . T2 and HT2 were produced according to a previously reported protocol  stock cultures were stored on glucose yeast malt (G-YM) agar plates at 4 °C. Twenty milliliters G-YM medium in a 50-mL Erlenmeyer flask were inoculated with approx. 0.75 cm2 of agar covered with S. viridifaciens. Following incubation for 3 days in the dark at 28 °C and 200 rpm on a horizontal shaker, 1 mL of the culture suspension was transferred to a 200-mL Erlenmeyer flask containing 100 mL G-YM medium and incubated again for 2 days in the dark at 28 °C and 200 rpm.B. muscicola are based on McCormick et al.  were prepared in 50:50 (v/v) glycerol and filter-sterilized glucose yeast nitrogen base (G-YNB) (Difico) medium and stored at − 80 °C. Prior to usage, B. muscicola cultures were grown on yeast malt (YM) agar plates in an incubator for 2 days in the dark at 28 °C. Unlike McCormick et al., the temperature was increased to 28 °C to facilitate bacterial growth in the following. These cultures were transferred to 15 100-mL Erlenmeyer flasks containing 20 mL G-YNB medium with an inoculation loop and grown for 4 days in the dark at 28 °C and 200 rpm. Cultures were centrifuged  and each pellet was resuspended in 80 mL G-YNB medium. One milliliter of the 2-day-old S. viridifaciens suspension in G-YM medium was added to every flask.Cultivation and inoculation conditions for k et al. . Stock cg). The pellets were resuspended in small amounts of water, centrifuged again, and the supernatants were combined. A liquid-liquid extraction using 400 mL tert-butyl methyl ether was applied three times to separate the hydrophilic glucosides from the aglycones. The volume of the aqueous layer was reduced on a rotary evaporator to approx. 100 mL and applied for solid phase extraction on a Strata® C18-E phase (10 g/60 mL)  to remove residual medium ingredients. The cartridge was equilibrated and loaded at an organic content of 10% methanol, washed with 10–20% methanol, and eluted with 100 mL of 70% methanol. To isolate glucosides in high purity, a preparative HPLC was performed on a ReproSil-Pur 120 C18-AQ column by Dr. Maisch GmbH  using a preparative HPLC-system  equipped with an UV detector operating at λ = 195 nm. Isocratic elution was applied at 28% ACN. Three fractions were isolated and evaporated to dryness yielding 2.8 mg (2.2%) HT2-3-O-α-Glcp, 3.8 mg (3.0%) 15-deacetyl-T2-3-O-α-Glcp, and 57 mg (42%) T2-3-O-α-Glcp.One hundred microliters of T2 in methanol  was added to the flasks. One hundred microliters methanol without T2 was added to two of the flasks acting as a growth control. Reaction monitoring was done by HPLC-MS/MS using calculated transitions in selected reaction monitoring mode (SRM). After 14 d of incubation in the dark at 28 °C and 200 rpm, the mixed culture suspensions were centrifuged  as starting material. Briefly, the four acetate groups were cleaved and replaced by triisopropylsilyl residues. Upon reaction of the glucose moiety with T2, the triisopropylsilyl ethers were cleaved yielding a mixture of α and ß configured glucosides. Twelve milligrams (13%) of T2-3-O-ß-Glcp were obtained after a cleanup via preparative HPLC, using the same conditions as applied for the purification of α-glucosides. Comparison of spectroscopic data and HPLC retention time to a reference of T2-3-O-ß-Glcp and published NMR data confirmed the structure of the ß-glucoside.The synthesis of T2-3-k et al.  using etO-ß-Glcp to obtain HT2-3-O-ß-Glcp and further deacetylation products was carried out in four 4-mL glass vials. Twelve milligrams (0.019 mmol) of T2-3-O-ß-Glcp were dissolved in 0.5 mL methanol, and 125 μL of this solution was added to each vial containing 1 mL 1 m ammonium hydroxide in methanol/water . The glass vials were closed and heated for 40 min at 60 °C with constant stirring. The reaction was stopped by the addition of 100 μL 10 m formic acid, the four vials were combined, and the reaction products isolated via preparative HPLC. Applying the chromatographic conditions described above for the purification of the α-glucosides resulted in three fractions that showed UV absorption at λ = 195 nm with corresponding m/z of 567.2403, 609.2518, and 651.2623. After solvent removal and NMR spectroscopy, these fractions could be identified as T2 triol-3-O-ß-Glcp , HT2-3-O-ß-Glcp , and T2-3-O-ß-Glcp .Alkaline hydrolysis of T2-3-d4-methanol with 0.03% tetramethylsilane  on an Agilent DD2 600 MHz NMR spectrometer . Chemical shifts δ are reported as parts per million (ppm) in relation to tetramethylsilane. Proton and carbon spectra and gCOSY, gHMBC, gHSQC, and zTOCSY correlation spectra were recorded. Data processing was done using the Mestrenova software (v. 10.0.0-14381) .NMR experiments were performed in m/z 150 to 700 with a resolving power of 30,000. Higher energy collision dissociation (HCD) spectra of the specific m/z of Na+ adducts were recorded with a relative collision energy of 35%. Source conditions were set as follows: capillary temperature, 275 °C; sheath gas flow rate, 5 arbitrary units; source voltage, 4.0 kV; capillary voltage, 40 V; and tube lens voltage, 130 V. Neither sweep gas nor auxiliary gas was applied. For the identification of T2 phase II metabolites HT2-3-O-α-Glcp, 15-deacetyl-T2-3-O-α-Glcp, T2-3-O-α-Glcp, T2 triol-3-O-ß-Glcp, HT2-3-O-ß-Glcp, and T2-3-O-ß-Glcp via exact mass measurements the following data were acquired:Exact mass measurements were done by direct infusion into a LTQ Orbitrap XL mass spectrometer  equipped with a heated electrospray ionization source. Mass spectra were acquired in positive full scan mode in mass ranges from O-α-Glcp: exact mass (FTMS), m/z 609.2512 for the [M + Na]+ ion ; MS/MS  m/z (%) 245.0631 (6.9), 285.1096 (16), 345.1307 (17), 447.1622 (79), 447.1984 (5.5), 507.1830 (83), 609.2512 (100).HT2-3-O-α-Glcp: exact mass (FTMS), m/z 609.2517 for the [M + Na]+ ion ; MS/MS  m/z (%) 285.1099 (5.2), 345.1310 (14), 417.1521 (9.6), 447.1626 (11), 447.19905 (3.4), 507.1835 (100), 609.2517 (77).15-deacetyl-T2–3-O-α-Glcp: exact mass (FTMS), m/z 651.2621 for the [M + Na]+ ion ; MS/MS  m/z (%) 327.1203 (10), 387.1414 (13), 407.1700 (24), 489.1729 (31), 489.2089 (10), 549.1940 (37), 651.2621 (100).T2-3-O-ß-Glcp: exact mass (FTMS), m/z 567.2403 for the [M + Na]+ ion ; MS/MS  m/z (%) 245.0630 (7.7), 405.1878 (0.03), 435.1621 (8.9), 465.1725 (100), 537.2301 (0.9), 567.2403 (64).T2 triol-3-O-ß-Glcp: exact mass (FTMS), m/z 609.2518 for the [M + Na]+ ion ; MS/MS  m/z (%) 245.0634 (8.5), 285.1101 (7.2), 447.1629 (93), 447.1981 (63), 507.1838 (100), 609.2518 (100).HT2-3-O-ß-Glcp: exact mass (FTMS), m/z 651.2623 for the [M + Na]+ ion ; MS/MS  m/z (%) 327.1206 (6.6), 407.1703 (33), 489.1731 (60), 489.2080 (1.5), 549.1944 (52), 651.2623 (100).T2-3-O-α-Glcp, HT2-3-O-ß-Glcp, T2-3-O-α-Glcp, and T2-3-O-ß-Glcp individually to the tap water used for moistening, the toxin level in 300-g batches of the flour mixtures was adjusted to 250 μg/kg, calculated on a dry weight basis (dwb). Non-fortified batches were prepared for extrusion as a blank reference and to calculate matrix effects and analyte recovery. Each batch was sieved through a 0.8-mm sieve to avoid clumping and to obtain a homogeneous toxin distribution, plastic bagged and stored for 1 h at room temperature for equilibration.A 3:2-mixture of oat and rice flour was used for extrusion experiments. This flour mixture was analyzed for the presence of T2, HT2, and their glucosides prior to extrusion with the method described below. T2 and HT2 were found at negligible levels of 1.1 and 1.9 μg/kg whereas T2 and HT2 glucosides signals remained below the limit of detection. The moisture level of this mixture was analyzed using a standard dry oven method . The overall barrel length was 48 cm and the length-to-diameter ratio was 25:1. The engine power was 1.5 kW with a maximum screw torque of 150 Nm. All experiments were done using a 4:1 core progressive screw operating at 150 rpm and a 1:1 feeding screw of the same diameter operating at 30 rpm. The temperatures of the four individual heating zones of the barrel were set to either 70, 100, 100, 120 °C (low temperature conditions) or to 70, 100, 160, 200 °C (high temperature conditions). A round strand die head with 2 mm inner diameter was used. Screw rotation speeds, pressure, temperature in each heating zone, and die temperature were monitored to ensure a stable process quality.To start the extrusion process, moistened maize grits (25% moisture level) were used. A constant pressure and temperature was awaited before blank flour mixtures were fed to the extruder. Sample collection (approx. 