Under pressures above 6 GPa and temperatures of 100–700 °C, β-FeOOH transformed into ε-FeOOH, as in the case of α-FeOOH. However, the established pressure–temperature phase diagram of β-FeOOH differs from that of α-FeOOH, probably owing to its open framework structure and partial occupation of Cl− ions. The OER activities of ε-FeOOH strongly depended on the FeOOH sources, synthesis conditions, and composite electrodes. Nevertheless, one of the ε-FeOOH samples exhibited a low OER overpotential compared with α-FeOOH and its parent β-FeOOH, which are widely used as OER catalysts. Hence, ε-FeOOH is a potential candidate as a next-generation earth-abundant OER catalyst.Research on materials under extreme conditions such as high pressures provides new insights into the evolution and dynamics of the earth and space sciences, but recently, this research has focused on applications as functional materials. In this contribution, we examined high-pressure/high-temperature phases of β-FeO Research on materials under extreme conditions such as high pressures provides new insights into the evolution and dynamics of the earth and space sciences, but recently, this research has focused on applications as functional materials. In the 20th century, it was examined in the context of the atmospheric rusting of iron and steel,3 solutions at 60–100 °C,1,15–20 before it was discovered as a natural product in the Akagane metal mine, Japan.216 octahedra form [2 × 2] tunnels via double corner-linked bands.22–26 A certain number of guest Cl− ions are necessary to maintain the [2 × 2] tunnels, leading to the specific chemical formula FeOx1−(OH)x1+Clx.1,2,16 Its structure was initially proposed to have tetragonal symmetry with the I4/m space group,22–24 but recent X-ray diffraction (XRD) measurements revealed monoclinic symmetry with the I2/m space group.25,26 The monoclinic distortion is extremely small, as understood by its lattice parameters: am = 10.5536(7) Å, bm = 3.03449(8) Å, cm = 10.5740(4) Å, and β = 90.086(5)°.26 β-FeOOH is isostructural to α-MnO2, which contains Na+ (manjiroite), K+ (cryptomelane), Ba2+ (hollandite), or Pb2+ (coronadite) ions as the guest in the [2 × 2] tunnels.27 A series of α-MnO2 has also been employed as the catalyst for the OER28–30 and as the electrode for nonaqueous or aqueous secondary batteries.31,32β-FeOOH was generally synthesized by hydrolyzing FeCl32ge2g → t52g) and hydrogen bond symmetrization occur at pressures exceeding 40 GPa.33–38 Moreover, α-FeOOH transforms into ε-FeOOH under HP/HT conditions at above 7 GPa and 300 °C.39–4147 and ε-FeOOH,39–41 respectively, wherein the latter adopts a distorted rutile structure with the Pmn21 space group, forming [1 × 1] tunnels. Among the three FeOOH polymorphs, β-FeOOH is significantly less compressible as a function of pressure (at room temperature), owing to its high bulk modulus of 284(1) GPa.42 The HP/HT phase of β-FeOOH is currently unknown, although Xu et al.43 reported that ε-FeOOH is a HP phase of β-FeOOH without any experimental evidence.Many studies have been performed on β-FeOOH, but mostly under ambient pressure. However, a high-pressure/high-temperature (HP/HT) study could provide insights into its fundamental structural and electronic properties to gain an in-depth understanding of the evolution and dynamics of earth and solar system. Indeed, in the case of α-FeOOH, a pressure-induced spin state transition  is generally accepted to play a crucial role in the OER performance.7 Furthermore, the O–H⋯O configuration is highly asymmetric under ambient pressure; for instance, the O–H and H⋯O lengths are 0.88 and 1.94 Å, respectively, for α-FeOOH47 and 0.86–0.98 and 2.10–2.49 Å, respectively, for β-FeOOH.26 Although β-FeOOH exhibited the maximum bulk modulus,42 the strength of the Fe–O bond and the asymmetric O–H⋯O configuration should be altered under HP/HT conditions and should significantly affect the OER performance of β-FeOOH. In the case of the quadruple perovskite oxide CaCu3Fe4O12, which was synthesized under HP/HT conditions, the OER performance was enhanced when the interatomic distance between the nearest neighboring OH adsorbates was ca. 2.6 Å.50Regarding the OER catalyst, materials synthesized from β-FeOOH under HP/HT conditions could offer a wide range of material options to enhance OER performance, because current studies on the FeOOH polymorphs focus on fabricating composite electrodes or heterodoping.57Fe Mössbauer spectroscopy were employed to clarify their structural and electronic properties. Consequently, β-FeOOH was found to transform into ε-FeOOH along a different pressure–temperature phase diagram from that of α-FeOOH. In addition, we found that the ε-FeOOH sample prepared at 8 GPa and 400 °C exhibited a lower OER overpotential than those of the parent β-FeOOH and α-FeOOH.In this contribution, we revealed HP/HT phases of β-FeOOH under pressures of 2–12 GPa and temperatures below 800 °C, as well as the OER activity of the obtained materials. Several methods such as synchrotron XRD measurements, X-ray absorption spectroscopy (XAS), and 22.11,15,18,20 We, however, used a recently developed method, namely, a combination of spontaneous peptization and Ostwald ripening, to obtain β-FeOOH colloidal solution at room temperature (ca. 25 °C).3–5 This method provides highly crystalline and uniform β-FeOOH nanorods with diameters of 3 nm and lengths of 15 nm,3–5 owing to the typical advantages of the Ostwald ripening method.51 As shown in 3·6H2O and ethylenediamine (CH2)2(NH2)2 . The pH of this mixture was maintained at 2.0–2.4. Then the solution was continuously stirred for 30 min and finally aged overnight at ca. 25 °C.β-FeOOH is usually prepared by aging solutions at 60–100 °C for several days, offering 100–500 nm-size particles.ca. 25 °C and rinsed with a 0.1 M KOH  solution. This powder was filtered with a cellulose acetate membrane with a pore size of 0.2 μm (Toyo Roshi Kaisha) and washed three times with distilled water. The powder was finally dried at 45 °C in vacuum for 8 h. The amount of Cl− ions, x in FeOx1−(OH)x1+Clx, was examined with a combination of inductively coupled plasma-atomic emission spectroscopy , and ion chromatography . The ICP-AES and IC methods revealed the atomic ratios of Fe and Cl in the sample, respectively. We recorded three independent measurements to confirm the reproducibility of the x value.A sample of the β-FeOOH nanorods was removed from the colloidal solution before the HP/HT treatment. The remaining colloidal solution was dried in a Petri dish at 2.250,52–54 Approximately 40 mg of the sample was sealed into a Pt capsule, and then packed into an octahedral O pressure medium with one side length of 14 mm (Mino Ceramics), together with a BN sleeve (Denka) and a graphite heater . The O octahedron was placed at the corners of eight WC anvils  and compressed at ca. 25 °C and 2, 4, 6, 8, 10, or 12 GPa  to compare the OER activities of α- and β-FeOOH sources. The applied pressure was 10 GPa, and the heating temperature was 400 or 500 °C. XX-YY”, where XX and YY correspond to the applied pressure and heating temperature, respectively. For example, sample 2-200 was treated with a pressure 2 GPa and 200 °C. The two samples synthesized with the α-FeOOH source are labeled with the prefix α, e.g., α-10-400 and α-10-500.The pristine β-FeOOH sample was heated below 800 °C and under pressures of 2–12 GPa using Walker-type equipment at Osaka Prefecture University. GPa see . After r2.3θ range.55 The wavelength of the X-rays was determined to be 0.799436(2) Å using standard silicon powder (NIST 640d). Rietveld analyses were performed using RIETAN-FP software,56 and crystal structures were drawn with the VESTA software.57The crystal structures were examined with powder XRD using Fe-Kα radiation  and synchrotron radiation sources. The synchrotron XRD patterns were measured at the BL5S2 beamline at the Aichi Synchrotron Radiation Center using a two-dimensional (2D) detector, PILATUS 100K (Dectris). Each sample was packed into a borosilicate glass capillary with a diameter of 0.3 mm (WJM-Glas) and then exposed to X-rays for 20 min to acquire XRD data over a wide 22O3  as a reference. Approximately 3 mg of α-Fe2O3 and BN powder  were mixed with a mortar and pestle and then pressed into a pellet with a diameter of 10 mm and a thickness of ca. 0.2 mm. XAS data were recorded at ca. 25 °C in transmission mode. Fe K-edge X-ray absorption near-edge structure (XANES) and the Fourier transform of the k3-weighted Fe K-edge extended X-ray absorption fine structure (EXAFS) spectra were analyzed using ATHENA and ARTHEMIS software.58 The experimental setup for XAS and spectral analyses are described in further detail elsewhere.4,59The local structures and electronic states of pristine β-FeOOH and sample 8-400 were investigated by XAS at the Toyota beamline (BL33XU) at SPring-8. XAS was also performed on Fe metal  and α-Feca. 25 °C and liquid-nitrogen temperature (ca. −196 °C), whereas sample α-10-400 was examined only at ca. 25 °C. The radiation source was a 57Co/Rh matrix with a radioactivity of 370 MBq for pristine β-FeOOH and sample 8-400, while 1.85 GBq for sample α-10-400. A multi-channel-analyzer with 512 channels  was employed to acquire spectral data. One channel was determined to be 0.055598 mm s−1 for pristine β-FeOOH and sample 8-400, while 0.0791871 mm s−1 for sample α-10-400, by calibrating the Doppler velocity with Fe foil. Spectral analyses were conducted using MossWinn 4.0i software.60We performed Mössbauer spectroscopy on pristine β-FeOOH and samples 8-400 and α-10-400 to examine the valence state and local environment of their respective Fe ions. Pristine β-FeOOH and sample 8-400 were measured (without a magnetic field) at x upon heating and the thermal stabilities of β-FeOOH and ε-FeOOH. TG curves were recorded with a heating rate of 20 °C min−1 up to 1000 °C under flowing air (100 mL min−1). After the TG analysis, XRD measurements were performed using an Fe-Kα radiation source .Thermogravimetric  analyses were conducted for pristine β-FeOOH and samples 8-100 through 8-500, to clarify changes in dave) were determined by using the ImageJ software.61Particles sizes and morphologies were examined in two different types of scanning electron microscope (SEM), that is, a conventional system with a tungsten filament  and a field-emission (FE) system using an emitter tip . For the conventional system, 1 mg of sample, which was attached to carbon tape, was coated with electrically conductive Au particles . Meanwhile, for the FE system, samples were coated with Os particles . The accelerating voltages were 15 and 1 kV for the conventional and FE-SEM, respectively. Particle distribution and average size (− ions in the particles of pristine β-FeOOH and sample 8-400 with an energy dispersive X-ray (EDX) spectrometer . The specimens were prepared through ion (Ar) milling under an accelerating voltage of 6 kV at −160 °C . We recorded FE-SEM images using Everhart–Thornley-type detector and backscattered electron (BSE) images using an in-lens detector . The accelerating voltage was 5 or 7 kV.We also investigated the distribution of Cl2.43–5i.e., pristine β-FeOOH, samples 8-200, 8-400, 12-500, pristine α-FeOOH, and samples α-10-400 and α-10-500. We chose these samples considering each crystal phase, crystallinity, particle size/morphology, and the FeOOH source employed. Approximately 0.3–0.5 mg of each sample was attached to carbon paper  with a geometrical surface area of 1 cm2 (1 cm × 1 cm) and used as a working electrode (WE). This procedure is slightly different from the method previously reported,3–5,48 where a colloidal solution of β-FeOOH nanorods was injected into CP. The total surface area of the electrode was 2 cm2 by adding both the front and back sides. The counter electrode (CE) was a Pt wire, whereas the reference electrode (RE) was Ag/AgCl. Linear sweep voltammetry  was performed at a scan rate of 5 mV s−1, and chronoamperometry was conducted at +0.7 V vs. Ag/AgCl for 