40 g each) was initiated 2 min after that to assure that no maize grits were collected unintentionally.Three samples were collected for each fortified flour mixture and every moisture and temperature combination and stored at − 20 °C prior to analysis. The extruder was purged with maize grits before another glucoside of T2 or HT2 was fed to the extruder to avoid unwanted carry-over. The moisture level of each extrudate was determined after cooling down, using the drying oven method. Thus, all calculations are based on dry weight.2O, and formic acid  using an Ultraturrax® T25 disperser , followed by centrifugation  and dilution by a factor of 8 in H2O.Preparation of all sample material for HPLC-MS/MS analysis was done according to Schmidt et al. . BrieflyTo determine the recovery rates of the four mycotoxin glucosides, non-spiked extrudates of each moisture and temperature combination were fortified individually to 25, 125, and 250 μg/kg (dwb) in triplicate by adding the toxins in small amounts of ACN to 5.00 ± 0.05 g sample material. These samples were left open overnight for solvent evaporation and analyzed as described above.O-α-Glcp, and HT2-3-O-α-Glcp and the other one containing 1 μg/mL of T2, HT2, T2-3-O-β-Glcp, and HT2-3-O-β-Glcp were prepared in ACN. From these stock solutions, for every moisture and temperature combination, two individual calibration curves were prepared resulting in a total of eight calibration curves. Therefore, aliquots of the standards were transferred into 1.5-mL glass vials, the solvent evaporated, and the residue dissolved in 125 μL blank matrix extract, followed by 875 μL of water (1:8 dilution). The calibration curves comprised the range from 0.1 to 10 ng/mL (3.2–320 μg/kg) in seven dilution steps. The slope of the matrix-matched calibration curves was used to calculate the sensitivity.Quantitation was done via matrix-matched calibration curves. Two stock solutions, one containing 1 μg/ml of T2, HT2, T2-3-2O . MS/MS fragment spectra and enhanced product ion spectra (EPI) of the NH4+ and the Na+ adducts were acquired for comparative purposes.HPLC-MS/MS analysis of all sample extracts and references was done on an Agilent 1260 Infinity series HPLC (Agilent Technologies) coupled to a QTrap® 6500 mass spectrometer with an electrospray ionization (ESI) source . The Analyst® software  was used for data acquisition and processing. The mass spectrometric parameters for all analytes were optimized by syringe pump infusion of the analytes solved in ACN/H18 Gravity-SB column  equipped with a guard column (4 × 2 mm) of the same material was used as a stationary phase. Column oven temperature was set to 40 °C and the thermostatted sample rack was set to 8 °C. The sample injection volume was 5 μL. A binary gradient with methanol (eluent A) and H2O (eluent B) at a constant flow rate of 500 μL/min was applied as follows: 0 min, 45% A; 3 min, 45% A; 6 min, 55% A; 11.5 min, 100% A; 12.5 min, 100% A; 12.6 min, 45% A; and 14.0 min, 45% A. The ESI interface was operated in positive ionization mode with the following settings: ionization voltage, 5.5 kV; curtain gas, 35 psi; nebulizer gas, 35 psi; heater gas, 45 psi; source temperature, 450 °C; and entrance potential, 10 V. Declustering potential (DP) and collision energy voltage (CE) were adjusted to each analyte (NH4+ and Na+ adducts) and SRM transition. The sodium adducts gave the highest signal-to-noise ratios (S/N) and were used for quantitation. SRM transitions were: T2 tetraol, DP = 100 V, m/z (CE) 321 → 285 (22 V), 321 → 102 (26 V); neosolaniol, DP = 44 V, m/z (CE) 400 → 245 (15 V), 400 → 215 (21 V); T2 triol, DP = 116 v, m/z (CE) 405 → 303 (22 V), 405 → 125 (23 V); T2 triol-3-O-ß-Glcp, DP = 57 V, m/z (CE) 567 → 465 (37 V), 567 → 435 (40 V), 567 → 245 (41 V); HT2, DP = 110 V, m/z (CE) 447 → 345 (27 V), 447 → 285 (30 V); HT2-3-O-Glcp, DP = 210 V, m/z (CE) 609 → 507 (39 V), 609 → 345 (44 V), 609 → 285 (46 V); T2, DP = 145 V, m/z (CE) 489 → 327 (32 V), 489 → 245 (36 V); T2-3-O-Glcp, DP = 150 V, m/z (CE) 651 → 549 (43 V), 651 → 489 (47 V), 651 → 407 (49 V), 651 → 387 (50 V).A Nucleodur® CNaturally contaminated oat kernels and the glucoside references were analyzed with the abovementioned method. All measurements were done in triplicate. References of α- and ß-glucosides were analyzed separately in every case. For confirmation, sample extracts were also fortified with the assigned glucosides, resulting in an increase of peak area and no additional signals.Mycotoxins and mycotoxin stock solutions were handled with special care using safety gloves and protective clothing throughout the experiments. Toxin waste including the liquids as well as single-use tubes and vials was properly disposed.in planta formed glucosides, are of increasing interest due to their possible health concern. Although they were discovered more than a half decade ago, the structures of the food-relevant glucosides of T2 and HT2 are not fully characterized and some of the reports in literature are conflicting. For this reason, the full set of T2 and HT2 glucosides in α and ß configuration was prepared and characterized.Modified forms of T2 and HT2, especially the O-α-Glcp was prepared by biotransformation using the yeast Blastobotrys muscicola as previously described , the loss of oxygen, or the addition of C2H2O. If done properly, this biotransformation approach is a powerful and time-saving tool for the production of mg-amounts of T2 and HT2 glucosides in one single culture flask that does not require complex purification methods.The fact that hardly any HT2 was found in the culture extract suggests that the glucosylation by O-β-Glcp was already synthesized and purified, a chemical hydrolysis was used to prepare HT2-3-O-β-Glcp. Furthermore, coupling of HT2 with glucose following the abovementioned protocol was not favored as selectivity directing the pyranose towards position 3 is not given for HT2. Exposure of the prepared T2-3-O-ß-Glcp for 40 min to a mild hydrolysis by 1 m ammonium hydroxide at 60 °C yielded HT2-3-O-ß-Glcp and T2 triol-3-O-ß-Glcp next to the unaltered T2-3-O-ß-Glcp. The experiment was performed in small scale in advance and monitored via HPLC-MS/MS. This revealed that 40 min is the optimum reaction time as the amount of HT2 glucoside started to decrease with longer reaction time. Upon preparative HPLC cleanup, all products were obtained in mg-amounts sufficient for NMR characterization. No likely products  were observed after 40 min of hydrolysis, indicating that the glucosidic linkage is stable under these conditions. Increasing the temperature to 60 °C correlated with a shorter reaction time (approx. 90%) compared to the T2 hydrolysis at 30 °C  but is diminished for an equatorial-axial dihedral angle (α) as predicted by the Karplus equation. Comparing the obtained α and ß configured glucosides, we observed the expected signals and coupling constants that are characteristic for their anomericity. The J1′-2′ coupling constant used to distinguish between α or ß configuration showed J1′-2′ of 3.71 ± 0.01 Hz for α-glucosides and 7.77 ± 0.05 Hz for ß-glucosides. Furthermore J2′-3′ was also clearly influenced by the glucose configuration of 9.81 ± 0.02 Hz (α-glucosides) and 9.10 ± 0.11 Hz (ß-glucosides). Additional differences in proton shifts of H-1′, H-2′, H-3′, and H-5′ and carbon shifts for C-2, C-1′, C-3′, and C-5′ were observed. This indicates that a differentiation between α and ß configuration cannot only be made by the J1′-2′ coupling constant but also by chemical shifts in the respective proton or carbon spectrum.Proton NMR data of the isolated glucosides are reported in Table The method used for simultaneous detection of T2 and HT2 and modified mycotoxins is usually HPLC-MS/MS  was one of the most intense transitions in most cases. The loss of the glucose moiety (m/z 162.0528) was observed for all glucosides but appeared less prominent than the loss of C7H14O4 .FTMS fragmentation spectra did not allow us to distinguish between both configurations because no consistent fragmentation pattern was observed. The loss of isovaleric acid . All spectra are shown in Fig. m/z 527 and m/z 485 fragments of ammonium adducts of T2 and HT2 glucosides that are formed by the loss of ammonia and isovaleric acid enable a rough distinction between α and ß form. Regarding T2 glucoside, this was first described by McCormick et al. . This was in contrast to our expectation that an additional glucosidic bond that might be cleaved inside the molecule presumably enhances the potential of chemical hydrolysis. We investigated if the loss of glucosides leads to the formation of the respective aglycones by analyzing the extrudates for the presence of T2, HT2, T2 triol, neosolaniol, and T2 tetraol. Though a cleavage of the glucosidic bond would be a plausible explanation for the loss of the glucosides, the aglycone signals remained on the same level as the blank reference. This demonstrated clearly that no T2 or HT2 is released from T2 or HT2 glucoside during extrusion cooking of oats. The question whether and how these glucosides are cleaved during processing that involves fermentation steps  was not addressed in this study and needs to be further investigated in future work.The unavailability of plant-derived phase II metabolites of mycotoxins often hampers their investigation in terms of occurrence, analytical determination, toxicity, and toxicokinetics. In this work, we report two efficient synthesis strategies for the production of relevant glucosides of T2 and HT2. Characterizing data on six different glucosides of T2 and T2 derivatives are reported in order to contribute to a better understanding of the phase II metabolism of these mycotoxins in plants. A first investigation on their thermal stability during food extrusion revealed that the majority withstands heat, pressure, and shearing and might therefore be present in the final food product. These findings clearly indicate that modified T2 and HT2 should be considered in the toxicity evaluation and human health risk assessment of T2 and HT2.


M, more commonly known under its commercial names Meldonium or Mildronate) co-crystalizes with sodium bromide and sodium iodide forming polymeric hydrates. Metal ions and M zwitterions are assembled into infinite layers via electrostatic interactions and hydrogen-bonded networks. These layers are connected via electrostatic attraction between halogenide ions and positive trimethylhydrazinium groups into a three-dimensional structure.3-propanoate  co-crystalizes with sodium bromide and sodium iodide forming polymeric hydrates poly[[tetra-μ-aqua-diaquabis[3-propanoate]disodium] dibromide tetrahydrate], [Na2(C6H14N2O2)2(H2O)6]Br2·4H2O, and poly[[di-μ-aqua-diaqua[μ-3-propanoate]disodium] diiodide], [Na2(C6H14N2O2)2(H2O)4]I2. The coordination numbers of the sodium ions are 6; the coordination polyhedra can be described as distorted octahedra. Metal ions and M zwitterions are assembled into infinite layers via electrostatic interactions and hydrogen-bonded networks. These layers are connected via electrostatic attraction between halogenide ions and positive trimethylhydrazinium groups into a three-dimensional structure.3-propanoate (C M), more commonly known under its commercial names such as Meldonium or Mildronate, was introduced by Grindeks (Latvia) as an anti-ischemic medication propanoate ; their high stability was a challenge that was necessary to overcome for the preparation of pharmaceutically pure forms of M. The stability of a sodium iodide binary compound was given as an example in Silva (2013Binary compounds of ilva 2013. The cryThe labelling schemes for structures (I)The coordination polyhedra of the sodium ions in (II)The octahedral environment around Na2 in (II)M have approximately the same geometry (the two pseudo-inversion-symmetric zwitterions in the structure of (I)M monohydrate and dihydrate  direction. Water molecules O11 and O13 form bridges between the cation chain and the ‘bromide’ chains as hydrogen-bond donors; they are also acceptors of four hydrogen bonds from the water molecules O5 and O10, and O8 and O9 respectively. These hydrogen bonds connect chains into a two-dimensional network. Two more enhanced hydrogen bonds Table 3, O7—H7A⋯de Fig. 7. These lvia the bidentate carboxylate group to form an infinite chain along the [001] axis is Fig. 8. The watis Fig. 8. An arrais Fig. 8 additionet al., 2016et al., 1983M also were also studied using powder X-ray diffraction , has a very rich crystal chemistry: the CSD (Version 5.39) contains 217 different structures of its compounds. There are several known crystal structures of TMG binary compounds with potassium iodide  and recrystallized from propanol-2. Equimolar amounts of it were mixed with sodium iodide and sodium bromide in aqueous ethanol; subsequent slow evaporation yielded crystals suitable for single–crystal X-ray experiments. IR spectra  are very similar to those of M dihydrate. (I)2O), 1571, 1483, 1402, 1320; (II)2O), 1568, 1480, 1405, 1317, 1088, 816; M dihydrate: 3201 (H2O), 1577, 1484, 1404, 1320, 1090, 816.Preparation and properties of binary compounds of  al. 1975 and Silv al. 1975. CommercCrystal data, data collection and structure refinement details are summarized in Table 5CELL_NOW; Sheldrick, 2008Structure (I)A, O6—H6B, O7—H7A, O7—H7B, O8—H8A, and O8—H8B; O11—H11D, O11—H11E, O12—H12E, O12—H12D, O13—H13A, and O13—H13B; O14—H14A and O14—H14B were restrained to be equal with an effective standard deviation of 0.02 Å. Distances N1—H1 and N3—H3 were also restrained to be equal with an effective standard deviation of 0.02 Å; Uiso(H) = 1.5Uiso(N).In the structure of (I)In the structure of (II)Uiso(H) = 1.2 Uiso(C); methyl hydrogen atoms were refined as rotating idealized methyl groups and with Uiso(H) = 1.5Uiso(C). Hydrogen atoms of water molecules were refined in an isotropic approximation with Uiso(H) = 1.5Uiso(O).In both structures, methylene hydrogen atoms were refined with riding coordinates and with 10.1107/S2056989018006977/zl2729sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989018006977/zl2729Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018006977/zl2729Isup4.cdxSupporting information file. DOI: 10.1107/S2056989018006977/zl2729IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989018006977/zl2729IIsup5.cdxSupporting information file. DOI: 1841872, 1841871CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


This probe demonstrates increased tumor specificity, compared to existing tumor-specific PET probes . Evaluation of probes by in vivo PET imaging, 18F-FIMP showed intense accumulation in LAT1-positive tumor tissues, but not in inflamed lesions, whereas intense accumulation of 18F-FDG was observed in both tumor tissues and in inflamed lesions. Metabolite analysis showed that 18F-FIMP was stable in liver microsomes, and mice tissues . Investigation of the protein incorporation of 18F-FIMP showed that it was not incorporated into protein. Furthermore, the expected mean absorbed dose of 18F-FIMP in humans was comparable or slightly higher than that of 18F-FDG and indicated that 18F-FIMP may be a safe PET probe for use in humans. 18F-FIMP may provide improved specificity for tumor diagnosis, compared to 18F-FDG, 18F-FET, and 11C-MET. This probe may be suitable for PET imaging for glioblastoma and the early-phase monitoring of cancer therapy outcomes.Positron emission tomography (PET) imaging can assist in the early-phase diagnostic and therapeutic evaluation of tumors. Here, we report the radiosynthesis, small animal PET imaging, and biological evaluation of a L-type amino acid transporter 1 (LAT1)-specific PET probe,  Hence, 18F-FDG PET cannot distinguish between tumor tissue and inflamed lesions2, and therefore results in false-positive results for tumor diagnosis3.Positron emission tomography (PET) imaging can assist in early-phase clinical evaluations of tumors. 2-Deoxy-2-11C]-methionine (MET), were developed to overcome the disadvantages of 18F-FDG4. 11C-MET was predicted to have a higher specificity for tumors; however, this probe was found to accumulate in tumor, normal, and inflamed tissues6. In order to improve the selectivity for tumor tissues, PET probes with tumor-specific molecular targets have developed9.Radiolabeled amino acid PET probes, such as L--2-methylpropanoic acid , was identified as a potentially effective PET probe for LAT1. Here, we report the radiosynthesis, small animal PET imaging, and biological evaluation of 18F-FIMP.The L-type amino acid transporter 1 (LAT1) is a sodium-independent L-type amino acid transporter (LAT), with four isoforms: LAT1, LAT2, LAT3, and LAT4As a result of screening our α-methyl amino acid chemical library using hLAT1 and hLAT2 overexpression cell lines, FIMP was found to have higher affinity for LAT1 than 2-aminobicyclo--heptane-2-carboxylic acid (BCH), a classical inhibitor of L-type amino acid transporters, which is also transported into cells as a substrate of LAT1. The half-maximal (50%) inhibitory concentration (IC50) value of FIMP was significantly lower than that of BCH .High expression levels of LAT1 and CD98 were observed for T24 and LNZ308 tumor cells. Conversely, WI-38 normal human fetal lung fibroblast showed low to moderate expression of both proteins Fig. .Figure 1We also evaluated LAT1 and CD98 expression in tumor xenografts and normal muscle tissue. Moderate to high LAT1 and CD98 expressions were observed for T24 and LNZ308 xenografts Fig. . Convers18F-FIMP accumulation by PET imaging in tumor, muscle and inflamed tissues. Time-activity curve of 18F-FIMP accumulation in all tissues plateaued by 90 min after probe injection  were significantly higher than that of 18F-FET (SUV 1.14 ± 0.20) and 11C-MET (SUV 0.79 ± 0.14) , comparable to that of 18F-FDG (SUV 2.55 ± 0.59) without significant difference (P = 0.29). However, accumulation of 18F-FIMP in inflamed lesions (SUV 0.96 ± 0.04) was comparable and significantly higher than that of 18F-FET (SUV 1.14 ± 0.20) and 11C-MET (SUV 0.79 ± 0.14) , significantly lower than that of 18F-FDG (SUV 1.73 ± 0.36) (P < 0.01)  Fig. .Figure 218F-FIMP (3.29 ± 0.16) was significantly higher than that of 18F-FET (1.38 ± 0.25) and 11C-MET (2.11 ± 0.40) , significantly lower than that of 18F-FDG (5.52 ± 1.65) (P < 0.01). INR of 18F-FIMP (1.36 ± 0.08) was comparable to that of 18F-FET (1.07 ± 0.92) and 11C-MET (1.10 ± 0.32) , significantly lower than that of 18F-FDG (3.77 ± 1.10) (P < 0.01)  further supported these findings. TMR of 01) Fig. . These r18F-FIMP uptake in the pancreas (33.99 ± 2.39%ID/g). Uptake of 18F-FIMP was higher in tumor (7.78 ± 1.11%ID/g) than inflamed tissue, muscle tissue, and blood . 18F-FET uptake was highest in the pancreas (22.73 ± 2.78%ID/g). Tumor uptake of 18F-FET (3.64 ± 0.77%ID/g) was higher compared to inflamed tissue and normal muscle, but equivalent to that of blood . 