1800 seconds to clarify the stability of the anodic currents. Electrochemical impedance spectroscopy  was performed on pristine β-FeOOH and samples 8-200, 8-400, and 12-500 in the frequency range from 0.1 to 105 Hz under flowing Ar (20 mL min−1).We examined the OER activity of seven samples in an Ar-purged 1 M KOH solution (pH = 13.6) with a three-electrode configuration see ;3–5i.e.,33.122–26 We first analyzed the data using the tetragonal symmetry with I4/m space group,26 providing reliability indices of Rwp = 5.99% and S = 1.09. However, employing the monoclinic symmetry with I2/m space group improved these reliability indices to Rwp = 4.11% and S = 0.74. The tetragonal symmetry only has one Fe site, whereas two Fe sites exist in monoclinic symmetry. As described later, the presence of two Fe sites was confirmed by Mössbauer spectroscopy. The lattice parameters were determined to be am = 10.4758(20) Å, bm = 3.0282(2) Å, cm = 10.5427(22) Å, and βm = 89.51(2) ° by ignoring the presence of H+ ions. The results of the Rietveld analysis are summarized in Fig. S1dFe–O) were calculated to be 1.867(13) Å × 1, 2.175(14) Å × 1, 1.948(6) Å × 2, and 2.242(7) Å × 2, whereas that between Fe and Fe ions (dFe–Fe) was 3.028(1) Å. The lattice parameters were similar to previously reported values,22–24 but two of the six dFe–O values [1.867(13) and 2.175(14) Å] slightly differ from those in the literature (1.90 and 2.05 Å).26 The relatively variable dFe–O value probably originated from the low aspect ratio of pristine β-FeOOH; i.e., the diameter and length are 3 and 15 nm, respectively.3–5Pmn21 space group, i.e., an ε-FeOOH-type structure. The Rietveld analysis revealed orthorhombic lattice parameters of ao = 2.9688(7) Å, bo = 4.4558(10) Å, and co = 5.0077(12) Å (not shown). The XRD pattern of 8-150 exhibits broad and weak diffraction lines, indicating low crystallinity and/or a small crystallite size. By contrast, sample 8-200 shows sharp and strong diffraction lines with the ε-FeOOH-type structure (2O3 (hematite) phase, and sample 8-800 consisted of α-Fe2O3 and Fe3O4 (magnetite) phases, wherein the weight fraction of the latter phase was estimated to be 92.8%. The main phase for each sample is summarized in 2O3 is represented by3O4 is represented bySample 8-100 also maintained the β-FeOOH-type structure, whereas sample 8-150 possessed a different crystal structure from pristine β-FeOOH. After careful examination, the crystal structure of sample 8-150 was assigned to the orthorhombic symmetry with the tructure . Furtherw) were calculated to be −10.14% for 3O4 significantly differs from the situation under ambient pressure, where α-Fe2O3 remains stable up to 1000 °C.1,16,21,22The weight losses , 4.4570(1), and 4.9527(1) Å, respectively, which slightly differ from those for sample 8-150 owing to differences in the crystallinity. The dFe–O values were calculated to be 1.942(3) Å × 1, 2.137 Å × 1, 1.970(1) Å × 2, and 2.097(1) Å × 2, whereas dFe–Fe was 3.002(1) Å. These structural parameters are comparable with those for previous ε-FeOOH compounds that were synthesized from an α-FeOOH source.40,41,54 Based on the geometry of polyhedral distortions, the distortion index in the FeO6 octahedron [DI(FeO6)] is defined as62di and dave are the ith and average dFe–O, respectively. Because the DI(FeO6) values for pristine β-FeOOH and sample 8-400 (ε-FeOOH) were calculated to be 0.100 and 0.037, respectively, the HP/HT environment evidently decreased the distortion in the FeO6 octahedron. Similar results were obtained for rhombohedral-structured LiCo0.64Mn0.36O2  values. As understood by the Fe K-edge XANES spectrum of α-Fe2O3  of approximately 1.5 and 2.7 Å, which were assigned to dFe–O and dFe–Fe, respectively. More specifically, dFe–O = 1.48(1) Å and dFe–Fe = 2.66(1) Å, for pristine β-FeOOH, whereas dFe–O = 1.54(1) Å and 2.65(1) Å for sample 8-400. The EXAFS spectrum of pristine β-FeOOH is similar to the spectrum that we recently reported for β-FeOOH,3–5 although to the best of our knowledge, the EXAFS spectrum of ε-FeOOH has never been reported thus far.Two major peaks appear in the EXAFS spectra centered at phase-uncorrected radial distances  and temperatures (ca. 200 °C) in the case of β-FeOOH. As mentioned above, the bulk modulus of β-FeOOH [=284(1) GPa] at room temperature is the highest among the FeOOH polymorphs.42 Considering this result, the required pressure for the transition to ε-FeOOH would be higher in the β-FeOOH, but the obtained fact was contrary to the expectation. As illustrated in − ions. The lower pressure-induced transition indicates a fragility of β-FeOOH at high temperatures above 100 °C. Indeed, β-FeOOH was reported to decompose into α-Fe2O3 and H2O at lower temperatures (under ambient pressure) compared to α-FeOOH, due to its strengthening of the hydrogen bond environments.65 Note that the structural phase transition from β-FeOOH to ε-FeOOH should be accompanied by breaking and reforming the Fe–O bonds. This is because the Pmn21 (ε-FeOOH) space group is a maximal nonisomorphic subgroup of the Pnma (α-FeOOH) space group, whereas I2/m or I4/m (ε-FeOOH) space group is not a minimal nonisomorphic supergroup of the Pmn21 space group, based on the crystallographic relationship.66 Further HP/HT studies for various amounts of Cl− ions and theoretical calculations would reveal detailed mechanisms of the structural phase transition from β-FeOOH and ε-FeOOH.3.216,17,19,22,24–26x in FeOx1−(OH)x1+Clx ranges from 0.003 to 0.37. Moreover, Cai et al.16 reported a decrease in x upon heating accompanying the structural phase transition to α-Fe2O3 (under ambient pressure). In this section, we discuss changes in x and the particle morphologies during the successive phase transitions of β-FeOOH → ε-FeOOH → α-Fe2O3 → Fe3O4. Only the composition of pristine β-FeOOH was analyzed, whereas TG was employed to analyze the HP/HT-treated samples because their weights were limited to ca. 40 mg.According to previous studies on β-FeOOH,x = 0.12 for pristine β-FeOOH, suggesting the chemical formula FeO0.88(OH)1.12Cl0.12. The decomposition reaction upon heating is described by26w is calculated to be −14.4%. However, as shown in w of pristine β-FeOOH approaches −21% at 1000 °C; specifically, the weight (w) gradually decreases until a slight drop at ca. 400 °C and then remains at 79% up to 1000 °C. The discrepancy between the observed and calculated Δw values is attributed to adsorbed water, residual ethylenediamine, and crystalline water in the sample. The gradual decrease in w and/or relatively large Δw were also observed in previous thermal studies on β-FeOOH.1,16,21,26The combined ICP-AES and IC analyses confirmed w value (=−15.9%), whereas the curves of samples 8-150, 8-200, 8-300, and 8-400 show different temperatures at which w rapidly decreases. For instance, sample 8-400 exhibits a rapid decrease in w at ca. 300 °C, which is ca. 100 °C lower than those of pristine β-FeOOH and sample 8-100. w at 1000 °C and the heating temperature during the HP/HT synthesis. Because the Δw values of samples 8-300 and 8-400 are close to the calculated Δw value (=10.14%) based on − ions. In other words, above 150 °C, the Cl− ions start to evaporate out of the β-FeOOH lattice, which is associated with the structural phase transition to ε-FeOOH. The disappearance of Cl− ions is strongly related to the mechanism underlying the structural phase transition; i.e., as described above, it inevitably induces the breaking and reforming of Fe–O bonds.The TG curve of sample 8-100 is similar to that of pristine β-FeOOH except for the Δ− ions in the ε-FeOOH lattice by comparing its TG curve with that of sample α-10-400 1.12Cl0.12 (at 8 GPa) are represented by3O4 above 800 °C is described by − ions on the surface of ε-FeOOH particles (sample 8-400) is discussed later.We confirmed the absence of Cl0 Fig. S8. Furthere Fig. S9. Accordi3O4 phase, exhibits a non-uniform shape with an average size of 50 μm. These changes indicate that the structural phase transitions under HP cause aggregation and change the particle size. Similar results were also observed for the phase transitions from α-FeOOH to ε-FeOOH54 and from spinel-structured Li[Li1/3Ti5/3]O4 to columbite-structured TiO2.6761 The dave values of β-FeOOH, 8-400, and 12-500 were estimated to be 0.14, 4.7, and 2.9 μm, respectively.We next investigated the effects of the applied pressure. Fig. S103.3j) was normalized by the total surface area of the electrode (2 cm2), while the potential vs. Ag/AgCl was converted into a potential vs. reversible hydrogen electrode (RHE). The anodic current of pristine β-FeOOH rapidly increases at ca. 1.60 V, which is consumed for the water oxidation accompanying with oxygen evolution.3,4 Sample 8-400 obviously exhibits a lower OER overpotential than that of pristine β-FeOOH; specifically, the anodic current rapidly increases above 1.55 V, in which threshold potential for OER is negatively shifted by ca. 60 mV compared to that of pristine β-FeOOH. For instance, the potential at which j = 1 mA cm−2 (Ej=1 mA cm−2) was 1.62 V for pristine β-FeOOH and 1.56 V for 8-400. Sample 12-500 indicates a similar threshold potential to pristine β-FeOOH, whereas sample 8-200 shows a higher threshold potential (ca. 1.75 V). The Ej=1 mA cm−2 values are summarized in vs. RHE. Sample 8-400 still maintains the highest anodic current density of ca. 15 mA cm−2, which is ca. 1.5 times greater than that of pristine β-FeOOH. To reveal the origins of this OER activity, Rtotal) of the electrode by the radius of the semicircle. The radius of sample 8-400 is apparently smaller than that of pristine β-FeOOH, indicating the decrease in Rtotal in the case for sample 8-400. More specifically, the EIS spectra were fitted to the equivalent circuit model shown in the inset of i.e., a solution resistance (Rsol), Rtotal, and a constant-phase element (CPE). Here, the CPE is represented by 1/(iwQ)α; when α = 1, Q has the dimension of capacitance (F); α < 1 indicates surface heterogeneity in the system, such as the roughness of the electrode. As summarized in Table S1,†Rtotal = 9.42 and = 5.11 Ω for pristine β-FeOOH and sample 8-400, respectively, which suggests that one cause for enhancing OER activity of sample 8-400 is due to its lower Rtotal.39–41 α-FeOOH was also transformed into ε-FeOOH under HP/HT conditions. We thus examined the OER activity of pristine α-FeOOH and samples α-10-400 and α-10-500 for comparison. Here, samples α-10-400 and α-10-500, which crystallized into the ε-FeOOH phase. Ej=1 mA cm−2 values of pristine α-FeOOH and samples α-10-400 and α-10-500 are listed in As reported previously,If we focus on the highest OER activity, this activity was enhanced in the order by α-FeOOH → β-FeOOH → ε-FeOOH. We next investigated the reproducibility of the OER activity among different electrodes prepared with the same sample. Fig. S133.4f(E) and B0 are the counts at the Doppler velocity (E) and the baseline count, respectively; Ii, Γi, and xi0, are the intensity, full-width at the half-maximum, and peak center at the ith peak, respectively; δ is the isomer shift; Δ is the quadruple splitting; and H is the internal field. The doublet peak consists of two components (i = 1 and 2), which are consistent with the two Fe sites, in the monoclinic symmetry. δ, Δ, Γ, H, and the component ratio. The Mössbauer parameters are comparable with previous results for β-FeOOH,24 although the origin of the two components was not fully understood in that investigation because the symmetry was wrongly concluded to be tetragonal.As sample 8-400 exhibited the best OER activity, we further characterized sample 8-400 using Mössbauer spectroscopy and EDX analysis. δ = 0.37 mm s−1, Δ = −0.24 mm s−1, Γ = 0.26 mm s−1, and H = 46.9 T at ca. 25 °C. Because these Mössbauer parameters are similar to previous results for ε-FeOOH ,68 the major contribution is attributed to the ε-FeOOH phase. This mixed nature of sample 8-400 differs from the Rietveld results shown in Fig. S2†Sample 8-400 exhibits sextet peaks at both 25 and −196 °C, indicating that the sample is in an (antiferro)magnetic state . A caref− ions in the ε-FeOOH lattice. However, we further investigated the distribution of Cl− ions, particularly on the surface. − ions are inhomogeneously distributed on a specific particle after the phase transition from β-FeOOH to ε-FeOOH. By contrast, we confirmed that the Cl− ions are homogeneously distributed in the particles of pristine β-FeOOH, as shown in Fig. S15.According to the TG analyses , sample 3.5dave values of sample 8-400 (=4.7 μm) and pristine β-FeOOH (= 0.14 μm), the active surface area of sample 8-400 should be smaller than that of pristine β-FeOOH. This indicates that the OER activity of sample 8-400 is attributed to an intrinsic and high OER performance of ε-FeOOH. In other words, a small Rbulk contributes to the relatively low Rtotal of sample 8-400 (= 5.11 Ω).Considering the 7O12 (A = La and Ca) which were synthesized by the HP/HT method,69 their OER activities relate with the nearest bond distance between O atoms (dO–O); the overpotential decreases with dO–O. Indeed, as shown in Fig. S17,dO–O value was decreased in the order by α-FeOOH (=3.035 Å) → β-FeOOH (=3.023 Å) → ε-FeOOH (=3.006 Å). Furthermore, it is interesting to compare the OER activities of ε-FeOOH and β-MnO2 (pyrolusite), as they are almost isostructural.27 Yan et al. clarified that the phase transition from β-MnO2 to α-MnO2 (cryptomelane) improved the OER activity,30 which is the opposite of the trend observed in this study. Note that both β-MnO2 and ε-FeOOH are classified as rutile, although ε-FeOOH is regarded as a distorted rutile structure, as in the case of columbite-type TiO2.67 Thus, there is a possibility that the superior OER activity of ε-FeOOH owes to its characteristic structural environments such as the shortest dO–O and slightly distorted FeO6 octahedra.Based on studies of quadruple manganese perovskites AMnet al.6 recently reported that using mixed phases of α- and β-FeOOH or β- and δ-FeOOH enhanced the OER activity owing to the formation of oxygen vacancies on the surface. As sample 8-400 contains a small amount (3%) of β-FeOOH, synergetic effects of the mixture of ε- and β-FeOOH could contribute to its OER activity.Another possibility would come from the mixed nature of sample 8-400. Hu Anyway, comparing the OER activities of samples 8-200 and 12-500 suggests that other factors besides the crystalline phase, such as the particle morphology and particle size, affect the OER activities. Further theoretical studies on FeOOH polymorphs would reveal details of the reaction mechanism underlying the OER activity of ε-FeOOH.3.6Ej=1 mA cm−2 = 1.56 V vs. RHE. This overpotential is still higher than those of other non-noble metal-based electrocatalysts; for instance, an exfoliated NiFe layered double hydroxide indicated Ej=1 mA cm−2 ≃ 1.52 V vs. RHE.72 The relatively large overpotential of ε-FeOOH is caused by its large particle size (dave > 2 μm), namely low active site for OER, as shown in ca. 0.05 V) overpotential.3–5,48 Therefore, investigations for decreasing particle size and optimizing composite electrodes could enhance OER activity of ε-FeOOH. The pressure-phase diagram shown in 70,71 although the present study is focused on the electrocatalytic properties.We revealed that ε-FeOOH exhibited the best OER activity among the FeOOH polymorphs investigated. However, as summarized in 4x1−(OH)x1+Clx with x = 0.12, and their OER activities. To our best knowledge, we firstly demonstrated that β-FeOOH transformed into ε-FeOOH above 6 GPa and 100 °C. In addition, Cl− ions, which were initially hosted in the [2 × 2] tunnels, evaporated from the surface during the structural phase transition. Above 400 °C, ε-FeOOH decomposed into α-Fe2O3 or Fe3O4, while releasing oxygen from the lattice. Above 6 GPa, ε-FeOOH was stabilized at higher temperatures; for instance, at 12 GPa, ε-FeOOH decomposed into α-Fe2O3 above 700 °C. The obtained pressure–temperature phase diagram of β-FeOOH differed from that of α-FeOOH, probably owing to the open framework of β-FeOOH and partial occupation of Cl− ions. In the viewpoint from earth and space sciences, this information is crucial for in-depth understanding of evolution and dynamics of the Earth interior particularly at the upper mantle.We examined HP/HT phases of β-FeOOH nanorods, more specifically, β-FeORtotal. Moreover, Mössbauer spectroscopy revealed that sample 8-400 contains a small amount (3%) of the β-FeOOH phase, possibly leading to synergetic effects from β- and ε-FeOOH. The residual Cl− ions were inhomogeneously distributed on the surfaces of particles in sample 8-400, resulting in relatively scattered OER activities along different composite electrodes fabricated from this sample. Nonetheless, ε-FeOOH is a possible candidate as a next-generation earth-abundant OER catalyst, particularly because material options are currently limited. The established pressure–temperature phase diagram is useful for further improving the OER activity of ε-FeOOH, because particle sizes, particle morphologies, and distribution of Cl− ions strongly depend on the HP/HT conditions.Sample 8-400 (ε-FeOOH), which was synthesized at 8 GPa and 400 °C, exhibited superior OER activity over the parent β-FeOOH or α-FeOOH. EIS results showed that this superior OER activity is due to its low There are no conflicts to declare.RA-010-D0RA09895G-s001


E)-N′--2-acetohydrazide ethyl acetate monosolvate, -N′--4-hydroxybenzohydrazide ethanol monosolvate and -N′-(6-methoxy-2-phenylchroman-4-ylidene)-2-acetohydrazide. All three hydrazones are in the E isomeric form and exhibit a pucker at the chiral carbon atom. The naringenin-derived hydrazones both show intramolecular hydrogen bonding between the hydrazone nitrogen atom and a hydroxy group on the chromane ring.The crystal structures of racemic mixtures of three new flavanone-hydrazones are reported: -N′--2-acetohydrazide ethyl acetate monosolvate, C27H22N2O5·C4H8O2, and of -N′--4-hydroxybenzohydrazide ethanol monosolvate, C22H18N2O6·C2H5OH, both exhibit an intramolecular O—H⋯N and multiple intermolecular O—H⋯O and C—H⋯O-type hydrogen bonds. The third structure, that of -N′-(6-methoxy-2-phenylchroman-4-ylidene)-2-acetohydrazide, C28H24N2O4, has only one intermolecular N—H⋯O-type hydrogen bond. In each of the three cases, the crystal packings are stabilized by π–π stacking interactions between various aromatic components of symmetry-related molecules. The chiral carbon atom of the substituted chromane ring system in each case is puckered away from rest of the ring system.The crystal structures of racemic mixtures of three new flavanone-hydrazones in the centrosymmetric space group ( Flavonoids include various subcategories – chalcones, flavones, flavanones, flavanols, isoflavones, anthocyanins – all of which have demonstrated differential health benefits such as anti-oxidative, anti-inflammatory, anti-mutagenic, and anti-carcinogenic properties , 0.397 (6), 0.331 (5) Å, and of θ = 57.9 (4), 53.9 (6), 58.1 (7)°, respectively. The chiral carbon (C8) in each case is displaced between 0.454 and 0.580 Å from the chromane ring planes. The puckering is similar to that seen in the previously reported structure  of the 4-hydroxyphenyl ring makes dihedral angles of 77.128 (5) and 83.872 (5)°, respectively, with the chromane ring system. An intramolecular O—H⋯N hydrogen bond exists between one of the hydroxy groups on the chromane ring and the nitrogen of the hydrazone group . Another hydroxy group on the chromane ring participates in a hydrogen bond with the carbonyl group of the solvent ethyl acetate molecule . The naphthalene ring system is close to perpendicular to the chromane ring system [dihedral angle 77.692 (5)°].In compound II, the 4-hydroxyphenyl ring of the hydrazone moiety is coplanar with the chromane ring [dihedral angle of 2.485 (3)° with the chromane ring system] whereas the other hydroxyphenyl ring is almost perpendicular [75.449 (5)°] to the chromane ring system. The chiral carbon of chromane ring (C8_1) and the methyl carbon (C2_1) of the solvent molecule show positional disorder. An intramolecular O—H⋯N hydrogen bond exists between one of the hydroxy groups on the chromane ring and the nitrogen of the hydrazone group .In III, the phenyl ring makes a dihedral angle of 86.17 (1)° with the chromane ring system, while the naphthalene ring system is perpendicular to the chromane ring system [dihedral angle = 89.65 (1)°].In 3.I and II -amino}benzamide  was performed in the CCDC’s free Access Structures online database  and 2-acetohydrazide  were dissolved in ethanol (10 mL). Acetic acid  was added and the resultant solution was heated at reflux for 21 h. The precipitate was isolated via vacuum filtration and recrystallized from ethyl acetate via slow evaporation at room temperature to furnish clear, plate-shaped crystals suitable for X-ray analysis.For the preparation of II, naringenin  and 4-hydroxybenzohydrazine  were dissolved in ethanol (20 mL). Acetic acid  was added and the resultant solution was heated at reflux for 48 h. The precipitate was isolated via filtration and recrystallized from ethanol via slow evaporation at room temperature to furnish transparent yellow, plate-shaped crystals suitable for X-ray analysis.For the preparation of III, 6-methoxyflavanone , 2-acetohydrazide , and p-toluenesulfonic acid  were dissolved in toluene (15mL). The resultant mixture was heated at reflux for 12 h with a Dean–Stark apparatus. The solvent was removed and the crude product was purified on an automated flash chromatography system using a normal phase silica gel column with a gradient of hexane:ethyl acetate (70:30 to 0:100). Recrystallization of the purified compound from ethanol via slow evaporation at room temperature furnished yellow, needle-shaped crystals suitable for X-ray analysis.For the preparation of 6.3), 0.97 Å (CH2), 0.86 Å (NH) or 0.82 Å (OH). Isotropic displacement parameters for these atoms were set to 1.2  or 1.5  times Ueq of the parent atom. In II, the positional disorder of the chiral carbon (C8) and phenoxy ring atoms (C10 through C15) refined to a percentage population ratio of 66/34, and that of the solvent (ethanol) molecule to 57/43, necessitating the use of a total of 136 restraints. The idealized Me of the ethanol molecule were refined as rotating group(s): C2_3 and C2_4 (H2A_3 through H2C_4).Crystal data, data collection and structure refinement details for all three structures are summarized in Table 410.1107/S2056989023001184/jy2026sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989023001184/jy2026Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026Isup5.molSupporting information file. DOI: 10.1107/S2056989023001184/jy2026IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026IIsup6.molSupporting information file. DOI: 10.1107/S2056989023001184/jy2026IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026IIIsup7.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026Isup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026IIsup9.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026IIIsup10.cmlSupporting information file. DOI: 10.1107/S2056989023001184/jy2026sup11.txthkl file for data_I. DOI: 10.1107/S2056989023001184/jy2026sup12.txthkl file for data_II. DOI: 10.1107/S2056989023001184/jy2026sup13.txthkl file for data_III. DOI: 10.1107/S2056989023001184/jy2026sup14.txtRES files for data_block I. DOI: 10.1107/S2056989023001184/jy2026sup15.txtRES file for data_block II. DOI: 10.1107/S2056989023001184/jy2026sup16.txtRES file for data_block III. DOI: 10.1107/S2056989023001184/jy2026sup17.txtmatched hkl file for data_III. DOI: 10.1107/S2056989023001184/jy2026sup18.txtmatched hkl file for data_II. DOI: 10.1107/S2056989023001184/jy2026sup19.txtmatched hkl file for data_I. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026sup20.zipSHELX data. DOI: Click here for additional data file.10.1107/S2056989023001184/jy2026sup21.zipSHELX data. DOI: 10.1107/S2056989023001184/jy2026sup22.txtStructure 1 res file. DOI: 10.1107/S2056989023001184/jy2026sup23.txtStructure I hkl file. DOI: 10.1107/S2056989023001184/jy2026sup24.txtStructure II res file. DOI: 10.1107/S2056989023001184/jy2026sup25.txtStructure II hkl file. DOI: 10.1107/S2056989023001184/jy2026sup26.txtStructure III res file. DOI: 10.1107/S2056989023001184/jy2026sup27.txtStructure III hkl file. DOI: 2240752, 2240751, 2240750CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