11C-MET uptake was highest in the liver and pancreas . Tumor uptake of 11C-MET (3.49 ± 0.05%ID/g) was higher compared to inflamed tissue and normal muscle, but equivalent to that of blood . In contrast, 18F-FDG uptake was highest in the heart (51.70 ± 12.52%ID/g); however, tumor uptake (8.71 ± 6.61%ID/g) was also higher compared to inflamed tissue, muscle, and blood   was significantly higher than that of 18F-FET (3.64 ± 0.77%ID/g) and 11C-MET (3.49 ± 0.05%ID/g) , comparable to that of 18F-FDG (8.71 ± 6.61%ID/g) without significant difference (P = 0.77). On the other hand, accumulation of 18F-FIMP in inflamed lesions (3.89 ± 0.17%ID/g) was significantly higher than that of 18F-FET (2.12 ± 0.37%ID/g) and 11C-MET (2.00 ± 0.55%ID/g) , not significantly difference than that of 18F-FDG (4.67 ± 1.91%ID/g) (P = 0.14)  was significantly higher and comparable than that of 18F-FET (1.37 ± 0.28) and 11C-MET (2.78 ± 0.29) , not significantly difference than that of 18F-FDG (17.5 ± 13.4) (P = 0.08). INR of 18F-FIMP (1.32 ± 0.16) was significantly lower and comparable to that of 18F-FET (0.80 ± 1.39) and 11C-MET (1.57 ± 0.30) , significantly lower than that of 18F-FDG (8.48 ± 2.44) (P < 0.01)  Fig. . 18F-FIM14) Fig. . TMR of 01) Fig. .Figure 418F-FDG, but not 18F-FIMP or 11C-MET, was observed in inflamed joints of CIA mice. No accumulation of any probe was detected in the joints of control mice  compared to control mice (2.31 ± 0.15) (P < 0.001). However, there was no significant difference in INR for 18F-FIMP and 11C-MET accumulation in CIA mice (0.91 ± 0.01 and 0.68 ± 0.20), compared to controls (0.86 ± 0.04 and 0.68 ± 0.11) . In vitro experiments using human, rat, and mouse liver microsomes, showed that FIMP was highly stable over 60 min, and the unchanged fraction were 87.1, 99.6, and 98.8% for human, rat and mouse microsomes, respectively  in the acid precipitation fraction of LNZ308 cells. This value significantly decreased (15.9 ± 1.8%) with pretreatment with cycloheximide (CHX), a protein synthesis inhibitor, was performed (P < 0.01). However, incorporation of 18F-FIMP was very low both with and without CHX pretreatment ; hence, suggesting that this probe may provide a more accurate approach to discriminate tumor tissues from inflamed tissues.Inflammation is an inseparable by-product in the pathophysiology of cancer. Inflammation is not only a side-effect of cancer treatments, such as radio- and chemotherapy, but also contributes to the development and progression of cancer. At the earliest stages of neoplastic progression, inflammation can promote the progression of incipient neoplasia into invasive cancers14. Most natural amino acid-derived PET probes, such as 11C-MET, are incorporated into the protein fraction, resulting in an increase in nonspecific accumulation in non-tumor tissues15. From our results, 18F-FIMP showed very high metabolic stability both under in vitro and in vivo conditions and is not incorporated into protein. These findings suggest that the high tumor accumulation value obtained using this tumor-specific PET probe maybe more reliable compared to that of other radiolabeled amino acids such as L-3-18F-fluoro-α-methyl tyrosine (FAMT)9, O-(2-18F-fluoroethyl)-L-tyrosine (FET)16, and anti-1-amino-3-18F-flurocyclobutane-1-carboxylic acid (FACBC)8. In order to demonstrate whether 18F-FIMP has superiority in tumor imaging, standard PET probes, such as 18F-FDG and 11C-MET, and 18F-FIMP need to be directly compared in the same cancer inoculation models by PET imaging studies.The efflux of radiolabeled amino acids and their metabolites from cells has been negatively correlated with their accumulation in tumors18F-FIMP in humans. The effective doses of 18F-FIMP were determined to be 25.1 ± 5.3 and 30.8 ± 6.7 μSv/MBq for males and females, respectively. These doses are comparable or slightly higher than that of 18F-FDG  and 18F-fluroethyltyrosine 17 but still indicate that 18F-FIMP is a safe PET probe for use in humans.An accurate estimation of radiation exposure is indispensable for defining a safe clinical PET study protocol. According to the biodistribution data from our PET imaging studies, we were able to estimate the expected mean absorbed dose of 18F-FIMP in humans. For example, additional comparisons between 18F-FIMP and other LAT1-specific PET imaging probes would provide a valuable assessment of its potential as a cancer diagnosis tool.However, additional studies are required to evaluate further applications of 18F-FIMP accumulated in LAT1-positive tumor tissue, but not in inflamed lesions. This markedly high discrimination between tumors and inflamed lesions is important for effective diagnosis and treatment. Hence, 18F-FIMP may have advantages over existing PET imaging probes, such as 18F-FDG and 11C-MET.In this study, we developed a tumor imaging PET probe with a high affinity for LAT1. PET imaging studies revealed that 18F-fluoride was produced via the 18O  18F nuclear reaction, by irradiation of a 98% 18O-enriched water target with a 12 MeV proton beam at the HM-12S cyclotron . The aqueous solution containing 18F-fluoride ion was passed through Sep-Pak Accell Plus QMA Plus Light Cartridge  and adsorbed 18F-fluoride ions were eluted with acetonitrile (0.7 mL), Kryptofix 2.2.2 (14 mg), and aqueous potassium carbonate (0.2 mL of 0.21 mol/L). The solvent was removed by azeotropic distillation with acetonitrile under He atmosphere.Radiochemical syntheses were carried out in a hot cell, using a computer controlled automated radiochemical synthesis unit . 18F-FIMP was prepared via a three-step reaction that consisted of l8F-fluorination of a tosyl precursor, deprotection, and neutralization  at 120 °C for 10 min, then the solution was neutralized by adding 2 N sodium hydroxide (500 μL). The solution was adjusted to pH 6 by addition of acetic acid (100 µL) and H2O (500 µL) and allowed to equilibrate at room temperature for 1 min. The crude 18F-FIMP was purified by reverse-phase HPLC ; flow rate, 10 mL/min; detection, UV absorption by 235 nm and gamma-ray detector)  Fig. . After r18F-FIMP stability was estimated under in vitro conditions. 18F-FIMP showed high stability (97.6%) standing at room temperature for 24 h after the synthesis  was produced by phase transfer–mediated nucleophilic 18F fluorination of N-trityl-O-(2-tosyloxyethyl)-L-tyrosine-tert-butyl ester and subsequent deprotection with a radiochemical purity of >99.5% determined by HPLC. 11C-Methionine (11C-MET) was synthesized as described previously18, with a radiochemical purity of >99.5% determined by HPLC. 18F-FDG for human diagnostic grade was provided by the Institute of Biomedical Research and Innovation (IBRI) hospital, Kobe, Japan.The LNZ308 human glioblastoma cell line was a kind gift from Prof. Motoo Nagane of Department of Neurosurgery, Kyorin University, Japan. T24 (RCB0431) human bladder transitional-cell carcinoma and WI-38 (RCB0702) human lung normal fibroblast cells were obtained from the RIKEN BRC through the National Bio-Resource Project of the MEXT/AMED, Japan. LNZ308 and WI-38 cells were cultured in Dulbecco’s modified Eagle’s medium  supplemented with 10% fetal bovine serum (FBS), 100 units/mL penicillin, and 100 μg/mL streptomycin. T24 cells were cultured in McCoy’s 5 A (Modified) medium  supplemented with 10% FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin.All animal experimental protocols were approved by the Animal Care and Use Committee of RIKEN and performed in accordance with the National Institutes of Health Principles of Laboratory Animal Care (Approved No. MAH28-02). All applicable institutional and/or national guidelines for the care and use of animals were followed.via subcutaneous injection of 5 × 106 cells in 100 μL phosphate buffered saline (PBS). Tumor growth was monitored twice weekly using a caliper. Acute-phase inflammation was induced by subcutaneous injection of 50 μL turpentine oil into the left forepaw of tumor-bearing mice 72 h before PET imaging19. We also used collagen-induced arthritis (CIA) model mice for evaluation of developed radiolabeled probes (see Supplementary Methods).LAT1-positive human glioblastoma (LNZ308) cells were inoculated into the right forepaws of female BALB/cAJcl-nu/nu nude mice  18F-FDG administration. 18F-FIMP and 11C-MET-PET were administered to unfasted mice. All mice were anesthetized with 1.5% isoflurane and placed on the bed of a microPET Focus 220 scanner . Radiolabeled probes were dissolved in saline (0.1 mL) and administered via a cannula inserted into the tail vein. Emission data were acquired for 90 min after administration using a 3-dimensional (3D) list-mode method for 18F-FIMP and 11C-MET, and for 45 min from 55 min after administration using a 3D list-mode method for 18F-FDG. Data were reconstructed using 2-dimensional filtered back projection (FBP) for quantification and a 2-dimensional ordered subset expectation maximization (OS-EM) algorithm for region of interest (ROI) definition. For ROI definition and further analysis, summed images (0–90 or 55–100 min post injection) were reconstructed. ROIs were drawn on several areas of tumor, muscle, and inflamed tissues. Regional uptake of radioactivity in organs were decay-corrected based on injection times and expressed as the standardized uptake value (SUV), where SUV = tissue radioactivity concentration (MBq/cm3)/injected radioactivity (MBq) × body weight (g). Quantitative analysis of PET imaging data also represented as tumor-to-muscle and inflamed lesion-to-normal muscle ratios . PET imaging was also performed with CIA mice using 18F-FIMP, 11C-MET and 18F-FDG.Mice were fasted for 14 h before After PET imaging, mice were sacrificed and their organs quickly removed and washed with saline. Blood samples were obtained immediately before dissection by cardiac puncture. Excised organs and blood samples were weighed and their radioactivity determined using a Wallac Wizard 1480 scintillation counter . Results were expressed as %injected dose per gram of tissue, TMR, tumor-to-blood ratio (TBR), and INR.18F-FIMP (μSv/MBq) in humans were estimated on the basis of PET imaging data from mice (n = 4). The mean %ID/g values for mouse livers, kidneys, pancreases, urinary bladders, and remainder of the body were extrapolated to estimate expected uptake in organs for a 73 kg human adult male. The organ fractions of total body mass for mice (25 g), human males (73 kg), and human females (53 kg) required for this extrapolation were obtained from Hui et al.20 for mice and ICRP Publication 6017 for humans, respectively. Dosimetry estimations were calculated using the OLINDA/EXM version 1.1 software  based on standard human male and female models21.Mean absorbed doses of Data are presented as mean ± standard deviation (SD). All statistical analyses were performed using Microsoft Excel 2010 version 14.0  using Student’s t test. P-values less than 0.05 were considered significant.Supplementary information


In the crystals, the isophthalate counter-ions form layers as a result of O—H⋯O hydrogen bonds with the water molecules, which are pillared with the macrocyclic cations and lie parallel to the  and (II), contain tetragonally distorted CuN trans-diaquacopper(II) isophthalate monohydrate, [Cu(C16H29N5)(H2O)2](C8H4O4)·H2O, (I), and trans-diaquacopper(II) isophthalate 0.9-hydrate, [Cu(C15H28N6)(H2O)2](C8H4O4)·0.9H2O, (II) consist of one diaqua macrocyclic cation, one dicarboxylate anion and uncoordinated water molecule(s). In each compound, the metal ion is coordinated by the four secondary N atoms of the macrocyclic ligand and the mutually trans O atoms of the water molecules in a tetragonally distorted octahedral geometry. The average equatorial Cu—N bond lengths are significantly shorter than the average axial Cu—O bond lengths . The coordinated macrocyclic ligand in the cations of both compounds adopts the most energetically favorable trans-III conformation. In the crystals, the complex cations and counter-anions are connected via hydrogen-bonding interactions between the N—H groups of the macrocycles and the O—H groups of coordinated water molecules as the proton donors and the O atoms of the carboxylate as the proton acceptors. Additionally, as a result of O—H⋯O hydrogen bonding with the coordinated and water molecules of crystallization, the isophthalate dianions form layers lying parallel to the (The asymmetric units of the title compounds,  The complexes of N3,N10-disubstituted diazacyclam , readily obtainable via template-directed Mannich condensation of bis(ethylenediamine) complexes with formaldehyde and primary amines  – a middle member of this series of ligands – have attracted considerably less attention, presumably because of the necessity of using a more sophisticated non-cyclic precursor, i.e. 3,7-diazanonane-1,9-diamine, in the Mannich condensation  are popular metal-containing building units for the construction of metal–organic frameworks (MOFs) possessing many promising applications  dianion is often used as bridging ligand in the construction of MOFs, a very limited number of its compounds with azamacrocyclic cations have been described to date and all they are complexes of the NiII complexes with azacyclam ligands and an isophthalate dianion, namely, trans-diaquacopper(II) isophthalate hydrate, [Cu(L1)(H2O)2](ip)·H2O, (I)trans-diaquacopper(II) isophthalate 0.9-hydrate, [Cu(L2)(H2O)2](ip)·0.9(H2O), (II)Herein, we describe the syntheses and crystal structures of the title CuII ion in the complex cations in the title compounds (I)Each Cuversus 2.495 (12) Å for (I)versus 2.507 (7) Å for (II)], which can be attributed to a large Jahn–Teller distortion. The CuII ions are displaced from the nearly planar (r.m.s. deviations less than 0.01 Å) mean planes of the N4 donor atoms towards the O1W water molecule by 0.024 and 0.033 Å in (I)trans-III  conformation  for (I)chair [bite angles 93.7 (2) for (I) and 93.7 (9)° for (II)] conformations. The methylene group of the substituent at the non-coordinated nitrogen atoms N3 in the six-membered chelate rings is axially oriented and the sum of the C—N—C angles around these atoms  and O2W—H2WA⋯O2}, as well as to the O3W molecule (ClO4)2 and [Cu(L2)](ClO4)2, were prepared by a method reported in the literature ](ClO4)2  in 8 ml of DMF were added 3 ml of an aqueous solution of Na2ip . A violet precipitate formed in 24 h; this was filtered off, washed with diethyl ether and dried in air. Yield: 27 mg (19%). Analysis calculated for C24H39N5CuO7: C 50.29, H 6.86, N 12.22%. Found: C 50.42, H 6.96, N 12.02%.Compound (I)L2)](ClO4)2. Yield: 30 mg (21%). Analysis calculated for C23H37.8N6CuO6.9: C 48.12, H 6.67, N 14.64%. Found: C 48.31, H 6.84, N 14.32%. Violet plates of (I)Compound (II)Safety note: perchlorate salts of metal complexes are potentially explosive and should be handled with care.Uiso(H) values of 1.2 or 1.5Ueq of the parent atoms. Water H atoms in (II)Uiso(H) = 1.5Ueq(O). All other H atoms were freely refined.Crystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989019008387/hb7828sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989019008387/hb7828Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019008387/hb7828IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1922733, 1922732CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


III–oxalate–carboxylate framework structure with [Eu(NH2—BDC)(ox)(H3O)] topology is reported. The non-porous three-dimensional structure is constructed from two-dimensional layers of EuIII–carboxylate–oxalate, which are pillared by NH2—BDC2− pillars. The basic structural unit of the layer is an edge-sharing dimer of TPRS-{EuIIIO9}, which is assembled through the ox2− moiety. The intralayer void is partially occupied by TPR-{EuIIIO6} motifs.An organically pillared Eu 5-2-aminobenzene-1,4-dicarboxylato)(oxomium)europium(III)], [Eu(C8H5NO4)(C2O4)(H3O)]n or [EuIII(NH2–BDC)(ox)(H3O)]n  is constructed from two-dimensional layers of EuIII–carboxylate–oxalate, which are connected by NH2–BDC2− pillars. The basic structural unit of the layer is an edge-sharing dimer of TPRS-{EuIIIO9}, which is assembled through the ox2− moiety. The intralayer void is partially occupied by TPR-{EuIIIO6} motifs. Weak C—H⋯O and strong, classical intramolecular N—H⋯O and intermolecular O—H⋯O hydrogen-bonding interactions, as well as weak π–π stacking interactions, affix the organic pillars within the framework. The two-dimensional layer can be simplified to a uninodal 4-connected sql/Shubnikov tetragonal plane net with point symbol {44.62}.The non-porous three-dimensional structure of poly[(μ To enhance interactions with guest species, additional functional groups can be introduced onto the phenyl ring of BDC2−, e.g. 2-amino-1,4-benzenediarboxylic acid (NH2-H2BDC) in NH2-MIL-53(Al) enhanced the carbon dioxide capture capacity , on the other hand, may not facilitate the porous framework  have emerged as authentic multifunctional materials finding potential in various applications, 2-H2BDC and H2ox were employed as mixed linkers in the synthesis of a new three-dimensional framework of europium, i.e. [Eu(NH2-BDC)(ox)(H3O)] (I). The crystal structure of I, which exhibits site disorder at both the EuIII ion and the amino group, is reported. Weak intermolecular interactions and the framework topology are also described.Herein, NH2–BDC)(ox)(H3O)] crystallizes in the monoclinic space group P21/c. Its asymmetric unit comprises one EuIII ion, which is disordered over two crystallographic sites with an occupying ratio of 0.86 (Eu1A): 0.14 (Eu1B) and whole molecules of NH2–BDC2−, ox2− and H3O+  and 2.562 (2) Å, are consistent with the values observed for other EuIII coordination frameworks, e.g. [(CH3)2NH2]2[Eu6(μ3-OH)8(BDC–NH2)6(H2O)6] 3(DEF)2]n where ATPA2− = 2-aminoterepthalate and DEF = diethylformamide , 0.44 (N2) and 0.31 (N3), respectively.