However, these cells represent a small fraction of the peripheral T-cell pool and require activation and proliferation for clinical benefits. In cord blood, there are some γδ T cells, which exhibit a naïve phenotype, and mostly include Vδ1+ T cells. In this study, we investigated the effect of CD3 signaling on cord blood γδ T-cell proliferation using K562-based artificial antigen presenting cells expressing costimulatory molecules. There were significantly more Vδ1+ T cells in the group stimulated with anti-CD3 antibody than in the group without. In cultured Vδ1+ T cells, DNAM-1 and NKG2D were highly expressed, but NKp30 and NKp44 showed low expression. Among various target cells, Vδ1+ T cells showed the highest cytotoxicity against U937 cells, but Daudi and Raji cells were not susceptible to Vδ1+ T cells. The major cytokines secreted by Vδ1+ T cells responding to U937 cells were Granzyme B, IFN-γ, and sFasL. Cytotoxicity by Vδ1+ T cells correlated with the expression level of PVR and Nectin of DNAM-1 ligands on the surface of target cells. Compared to Vδ2+ T cells in peripheral blood, cord blood Vδ1+ T cells showed varying cytotoxicity patterns depending on the target cells. Here, we determined the ideal conditions for culturing cord blood Vδ1+ T cells by observing that Vδ1+ T cells were more sensitive to CD3 signals than other subtypes of γδ T cells in cord blood. Cultured cord blood Vδ1+ T cells recognized target cells through activating receptors and secreted numerous cytotoxic cytokines. These results are useful for the development of tumor immunotherapy based on γδ T cells. ParticuRecently, γδ T cells have been applied for adoptive tumor immunotherapy. However, they require activation and propagation for clinical benefits as they are only a small fraction of the peripheral T-cell pool . A Vδ2+ Cord blood is whole blood obtained from the umbilical cord at childbirth and is an alternative source of stem cells useful for hematopoietic stem cell transplantation . Most γδHuman cord blood and peripheral blood were provided by the Catholic Hematopoietic Stem Cell Bank. The use of human material was reviewed and approved by the Institutional Review Board of the College of Medicine, Catholic University of Korea, Seoul, Republic of Korea (permit No. MC18SESI0111). All subjects provided written informed consent for sample donation in accordance with the Declaration of Helsinki.2.K562, U937, Daudi, and Raji cells were obtained from the American Type Culture Collection . aAPCs in which costimulatory molecules  were stably expressed in K562 cells , using a5 cells/mL, were co-cultured in a 24-well plate with irradiated (100 Gy) aAPCs at a 2:1 T cell:aAPCs ratio with 200 U/mL of IL-2  with/without 500 ng/mL of anti-CD3 antibody  and added with 200 U/mL of IL-2  on day 3. To culture total γδ T cells, cells were further cultured under the same conditions on day 7, refreshed with 200 U/mL of IL-2 in complete media  on day 10, and cultured until day 14.Cord blood mononuclear cells were isolated by Ficoll-Hypaque  density gradient centrifugation. γδ T cells were negatively isolated using magnetic microbeads  according to the manufacturer’s instructions. RPMI 1640 medium  supplemented with 10% fetal bovine serum , 1% L-glutamine , and 1% penicillin/streptomycin  was used as a basic culture medium for γδ T cells. γδ T cells, at 5 × 105 cells/mL, were co-cultured in a 24-well plate with irradiated (100 Gy) aAPCs at a 2:1 T cell:aAPCs ratio in the presence of 200 U/mL of IL-2  with 500 ng/mL of anti-CD3 antibody . On day 10, the media including 200 U/mL of IL-2  were refreshed, and cells were cultured until day 14. To calculate fold expansion, after sorting on day 7, seeding of 5 × 105 cells per mL was cultured for 1 week; the expanded fold compared to 7 days was calculated.To culture pure Vδ1+ T cells, cells were labeled with an anti-TCRVδ1-APC (TS8.2) antibody on day 7, and analyzed and sorted by a fluorescence-activated cell sorter FACS Aria Fusion  using FACSDiva software (BD Biosciences). Vδ1+ T cells, 5 × 105 cells/mL in a 24-well plate. From day 7, γδ T cells were treated with IL-2 (1000 U/mL) every 3–4 days, stimulated weekly with irradiated (100 Gy) aAPCs at a 2:1 T:aAPC ratio. On day 21, cells were frozen using a controlled-rate freezer  and cryopreserved in liquid nitrogen until further use. For cytotoxicity assays, frozen γδ T cells were thawed and cultured overnight in the presence of IL-2 (1000 U/mL).γδ T cells were maintained in RPMI 1640 medium  supplemented with 10% fetal bovine serum , 1% L-glutamine , and 1% penicillin/streptomycin . On day 0, PBMCs were stimulated with 3 μM of zoledronic acid  in the presence of 1000 U/mL of IL-2  and seeded at 5 × 10γδ T-cell analyses were performed on day 0, day 7, and day 14. Cells were harvested and stained with the following antibodies: anti-TCRVδ2-FITC (B6), anti-CD45RA-PE (HI100), anti-CD27-APC (M-T271), anti-CD226-Brilliant Violet 421 (11A8), anti-TIGIT-APC (A15153G), anti-CD152-Brilliant Violet 421 (BNI3), anti-CD314 (NKG2D)-APC/Cyanine 7 (1D11), anti-CD336 (NKp44)-PE/Cyanine 7 (P44–8), and anti-CD337 (NKp30)-PerCP/Cyanine5.5 (P30–15), purchased from BioLegend ; and anti-CD279-PE (MIH4), anti-CD274-APC-eFluor 780 (MIH1), anti-TCRVδ1-APC (TS8.2), and anti-CD45-PerCP-eFluor 710 (HI30), purchased from Invitrogen. Acquisition was performed on a FACSCANTO flow cytometry instrument  using the FACSDiva software. Acquired data were analyzed with FlowJo .2 in the dark and washed twice with phosphate-buffered saline and suspended in 10% RPMI at 1 × 105 cells/mL. Effector and target cells were co-cultured at the indicated effector:target ratios in a 96-well U-bottom plate. Triton X-100 was added to a final concentration of 2% for maximum fluorescence release and only target cells were cultured without effector cells for spontaneous release. After a 4 h incubation, plates were centrifuged at 2500 rpm for 5 min, and the supernatants carefully transferred to a black opaque plate. The fluorescence was measured with a Synergy H1 microplate reader . The specific release was calculated as  − (Spontaneous Fluorescence) / (Maximum Fluorescence) − (Spontaneous Fluorescence) × 100.For the cytotoxicity assay using calcein AM (Invitrogen), target cells were stained with 5 μM calcein AM and incubated for 20 min at 37 °C and 5% COFor secretome analysis, Vδ1+ γδ T cells and Daudi, U937 cells were co-cultured in a 96-well U-bottom plate at a 2:1 ratio for 4 or 24 h; plates were centrifuged at 2500 rpm for 5 min, and the supernatants were carefully transferred to 5 mL round tubes. The supernatants were analyzed using a LEGENDplex human CD8/NK panel (BioLegend) according to the manufacturer’s protocol.t-tests, and one-way and two-way analysis of variance. All analyses were performed using Prism 5.01 software .Statistical significance was determined by paired To investigate the effect of CD3 signals on the γδ T-cell subtypes of cord blood, γδ T cells negatively isolated with magnetic beads were stimulated for two weeks using irradiated K562-based aAPCs expressing costimulatory molecules, in the presence or absence of anti-CD3 antibody, respectively. The subtypes of γδ T cells negatively isolated from 31 cord blood samples were 11.2% ± 12.5% of Vδ1+ γδ T cells, 2.6% ± 2.3% of Vδ2+ γδ T cells, and 86.1% ± 14% of Vδ1-Vδ2- γδ T cells a. A reprThe isolation and culture of pure Vδ1+ T cells isolated directly from cord blood were not efficient due to low cell recovery and viability. Therefore, after culturing negatively isolated γδ T cells with aAPCs and anti-CD3 antibody for one week, Vδ1+ T cells were separated using a FACS cell sorter and cultured for one more week. The proportions of Vδ1+ T cells before and after sorting on day 7 were 12.2% ± 9.4% and 90.6% ± 8%, respectively. After one week of additional culture, Vδ1+ T cells accounted for 93.2% ± 4.4% of all cells a,b. The After sorting on the 7th day, about 93% of Vδ1+ T cells were harvested on day 14. To investigate the characteristics of these expanded Vδ1+ T cells, both the differentiation status and surface expression of relevant molecules were investigated. The differentiation markers of Vδ1+ T cells harvested on day 14 were analyzed using flow cytometry. Most cultured Vδ1+ T cells had differentiated into central memory cells  and effector memory cells . There were very low frequencies of naïve  and CD45RA+ terminal effector memory cells  Vδ1+ T cells a.n = 8). Among the activating receptors, DNAM-1 (97.7% ± 2.1%) and NKG2D (53.3% ± 11.1%) showed high expression whereas NKp30 (5.1% ± 7.4%) and NKp44 (0.2% ± 0.1%) showed low expression. The expression of inhibitory molecules such as PD-1 (8.3% ± 4.5%), CTLA-4 (7.9% ± 11.6%), and PD-L1 (0.1% ± 0.1%) were also very low , and with PVR and Nectin of DNAM-1 ligands   . These r= 0.032) a,b. ReprAfter co-culturing Vδ1+ T cells and target cells at an E:T ratio of 2:1 for 24 h, supernatants were harvested, and the molecules and cytokines secreted by Vδ1+ T cells were measured by a bead array. When co-culturing Vδ1+ T and U937 cells, the concentrations of Granzyme B, IFN-γ, and sFasL were significantly higher than those in Daudi cells .Additionally, we compared the cytotoxicity of cord blood Vδ1+ T cells and adult Vδ2+ T cells to U937, K562, and Daudi cells . Vδ2+ T Although αβ and γδ TCRs both require the CD3 complex for signaling and show similar structures, these receptors had different characteristics in terms of complex geometry, glycosylation pattern, plasma membrane organization, and the accessibility of signaling motifs in the CD3 intracellular tails . In the Cord blood Vδ1+ T cells showed high cytotoxicity in K562 and U937 cells, but not in Raji or Daudi cells. The anti-γδ TCR antibody could expand both the Vδ1 and Vδ2 subsets of peripheral blood γδ T cells. In the comparison of the cytotoxic activity between them, purified Vδ1 and Vδ2 subsets exhibited similar cytotoxicity against Daudi and K562 cells . HoweverDaudi cells showed the greatest difference in cytotoxicity of cord blood Vδ1+ T cells proliferated compared to adult peripheral blood Vδ2+ T cells. Vδ2+ T cells are activated by cells that accumulate HMBPP and/or IPP and can also be stimulated through receptors NKG2D and DNAM-1 of various stress-induced molecules expressed on tumor cells including MICA/B, UL16-binding proteins (ULBP1–6), Nectin-2, and PVR, respectively ,40. DaudIn a co-culture of Vδ1+ T cells and U937, the concentrations of Granzyme B, IFN-γ and sFasL were significantly higher than those of Daudi cells . PerforiIn summary, we established the conditions for culturing cord blood Vδ1+ T cells by observing that, among cord blood γδ T cells, Vδ1+ T cells were more sensitive to CD3 signals than other subtypes, and thus effectively proliferated. Cultured cord blood Vδ1+ T cells expressed high DNAM-1 and NKG2D, and showed strong cytotoxicity and high secretion of Granzyme B, sFasL, and IFN-γ by recognizing target cells with their ligands. Although Vδ2+ T cells derived from adult blood were compared with cord blood Vδ1+ T cells in this study, a direct comparison with Vδ1+ T cells derived from adult blood or Vδ1+ derived from other tissue sources is necessary to investigate the biology of human γδ T cells. Since the stimulation of Vδ1+ T cells with aAPC and anti-CD3 antibody used in this study was the same as the αβ T cell stimulation conditions, and the proliferation rate was similar, further study is needed to prove whether the structural properties of the Vδ1 TCR and CD3 complex would have similar structural properties to those of the αβ TCR.