[Eu(atpt)2.5(phen)2]n·1.75nH2O where atpt2− = 2-aminoterephthalate and phen = 1,10-phenanthroline 2(μ4-ATA)(H2O)4]·2DMF·0.5H2O}n where ATA2− = 2-aminoterephthalate . It exhibits none of the characteristic f–f emission of EuIII. Even the broad emission characteristic of the ligand-centered π–π emission was not observed. This may be attributed to a proton-induced fluorescence-quenching mechanism facilitated by the presence of H3O+ in close proximity to the phenyl ring of NH2-BDC2− tetraaquabis(nitrato)lanthanum] diaquapraseodymium(III)] dichlorotris(tetrahydrofuran-O)gadolinium tetrahydrofuran solvate , oxalic acid  and 1,4-diazabicyclo[2.2.2]octane  were dissolved in 8.0 mL of DMF/H2O (1 m:7 mL) to prepare solution A. Separately, solution B was prepared by dissolving Eu2O3  in 1.0 mL of concentrated HNO3 aqueous solution, which was then adjusted to pH 7 using a 10 M NaOH aqueous solution. Solution B was then gradually introduced into solution A, and the mixture was then transferred to a 22 mL Teflon-lined stainless-steel autoclave. The reaction was carried out under an autogenous pressure generated at 393 K for 7 days. Yellow crystals of I were then recovered by filtration. FT–IR of I : 3361, 2987, 1617, 1313, 1053, 798.To synthesize III ion was refined as being disordered over two crystallographic sites resulting in refined occupancies of 0.855 (Eu1A) and 0.145 (Eu1B). The disorder of the amino group over three crystallographic sites could be clearly seen in the electron-density map and was refined using the SUMP command providing occupancies of 0.259 (N1), 0.440 (N2) and 0.305 (N3). EADP constraints were necessary to make the anisotropic refinements of the disordered N atoms stable. The three H atoms on the ligated H3O+ (O9W) were evident in the electron-density map and therefore assigned as such. The SADI restraint was nonetheless applied on the refinements of the three O—H bonds. H atoms could be positioned from the electron-density maps and were refined as riding with Uiso(H) = 1.2Ueq or 1.5Ueq(O). Bond restraints o N—H and O—H were applied in the refunements.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989019014713/jj2217sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019014713/jj2217Isup2.hklStructure factors: contains datablock(s) I. DOI: 1962371, 1962371CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


HEK-293 cells, stably transfected with M1Rs, were used to transiently transfect D2Rs and CaV1.3b with different CaVβ-subunits, allowing for whole-cell current measurement from a pure channel population.We examined whether two G protein-coupled receptors (GPCRs), muscarinic M1R activation with Oxotremorine-M inhibited currents from CaV1.3b coexpressed with α2δ-1 and a β1b, β2a, β3, or β4-subunit. Surprisingly, the magnitude of inhibition was less with β2a than with other CaVβ-subunits. Normalizing currents revealed kinetic changes after modulation with β1b, β3, or β4, but not β2a-containing channels. We then examined if D2Rs modulate CaV1.3b when expressed with different CaVβ-subunits. Stimulation with quinpirole produced little inhibition or kinetic changes for CaV1.3b coexpressed with β2a or β3. However, quinpirole inhibited N-type Ca2+ currents in a concentration-dependent manner, indicating functional expression of D2Rs. N-current inhibition by quinpirole was voltage-dependent and independent of phospholipase A2 (PLA2), whereas a PLA2 antagonist abolished M1R-mediated N-current inhibition. These findings highlight the specific regulation of Ca2+ channels by different GPCRs. Moreover, tissue-specific and/or cellular localization of CaV1.3b with different CaVβ-subunits could fine tune the response of Ca2+ influx following GPCR activation.MThe online version of this article (10.1186/s13104-018-3783-x) contains supplementary material, which is available to authorized users. CaVβ1b (#X61394), CaVβ2a (#M80545), and CaVβ4 (#L02315) constructs were provided by Edward Perez-Reyes (University of Virginia). The D4.4R (#AF1199329) construct was provided by Hubert H. M. Van Tol (University of Toronto). D2R cDNA (#NM_000795) was obtained from the UMR cDNA Resource Center (https://www.cdna.org). Per well, a total of 0.5 μg of DNA (of which GFP cDNA was less than 10%) was used following the methods of Roberts-Crowley and Rittenhouse (2009) [HEK-M1 cells, grown in 12-well plates (~ 60–80% confluent), were transfected with a 1:1:1 molar ratio of Cant CaVβs , using LaVβ1b #X694, CaVβ22+ Tyrode’s (in mM): 5 MgCl2, 145 NaCl, 5.4 KCl, and 10 HEPES, brought to pH 7.50 with NaOH. Once a seal was established and the membrane ruptured, the Tyrode’s solution was exchanged for external bath solution (in mM): 125 NMG-aspartate, 20 Ba-acetate, 10 HEPES, brought to pH 7.50 with CsOH. Only cells with ≥ 0.2 nA of current were used. Data were acquired using Signal 2.14 software (CED) and stored for later analysis on a personal computer. Linear leak and capacitive currents were subtracted from all traces.Whole-cell currents were recorded following the methods of Liu et al. . High reAll chemicals were purchased from Sigma unless otherwise noted. FPL 64176 (FPL), nimodipine (NIM), and oleoyloxyethyl phosphorylcholine  were prepared as stock solutions in 100% ethanol. Quinpirole (quin) and Oxotremorine-M  were dissolved in DDW and stored as 10 mM stock solutions at − 70 °C. Stocks were diluted daily to the final concentration by at least 1000-fold with external solution. For ethanol-prepared stocks, the final ethanol concentration was less than 0.1%.t-test for two means, or a one-way ANOVA followed by a Tukey multiple-comparison post hoc test. Statistical significance was set at p < 0.05 or < 0.001. Analysis programs included Signal (CED), Excel (Microsoft), and Origin (OriginLab).Data are presented as the mean ± s.e.m. Data were analyzed for significance using a Student’s paired 3-containing L-channels, elicited from a holding potential of − 60 mV to a test potential of − 10 mV, averaged − 4699 ± 279 pA (n = 3) compared to − 9 ± 1 pA for HEK-M1 cells transfected with only accessory subunits . Lack of current from cells transfected without CaV1.3b, confirmed that HEK-M1 cells exhibit little endogenous Ca2+ current and transfection of accessory subunits does not upregulate endogenous Ca2+ channels. Recombinant current was confirmed as L-type by showing sensitivity to the L-VGCC antagonist NIM. NIM inhibited β3-containing currents . These differences in the magnitude of current inhibition and kinetics suggest that the CaVβ-subunit affects M1R modulation of CaV1.3b.In MSNs, M animals . Only Cain brain , implyin1Rs and D2Rs activate pathways involving G proteins, PLC, and AA release  was unaffected by activation of D2Rs, expressed in HEK-293 cells, similar to a previous report on CaV1.3a, which has a longer C-terminus [2R-long inhibited CaV1.3a [2R-short inhibited CaV1.3b . After 2 h, cells were washed with supplemented media and washed a final time 2 h later. 10 mM MgSO4 was added to the medium to block basal activity of CaV1.3b, which helped minimize excitotoxicity of transfected cells. Cells were transferred 24–72 h post-transfection using 2 mM EDTA in 1X PBS, to poly-l-lysine-coated coverslips. Recording began 1 h after transfer to coverslips. A Individual traces of CaV1.3b-β3 current before (CTL) and after exposure to 0.3 µM NIM. B Concentration–response curve of L-current inhibition to NIM (n = 4–8). C CaV1.3b-β2a currents before and after exposure to FPL (1 µM). Cells were stepped to a test potential of − 10 mV from a holding potential of − 90 mV followed by repolarization to − 90 or − 50 mV. Control (CTL) currents from β2a-containing L-VGCCs show little to no inactivation as observed previously [D CaV1.3b-β3 currents before and after FPL. Cells were stepped to a test potential of − 10 mV from a holding potential of − 60 mV followed by repolarization to − 60 mV. Following FPL, both β2a- and β3-containing channels exhibited slower activation and deactivation kinetics, hallmarks of agonist action on L-current [E FPL enhancement of the CaV1.3b-β2a current–voltage plot from a holding potential of − 90 mV . F Concentration–response curve of CaV1.3b-β3 tail current enhancement to FPL (n = 4–8). Currents inhibited by NIM and enhanced by FPL fully recovered by washing with bath solution (data not shown).eviously . D CaV1.-current . E FPL eAdditional file 2. D4.4Rs do not inhibit recombinant L-current. A Summary bar graph of CaV1.3b-β3 current inhibition by 0.5 μM quin (n = 5). B & C Summary bar graphs of TTP and r40 kinetic analysis.


The conformation of synthesized cyclo(GSPE) fixes the coordination to lead ion in a 1:1 ratio. This cyclo(GSPE)-Pb complex was constructed as an asymmetric 3D network in the crystalline state. The polymerization of a heavy metal ion with a rigid asymmetric cyclic tetrapeptide represents the first example of a new class of macrocyclic complexes.The crystal structure of a naturally occurring cyclic tetrapeptide  It was first isolated from the Ruegeria strain of marine bacteria ·2H2O}n2 are reported for the first time. Further {[cyclo(Gly-l-Ser-l-Pro-l-Glu)Pb(NO3)]·2H2O}n2 was constructed as an asymmetric 3D network in the crystalline state. Coordination relied on the prearranged conformation of synthesized cyclo(GSPE) 1. The polymerization of a heavy metal ion with a rigid asymmetric cyclic tetrapeptide represents the first example of a new class of complexes. bacteria  and was guration . HoweverCyclo(GSPE) 1 was synthesized by using standard solution-phase peptide coupling protocols. Its spectral data were identical with those of natural 1 ·2H2O}n2 complex was obtained. Crystals were prepared by slow cooling to room temperature and resulted in a 3D coordination polymer. Several coordination polymers of lead with a variety of synthetic organic ligands have been reported earlier  direction (The geometry around the Pb center appears holodirected . The bidirection . cyclo(GSPE) 1 binds with three Pb(II) ions to form one cyclic ring and the same unit repeats along b-axis to form a 2D layer structure °). Two adjacent Pb(II) ions in molecule  to buildmolecule , left ancyclo(GSPE)-Pb complex 2 is unique as all the complexes reported to date are with higher cyclic peptides or β-cyclic tetrapeptides and are sandwich complexes +, ESI-MS: [M−H]− calculated m/z 369.1488, obtained m/z 369.1426.achieved . The linachieved . Deprote{[cyclo(Gly-l-Ser-l-Pro-l-Glu)Pb(NO3)]2O}n·2H (2). Pb(NO3)2  was dissolved in water (1 mL). cyclo(GSPE) 1  was dissolved in water (1 mL) and added to the metal solution. The pH of solution was adjuted to 5 with diluted HNO3 solution. The reaction mixture was stirred at RT for 10 minutes and then at 70 °C for 4 days. Crystals suitable for X-ray diffraction were obtained from saturated solution by slow cooling. Anal. Calc. for C15H25N5O12Pb (674.6): C 26.70, H 3.73, N 10.38, O 28.46. Found: C 26.23, H 2.94, N 10.64, O 28.17. IR : 3433, ν(NH); 1645, ν(C=O vs amide); 1384, ν(NO3). 1H-NMR: (D2O) 1.86–1.99 ; 2.04–2.31 ; 2.35–2.53 ; 3.45–3.55 ; 3.59 ; 3.65–3.71 ; 3.73–3.81 ; 3.82 ; 3.87–4.00 ; 4.19–4.26 ; 4.38–4.45 ; 4.76 . MALDI-TOF: m/z 577.69 [M+Pb–H]+ , 595.71 [M+Pb+H2O–H]+ and 719.69 [M+Pb+2NO3+H2O–H]+. ESI-MS: [M+Pb–H]+ calculated m/z 577.1177, obtained m/z 577.1183; [M+Pb+NO3–H]+ calculated m/z 639.1055, obtained m/z 639.1049; [M+Pb+2NO3–H]+ calculated m/z 701.0933, obtained m/z 701.0973; [M+Pb+2NO3+2H2O+H]+ calculated m/z 739.1301, obtained m/z 739.2925. 1 and rod-like crystals of 2 were obtained by slow solvent evaporation at room temperature, respectively. Crystals suitable for X-ray diffraction analysis were selected with size of 0.13 × 0.08 × 0.05 mm for 1 and 0.44 × 0.09 × 0.05 mm for 2. Crystals were mounted on a glass fiber and used for data collection. For both compounds 1 and 2 diffraction data were preliminarily collected with a Bruker APEX-II CCD diffractometer using graphite monochromated MoKα radiation (λ = 0.71073 Å). Absorption corrections for the area detector were performed with the program SADABS. Structures were solved by direct methods and were refined against the least-squares methods on F2 with the SHELXL-97 package, incorporated in SHELXTL/PC V5. Anisotropic thermal factors were used only for all non-hydrogen atoms. Colorless prism-like crystals of www.ccdc.cam.ac.uk/conts/retrieving.html .CCDC 831318 and 831319 contains the supplementary crystallographic data for this paper. These data can be obtained free of charge via cyclo(GSPE) 1. The synthetic building blocks were coupled through hydroxyl, carboxylate and two amide bonds to lead ion and thus created water-soluble, coordination polymers (tris-CTPs) with well-defined three-dimensional structures. This is the first report of a naturally occurring CTP as the host for metal complexation. The CTP retains its conformation through the binding process with metals. The unique crystal structure of this rigid molecule suggests the formation of a designated complex with nanotube properties. We have developed the synthesis of a CTP


H-benzimidazole acetone disolvate exhibits O—H⋯N hydrogen bonds between benzimidazole units and O—H⋯O hydrogen bonds with one of the acetone solvate molecules as the acceptor. Density functional theory is used to estimate the strength of the interactions.The solid-state structure of 2-(4-hydroxyphenyl)-1-[(4-hydroxyphenyl)methyl]-5,6-dimethyl-1 H-benzimidazole was isolated as the acetone disolvate, C22H20N2O2·2C3H6O. O—H⋯N hydrogen bonding between benzimidazole molecules results in chains parallel to [010]. One of the acetone solvate molecules participates in O—H⋯O hydrogen bonding with the benzimidazole derivative. C—H⋯π interactions are observed in the extended structure. Hirshfeld surface analysis was used to explore the intermolecular interactions and density functional theory was used to estimate the strength of the hydrogen bonds.Hydrogen bonding plays an important role in the design of solid-state structures and gels with desirable properties. 1-(4-Hydroxybenzyl)-2-(4-hydroxyphenyl)-5,6-dimethyl-1 The study of the gelation properties of small organic compounds (organogelators) is of importance in soft-matter research because of possible biomedical applications , and we report herein its structural characterization and an exploration of its three-dimensional superstructure, including an examination of hydrogen-bond strengths.In our continuing efforts to exploit benzimidazole as a gelator core, we synthesized 1-(4-hydroxybenzyl)-2-(4-hydroxyphenyl)-5,6-dimethyl-1c.f. Geiger & DeStefano, 2016A view of the molecular structure of (1) with the atom-labeling scheme employed is seen in Fig. 1versus 1.192 (3) Å]. This observation is consistent with previous results  contains two acetone molecules, one of which uses its carbonyl oxygen atom as acceptor in an O—H⋯O hydrogen bond  functional/basis set combination (see Section 7 for details). The results of the calculations are reported in Table 2The interaction energies were calculated using density functional theory with the CE-B3LYP/6-31G functional/basis set combination was also used to calculate the interaction energies. The results are found in Table 2The M06 suite of density functionals are reported to outperform B3LYP for dispersion and ionic hydrogen-bonding interactions ° with the benzimidazole moiety ° between the benzimidazole core and the 2-hydroxyphenyl substituent phenol is essentially planar and exhibits an intramolecular hydrogen bond methyl]phenol benzene hemisolvate phenol phenol -15,6-Dimethyl-2-(4-hydroxyphenyl)-1-[(4-hydroxyphenyl)methyl]-1H-benzimidazole: 1.96 g (14.37 mmol) of 4,5-dimethyl-1,2-diaminobenzene were dissolved in 50 mL of ethanol and stirred under nitrogen. 3.65 g (29.9 mmol) of 4-hydroxybenzaldehyde were dissolved in ethanol, purged with nitrogen for 5 min., and then added dropwise to the solution. The solution was refluxed for 24 h and cooled, after which a yellow solid formed. This solid was filtered and washed with cold ethanol. 4.23 g  of the yellow solid was obtained. Rf (3:1 acetone/hexane) = 0.64. 1H NMR : δ 2.23 ppm , δ 2.28 ppm , δ 5.33 ppm , δ 6.64 ppm , δ 6.78 ppm , δ 6.85 ppm , δ 7.15 ppm  δ 7.40 ppm , δ 7.50 ppm , δ 9.34 ppm , δ 9.88 ppm .Single crystals of (1) were obtained by slow evaporation of a dilute acetone solution of the product.Uiso(H) = kUeq(C), where k = 1.2 for H atoms bonded to aromatic and methylene C atoms and 1.5 for H atoms bonded to methyl C atoms. H atoms bonded to oxygen were refined freely, including isotropic displacement parameters.Crystal data, data collection and structure refinement details are summarized in Table 3CrystalExplorer17  functional/basis set combination and are corrected for basis set superposition energy (BSSE) using the counterpoise (CP) method  basis set were employed for the determination of interaction energies, which were corrected for BSSE employing the CP method  functional/basis set combination.Interaction energy calculations were also performed on molecules in the gas phase using 10.1107/S2056989019001270/zl2750sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019001270/zl2750Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019001270/zl2750Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019001270/zl2750Isup4.cmlSupporting information file. DOI: 1893078CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