P21) symmetry at room temperature. The two molecules in the asymmetric unit of the title compound, C16H14FNO3, exhibit different torsion angles along the central sp3 C—N bonds and are linked together through two N—H⋯O hydrogen-bonding interactions.The crystal structure exhibits monoclinic  symmetry at room temperature. The two independent molecules that comprise the asymmetric unit display distinct torsion angles of 173.2 (2) and 72.6 (2)° along the central sp3 C—N bond. In the crystal, hydrogen bonding through N—H⋯O contacts couples the asymmetric unit molecules into pairs that align in layers extending parallel to (100) via additional O—H⋯O interactions. The phenyl ring of one independent molecule was found to be disordered over two sets of sites in a 0.55 (3):0.45 (3) ratio.The title compound, C The resin was rinsed with three 3 ml aliquots of N-methyl-2-pyrrolidone (NMP) and three 2 ml aliquots of NMP:piperidine (4:1). The bottom of the vial was capped to prevent the acylating reagents from draining from the vial. To the resin was added 1.0 ml of a solution of p-fluorobenzoic acid (0.25 M) and hydroxybenzotriazole  in NMP and 0.5 ml of 0.5 M diisopropylcarbodiimide in NMP. The vial was capped, inverted three times, and allowed to sit. After four days, the vial was uncapped at both ends, and the resin was washed sequentially with two 3 ml portions of NMP, two 3 ml portions of tetrahydrofuran, and three 3 ml portions of dichloromethane. The reaction vial was placed over a collection vial, and the resin was washed twice with 2.0 ml of trifluoroacetic acid–dichloromethane–water (35:60:5) and once with 2.0 ml of dichloromethane. The collection vial containing the combined rinses was placed in a vacuum chamber to remove the solvents. Column chromatography  afforded 12.8 mg of the title compound . The purified product was crystallized from a dichloromethane solution layered with a solution of heptane and benzene. 1H NMR  δ 7.73 , 7.13 , 7.10 , 5.05 , 3.27 , 3.21 . 13C NMR  δ 172.23, 166.11, 164.09, 135.77, 129.49, 129.43, 129.29, 128.67, 127.26, 115.78, 53.55, 37.78.50 µmol of Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2414314620008974/wm4133sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620008974/wm4133Isup2.cmlSupporting information file. DOI: 1975451CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Abordagem Fistula First: ainda válida?”, com o DOI: https://doi.org/10.1590/2175-8239-JBN-2020-U001, publicado no BrazilianJournal of Nephrology, 2021;43(2):263-268 na versão em português:No artigo “Onde está escrito:Leia-se:


Two new salts of the 4-(4-nitrophenyl)piperazin-1-ium cation have been prepared by co-crystallization with aromatic carboxylic acids. The supramolecular assembly in the benzoate salt, which crystallizes as a mono-hydrate, is two dimensional, while that in the 2-carboxy-4,6-dinitrophenolate salt is three dimensional. N-(4-nitrophenyl)piperazine with benzoic acid gives the benzoate salt, which crystallizes as a monohydrate, C10H14N3O2·C7H5O2·H2O, (I), and similar co-crystallization with 3,5-dinitrosalicylic acid yields the 2-carboxy-4,6-dinitrophenolate salt, C10H14N3O2·C7H3N2O7, (II). In the structure of (I), a combination of O—H⋯O, N—H⋯O and C—H⋯O hydrogen bonds links the components into sheets, while in the structure of (II), the supramolecular assembly, generated by hydrogen bonds of the same types as in (I), is three dimensional. Comparisons are made with the structures of some related compounds.Crystal structures are reported for two molecular salts containing the 4-(4-nitrophenyl)piperazin-1-ium cation. Co-crystallization from methanol/ethyl acetate solution of  The second sub-structure, which includes the C—H⋯O hydrogen bond  alternate with n, n, 0.5), where n again represents an integer, so forming a chain of rings running parallel to the [110] direction d Table 1, takes ton Fig. 4. The comon Fig. 4 lies witx, y, z) and atom N24 at : this probably represents a dipolar attraction between negatively charged O and positively charged N atoms.The supramolecular assembly for compound (II)gs Fig. 7, and thegs Fig. 7. The com4.N-(4-nitrophenyl)piperazine concerned the chloride salt, which crystallizes as a monohydrate piperazine with aromatic carboxylic acids have recently been reported piperazine piperazine piperazine  in methanol (10 ml) was mixed with a solution of either benzoic acid  for (I)v/v, 20 ml). The solutions of the base and the corresponding acid were mixed, stirred at ambient temperature for 15 min, and then set aside to crystallize at ambient temperature and in the presence of air. After one week, crystals suitable for single-crystal X-ray diffraction were collected by filtration and dried in air: compound (I)For the preparation of compounds (I)6.2), and with Uiso(H) = 1.2Ueq(C). For the H atoms bonded to N or O atoms, the atomic coordinates were refined with Uiso(H) = 1.2Ueq(N) or 1.5Ueq(O), giving the N—H and O—H distances shown in Tables 1Crystal data, data collection and refinement details are summarized in Table 310.1107/S2056989022007472/hb8030sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989022007472/hb8030Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022007472/hb8030IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989022007472/hb8030Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022007472/hb8030IIsup5.cmlSupporting information file. DOI: 2191691, 2191690CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


X , C—H⋯π(Cp) and C—Cl⋯π(phenyl) interactions, building a three-dimensional network. The cation has planar chirality with Sp(Fc) absolute configuration. The structure of the title compound is compared with related disubstituted (trimethylammonio)methyl ferrocenes.The packing of the title disubstituted ferrocene derivative is stabilized by weak C—H⋯ 5H5)(C15H19N)]I·CH2Cl2, is described. The cation molecule is built up from a ferrocene disubstituted by a trimethylammonium methyl group and a phenyl ring. The asymmetric unit contains the iodide to equilibrate the charge and a disordered dichloromethane solvate. The disordered model results from a roughly statistical exchange (0.6/0.4) between one Cl and one H. The packing of the structure is stabilized by weak C—H⋯X , C—H⋯π(Cp) and C—Cl⋯π(phenyl) interactions, building a three-dimensional network. The cation has planar chirality with pS(Fc) absolute configuration. The structure of the title compound is compared with related disubstituted (trimethylammonio)methyl ferrocenes.As a follow-up to our research on the chemistry of disubstituted ferrocene derivatives, the synthesis and the structure of the title compound, [Fe(C You and co-workers -(2-phenylferrocenyl)methyl]trimethylammonium iodide, should then be favoured and should provide an efficient access to a wide range of various enantiomerically pure ferrocene derivatives of type C including ligands, by reaction with various nucleophiles   Fig. 1.2.The molecular structure is based on a ferrocene moiety in which one of the Cp rings is disubstituted in the 1,2 position by a tri-methylammonium-methyl and a phenyl substituent. The molecule has a positive charge, which is counterbalanced by an iodide Fig. 2. MoreovepS, which is confirmed by the refinement of the Flack parameter , C—H⋯π and C—Cl⋯π interactions  and atom C23 of the phenyl group with the centroid of the substituted Cp ring . Finally, there is also a C—Cl⋯π interaction involving the Cl1 atom of the solvate . All these interactions build up a three-dimensional network.The crystal packing is governed by the occurrence of weak C—H⋯s Table 1. The iodis Fig. 4. Then thne Fig. 4. Moreove4.et al., 20163)3 group is always above the Cp ring to which it is attached. The dihedral angles between the Cp and the C–C–N plane range from 69.8 to 89.7° for the title compound.A search in the Cambridge Structural Database -1-dimethylaminomethyl-2-phenylferrocene [(pS)-A]: To a solution of phenylboronic acid  in DMA (8 mL) were added Boc–L–Val–OH , Pd(OAc)2 , K2CO3 , TBAB  and N,N-dimethylferrocenylmethylamine  successively. The mixture was stirred at 333 K under air (open flask). When the reaction was complete (TLC monitoring), the mixture was quenched with saturated aqueous NaHCO3 and the organic phase was extracted three times with EtOAc. The combined organic layers were washed with H2O and brine successively, dried (Na2SO4) and filtered. The solvent was removed under reduced pressure and the residue purified by column chromatography  to afford the desired product A as a yellow oil . The results are in agreement with published analytical data  δ ppm 7.79–7.71 , 7.41–7.31 , 7.30–7.21 , 4.53–4.47 , 4.33 , 4.26 , 4.08 , 3.67 , 3.18 , 2.21 . 13C NMR  δ ppm 138.91 , 129.36 (CH Ph), 127.93 (CH Ph), 126.06 (CH Ph), 88.17 (Cq subst Cp), 82.24 (Cq subst Cp), 71.56 (CH subst Cp), 70.06 (CH Cp), 69.94 (CH subst Cp), 67.10 (CH subst Cp), 57.93 (CH2), 45.08 (CH3).Synthesis of [(Sp)-(2-phenylferrocenyl)methyl]trimethylammonium iodide salt [(pS)-B]: An excess of MeI  was added to a solution of A  in Et2O (3 mL). The reaction mixture was stirred for 4 h at RT. An abundant yellow solid precipitated. The yellow solid was filtered, washed with Et2O and dried to yield B as a yellow solid , which was crystallized in dichloromethane.1H NMR  δ ppm 7.58–7.50 , 7.46–7.37 , 7.37–7.26 , 5.33 , 4.91 , 4.83 , 4.56 , 4.52 , 4.34 , 3.02 . 13C NMR  δ ppm 136.54 , 129.86 (CH Ph), 129.05 (CH Ph), 127.72 (CH Ph), 90.61 (Cq subst Cp), 73.44 (CH subst Cp), 72.15 (Cq subst Cp), 70.96 (CH Cp), 70.44 (Cq subst Cp), 70.04 (Cq subst Cp), 65.21 (CH2), 52.76 (CH3).6.Uiso(H) = 1.2Ueq or Uiso(H) = 1.5Ueq(CH3).Crystal data, data collection and structure refinement details are summarized in Table 3SHELXL gave a ratio of 0.6/0.4. So the disordered model is based on an exchange between one H and one Cl . Their coordinates and thermal parameters were constrained to be identical using the EXYZ and EADP commands available in SHELXL. This disordered model is not perfect, as suggested by a large residual electron density in the vicinity of the atom H30B.The occurrence of three large residual densities around the C atom of the solvate with distances around 1.76 Å initially suggested the presence of a chloroform solvate molecule. However, if one of the Cl atoms (Cl1) could be refined correctly with full occupancy, the two others display large and elongated ellipsoids. Refining their occupancy factors using the restraints available in Cl Fig. 3. The mod10.1107/S2056989022006053/dj2043sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989022006053/dj2043Isup2.hklStructure factors: contains datablock(s) I. DOI: 2177602CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Introduciton: The α,ω-diamines (NH2-(CH2)n-NH2) and ω -amino fatty acids (NH2-(CH2)n-COOH) have been widely used as building blocks in polymerindustries. Medium- to long-chain (C8 to C18) fatty acid monomers with amino residues are almost exclusively produced via chemical processes that generate hazardous waste and induce severe environmental problems, such as global warming and pollution. Here, we present the construction platformstrains of Yarrowia lipolytica a cheese-ripening yeast, for direct biotransformation of hydrocarbons into medium- to long-chain α,ω-diamines and ωamino fatty acids using metabolic engineering of endogenous fatty acid ω- and β-oxidation pathways and introducing heterologous ω-transaminase in Y. lipolytica.Methods: We deleted six genes encoding the acyl-CoA oxidase (ACO1–6) and four fatty aldehyde dehydrogenase genes (FALDH1-4), which catalyze fatty acid β-oxidation and downstream oxidation of fatty aldehydes in Y. lipolytica, respectively. The ω-transaminase from Chromobacterium violaceum DSM30191 was introduced into the genome of the ΔPOX ΔFALDH strain under the control of Y. lipolytica-derived EXP1 promoters.Results and Discussion: The ΔPOX ΔFALDH strains with ω-CvTA successfully accumulated the corresponding C12 αω-diamines into a shaking culture medium with dodecane or dodecanol. In addition, these strains accumulated C12 ω-amino fatty acids from dodecanoic acid. With the commercially available α,ω-diacid bioprocess, this yeast biosynthesis producing medium- and longchain α,ω-diamines and ω-amino fatty acids could complete the yeast platform technology generating all medium- and long-chain aliphatic polyamide monomers, α,ω-biofunctionalized with one or both carboxylic acid and amino residues. Plastics play essential and ubiquitous roles in modern life. Synthetic plastics are commonly produced oduction .2–C6) plastics, medium-chain (C8–C14) repeating monomers exhibit high-performance properties, including flexibility, chemical resistance, low moisture absorption, and resistance to stress cracking  and polyamide (PA) plastics are the two most important classes of commercial plastics, and the monomers of each plastic are joined by an ester (-O-CO-) and an amide (-CO-NH-) linkage, respectively. Both plastics can be synthesized as homopolymers  or copolymers . For PA, the monomers are α,ω-fatty dicarboxylic acids  and α,ω-fatty diamines  or ω-amino fatty acids (ω-AFA), while for PE, the monomers are α,ω-diacids and α,ω-fatty dialcohols  or ω-hydroxy fatty acids (ω-HFAs) . Comparecracking   and can utilize the ω-oxidation pathway to oxidize hydrophobic substrates, generating corresponding α,ω-bifunctional metabolites  that can be used as polymer building blocks  carbons of medium- to long-chain  pathway , therebyg blocks . For plasaminase . Sung etY. lipolytica, a cheese-ripening yeast, for direct biotransformation of hydrocarbons into medium- to long-chain α,ω-fatty diamines [NH2-(CH2)n-NH2] and ω-amino fatty acids [NH2-(CH2)n-COOH], which are building blocks for high-performance polymers (via metabolic engineering of endogenous fatty acid ω- and β-oxidation pathways and introducing heterologous ω-transaminase (ω-TA).Here, we present the construction of platform strains of polymers , via metY. lipolytica strains, plasmids, and oligonucleotides used in this study are listed in Y. lipolytica was based on the homologous recombination and 5-fluoroorotic acid (5-FOA) selection using URA3 as a recyclable selection marker  from C. violaceum DSM30191 was expressed under the control of Y. lipolytica-derived EXP1 promoters . A Biometra T3000 thermocycler with Prime Star DNA polymerase  was used for PCR amplification. Amplified fragments were purified using the QIAgen purification kit , and DNA fragments were recovered from the agarose gels using the QIAquick gel extraction kit .Genome modification of n marker . URA3 wag vector  using th primers . These hromoters  with or nchoring  (atgtccY. lipolytica strains was pre-grown in a test tube containing 3 ml yeast extract Bacto-pepton dDextrose (YPD) medium overnight at 30°C with shaking at 200 rpm. One milliliter of pre-cultured broth was inoculated into a 500 ml baffled flask containing 100 ml of growth medium (pH 6.0) containing 50 g/L glucose, 10 g/L yeast extract, 6.7 g/L yeast nitrogen base without amino acids, 5 g/L (NH4)2SO4, and .05 g/L uracil. The cultures were incubated overnight at 30°C with rotation at 200 rpm. When an OD600 of 40–50 was reached, 50 ml of the culture broth was transferred to 250 ml baffled flasks containing 50 ml of the conversion medium (pH 7.6) containing 30 g/L glucose, 3 g/L yeast extract, 6.7 g/L YNB w/o amino acids, 5 g/L (NH4)2SO4, .05 g/L uracil, and 1 g/L substrate  with .05% Tween 80. This biotransformation was performed for 2 days at 30°C in a shaking incubator (200 rpm). To investigate the effect of multiple gene knock-out on cell growth, Y. lipolytica was incubated in a yeast extract-peptone medium (YPD) containing 10 g/L yeast extract and 20 g/L Bacto-peptone in sterile 24-well PS plates . Each well contained 400 µl of YP media with 2 µl of the pre-cultured Y. lipolytica. The Y. lipolytica culturing was done in 20 ml of YPD medium in 250 ml baffled flask incubated overnight at 30°C. The plate was incubated on a shaking plate reader  at 30°C at 200 RPM, and the turbidity of each well was measured every 15 min (no sampling of the culture).A single colony of the engineered CvTAER and ω-CvTAcytosol introduced into Y. lipolytica, cells were prepared, and fluorescence microscopy observations were performed. PDI monoclonal antibody (MA1-10032) was used as the primary antibody for ω-CvTA  for the ER marker, and the goat anti-mouse IgG1  was used as the secondary antibody. Substrates and products were identified and quantified using gas chromatography-mass spectrometry (GC-MS) (Agilent GC/MSD 5975 coupled with an Agilent 7890A) equipped with a quadrupole mass selective detector on electroionization (EI) operated at 70 eV. An Agilent HP-5MS column  was used at a 10:1 split ratio, helium was used as a carrier gas, and the oven temperature was 150°C–172°C. The substrates and products were extracted and prepared for GC-MS analysis according to the corresponding methods shown in 2SO4 to convert the ionized ω-amino fatty acid to ω-amino fatty acid and directly extracted with diethyl ether. For sialylation, N,O-bis(trimethylsilyl)trifluoroacetamide was mixed with the extracted samples  and the mixture was incubated for 20 min at 60°C. Tetradecane was used as an internal standard.The biomass was measured using a UV spectrophotometer  at 600 nm absorbance. To analyze the subcellular localization of ω-Y. lipolytica suitable to produce α,ω-bifunctional monomers with amino residues, aliphatic polymeric monomers with amino residues, and hydrophobic substrates. In Y. lipolytica, non-homologous end-joining (NHEJ) is used more than homologous recombination (HR) for the repair of double-strand breaks in DNA , which catalyzes the amination between the ω-positioned ketone and amine groups. However, as shown in Y. lipolytica oxidizes hydrocarbons substrates such as alkanes, fatty alcohols and acids to their corresponding terminal carboxy function  encoding the acyl-CoA oxidase. The ΔPOX strain did not grow on fatty alkanes, fatty alcohols or acids as sole carbon sources  was exclusively localized to the ER and the other  to the cytosol  and lag time (λ) of each mutant. Compared to Pol1g strain, all mutant showed the lower μmax, representing a theoretical maximum growth rate based on the inflection point of the curve, and more increased λ, the adaptation time. However, a remarkable toxic effect on all mutants was not observed.The effect of multiple gene knock-out on cell growth was investigated by monitoring the growth of each mutant using a plate reader. Time-profiles of the monitored cell growth  were fitPOXΔFALDH strains with ω-CvTAER or ω-CvTAcytosol successfully accumulated the corresponding 1,12-diaminododecanes into the culture medium in a shaking flask culture with 1 g/L of dodecane or dodecanol as C12 hydrocarbon substrates  than those with ω-CvTAcytosol . To examine whether the strains could convert FAs to the corresponding ω-AFAs, these strains were cultivated in a shaking flask with 1 g/L of dodecanoic acid. These strains successfully accumulated 12-aminododecanoic acids in the culture medium  than those with ω-CvTAcytosol (1.91 ± .65 mg/L). Surprisingly, 12-aminododecanoic acids were produced from dodecane (1.91 ± .18 mg/L for ω-CvTAER and .77 ± .28 mg/L for ω-CvTAcytosol) and dodecanol (1.11 ± .58 mg/L for ω-CvTAER and .48 ± .16 mg/L for ω-CvTAcytosol)  , which mcytosol) . These s11) or long chain (C16), both ΔPOX and ΔFALDH strains with ω-CvTAER or ω-CvTAcytosol also produced the corresponding ω-AFAs (1.91 ± .18 mgC11 ω-AFAs/L and .77 ± .28 mgC16 ω-AFAs/L for ω-CvTAER and 1.91 ± .18 mgC11 ω-AFAs/L and .77 ± .28 mgC16 ω-AFAs/L for ω-CvTAcytosol)  . To incrty acids . These fty acids .E. coli by introducing heterologous CYP genes. However, attempts encountered some problems, such as CYP expression and hydrophobic substrate uptake in E. coli-based whole-cell biotransformation. In particular, the correct expression of CYP in E. coli requires a heme precursor such as expensive δ-ala-leuvenic acid (n-alkane-assimilating yeasts with an endogenous CYP belonging to the CYP52 family and a reductase provide an advantageous approach for ω-oxidation of hydrocarbons. The diploid yeast C. tropicalis produces 174 g/L C14 diacid from methyl ester C14 fatty acid and 153 g/L of C13 α,ω-diacids from tridecane (C. tropicalis to industrially produce C11–C16 α and ω-diacids from petroleum-based n-paraffin. C. tropicalis through a modification of the α- and β-oxidation pathways.Biotransformation of hydrocarbons into polymeric monomers with amino residues has mainly been attempted using whole-cell systems of nic acid . The n-aridecane . Cathay n-alkane-assimilating haploid Y. lipolytica as a starting strain since C. tropicalis is an obligate diploid (via ω-oxidation and converting an aldehyde group into an amino group by ω-transaminase, which catalyzes the amination between the ω-positioned ketone and amine group. To the best of our knowledge, this study is the first to establish a Y. lipolytica-based biocatalytic system for the production of α,ω-diamines or ω-amino fatty acids from n-alkanes, fatty alcohols and acids. Although whole-cell catalytic systems using E. coli has been recently developed (E. coli requires a heme precursor, which supplements culture medium with δ-ala-leuvenic acid, an expensive commodity (E. coli shows limited uptake of hydrophobic substrates, which can lower the productivity.In this study, we needed genome-wide modification and thus selected an alternative  diploid . We succeveloped , the sysommodity . FurtherIn conclusion, with the commercially available α,ω-diacid bioprocess, this yeast biosynthesis that produces medium- and long-chain α,ω-diamines and ω-amino fatty acids could complete the yeast platform technology generating all medium- and long-chain aliphatic polyamide monomers, α,ω-biofunctionalized with one or both of carboxylic acid or amino residues . Therefo


The title molecule is nearly planar with a slight structural bend involving the carbonyl group. The molecules pack in the crystal by intermolecular C—H⋯O/F hydrogen bonding, π–π stacking, and H–π interactions. 17H15FO2, was synthesized by a Claisen–Schmidt condensation with 4-fluorobenzaldehyde and 4′-ethoxyacetophenone. The torsion angles between the 4-fluorophenyl ring and the alkene and the 4′-ethoxyphenyl ring and the 2-propen-1-one are −1.2 (4) and 1.2 (3)°, respectively; however, there is a larger torsion between the bonds comprising the 2-propen-1-one unit of 12.0 (4)°. The crystal packing is stabilized by intermolecular C—H⋯O/F hydrogen bonding, π–π stacking, and H–π interactions.The title molecule, C However, the torsion angle of the chalcone between the O1—C9 and C7—C8 bonds is 12.0 (4)°, indicating a break in planarity. This single deviation causes a slight concave bend in the molecule. The title compound crystallized as a racemic mixture in the space group Pca21; thus, a clockwise and anticlockwise torsion of the chalcone are present with a 1:1 ratio in the unit cell.This chalcone has aromatic rings with substitutions in the 4 position on both ends of the molecule, where the phenyl on the alkene is fluorinated, and the phenyl on the carbonyl contains an ethoxide Fig. 1. Both phX) is maintained as a fluorine, the other substituent (–R) varies as either a methyl, hydroxyl, methoxy, or ethoxy group. Examination of the three torsion angles described above suggests that there is a trend in the degree of distortion from planarity, with an order of methyl, methoxy, ethoxy, to hydroxy by increasing planarity. While there are no direct examples that contain a halogen and an ethoxy, we felt comparison of our compound to the nearest chloro- and bromo-substituted compounds was warranted. The closest examples are a bromo/methoxy and a chloro/methoxy 4,4′ -substituted chalcone. Both cases are more distorted from planar than our fluoro/ethoxy chalcone. Lastly, we found a set of chalcones with an ethoxy substituent, where there are chlorine atoms in the 2 and 3 position of the respective phenyl ring. Both of these cases are more planar than our chalcone.There are several other chalcones with a comparable 4 and 4′ set of substitutions that are summarized in Table 13.E)-1-(4-Ethoxyphenyl)-3-(4-fluorophenyl)-2-propen-1-one crystallizes in the orthorhombic space group Pca21, with four molecules occupying one unit cell. The molecules pack using hydrogen bonding, π–π stacking, and H–π interactions -3-(4-fluorophenyl)-1-(4-methylphenyl)prop-2-en-1-one -3-(4-fluorophenyl)-1-(4-hydroyxlphenyl)prop-2-en-1-one -3-(4-fluorophenyl)-1-(4-methoxylphenyl)prop-2-en-1-one -3-(4-chlorophenyl)-1-(4-methoxyphenyl)prop-2-en-1-one -3-(4-bromophenyl)-1-(4-methoxyphenyl)prop-2-en-1-one -3-(2-chlorophenyl)-1-(4-ethoxylphenyl)prop-2-en-1-one -3-(3-chlorophenyl)-1-(4-ethoxylphenyl)prop-2-en-1-one  was added to the mixture dropwise. The mixture was allowed to stir at room temperature for 45 min. Cold distilled H2O (4 mL) was added and the mixture was cooled in an ice bath before isolating the solid product by vacuum filtration. The chalcone was purified by recrystallization with dichloromethane/hexane (4:1) to yield colorless crystals (77% yield). High-quality crystals for diffraction were grown from slow evaporation of 190 proof ethanol at room temperature, m.p. 392–394 K; IR (ATR) νmax 3067, 2936, 1653, 1596, 1572, 1504, 1157, 1033 cm−1; 1H NMR  δ 8.03 , 7.76 , 7.62 , 7.46 , 7.10 , 6.79 , 3.88  ppm; 13C{1H} NMR  δ 188.4, 163.5, 163.9 , 142.6, 131.3 , 131.0, 130.8, 130.2 , 121.5 , 116.0 , 113.8, 55.5 ppm. 1H NMR data have previously been reported  and 4-ethoxyacetophenone (3 mmol) were mixed in 95% EtOH (2.5 mL). An aqueous solution of sodium hydroxide  was 1.2 times Ueq(C) for aromatic and methylene hydrogen atoms, and 1.5 times Ueq(C) for methyl hydrogen atoms. There is minor whole molecule disorder visible in the residual peaks that was not refined since these peaks are rather small (< 0.26 e Å−3) and there was little improvement in the model.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989022007423/zl5033sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022007423/zl5033Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022007423/zl5033Isup3.cmlSupporting information file. DOI: 2191384CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Coleoptera) of the Farasan Archipelago of the southern Red Sea, Kingdom of Saudi Arabia. ZooKeys 959: 17–86.In Abdel-Dayem MS, Abu El-Ghiet UM, Elsheikh TM, Elgharbawy AA, Al-Fifi ZIA, Aldhafer HM (2020) The first survey of the beetles . The correct identification for this species is Orbeolahirsutulum . Other corrections are as follows:After the publication of the work referenced above, our attention was drawn to misidentified species of Page 1, “Abstract” line 13: “… Arabian and Afrotropical elements .” should be“….. Arabian and Afrotropical elements .”Page 34, line 25: “Attagenusobtusus 41. ” should beOrbeolahirsutulum 41. ”“Page 34, line 28: The paragraph under “Literature records” should read:“Riyadh (Mroczkowski 1979).”Page 34–35, line 32: The paragraph under “General distribution” should read:“AFR-SAR species, distributed in East Africa (Eritrea); Levant, Turkmenistan and Arabian Peninsula .”Page 77, line 10: “that 41.3% of the species belong to Saharo-Arabian …..“ should be“that 41.9% of the species belong to Saharo-Arabian ….”Page 79, lines 4–6: “… whereas only 28 of Farasan Archipelago’s 179 species (15.6%) are shared with the 645 species currently known from the Socotran fauna, half  of which ….” should be“… whereas only 29 of Farasan Archipelago’s 179 species (16.2%) are shared with the 645 species currently known from the Socotran fauna, about half  of which ….”Page 79, line 16: “Only 15.6% of its species are shared …..” should be“Only 16.2% of its species are shared …..”Page 79, Table 1: In 20th row 20 “Dermestidae”: 4th column: “1 (3)” should be “2 (3) and 5th column: “0.6% (2.1%)” should be “1.1% (2.1%)”


The 1,2-diazole ring in the title compound is fused to a non-aromatic six-membered ring and bears an N-bound phenyl ring. In the crystal, weak C—H⋯O, C—H⋯π and π–π interactions contribute to the three-dimensional architecture. 21H18N2O, the non-aromatic six-membered ring adopts a distorted envelope conformation with one of the methylene-C atoms being the flap atom. The dihedral angle between the phenyl and 4-tolyl rings is 75.3 (1)°. The 1,2-diazole ring forms dihedral angles of 41.9 (1) and 65.5 (1)° with the phenyl and 4-tolyl rings, respectively. In the crystal, stabilizing C—H⋯O, C—H⋯π and π–π interactions are evident. The calculated Hirshfeld surfaces highlight the prominent role of C—H⋯O interactions (8.6%), along with H⋯H (51.7%) and C⋯H/H⋯C (29.2%) surface contacts.In the title compound, C In (I), the non-aromatic six-membered ring adopts a distorted envelope conformation with the methylene-C9 atom being the flap atom, Fig. 1The molecule of (I) and the recently reported 4-chlorobenzylidene derivative (II)  motif along the c-axis direction of the unit cell  through white to blue . The different types of intermolecular interactions can be identified by colour coding the distances from the surface to the nearest atom exterior (de) or interior (di) plots to the surface. The 2-D fingerprint plots from the surface analysis and the dnorm surface were analysed for (I) to further explore the packing modes and intermolecular interactions. The 3-D Hirshfeld surfaces and 2-D fingerprint plots with percentage contributions are shown in Fig. 5As a quantitative approach to analyse the intermolecular interactions, the Hirshfeld surfaces and two-dimensional (2-D) fingerprint plots were generated by employing the H-indazol-4-one (1 mmol) and 4-methylbenzaldehyde (1 mmol) was dissolved in ethanol followed by the addition of alcoholic NaOH. The mixture was stirred at room temperature for 1 h to afford (E)-5-(4-methylbenzylidene)-1-phenyl-1,5,6,7-tetrahydro-4H-indazol-4-ones as a precipitate, which was filtered, dried and recrystallized from ethanol: yield: 99%, m.p. 172–175°C.A mixture of 1-phenyl-1,5,6,7-tetrahydro-4Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2414314622002838/tk4075sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622002838/tk4075Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622002838/tk4075Isup3.cmlSupporting information file. DOI: 2158365CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Linum is the largest genus of the Linaceae family; the species of this genus are known to have anticancer activity.L. numidicum Murb. (EAELN) and L. trigynum L. (EAELT) were examined, for the first time, for their anticancer capacity. The secondary metabolites compositions were analysed by LC-HRMS/MS.In this study, ethyl acetate extracts of The antiproliferative effect of EAELN and EAELT (0–10.000 μg/mL) against PC3 and MDA-MB-231 cell lines were  evaluated by the MTT assay after 72 h of treatment. Flow cytometer analysis of apoptosis (Annexin V-FITC/PI) and cell cycle (PI/RNase) was also performed after treatment with EAELN and EAELT at 250, 500, and 1000 μg/mL, for 24 h.50 133.2 ± 5.73 μg/mL) and MDA-MB-231 (IC50 156.9 ± 2.83 μg/mL) lines, EAELN had also shown better apoptotic activity with 19 ± 2.47% (250 μg/mL), 87.5 ± 0.21% (500 μg/mL), and 92 ± 0.07% (1000 μg/mL), respectively, causing cell cycle arrest of PC3 cells in G2/M phase, whereas arrest in G0/G1 and G2/M phases was observed after treatment with EAELT. LC-HRMS/MS profiling of the extracts revealed the presence of known compounds that might be responsible for the observed anticancer activity such as chicoric acid, vicenin-2, vitexin and podophyllotoxin-β-d-glucoside.EAELN had the highest antiproliferative activity against PC3 (ICWe have shown, for the first time, that EAELN and EAELT exert anticancer activity through cell cycle arrest and induction of apoptosis. EAELN can be considered as a source to treat cancer. Further studies will be required to evaluate the effect of the active compounds, once identified, on other cancer cell lines. Cancer is the second main cause of death in the world and remains one of the most difficult diseases to combat  includes more than 200 species distributed throughout the world  of two Algerian According to the World Health Organisation (WHO) 2020, the highest incidences in the world are observed for breast and prostate cancers. We, therefore, decided, as proof of concept, to test the effect of our extracts on breast and prostate cancer cell lines.L. numidicum was collected in June 2018 from Djebel Babor in Sétif (North-Eastern Algeria), while the collection of L. trigynum was made in June 2017 from the region of Djebel El-Ouahch in Constantine (North-Eastern Algeria). The two plants used in the study were authenticated by Prof. Gérard de Bélair . They are registered in the herbarium of the Laboratory LOST at University Mentouri Constantine 1 with the respective Voucher codes LOST.LN.06.18 and LOST.LT.06.17.https://gdebelair.com/tax/zzeo.html#Linaceae, Linum numidicus Murbeck, Coordinates: x = 5.455072, y = 36.493397, z = 1855 m, Locality: Djebel Babor (Sétif)Id: 077_44, Taxon: Linaceae, Linum trigynum L., Coordinates: x = 6.66415, y = 36.403014, z = 970 m, Locality: Djebel El Ouahch (Constantine)Id: 011_42, Taxon: LinaceaeThey are also registered at the national herbarium GdB: L. numidicum and L. trigynum (900 g), previously dried and pulverised, were individually macerated at room temperature in a hydroalcoholic mixture  for a period of 72 h. The crude extracts obtained were taken up with distilled water (1000 mL). The latter underwent liquid/liquid type extractions, using solvents of increasing polarity . The obtained organic phases were concentrated under reduced pressure to dryness.The aerial parts of Dried acetate extracts from both plants (EAELN and EAELT) were dissolved in dimethylsulphoxide  followed by an RPMI medium. The DMSO content in the solution did not exceed 0.1%. The mixture was then centrifuged to remove insoluble ingredients and the supernatant was passed through sterilisation filters. The solution was diluted with an RPMI medium and prepared at different concentrations.2 and 95% humidity.MDA-MB-231 cells, derived from pleural effusion of breast adenocarcinoma, from ATCC, were cultured in DMEM  supplemented with 10% foetal bovine serum  and 1% penicillin/streptomycin. The PC3 prostate cancer line from ATCC was cultured using RPMI 1640 medium  supplemented with 10% FBS and 1% penicillin/streptomycin. The cell lines were grown in an incubator at 37 °C with 5% CO2. After that, the medium was carefully aspirated and the formed formazan crystals were dissolved in DMSO. The absorbance in each well was measured at 570 nm using a microplate reader .The antiproliferative activity of EAELN and EAELT against the two selected cell lines was evaluated by the MTT assay , according to Mossman (1983). PC3 and MDA-MB-231 lines (5000 cells/well) were seeded in 96-well plates and incubated with different concentrations of EAELN and EAELT . Cells without treatment were used as controls. After 72 h of exposure to the extracts, the wells were emptied, washed with PBS and then 100 µL of MTT (5 mg/mL) was added to each well. The plates were then incubated for an additional 2 h at 37 °C and 5% CO5 cells/well) were treated with EAELN and EAELT after seeding in 96-well plates with different concentrations  for 24 h. Cells were then washed with PBS buffer, centrifuged, and stained with 5 μL of Annexin V and 5 μL of propidium iodide (PI). After 15 min of incubation at room temperature, 400 µL of binding buffer was added. Stained cells were determined using Accuri TM-C6 flow cytometry (BD-Biosciences).Cell apoptosis induced by EAELN and EAELT was detected by Annexin V-FITC/PI staining according to the manufacturer's instructions (BD Pharmingen TM FITC Annexin V Apoptosis Detection kit I). PC3 cells (106 cells/well) were treated with EAELN and EAELT after seeding in 6-well plates with different concentrations  for 24 h. At the end of the treatment, cells were collected and washed with cold PBS and fixed in cold 70% ethanol (−20 °C) for 2 h. The cells were then centrifuged and the cell pellet was resuspended with 0.5 mL of a mixture containing propidium iodide and RNase. After 15 min of incubation at room temperature, the DNA content of the cells was quantified using an Accuri TM-C6 flow cytometer (BD-Biosciences).The cell cycle was determined using a cell cycle detection kit (BD Pharmingen TM PI/RNase Staining Buffer). PC3 cells .LC − ESI − DDA − HRMS/MS spectra were obtained using a Dionex Ultimate 3000 liquid chromatography (HPLC) system equipped with an Ultimate 3000 RS pump hyphenated to a Thermo Instruments MS system (LTQ Orbitrap XL). The HPLC analysis was performed on a Water BEH C18 column  and using the following solvents: Solvent A 0.1% formic acid Hpost hoc comparisons was used as indicated in the legends. Data are presented as mean ± SD. Significant differences were set at a p-value < 0.05.Data analyses were performed using GraphPad PRISM software (version 8.0.2). All experiments were performed in triplicate. One-way or 2-way ANOVA with Bonferroni’s To study the effect of EAELN and EAELT on the growth of human prostate and breast cancer cells, PC3 and MDA-MB231 cells were treated with different concentrations of extracts for 72 h, and cell viability was determined using the MTT assay.50) values of 133.2 ± 5.73 μg/mL and 156.9 ± 2.83 μg/mL, respectively, for EAELN and 415.8 ± 2.33 μg/mL and 307.5 ± 2.97 μg/mL, respectively, for EAELT .p < 0.0001) compared with the negative control  after treatment with EAELN increased from 8.3 ± 2.26% (control) to 19 ± 2.47% (250 μg/mL), 87.5 ± 0.21% (500 μg/mL), and 92 ± 0. 07% (1000 μg/mL), respectively, and after treatment with EAELT, the proportion of total apoptotic cells increased from 8.2 ± 2.33% (control) to 19.1 ± 0.35% (250 μg/mL), 50.4 ± 1.77% (500 μg/mL), and 74.4 ± 1.27% (1000 μg/mL), respectively.p < 0.0001).To better understand the mechanism by which EAELN and EAELT inhibit cell proliferation, flow cytometry analysis using PI staining was performed to assess cell cycle distribution in the PC3 cell line after 24 h of exposure to 250, 500, and 1000 μg/mL of extract. After treatment with EAELN, the percentage of cells in the G0/G1 and S phase was reduced, while that in the G2/M phase was significantly increased (****p < 0.001) for 250, 500, and 1000 μg/mL, respectively, while that of the control was 74.46 ± 0.49%. The percentage of cells in S phase increased from 19.29 ± 1.39% (control) to 5.66 ± 0.53% (250 μg/mL), 7.96 ± 0.20% (500 μg/mL), and 10.55 ± 0.55 (1000 μg/mL), all ****p < 0.0001 compared with control, while that in G2/M phase went from 7.50 ± 1.36% (control) to 18.73 ± 0.67% (250 μg/mL), 18.29 ± 0.12% (500 μg/mL), and 17.72 ± 0.64% (1000 μg/mL), all p < 0.0001 compared with control , 77.36 ± 1.38%  and 78.58 ± 0.37% , respectively, and that of the G2/M phase increased from 6.25 ± 0.92% (control) to 16.23 ± 0.97% (250 μg/mL), 16.39 ± 0.79% (500 μg/mL), and 15.80 ± 1.32% (1000 μg/mL), respectively, all ****p < 0.0001 compared with control, whereas the percentage of cells in S phase was significantly reduced (****p < 0.0001) from 19. 29 ± 1.39 (control) to 11.42 ± 0.44 (250 μg/mL), 6.25 ± 0.68 (500 μg/mL), and 5.62 ± 1.1 (1000 μg/mL) (The percentage of cells in G0/G1 phase after EAELT treatment increased from 74.46 ± 0.49% (control) to 72.35 ± 1.20%  . The resWith the aim to analyse the phytochemical composition of EAELN and EAELT we conducted UHPLC-DAD-ESI/HRMS-MS. The latter revealed the presence of 75 compounds  in each 4), vitexin (37), trihydroxyflavone (74), homoorientin (27), vicenin-2 isomer 4 (21), vicenin-2 isomer 2 (12), vicenin-2 isomer 3 (16), cirsiliol (64), tricin 5-glucoside isomer 2 (66), 5,3′-dihydroxyflavone (17), luteolin (51), orientin isomer 1 (25), with respective percentages 8.08, 7.53, 6.70, 5.58, 5.17, 4.75, 4.67, 3.52, 3.38, 2.80, 2.61, and 2.54%. Four lignans, lanicepside B (14), olivil 4′-O-glucoside (15), a phenylnaphthalene (69), and the major compound podophyllotoxin-β-d-glucoside (71) with a percentage of 4.41%, were detected. In addition, 19 phenolic acids, from which 17 hydroxycinnamic acids and two hydroxybenzoic acids were detected with chicoric acid (24) as the major compound (8.19%). The analyses also showed the presence of one phenylethanoid (30) and one coumarin derivative (55), besides other compounds including one carbohydrate, melibiose (1), one alkaloid (26), two polyketides (50 and 75), three terpenoids , and three fatty acids   were detected. The major compounds were vicenin-2 isomer 1 ( and 78) .27), cirsiliol (64), tetrahydroxyflavone (72), 2ʺ-O-β-d-xylopyranosylorientin (8), hyperoside (43), vicenin-2 isomer1 (4), quercitrin (58), and vitexin (37) as major compounds with respective percentages of 9.21, 8.24, 4.36, 4.05, 3.87, 3.50, 3.35, and 2.93%. The same lignans  as those detected in EAELN, were also found in EAELT together with nineteen phenolic acids (17 hydroxycinnamic acids and two hydroxybenzoic acids) namely, 4-caffeoylquinic acid (10), chlorogenic acid (2), 3-O-coumaroylquinic acid isomer 2 (19), 3-O-feruloylquinic acid isomer 2 (23), 4,5-dicaffeoylquinic acid isomer 2 (44), 4,5-dicaffeoylquinic acid isomer 1 (33), and methyl chlorogenate isomer 1 (9) with respective percentages of 10.51, 7.44, 4.63, 3.85, 3.55, 3.05, and 2.97%. The same other compounds   were detected with homoorientin ( and 78) , belongiMedicinal plants are important sources for the development of effective anticancer agents. Currently, many drugs available on pharmacy shelves are either natural products or their derivatives  and MDA-MB-231 lines (IC50 156.9 ± 2.83 μg/mL) which could be due to the difference in chemical contents and their concentration in the two extracts.The results of our study showed, for the first time, that EAELN and EAELT inhibited the proliferation of PC3 and MDA-MB-231 cells significantly in a concentration-dependent manner  and 2. TApoptosis plays an essential role in cellular homeostasis; nevertheless, abnormal apoptosis is a pathological process , melibiose (3.97%), cirsiliol (3.52%), tricin 5-glucoside isomer 1 (3.38%), 5,3′-dihydroxyflavone (2.80%), luteolin (2.61%), and orientin isomer 1 (2.54%) , vicenin-2 isomer 1 (8.08%), vitexin (7.53%), trihydroxyflavone (6.70%), homoorientin (5.58%), vicenin-2 isomer 4 (5.17%), vicenin-2 isomer 2 (4.75%), vicenin-2 isomer 3 (4.67%), podophyllotoxin-β- (2.54%) .O-coumaroylquinic acid isomer 2 (4.63%), tetrahydroxyflavone (4.36%), 2ʺ-O-β-d-xylopyranosylorientin (4.05%), hyperoside (3.87%), 3-O-feruloylquinic acid isomer 2 (3.85%), 4,5-dicaffeoylquinic acid isomer 2 (3.55%), vicenin-2 isomer 1 (3.50%), quercitrin (3.35%), 4,5-dicaffeoylquinic acid isomer 1 (3.05%), methyl chlorogenate isomer 1 (2.97%), and vitexin (2.93%) , homoorientin (9.21%), cirsiliol (8.24%), chlorogenic acid (7.44%), 3- (2.93%) .d-glucoside (4.41%) compared to EAELT, which presented lower levels of these compounds known for their anticancer activities. Indeed, it has been reported that chicoric acid has a strong growth inhibitory effect against HCT-116 colon cancer cells and effectively induces apoptosis, characterised by DNA fragmentation, caspase-9 activation, PARP cleavage, and β-catenin downregulation  in various cancer cell lines  , vicenin-2 isomer 1 (8.08%), vitexin (7.53%), and podophyllotoxin-β-Linum species, L. numidicum (EAELN) and L. trigynum (EAELT), inhibited the proliferation of PC3 and MDA-MB-231 cells in a dose-dependent manner. EAELN had the highest antiproliferative activity against both lines tested. EAELN and EAELT induced apoptosis of PC-3 cells; the apoptotic effect of EAELN was higher than that of EAELT extract. EAELN induced a significant cell cycle arrest in the G2/M phase, while an arrest in the G0/G1 and G2/M phases was observed after treatment with EAELT. In this study, we showed, for the first time, that EAELN and EAELT exert anticancer activity by inducing apoptosis and blocking the cell cycle. This could be due to the presence of phenolic compounds such as flavonoids, lignans and phenolic acids which were detected by LC-HRMS/MS. The anticancer effect of EAELN was higher than that of EAELT. The effect induced by EAELN could be attributed to its major bioactive compounds such as chicoric acid, vicenin-2, vitexin, and podophyllotoxin-β-d-glucoside and other compounds present in the extract by a direct and/or synergistic effect. EAELN can be consered as a source of phytochemicals to treat cancer. Owing that most of the detected compounds can be obtained from commercial sources, it is intended to test those of them that are known for their anticancer activity.Our results demonstrated for the first time that ethyl acetate extracts of two