H-furopyrazole ring system is still quite limited, and their chemistry and functional properties remain largely underexplored. The current study investigates routes to construct this system from easily accessible starting materials using metal-catalyzed reactions. A simple and efficient procedure to access the 2H-furopyrazole ring system was developed by employing the silver(I) ion-mediated ring-closure reaction of 4-alkynyl-3-hydroxy-1-phenyl-1H-pyrazoles as a key step. The required intermediate hydroxyalkynyl substrates for this reaction were prepared by a Pd-catalyzed coupling of 4-iodo-1-phenyl-1H-pyrazol-3-ol with ethyne derivatives. The structures of the obtained target compounds were unequivocally confirmed by detailed 1H, 13C and 15N NMR spectroscopic experiments, HRMS and a single-crystal X-ray diffraction analyses. This silver(I)-mediated 5-endo-dig cyclization of readily available 4-alkynyl-3-hydroxy-1H-pyrazoles can be used as an efficient method to access many novel 2,5-disubstituted 2H-furopyrazoles.Fused pyrazole ring systems are common structural motifs of numerous pharmaceutically important compounds. Nevertheless, access to derivatives of the aromatic 2 Huang et al. have reported the synthesis and the evaluation of antiplatelet and anti-allergic activities of 2,3,4-trisubstituted 2H-furopyrazole-5-carboxylic acids and related carboxamides [c]pyrazol-6(1H)-one derivatives followed by heating of the obtained 5-bromo derivatives in the presence of sodium alkoxide pyrazole ring system is structurally similar to the benzo[b]furan system, which is a privileged motif in natural products and biologically active compounds [b]furan and its derivatives [b]furans with good yields by the base-promoted cyclization of easily accessible 2-alkynylphenols [b]furans were conveniently synthesized by the one-pot tandem Hiyama alkynylation/cyclization reaction between 2-iodophenol and (triethoxysilyl)alkynes [b]furan ring system through intramolecular cyclization of 2-alkynylphenol substrates [b]furans furans, where AuCl3 or a mixture thereof with AgOTf has been used to promote the appropriate cyclization furans  in the plization .H-furopyrazole ring system by a Sonogashira-type alkynylation of 4-iodopyrazol-3-ol and subsequent intramolecular 5-endo-dig cyclization of the obtained hydroxyalkynyl substrate mediated by a Ag(I) catalyst.In the present work, we describe a method for the construction of the 2H-furopyrazole ring system employs a hydroxyethynyl substrate that contains adjacent ethyne and hydroxy groups on the pyrazole core (H-pyrazol-3-ol (1), which is readily accessible from the oxidation of 1-phenyl-3-pyrazolidinone pyridine [f]triazolooxazepine [a]benzimidazole BF4 was applied in the regioselective intramolecular cyclization of alkynols to construct bicyclic ethers pyrazole ring system in the regions of δ 115.0–116.3 for C-3, δ 113.1–113.9 for C-3a, δ 94.5–96.8 for C-4, δ 156.1–163.8 for C-5 and δ 168.4–169.0 ppm for C-6a. The 15N spectra of 4b–e exhibited signals for two nitrogen atoms in the regions of δ −169.4 to −172.6 for N-2 and δ −127.5 to −128.0 ppm for N-1.The structural assignments of 4d was investigated additionally by single crystal X-ray analysis  and 4d(B) held together by the weak CH···N type hydrogen bonds C19–H···N51 (C···N = 3.470(11) Å, H···N = 2.60 Å, C–H···N = 151°) and C69–H···N1 (C···N = 3.355(11) Å, H···N = 2.51 Å, C–H···N = 144°), respectively, whose length and angles are typical for crystals of aza-heterocycles pyrazole systems in 4d(A) and 4d(B) are given in The asymmetric unit of the crystal consists of two molecules rocycles . Althoug4d(A) and 4d(B) are 1.356(10) Å and 1.392(11) Å, respectively. All ring atoms of the 2H-furopyrazole moiety lie in almost one plane, but the phenyl substituents are slightly turned in relation to this plane  catalyst.In conclusion, we have demonstrated a new, three-step synthetic route to 2File 1Experimental details and characterization data.


All analogs exhibited a variable degree of β-glucuronidase inhibitory activity with IC50 values ranging in between 0.8 ± 0.1–42.3 ± 0.8 μM when compared with the standard d-saccharic acid 1,4 lactone (IC50 = 48.1 ± 1.2 μM). Structure activity relationship has been established for all compounds. Molecular docking studies were performed to predict the binding interaction of the compounds with the active site of enzyme.Chromen-4-one substituted oxadiazole analogs  Water molecules, ligands and Chain-B were removed from the downloaded PDB file by means of Discovery Studio Visualizer [In silico binding interactions of synthesized derivatives against human-software . The X-rsualizer . Two-dimsualizer . Other psualizer  is entir


The synthesis and structural properties of a series of chromium tricarbonyl ‘piano-stool’ complexes bearing substituted pentafulvene ligands were studied. Significant deviation of the exocyclic fulvene double bond from the cyclopentadiene plane accompanies coordination. Evidence of non-covalent π-π interactions was observed. 24H18)(CO)3]·0.5C6H6 (I), tricarbonylchromium(0), [Cr(C26H20)(CO)3] (II), and tricarbonylchromium(0), [Cr(C34H22)(CO)3] (III), each have a distorted octahedral geometry, with the fulvene coordinated in a π–η2:π–η2:π–η2 fashion. Significant deviation of the exocyclic fulvene double bond from the cyclopentadiene plane accompanies coordination. Evidence of non-covalent π–π interactions was observed in both (I) and (III), with centroid-to-centroid distances ranging from 3.330 (8) to 3.494 (8) Å.The synthesis and structural properties of a series of chromium tricarbonyl ‘piano-stool’ complexes bearing substituted pentafulvene ligands were studied. The complexes, tricarbonylchromium(0) benzene hemisolvate, [Cr(C Analysis of the fulvene bond lengths when compared to the previously reported uncomplexed fulvenes reveals nearly unchanged C—C single bonds  with slight elongation of the C=C double bonds   results in the bending of this bond from the cyclopentadiene plane by 33.22 (18) to 34.2 (3)°. This is in agreement with a previously reported chromium complex with 6,6-dimethylfulvene  Table 1. This doI and III. In both I and III, the molecules are arranged in layers in which the π system composed of the cyclopentadiene core (head) of the fulvene and the 3-phenyl substituent (tail) adopt a head-to-tail  Å end-to-end and 1.505 (17) Å edge-to-edge with respect to the opposing 3-phenyl substituent centroid. This results in a near perfect alignment of the fulvene C2 atom over the centroid of the opposing phenyl ring, with angles betwen the cyclopentadiene-phenyl ring normal and the C2 to phenyl ring centroid vector of only 2.05 (2)° end-to-end and 5.85 (3)° edge-to-edge. In complex III, the centroid of each cyclopentadiene ring is by slipped 0.286 (8) Å end-to-end and 0.761 (7) Å edge-to-edge with respect to the opposing 3-phenyl substituent centroid. Again, the C2 fulvene atom is brought into near perfect alignment over the centroid of the opposing phenyl ring, with angles betwen the cyclopentadiene-phenyl ring normal and the C2 to phenyl ring centroid vector of 7.67 (9)° end-to-end and 6.16 (9)° edge-to-edge.Evidence for π–π interactions in the solid state is observed in 9I Fig. 4 and 3.33II Fig. 5, both weIII between the pyrene units. The interplanar contact distance is 3.494 (8) Å fulvene, and 1,3-diphenyl-6-(1-pyrene)fulvene were prepared in accordance with literature procedures tricarbonylchromium(0) (I) A solution of 1,3,6-triphenylfulvene  in THF (10 mL) was added to a stirred suspension of Cr(CO)3(MeCN)3  in THF (15 mL) under N2. The solution quickly turned from pale yellow to dark red. The reaction mixture was allowed to stir at room temperature for 24 h before removal of the solvent in vacuo. The residue was dissolved in diethyl ether (100 mL), filtered under ambient conditions, and the solvent removed in vacuo. Crystals suitable for single-crystal X-ray diffraction were obtained by dissolving the crude product in benzene and layering with pentane.{1,3-Diphenyl-6-(3-vinylphenyl)fulvene}tricarbonylchromium(0) (II) 1,3-Diphenyl-6-(3-vinylphenyl)fulvene  and Cr(CO)3(MeCN)3  were used employing the procedure outlined for the preparation of I. Crystals suitable for single-crystal X-ray diffraction were obtained by dissolving the crude product in benzene and layering with pentane.{1,3-Diphenyl-6-(1-pyrene)fulvene}tricarbonylchromium(0) (III) 1,3-Diphenyl-6-(1-pyrene)fulvene  and Cr(CO)3(MeCN)3  were used employing the procedure outlined for the preparation of I. Crystals suitable for single-crystal X-ray diffraction were obtained by vapor diffusion of diethyl ether into a chloroform solution of the crude product.Uiso(H) = 1.2 Ueq(C). In I, an outlier (101) was omitted in the last cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989018010794/rz5242sup1.cifCrystal structure: contains datablock(s) global, I, II, III. DOI: 10.1107/S2056989018010794/rz5242Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018010794/rz5242IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989018010794/rz5242IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 1858307, 1858306, 1858305CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: