The Log10 Kw>c, values, of the complementary amino acid pairs are strongly correlated to the central (2nd) purine base of the mRNA codon and the complementary pyrimidine base of the tRNA anticodon. Clustering of amino acids is temperature independent with regard to the direction of translation (3′ → 5′ or 5′ → 3′). The Log10 Kw>c discriminate between artificial Hecht α- and β-protein datasets at 25 °C and 100 °C. Interpretation of this data may be found in the research article entitled “Determining amino acid scores of the genetic code table: complementarity, structure, function and evolution” .We present the data concerning the clustering of sense and antisense amino acid pairs into polar, nonpolar and neutral groups, as measured using hydrophobicity parameter—logarithmic equilibrium constants (Log Specifications table•10 Kw>c) discriminates between artificial α- and β-protein datasets at 25 °C and 100 °C.The data are useful since it is shown that the nucleobase coding of amino acid hydrophobicity, specified by the 2nd codon base, is temperature independent at 25 °C and 100 °C. The hydrophobicity parameter—logarithmic equilibrium constant  nucleobases.•The data presented can be used for theoretical analyses of proteins, experiments with sense and antisense peptide binding, and research of biological systems at different temperature conditions.110 Kw>c10 Kw>c are logarithmic equilibrium constants for the transfer of amino acid side-chains from neutral solution to cyclohexane at 25 °C and 100 °C The data presented here describe the analysis of temperature dependence of amino acid hydrophobicity parameter—Log10 Kw>c, in a temperature independent manner.10 Kw>c values of the complementary, i.e. sense and antisense, amino acid pairs depend strongly on the central (2nd) purine base of the mRNA codon and the complementary pyrimidine of the tRNA anticodon. All calculated correlations are strong (r ≥ 0.85). With respect to the Log10 Kw>c measurements observed, temperatures of 25 °C and 100 °C do not affect the result  specify polar-nonpolar and neutral-neutral clusters for all possible complementary codon pairs irrespective of temperature value and direction of sequence translation.In 10 Kw>c) accurately predict the α- and β-artificial protein class at 25 °C and 100 °C  and Phase 2 (secondary) amino acids are clearly separated based on temperature independence of LogPART decision list (25 °C)2 AND Log10 Kw>c at 25 °C <= 2.64 AND Mean Buried Area > 113.9 Å2IF Mean Buried Area > 97.8 ÅTHEN aa Prebiotic Phase 2 1: L,I,V,S,P,T,A,N,D,E,G)ELSE aa Prebiotic Phase 1 (11/———————————PART decision list (100 °C)2 AND Log10 Kw>c at 100 °C <= 2.60 AND Mean Buried Area > 113.9 Å2IF Mean Buried Area > 97.8 ÅTHEN aa Prebiotic Phase 2 1: L,I,V,S,P,T,A,N,D,E,G)ELSE aa Prebiotic Phase 1  = +1,•polar or hydrophilic amino acid (2nd A codon score) = -1,•neutral or intermediate amino acid (2nd C or G codon score) = 0.The agglomerative hierarchical clustering (HAC) method in High agglomerative coefficients of 0.96 were observed for 25 °C and 100 °C measurements.2.2k = 3 clusters, Manhattan metric and standardized variables). The first variable involved the amino acid logarithmic equilibrium constants (Log10 Kw>c), and the second variable was amino acid codon scoring according to Davis Section 2.1.). The values of average silhouette widths for 25 °C (0.67) and 100 °C (0.65) confirm the validity of the model Two-variable fuzzy partitioning was produced using S-Plus 2000 software (2.310 Kw>c) for transfer of amino acid side-chains from neutral solution to cyclohexane at 25 °C and at 100 °C. The correlations are presented in https://folk.uio.no/ohammer/past/): x = free energy ligandaa, and y = |ligandaa − receptoraa| free energy absolute difference (aa = amino acid).Correlations of complementary pairs of polar-nonpolar residues and neutral-neutral residues in a 3′ → 5′ and 5′ → 3′ translation directions, with respect to the logarithmic equilibrium constants  Primary amino acid sequences of 15 artificial Hecht α- and 17 β-protein folds were converted into a numerical series by assigning the Log2.610 Kw>c values, and Mean Buried Area parameter https://www.cs.waikato.ac.nz/ml/weka/) The prediction of Phase 1 (primary) and Phase 2 (secondary) amino acids based on temperature independence of Log


This structure features C—H⋯O hydrogen bonds (both intra- and intermolecular) and intermolecular C—H⋯π interactions.The synthesis and crystal structure of a diisopropyl-substituted  13H21NO2S, is reported here along with its crystal structure. This compound crystallizes with two molecules in the asymmetric unit. The sulfonamide functional group of this structure features S=O bond lengths ranging from 1.433 (3) to 1.439 (3) Å, S—C bond lengths of 1.777 (3) and 1.773 (4) Å, and S—N bond lengths of 1.622 (3) and 1.624 (3) Å. When viewing the molecules down the S—N bond, the isopropyl groups are gauche to the aromatic ring. On each molecule, two methyl hydrogen atoms of one isopropyl group are engaged in intramolecular C—H⋯O hydrogen bonds with a nearby sulfonamide oxygen atom. Intermolecular C—H⋯O hydrogen bonds and C—H⋯π interactions link molecules of the title compound in the solid state.The synthesis of the title compound, C We attribute this relatively small torsion angle to the presence of intramolecular C-H⋯O interactions, which are described in more detail below. The torsion angles (O2—S1—N1—C11 and O2A—S1A—N1A—C11A) between the methine carbon atom of the other isopropyl group and the other sulfonamide oxygen are 46.7 (3) and 46.8 (3)°, respectively. Both sulfur atoms adopt a slightly distorted tetrahedral geometry with τ4 descriptors for fourfold coordination of 0.94 for both S1 and S1A , we describe them here as potential C—H⋯O hydrogen bonds. Specifically, O1 interacts with C9(H9A) and C10(H10B), while the equivalent atom O1a interacts with C9A(H9AC) and C10A(H10F). These interactions have D⋯A distances ranging from 3.039 (5) to 3.157 (5) Å and D—H⋯A angles ranging from 117 to 121°  x, −1 + y, z] with a D⋯A distance of 3.465 (4) Å and a D—H⋯A angle of 150.8°  to a stirring mixture of diisopropylamine , pyridine  and 10 mL of degassed dichloromethane under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 24 h under a nitrogen atmosphere. After acidification with 5 M HCl and dilution with 15 mL of dichloromethane, the organic layer was washed with water and brine. The aqueous layers were back extracted with 10 mL of dichloromethane. The combined organic layers were then dried over anhydrous sodium sulfate and evaporated to dryness. The resulting solid was dissolved in hot ethanol and filtered. The filtrate was placed in a freezer for two days and the product was isolated via vacuum filtration to give colorless crystals .The title compound was prepared by the dropwise addition of Uiso(H) = 1.2Ueq(C) for methine groups and aromatic hydrogen atoms, and Uiso(H) = 1.5Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989020007185/pk2628sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020007185/pk2628Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020007185/pk2628Isup3.cmlSupporting information file. DOI: 2006237CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The 4-iodobenzene ring is inclined to the phenol ring by 39.1 (2)°. The configuration about the C=N bonds is E. The crystal structure features C—H⋯O hydrogen-bonding interactions.In the title Schiff base compound, C 13H9IN2O3, was synthesized by a condensation reaction between 2-hydroxy-5-nitrobenzaldehyde and 4-iodoaniline, and crystallizes in the orthorhombic space group Pna21. The 4-iodobenzene ring is inclined to the phenol ring by a dihedral angle of 39.1 (2)°. The configuration about the C=N double bond is E. The crystal structure features C—H⋯O hydrogen-bonding interactions. A Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the packing arrangement are O⋯H/H⋯O (26.9%) and H⋯H (22.0%) interactions.The title compound, C The imine group displays a C8—C7—N1—C4 torsion angle of 174.5 (6)°. The 4-iodobenzene ring (C1–C6) is inclined by a dihedral angle of 39.1 (2)° to the phenol ring (C8–C13), which renders the molecule non-planar. The configuration of the C7=N1 bond of this Schiff base is E, and the intramolecular O1—H1⋯N1 hydrogen bond forms an S(6) ring motif if Fig. 1. The 4-ni  hydrogen bonds between screw-related molecules, which form helical chains propagating along the crystallographic screw axis parallel to c  x + y, −1 + z)], which makes a zigzag tape motif  c Fig. 2. The shoif Fig. 3. There aCrystal Explorer 17.5  or longer than the van der Waals radii sum, respectively rm Fig. 4, white set al., 2016E)-2-{[(4-iodophenyl)imino]methyl}-phenol fragment. Of these 26, the most similar to (I)p-iodo-N-(p-cyanobenzylidene)aniline -5-(diethylamino)-2-[(4-iodophenylimino)methyl]phenol -4-iodobenzene; imino]methyl}-6-methoxyphenol -4-iodoaniline  ring motif.A search of the Cambridge Structural Database  and 4-iodoaniline  in ethanol (15 ml). After the mixture had refluxed for about 15 h, the orange product was washed with ether and dried at room temperature . Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution.Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989020008191/pk2635sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020008191/pk2635Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020008191/pk2635Isup3.cmlSupporting information file. DOI: 1922980CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The title compound, containing a tetra-substituted pyrrolidine ring, has an N-bound  4-nitrophenyl)ethylcarboxylate group with an adjacent C-bound 4-methoxyphenyl  and then two acetyloxy subtituents in equatorial and axial positions, respectively. The five-membered ring is twisted about the bond bearing the acetyloxy subtituents. 23H24N2O9, is a tetra-substituted pyrrolidine derivative with a twisted conformation, with the twist evident in the C—C bond bearing the adjacent acetyloxy substituents. These are flanked on one side by a C-bound 4-methoxyphenyl group and on the other by a methylene group. The almost sp2-N atom [sum of angles = 357°] bears a 4-nitrobenzyloxycarbonyl substituent. In the crystal, ring-methylene-C—H⋯O(acetyloxy-carbonyl) and methylene-C—H⋯O(carbonyl) interactions lead to supramolecular layers lying parallel to (The title compound, C The conformation of the five-membered ring is twisted about the C2—C3 bond with the C1—C2—C3—C4 torsion angle being 39.70 (16)°, consistent with a (+)syn-clinal configuration. The sum of the angles about the N1 atom is 356.7°, indicating an approximate sp2 centre. The N1-bound group occupies an equatorial position with those at the C1–C3 centres being bisectional, equatorial and axial, respectively °.The molecular structure of (I)With respect to the least-squares plane through the pyrrolidine ring, the nitrobenzene and methoxybenzene rings are splayed, as seen in the dihedral angles of 58.58 (8) and 77.65 (6)°, respectively; the dihedral angle between the benzene rings is 50.56 (5)°. There is a twist in the nitrobenzene ring as seen in the value of the C11—C10—N2—O4 torsion angle of 17.7 (3)°. By contrast, the methoxy group is co-planar with the ring to which it is connected, as shown by the C15—C16—O5—C19 torsion angle of 176.2 (2)°.b-axis direction, being propagated by 21 symmetry. The other interactions falling within the distance criteria of PLATON . A view of the unit-cell contents is shown in Fig. 2b), highlighting the stacking of layers, without directional interactions between them.The only directional non-covalent interactions of note in the crystal of (I)Supramolecular features were also evaluated by calculating non-covalent interaction plots  with the green isosurface between the interacting atoms and the distinctive blue feature in the reduced density gradient (RDG) versus sign(λ2)ρ(r) plot in the lower view, i.e. indicating the density value is less than 0.0 a.u., suggest these interactions are weakly attractive. The same is true for the ring-methylene-C4—H⋯O7(acetyloxy-carbonyl) interactions that lead to the helical chain, Fig. 3b).The aforementioned weak C—H⋯O contacts identified in dnorm-surface plots, electrostatic potential  and two-dimensional fingerprint plots following literature procedures B as well as the O5 and O7 atoms  and O2⋯O5 short contacts, being ∼0.02 Å shorter than their respective sums of the van der Waals radii, Table 2The Hirshfeld surface analysis of (I)B), benzyl (C15 and H9), methyl (C21) and nitro (O4) atoms correspond to long-range intra-layer methylene-C6—H6B⋯C15(benzyl), benzyl-C9—H9⋯C21(methyl) interactions and inter-layer O4⋯O4 short contacts, Table 2In the views of Fig. 5a), the two-dimensional fingerprint plot for the Hirshfeld surface of (I)de and di diagonal axes, and those delineated into H⋯H, H⋯O/O⋯H, H⋯C/C⋯H, O⋯O and O⋯C/C⋯O contacts are illustrated in Fig. 7b)–(f), respectively. The percentage contributions from different interatomic contacts are summarized in Table 3de = di ∼2.4 Å, Fig. 7b), corresponding to the H17⋯H23B inter-layer contact listed in Table 2de + di ∼2.5 Å in Fig. 7c). The H⋯C/C⋯H contacts that match the long-range C—H⋯C interactions discussed above are shown as a pairs of forceps-like tips at de + di ∼2.7 Å in the fingerprint plot delineated into H⋯C/C⋯H contacts, Fig. 7d). Although both O⋯O and O⋯C/C⋯O contacts appear at de + di ∼3.0 Å in the respective fingerprint plots, Fig. 7e) and (f), their contributions to the overall Hirshfeld surface are only 2.1 and 1.2%, respectively. The other interatomic contacts have a negligible effect on the molecular packing as their accumulated contribution is about 2.2%.As illustrated in Fig. 7Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies as per the literature  basis set with the B3LYP function. The individual energy components as well as the total interaction energies are collated in Table 4Hirshfeld surface analysis as well as two additional C—H⋯O interactions, i.e. methylene-C4—H4A⋯O4(nitro) and methyl-C21—H21C⋯O4(nitro) with H⋯O separations of 2.63 and 2.77 Å, respectively.The pairwise interaction energies between the molecules in the crystal of (I)Edis terms associated with the H⋯H contacts as well as the long-range C—H⋯O interactions (−14.4 kJ mol−1). For the former, the maximum energy is not found for the shortest H17⋯H23B contact (−7.1 kJ mol−1), Table 2b), but rather for a pair of benzene-H⋯H(methyl) interactions occurring in close proximity in a hydrogen-rich region but at longer separations (−34.2 kJ mol−1). For the inter-layer O4⋯O4 contact mentioned above, there are almost equal contributions from Eele and Edis, Table 4−1. The magnitudes of intermolecular energies are represented graphically in Fig. 8Edis in the molecular packing.The stabilization energies in the inter-layer region are also dominated by the et al., 2016et al., 2004There are relatively few related structures having a similar substitution pattern to the tetra-substituted pyrrolidine ring of (I)S,3S,4R)-3,4-dihydroxy-2-(4-methoxyphenyl)pyrrolidine-1-carboxylate  in CH2Cl2 (15 ml) were added pyridine , acetic anhydride  and N,N-dimethyl-4-aminopyridine . The solution was stirred for 2 h at room temperature, concentrated in a rota-evaporator and the residue dissolved in EtOAc (10 ml). The resulting solution was washed with a HCl 5% solution (3 × 5 ml) and with saturated solutions of NaHCO3 (2 × 5 ml) and of NaCl (5 ml). The phases were separated and the organic phase was dried with anhydrous Na2SO4, filtered and concentrated in vacuo.To a solution of 4-nitrobenzyl (2n-hexane elution gradient (1:3 and 1:2). Yield: 716 mg (98%). Colourless irregular crystals for the X-ray analysis were obtained by the slow evaporation of its n-hexane solution. M.p. 409.5–410.5 K. The 1H and 13C{1H} NMR reflect the presence of two conformational rotamers in solution. 1H NMR : δ = 7.75 ; 7.65 ; 7.18 ; 6.99 ; 6.76 ; 6.72 ; 6.65 ; 6.37 ; 5.42 ; 5.33 ; 5.00 ; 4.92 ; 4.74 ; 4.44 ; 3.89 ; 3.72 ; 3.29 ; 3.35–3.23 ; 1.61–1.60 . 1H NMR : δ = 8.23 ; 8.00 ; 7.53 ; 7.16 ; 6.96 ; 6.88 ; 5.45–5.32 ; 5.31–5.18 ; 5.01–4.87 ; 4.13 ; 4.06 ; 3.85–3.67 ; 2.12-2.07 . 13C{1H} NMR : δ = 169.9; 169.8; 159.4; 159.2; 154.2; 154.1; 147.6; 147.2; 143.6; 143.4; 130.6; 129.4; 128.1; 127.5; 126.8; 126.7; 123.7; 123.4; 114.2; 78.2; 69.2; 68.7; 65.7; 65.5; 64.7; 64.1; 55.3; 55.2; 49.0; 48.4; 20.8; 20.7; 20.6.The residue was purified by flash column chromatography in silica gel, using an EtOAc/Uiso(H) set to 1.2–1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 510.1107/S2056989020007914/hb7923sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020007914/hb7923Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020007914/hb7923Isup3.cmlSupporting information file. DOI: 2009242CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In a palliative situation like metastatic spinal cord compression (MSCC), overall treatment time of radiotherapy should be as short as possible. This study compared 5 × 5 Gy in 1 week to 10 × 3 Gy in 2 weeks in a prospective cohort.Forty patients receiving 5 × 5 Gy in a phase II trial were matched 1:2 to 213 patients receiving 10 × 3 Gy in two previous prospective studies for tumor type, ambulatory status, time developing motor deficits, interval between tumor diagnosis and MSCC and visceral metastases. These factors were consistent in all three patients (triple) used for each 1:2 matching. Groups were compared for local progression-free survival (LPFS), motor function, ambulatory status, and overall survival (OS).p = 0.36), 6-month OS-rates 43% and 35% (p = 0.74). Improvement of motor function was achieved in 59% and 34% of patients (p = 0.028); overall response rates (improvement or no further progression of motor deficits) were 94% and 89% (p = 0.71). Post-treatment ambulatory rates were 81% after 5 × 5 Gy and 85% after 10 × 3 Gy (p = 0.61). Of non-ambulatory patients, 50% (6/12) and 46% (11/24) regained the ability to walk (p = 1.00).After matching, 32 triples remained for analyses . Six-month LPFS-rates were 94% after 5 × 5 Gy and 87% after 10 × 3 Gy (5 × 5 Gy in 1 week appeared similarly effective as 10 × 3 Gy in 2 weeks. These results may not be applicable to long-term survivors and should be confirmed in a randomized trial directly comparing 5 × 5 Gy and 10 × 3 Gy.Trial registration clinicaltrials.gov NCT03070431. Registered 27 February 2017. Metastatic spinal cord compression (MSCC) occurs in 5–10% of patients with malignant diseases . Many ofPatients in the phase II trial  received precision radiotherapy with 5 × 5 Gy in 1 week between 02/2017 and 03/2018 . ImproveSince the EQD2 of 5 × 5 Gy (31.3 Gy) for tumor cell kill  is similar to 10 × 3 Gy (32.5 Gy), it is assumed that both regimens are similarly effective . PatientThe 40 patients of the PRE-MODE trial were matched 1:2 to the 213 patients receiving 10 × 3 Gy in a previous trial , 10. MatComparisons for LPFS and OS were performed using Kaplan–Meier method and log-rank test. For the comparison regarding age, the Mann-Whiney U test was used. The comparisons for patient characteristics, improvement of motor function, overall response and ambulatory status were performed with the Fisher’s exact test.z-score was 0.933, and distribution was considered approximately normal.After 1:2 matching, 32 triples remained for analyses, corresponding to a total of 96 patients. The distribution of the patient characteristics was not significantly different (Table p = 0.36), and 6-month OS-rates 43% and 35% . Improvement of motor function was achieved in 59% (19/32) and 34% (22/64) of patients (p = 0.028). Overall response rates were 94% (30/32) and 89% (57/64) (p = 0.71). Deterioration of motor deficits occurred in 6% (2/32) and 11% (7/64) of patients (p = 0.71). Post-treatment ambulatory rates were 81% after 5 × 5 Gy and 85% after 10 × 3 Gy (p = 0.61). Of non-ambulatory patients, 50% (6/12) after 5 × 5 Gy and 46% (11/24) after 10 × 3 Gy, respectively, regained walking ability (p = 1.00). Of ambulatory patients, 100% (20/20) and 93% (37/40), respectively, remained ambulatory (p = 0.54).Six-month LPFS-rates were 94% after 5 × 5 Gy and 87% after 10 × 3 Gy  when compared to a historical control group receiving 5 × 4 Gy [Important endpoints of radiotherapy for MSCC include functional outcome and local control –3. In se5 × 4 Gy . EQD2 fo5 × 4 Gy . 5 × 5 GThis study compared 5 × 5 Gy to 10 × 3 Gy using data of three prospective studies. Forty patients receiving 5 × 5 Gy were matched 1:2 to 213 patients receiving 10 × 3 Gy. Matching criteria were selected according to previous studies , 12–14. 5 × 5 Gy resulted in a significantly higher rate of improvement of motor function than 10 × 3 Gy. For the other investigated endpoints including the main objective LPFS, no significant differences were found. Thus, 5 × 5 Gy appeared similarly effective as 10 × 3 Gy. When interpreting these results, the limitations of the study must be considered including the non-randomized design. To minimize the risk of hidden selection biases, we used only data from prospective studies. Further limitations include the facts that follow-up MRI was not performed at pre-defined time points and that for grading of MSCC radiological criteria were not considered –10, 15. 5 × 5 Gy in 1 week appeared similarly effective as 10 × 3 Gy in 2 weeks for LPFS, functional outcome and OS. These results may not be applicable to long-term survivors and should be confirmed in a randomized trial that directly compares 5 × 5 Gy and 10 × 3 Gy.


The aim of the study was to explore genotype distribution thalassemia and G6PD deficiency in Meizhou city, China.G6PD mutations by gene chip analysis. Genotypes and allele frequencies were analyzed.A total of 16 158 individuals were involved in thalassemia genetic testing. A total of 605 subjects were screened for common Chinese SEA (65.12%), ‐α3.7 (19.05%), and ‐α4.2 (8.05%) deletion were the main mutations of α‐thalassemia, while IVS‐II‐654(C → T) (40.39%), CD41‐42(‐TCTT) (32.72%), ‐28(A → G) (10.11%), and CD17(A → T) (9.32%) mutations were the principal mutations of β‐thalassemia in Meizhou. There were significant differences in allele frequencies in some counties. Genetic testing for G6PD deficiency, six mutation sites, and one polymorphism were detected in our study. A total of 198 alleles with the mutation were detected among 805 alleles (24.6%). G6PD Canton (c.1376 G → T) (45.96%), G6PD Kaiping (c.1388 G → A) (39.39%), and G6PD Gaohe (c.95 A → G) (9.09%) account for 94.44% mutations, followed by G6PD Chinese‐5 (c.1024 C → T) (4.04%), G6PD Viangchan (c.871G → A) (1.01%), and G6PD Maewo (c.1360 C → T) (0.51%). There were some differences of the distribution of G6PD mutations among eight counties in Meizhou.A total of 5463 cases carried thalassemia mutations were identified, including 3585 cases, 1701 cases, and 177 cases with α‐, β‐, and α + β‐thalassemia mutations, respectively. ‐‐SEA, ‐α3.7, and ‐α4.2 deletion were the main mutations of α‐thalassemia, while IVS‐II‐654(C → T), CD41‐42(‐TCTT), ‐28(A → G), and CD17(A → T) mutations were the principal mutations of β‐thalassemia in Meizhou. G6PD c.1376 G → T, c.1388 G → A, and c.95 A → G were the main mutations of G6PD deficiency. There were some differences of the distribution of thalassemia and G6PD mutations among eight counties in Meizhou.The ‐‐ As one of the commonest monogenic disorder in the world, thalassemia assumes diversity in clinical phenotypes varying from almost asymptomatic to lethal hemolytic anemia.G6PD gene is located on chromosome Xq28 which consists of 13 exons and 12 introns, encoding 515 amino acids. The deficiency is widely distributed and occurs in about 400 million people worldwide.Hereditary G6PD deficiency is one of the most common genetic enzyme deficiency diseases in the world. G6PD deficiency is an X‐linked incomplete dominant inherited disease. The Meizhou is a city located in the northeast of Guangdong Province, and most of the residents living in this area are Hakka peoples. Hakka is an intriguing Han Chinese population that mainly inhabit in southern China who migrated to south originally from northern China.Population screening and genetic counseling are important to prevent the birth of children with thalassemia major. Using genetic analysis for prenatal diagnosis can diagnose thalassemia major fetuses in early pregnancy and terminate pregnancy in time, so as to avoid the birth of thalassemia major patients, which is an effective method to prevent this disease at present. Precise mutation frequencies studies in different populations will help healthcare programs to control thalassemia.22.1A total of 16 158 individuals who visited Meizhou People's Hospital  from January 2015 to May 2018 were involved in thalassemia genetic testing in this study. From February 2016 to May 2018, 605 subjects were screened for common Chinese G6PD mutations by gene chip analysis. The subjects included patients who went to cardiovascular disease center, prenatal diagnosis center, reproductive medicine center, physical examination center, pediatrics, gynecology, and other professional departments of our hospital, excluding patients with blood diseases. Figure 2.22.2.12 (<2.5%) were considered probable α‐thalassemia carriers, whereas with high HbA2 (>3.5%) were deemed possible β‐thalassemia carriers.Samples were obtained via venipuncture of an antecubital vein, and then, 2 mL of peripheral blood was collected in EDTA anticoagulant tube. Sysmex XE‐2100 blood analyzer  was used to determine erythrocyte correlative indices following the standard operating procedures. Hemoglobin electrophoresis analysis was performed by Sebia capillary electrophoresis system  compiling with standard operating procedures. Subjects detected to low mean corpuscular volume (MCV) values (<82 fL) and (or) lower mean corpuscular hemoglobin (MCH) values (<27 pg) were thought as suspicious thalassemia carriers. Subjects with low HbA2.2.2SEA, ‐α3.7, and ‐α4.2) and non‐deletional mutations (Hb Constant Spring (αCSα) , Hb Quong Sze (αQSα) , and Hb Westmead (αWSα) ). Sixteen common non‐deletional mutations in β‐globin gene were detected by PCR and flow‐through hybridization technology : CD41‐42(‐TCTT), CD43(G → T), IVS‐II‐654(C → T), CD17(A → T), CD14‐15(+G), ‐28(A → G), −29(A → G), CD71‐72(+A), CD26(G → A), IVS‐I‐1(G → T), IVS‐I‐1(G → A), CD27‐28(+C), IVS‐I‐5(G → C), Cap + 40‐43(‐AAAC), initiation codon (T → G), and CD31(‐C).Peripheral blood was collected with EDTA anticoagulant tube, in which genomic DNA was extracted from leukocytes. DNA concentration was quantified using NanoDrop 2000™ Spectrophotometer (Thermo Fisher Scientific). Gap‐polymerase chain reaction (gap‐PCR) and flow‐through hybridization technology  were used to detect α‐thalassemia mutations, both deletional mutations (‐‐2.3G6PD Gene Typing Detection Kit (gene chip assay) (Sinochips Bioscience Co.). We detected the six mutation and one polymorphism sites in G6PD gene most commonly seen in the Chinese population by gene chip assay, including G6PD Gaohe , G6PD Viangchan , G6PD Chinese‐5 , G6PD Maewo , G6PD Canton , G6PD Kaiping , and one polymorphism .Genomic DNA was extracted from EDTA anticoagulant blood of subjects using the QIAamp DNA Blood Mini Kit (Qiagen). Amplification was performed using the 2.4P < .05 was considered to statistical difference.This study used SPSS statistical software version 20.0  to analyze data, and the results would be displayed with corresponding proportion. Descriptive analysis and Pearson chi‐square test were used to compare the frequencies of genotype and allele among different counties in Meizhou region. 3A total of 16 158 blood samples were obtained and analyzed from eight counties of Meizhou area. A total of 8701 cases with microcytosis (MCV < 82 fL and/or MCH < 27 pg) were found, and corresponding percentages of microcytosis in Xingning, Wuhua, Meixian, Meijiang, Fengshun, Jiaoling, Pingyuan, and Dabu were 56.96% (1792/3146), 56.87% (1750/3077), 51.29% (1407/2743), 55.14% (1025/1859), 48.76% (1023/2098), 48.33% (652/1349), 55.54% (536/966), and 55.98% (515/920), respectively. The corresponding abnormal ratio of hemoglobin in Xingning, Wuhua, Meixian, Meijiang, Fengshun, Jiaoling, Pingyuan, and Dabu were 52.64% (1656/3146), 52.00% (1600/3077), 46.34% (1271/2743), 47.98% (892/1859), 47.47% (996/2098), 44.48% (600/1349), 48.96% (473/966), and 50.98% (469/920), respectively.P = .043) was lower than other counties in Meizhou area. However, the relative proportions of α‐ and β‐thalassemia did not differ greatly among eight counties. The prevalence of α‐thalassemia in Meijiang  was lower than other counties in Meizhou area. The prevalence of Hb H disease in Wuhua  was higher than other counties in Meizhou area, while in Jiaoling  was lower than other counties.The result of the incidence rate of α‐ and β‐thalassemia of eight counties in Meizhou area is shown in Table P = .032) and ‐α3.7 deletion , and higher frequencies in αWSα allele of Xingning , in ‐α4.2 deletion of Jiaoling , in αCSα allele of Dabu , and in IVS‐II‐654(C → T) allele of Pingyuan  were showed compared with other counties separately. IVS‐II‐654(C → T) mutation was found dominance in most of these counties, while Meixian and Fengshun had a higher percentage in CD41‐42(‐TCTT)  allele. The β‐thalassemia mutations in the order of allele frequency between eight regions of Meizhou, the mutations were IVS‐II‐654(C → T) > CD41‐42(‐TCTT) > ‐28(A → G) > CD17(A → T) in Xingning, Meijiang, Jiaoling and Pingyuan (similar to Jiangxi), IVS‐II‐654(C → T) > CD41‐42(‐TCTT) > CD17(A → T) > ‐28(A → G) in Wuhua and Dabu (similar to Fujian), CD41‐42(‐TCTT) > IVS‐II‐654(C → T) > CD17(A → T) > ‐28(A → G) in Meixian and Fengshun.Allele frequencies of α‐ and β‐thalassemia of eight counties in Meizhou area are exhibited in Table 4.2/αα, βN/βN of Jiaoling , in ‐‐SEA/‐α3.7, βN/βN  and αα/αα, βIVS‐II‐654/βN  of Pingyuan, in αCSα/αα, βN/βN of Meixian , and in ‐‐SEA/αCSα, βN/βN of Dabu  compared with other counties. There are lower frequencies in αCSα/αα, βN/βN of Xingning  and in αα/αα, β‐28/βN of Dabu  compared with other counties.The result of genotypes of different counties in Meizhou area is shown in Table G6PD mutations and 19 genotypes were detected, including six kinds of heterozygotes, six kinds of hemizygotes, and seven kinds of homozygotes. The result of allele frequencies of G6PD in different counties of Meizhou area is shown in Table G6PD mutations including G6PD Canton (c.1376 G → T)(45.96%), G6PD Kaiping (c.1388 G → A)(39.39%), and G6PD Gaohe (c.95 A → G)(9.09%) account for 94.44% of mutations, followed by G6PD Chinese‐5 (c.1024 C → T)(4.04%), G6PD Viangchan (c.871G → A)(1.01%), and G6PD Maewo (c.1360 C → T)(0.51%). There were higher frequency in G6PD c.1376 G → T allele of Wuhua  and lower frequency in G6PD c.1376 G → T allele of Meijiang  compared with other counties. The mutations were c.1376 G → T > c.1388 G → A > c.95 A → G in Xingning and Dabu, c.1376 G → T > c.1388 G → A > c.95 A → G in Wuhua and Meixian, c.1388 G → A > c.1376 G → T > c.1024 C → T in Meijiang, c.1388 G → A > c.1376 G → T > c.95 A → G in Fengshun, c.1388 G → A > c.1376 G → T > c.1024 C → T in Jiaoling, and c.1376 G → T > c.1388 G → A > c.1360 C → T in Pingyuan. In addition, we detected 55 patients (9.09%) with the polymorphism (c.1311 C → T).For G6PD deficiency, there were six Among these cases, 130 cases were simultaneously performed on genetic testing for thalassemia and G6PD deficiency, among them, 70 cases with G6PD deficiency mutations, 27 cases carried thalassemia mutations were identified, and 13 cases with both thalassemia and G6PD deficiency mutations and 46 cases did not carry thalassemia and G6PD deficiency mutations. Due to the small number of cases, we cannot analyze the relationship of thalassemia and G6PD deficiency. Of course, further studies are required to reveal the relationship, which is one of our next major research goals.4Thalassemia is an inherited autosomal recessive disease that is mainly prevalent in Guangdong, Guangxi, and Hainan provinces. Thalassemia major places a heavy financial burden on some families.In this study, the total incidence of thalassemia in Meizhou region was 33.80%, of α‐thalassemia was 22.17%, of β‐thalassemia was 10.53%, and of α‐compound β‐thalassemia was 1.10%. Compared with previous studies, our research presented relatively higher prevalence of thalassemia because our subjects were hospitalized patients.SEA/αα and 39.4% of β‐thalassemia with the main genotype of CD41‐42(‐TCTT) and IVS‐II‐654(C → T) (5.8%) ranked the fifth position.4.2, ‐α3.7 deletion, 3.83% of β‐thalassemia with lead genotype of CD41‐42(‐TCTT) mutation, while the Han people in Hainan Province presented 12.16% of α‐thalassemia with chief genotype of ‐‐SEA, ‐α3.7 deletion, 6.11% of β‐thalassemia with the main mutation is CD41‐42(‐TCTT).SEA and ‐α3.7 deletion were the main genotypes with frequencies 46.9% and 36.5%, respectively, while the carrier rate of β‐thalassemia was 2.54% in which CD41‐42(‐TCTT) (36.4%) and IVS‐II‐654(C → T) (24.8%) were the principal genotypes.SEA deletion (74.85% of mutations), 1.99% of β‐thalassemia in which CD41‐42(‐TCTT) and IVS‐II‐654(C → T) played a dominant role with frequencies 41.25% and 27.50%, respectively.SEA deletion was 3.44%, 3.97% of β‐thalassemia with chief genotype of CD41‐42(‐TCTT).SEA/αα was the most common α‐thalassemia genotype, accounted for 62.93% of α‐thalassemia. αWSα  accounted for 0.37% (120/32316) of α‐thalassemia and was relatively lower than the proportion of in Guangdong Province reported in the previous study. The main genotype of β‐thalassemia in Meizhou is IVS‐II‐654(C → T), whereas the counterpart in Guangdong is CD41‐42(‐TCTT); however, CD41‐42(‐TCTT) showed higher proportion in Fengshun and Meixian.The statistics and analysis of molecular characteristics and morbidity of thalassemia in different regions have increasingly enriched in many studies. For example, frequency of carriers for α‐thalassemia is 15% and β‐thalassemia carriers comprise 4.8% in Guangxi Province, while 46.7% of α‐thalassemia with the main genotype of ‐‐SEA deletion is potentially at risk of begetting infants with HbH disease or the Hb Bart's hydrops fetalis syndrome, depending upon the α‐globin genotype of his/her partner, which would exhibit range from hypochromic anemia to hydrops fetalis. According to previous researches, most of regions indicated a similar feature with higher genotype of ‐‐SEA deletion. Our results are consistent with those reported in most other regions. Nonetheless, the distributions and proportions in different areas regarding β‐thalassemia genotype seem to be subtly difference. Our research may not fully represent the epidemiology of thalassemia in this region related to the randomization and sample capacity of study subjects possibly as for most of our subjects were hospitalized patients.A person who is a carrier of ‐‐G6PD mutations in Hakka population of Guangdong Province studied by Yu et alG6PD gene mutations in Chinese population, which generally accounts for more than 50%. In this study, c.1376 G → T, c.1388 G → A, and c.95 A → G mutations accounted for 94.44% of the total mutation types, and the mutation distribution was consistent with the results in other places.G6PD deficiency gene carrying incidence rate was 5.41% in Guangdong Province.G6PD gene mutations, and next study may reveal the correlation between thalassemia and G6PD deficiency.It is reported that the G6PD enzyme activity in patients with G6PD deficiency compound thalassemia may be higher than that of G6PD deficiency patients, due to an increase in the number of newborn erythrocytes and increase in the activity of G6PD in the chronic hemolysis of the G6PD deficiency compound thalassemia patients.The genotype distribution and frequencies of thalassemia and G6PD deficiency in Meizhou area have regional characteristics, and there are also significant differences in different counties. Local governments can formulate corresponding measures and detection projects to prevent and control thalassemia major and G6PD deficiency according to the genotype distribution and frequencies, effectively saving costs and enhancing social benefits.5SEA, ‐α3.7, and ‐α4.2 deletion were the main mutations of α‐thalassemia, while IVS‐II‐654(C → T), CD41‐42(‐TCTT), ‐28(A → G) and CD17(A → T) mutations were the principal mutations of β‐thalassemia in Meizhou. Genetic testing for G6PD deficiency, c.1376 G → T, c.1388 G → A, and c.95 A → G were the main mutations of G6PD deficiency in this region. In addition, there were some differences of the distribution of thalassemia and G6PD mutations among eight counties in Meizhou.In conclusion, ‐‐


N atom generating an S(6) ring motif. The 3-chlorobenzene ring is inclined to the phenol ring by 9.38 (11)°. The configuration about the C=N bond is E.In the title Schiff base compound, the hydroxy group forms a intramolecular hydrogen bond to the imine  15H14ClNO, was synthesized by condensation reaction of 2-hydroxy-5-methylbenzaldehyde and 3-chloro-4-methylaniline, and crystallizes in the monoclinic space group P21/c. The 3-chlorobenzene ring is inclined to the phenol ring by 9.38 (11)°. The configuration about the C=N bond is E and an intramolecular O—H⋯N hydrogen bond forms an S(6) ring motif. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the packing arrangement are from H⋯H (43.8%) and C⋯H/H⋯C (26.7%) interactions. The density functional theory (DFT) optimized structure at the B3LYP/ 6–311 G level is compared with the experimentally determined molecular structure and the HOMO–LUMO energy gap is provided.The title compound, C RCH=N–R′) and are prepared by condensation reactions between amines and active carbonyl compounds. Schiff bases are employed as catalyst carriers . The imine group, which displays a C9—C8— N1—C5 torsion angle of −177.49 (18)°, contributes to the general non-planarity of the molecule. The chlorobenzene ring (C2–C7) is inclined by 9.38 (11)° to the phenol ring (C9–C14). The configuration of the C7=N1 bond of this Schiff base is E, and the intramolecular O1—H1⋯N1 hydrogen bond forms an S(6) ring motif A⋯N1 [H1A⋯N1 = 2.86 Å] interactions, forming sheets propagating along the a-axis direction . Weak C—H⋯π interactions [C1—H1C⋯Cg1 = 2.92 Å] are observed (Table 1b). Notably, weak π–π stacking interactions between chlorobenzene rings  along the a axis lead to the formation of a three-dimensional network.In the crystal packing of (I)on Fig. 2a. Weak d Table 1b. NotabCrystal Explorer 17.5  and shape index (e) surface mappings are shown in Fig. 3dnorm surface represent O1⋯Cl1 interactions  and C1—H1C⋯Cg1 interactions . Some additional interactions indicated by light-red spots are corresponding to contacts around phenolic and chlorobenzene rings . The red and blue triangles are absent on the shape-index surface, which indicates there are no strong π–π stacking interactions in the crystal structure.The intermolecular interactions were investigated quantitatively and visualized with ns Fig. 3b and C1ns Fig. 3c. Some gs Fig. 3d. The ra–f) indicates that the H⋯H (43.8%) interactions are the major factor in the crystal packing with C⋯H/H⋯C (26.7%) interactions making the next highest contribution. The percentage contributions of other weak interactions are: Cl⋯H/H⋯Cl (12.4%), O⋯H/H⋯O (6.6%) and N⋯H/H⋯N (3.8%).Analysis of the two-dimensional fingerprint plots Fig. 4a–f indivia density functional theory (DFT) using standard B3LYP functional and 6–311 G basis-set calculations , hardness (η), electrophilicity (ω), softness (σ) and fraction of electron transferred (ΔN). These data are recorded in Table 3σ is for the evaluation of both the reactivity and stability. The electron transition from the HOMO to the LUMO energy level is shown in Fig. 5E = ELUMO − EHOMO] of the molecule is 4.0023 eV, the frontier molecular orbital energies EHOMO and ELUMO being −5.9865 eV and −1.9842 eV, respectively. The dipole moment of (I)The optimized structure in the gas phase of compound (I)et al., 2016E)-4-methoxy-2-{[(4-methylphenyl)imino]methyl}phenol -(5-chloro-2-methylphenyl)iminomethyl]-4-methylphenol -[(3-chloro-4-methylphenyl)imino]methyl}-4-(trifluoromethoxy)phenol  ring. In 2-{(E)-[(3-iodo-4-methylphenyl)imino]methyl}-4-(trifluoromethoxy)phenol -2,4,6-trimethylaniline  in ethanol (15 ml) and 3-chloro-4-methylaniline  in ethanol (15 ml). The reaction mixture was stirred for 5 h under reflux. Single crystals of the title compound suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution .H atom was located in a difference-Fourier map and its positional parameters were refined freely with Uiso(H) = 1.5Ueq(O). Other H atoms were fixed geometrically and treated as riding with C—H = 0.96 Å (methyl) or 0.93 Å (aromatic), and Uiso(H) = 1.2Ueq(C) for aromatic H atoms or Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989020009421/vm2236sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020009421/vm2236Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020009421/vm2236Isup3.cmlSupporting information file. DOI: 2015356CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Ohydroxy—H⋯Owater, Ohydroxy—H⋯Ohydroxy, Owater—H⋯Ohydroxy, and Owater—H⋯Npyridine, result in the formation of a ribbon structure running along .In the title pyridine derivative, C In this paper, we report the crystal structure of 2,6-bis(3-methylbutyn-3-ol)pyridine monohydrate, 1·H2O.Pyridine derivatives with propargyl alcohol groups as substituents in the 2,6-positions are interesting compounds that have been used as synthons of many reactive compounds  and C≡C–C(OH) (C6≡C7—C8 and C11≡C12—C13) bond angles are 176.0 (2), 176.4 (2), 174.6 (2) and 178.5 (2)°, respectively. C6≡C7—C8 is slightly distorted from a linear structure compared to the other bonds. The two OH groups are oriented in directions opposite to each other with respect to the plane of the pyridine ring, and the pyridine ring makes dihedral angles of 50.50 (17) and 57.58 (15)°, respectively, with the C7/C8/O1 and C12/C13/O2 planes.The molecular structure of the title compound is depicted in Fig. 11·H2O along the c axis. The water molecules present as the crystallization solvent form intermolecular O—H⋯O and O—H⋯N interactions with the hydroxyl groups and the N atoms of the pyridine unit of molecule 1  −x + y + 1, z + 1  have been reported 2  as a catalyst in a THF (50 mL)–NEt3 (150 mL) solvent for 19 h at room temperature. The resulting dark-brown solution was quenched with an aqueous NH4Cl solution and the obtained solid was eliminated by celite filtration. The solution was extracted by AcOEt, and the organic phase was dried over MgSO4. After filtering off the desiccant, the filtrate was concentrated and subjected to silica-gel chromatography (eluent: AcOEt:hexane 3:2). Single crystals suitable for X-ray diffraction studies were obtained from an ethyl acetate solution via slow evaporation in air.2,6-Bis(3-methylbutyn-3-ol)pyridine was prepared by using a modified Potts method (Potts Uiso(H) = 1.2Ueq (aromatic-C) or 1.5Ueq (methyl-C).Crystal data, data collection and refinement details are summarized in Table 210.1107/S2056989020013304/is5553sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020013304/is5553Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020013304/is5553Isup3.cmlSupporting information file. DOI: 2035321, 2035321CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The bridged bimetallic compounds adopt Λ-shaped configurations, with the octahedrally coordinated copper(II) center linked to the fluorinated early transition metal via a fluoride linkage. The extended structures of these Λ-shaped compounds are organized through both intra- and intermolecular hydrogen bonds and intermolecular π–π stacking. The salt compound [Cu(phen)(H2O)2F]2[HfF6]·H2O displays an isolated square-pyramidal Cu(phen)(H2O)2F+ complex linked to other cationic complexes and isolated HfF62− anions through intermolecular hydrogen-bonding interactions.The crystal structures of three bridged bimetallic molecular compounds, namely, triaqua-2κ Although these compounds crystallize with inversion symmetry, the novel molecular building units are potential targets of future studies aimed to perturb their packing arrangement to form NCS structures. The salt compound [Cu(phen)(H2O)2F]2[HfF6]·H2O provides a point of comparison as an unbridged analogue of [Cu(phen)(H2O)3(HfF6)]·H2O.Lambda (Λ)-shaped molecules have been demonstrated as efficient building blocks in the synthesis of non-centrosymmetric (NCS) materials 2O)3(TiF6)]·H2O and crystallizes in the orthorhombic space group Pbca  to 1.9035 (13) Å. The Λ-shape, indicated by the Cu1—F1—Ti1 bond angle of 134.93 (6)°, is enforced by the two intramolecular O2—H2B⋯F5 and O3—H3B⋯F6 hydrogen bonds (Table 1Compound (I)ca Fig. 1. The strs Table 1.2O)3(ZrF6)]·H2O and crystallizes in the monoclinic space group P21/n  to 2.0430 (6) Å. The Λ-shape, indicated by the Cu1—F1—Zr1 bond angle of 132.59 (3)°, is enforced by an intramolecular O2—H2B⋯F6 hydrogen bond (Table 262− group significantly relative to the TiF62− group in compound (I)Compound (II)/n Fig. 2. The strd Table 2. The sin2O)3(HfF6)]·H2O and crystallizes in the monoclinic space group P21/n /n Fig. 3. Compoun2O)2F]2[HfF6]·H2O and crystallizes in the monoclinic space group P21/n 2F+ cations and octahedral HfF62− anions. The free HfF62− octahedron occupies an inversion center with three distinct bond lengths ranging between 1.9863 (10) and 1.9957 (9) Å.Compound (IV)/n Fig. 4. The strThe Λ-shaped building units in compounds (I)–(III) are arranged in head-to-tail chains via intermolecular hydrogen bonding with multiple hydrogen-bonding interactions and π–π stacking contacts to adjacent chains.2O)3(TiF6)] complex in compound (I)2O)3(TiF6)] complexes and three free water molecules 3(MF6)]  units in compound (II)2O)3(MF6)] complexes and three contacts to hydrating water molecules 3(MF6)] complexes participate in parallel displaced π–π stacking interactions (Table 5The [Cu(phen)2F+ complexes (H2O)2F+ complexes and O1—H1B⋯F4 hydrogen bonds with HfF62− groups (Table 4B⋯F1 hydrogen bond to an adjacent Cu(phen)(H2O)2F+ complex and a O2—H2A⋯O3 hydrogen bond with a free water molecule (Table 4MF62− group forms hydrogen bonds with four free water molecules and two Cu(phen)(H2O)2F+ complexes. The Cu(phen)(H2O)2F+ complexes pack with both face-to-face and parallel displaced π–π stacking interactions (Table 5In compound (IV)es Fig. 8. The equs Table 4. The apie Table 4. Each MFs Table 5.2O)5(VO(H2O)F4)]·H2O 5(VOF4(H2O))]·H2O contains a molecular Λ-shaped [Cu(H2O)5(VOF4(H2O))] molecule that is bridged via the Cu1—O8—V1 linkage with a bond angle of 142.88°. The Λ-shape of this complex is supported by a single intramolecular hydrogen bond as well as two hydrogen-bonding interactions with a free water molecule that serves as an intermolecular ‘bridging molecule’. In contrast, the hydrating water molecules in compounds (I)2O)5(VOF4(H2O))]·H2O is 88.42°, meaning that the complex has a small tilt similar to compound (I)Aside from compounds (I)2O)5(VO(H2O)F4)]·H2O are arranged in a polar NCS lattice containing head-to-head/tail-to-tail chains in which the polar moments of the Λ-shaped complexes are partially aligned perpendicular to the chain direction, with head-to-tail orientations between chains. In contrast, the Λ-shapes found in compounds (I)The Λ-shapes in [Cu2, 2.56 mmol of 1,10-phenanthroline, 1.0 mL (27.6 mmol) of HF(aq) (48%), and 0.1 mL (5.5 mmol) of deionized H2O. Compound (II)2, 2.56 mmol of phen, 1.0 mL (27.6 mmol) of HF(aq) (48%), and 0.1 mL (5.5 mmol) of deionized H2O. Compound (III)2, 2.56 mmol of phen, 1.0 mL (27.6 mmol) of HF(aq) (48%), and 0.1 mL (5.5 mmol) of deionized H2O. Compound (IV)2, 2.56 mmol of phen, 0.4 mL (11.03 mmol) of HF(aq) (48%), and 0.7 mL (38.85 mmol) of deionized H2O.The compounds reported here were synthesized by the hydrothermal pouch method (Harrison Uiso(H) = 1.2Ueq(C) within OLEX2 .Crystal data, data collection and structure refinement details are summarized in Table 610.1107/S2056989021000645/tx2034sup1.cifCrystal structure: contains datablock(s) I, II, III, IV, I, II, III, IV. DOI: 10.1107/S2056989021000645/tx2034Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989021000645/tx2034IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989021000645/tx2034IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989021000645/tx2034IVsup5.hklStructure factors: contains datablock(s) IV. DOI: 2045776, 2045775, 2045774, 2045773CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Retinal ischemia is a medical condition associated with numerous retinal vascular disorders, such as age-related macular degeneration, glaucoma, and diabetic retinopathy. This in vitro cell and in vivo animal study investigated not only the protective effect of S-allyl L-cysteine  against retinal ischemia but also its associated protective mechanisms. α, and vascular endothelium factor (VEGF) in the proposed anti-ischemic mechanism. Lastly, the safety of drug consumption was investigated for changes in the animal's body weight, ERG waves, and blood biochemical parameters .  Retinal ischemia was mimicked by raising the intraocular pressure to 120 mmHg for 1 hour in one eye. The effects of pre-/postischemic administration of vehicle vs. SAC 0.18 mg vs. SAC 0.018 mg vs. SAC 0.0018 mg treatments on retina cells were evaluated through cellular viability (MTT assay), flash electroretinograms (ERGs), and fluorogold retrograde labelling  counting). Also, protein immunoblot was utilized to assess the role of Wnt, hypoxia inducible factor (HIF)-1α/VEGF protein was ameliorated significantly by preischemic low dose of SAC. In terms of the animal safety, no significant body weight and electrophysiological differences were observed among defined different concentrations of SAC without following ischemia. In low SAC dosage and vehicle groups, various blood biochemical parameters were normal; however, high and medium concentrations of SAC significantly lowered the levels of uric acid, Hb, and MCHC.  The characteristic ischemic changes including significant reduction in ERG b-wave ratio and RGC number were significantly counteracted by pre- and postischemic low dose of SAC. Additionally, ischemia-induced overexpression of Wnt/HIF-1α/VEGF might presently explain SAC's anti-ischemic mechanism. This study shows that preischemic administration of low SAC dosage has been proved to be safe and most effective against rat retinal ischemia electrophysiologically and/or histopathologically. Moreover, counteracting the ischemia-induced overexpression of Wnt/HIF-1 Common retinal ischemic disorders include central/branch retinal artery occlusion, central/branch retinal vein occlusion, glaucoma, diabetic retinopathy, and neovascular age-related macular degeneration. These diseases are associated with visual impairment and even blindness in more severe cases. Characteristic retinal ischemic findings are known to affect electroretinogram's b-wave amplitude and retinal ganglion cell (RGC) number . There aβ-catenin expression. This is done by binding to frizzled and low-density lipoprotein receptor-related protein 5 or 6, which prevents the breakdown of β-catenin [β-catenin can work with lymphoid enhancer-binding factor 1, in order to regulate the target gene expression as vascular endothelial growth factor (VEGF) [α (HIF-1α), which consequently also increases VEGF levels in hypoxia [The current knowledge is that the canonical Wnt pathway is said to stabilize -catenin . Liu et r (VEGF) . Other s hypoxia –12. Our  hypoxia .−8 g/kg≈3∼150 ng of SAC has been proven to protect the human eye against retinal ischemia via its antioxidative and other neuroprotective properties [Therefore, it is important to carry out an investigation into the ischemic alterations associated with the Wnt pathway as well as provide an alternative and complementary treatment to it. Due to the limitations of current treatment , new strategies for retinal vascular disorders driven by persistent ischemia/hypoxia are needed . In thisoperties , 8. Lastoperties  makes itα, or VEGF (identified through western blotting assay). Finally, the safety of fortified SAC in animals was also evaluated through its effects on the animal's body weight (BW), electrophysiology, and biochemical alterations. Overall, it is hypothesized that SAC would be able to nullify ischemic-induced insults in the retina and with little side effects. Specifically, the addition of SAC would downregulate the level of Wnt, HIF-1α, and VEGF protein expression. Thus, it would lead to less pronounced retinal ischemia associated alterations, namely, decreased RGC number and ERG b-wave amplitude.In the following study, the effects and mechanisms of SAC against retinal ischemia-reperfusion (I/R) injury were explored, via cellular viability analysis, electroretinogram measurement along with retrograde fluorogold labelling RGC number count, and also its regulation on the defined proteins of Wnt, HIF-1μg (0.00018 mg) in a 0.2 kg rat and 10 μg of a 70 kg human. This means that the cellular concentration of the fortified (10-fold) SAC  is equivalent to 0.0018 mg (1.8 μg) per day in animal tests. In the present in vitro cellular viability tests, ten (1 mM), hundred (10 mM), and thousand folds (100 mM) of the original dose were prepared. These are equivalent to 0.0018 (low dose), 0.018 (medium dose), and 0.18 mg (high dose) used in the in vivo animal studies.SAC was purchased from Sigma-Aldrich  and prepared by dissolving it in distilled water. Based on our previous publication , the maxFor the animal study, defined administration by oral gavage of daily dose of SAC was carried out in these following groups, namely, a preischemic administration (SAC 0.0018 mg + I/R and SAC 0.018 mg + I/R) for 4 weeks or a postischemic administration (I/R + SAC 0.0018 mg) for seven days. In contrast, the rats in the Vehicle + I/R group subjected to ischemia were preadministrated with the same volume of vehicle as that of the pre-/postadministrated SAC.The RGC-5 cell lines for the dosing regimen were prepared using the protocol described by Chao et al.  but withμL of experimental cells and allowed to react for a total of 3 h at 37°C. Then, the reduced MTT was solubilized by the addition of 100 μL of dimethyl sulfoxide [The MTT assay was carried out, in order to test the efficient concentrations of SAC. When the NAD(P) H-dependent cellular oxidoreductase enzymes reduces MTT, it forms a purple-colored substance called formazan. In this case, a more purple solution reflects a higher number of viable cells. In order to conduct this test, MTT  was placed into 96-well plate that contains 100 This animal study followed the regulations, which is set by ARVO Statement for the Use of Animals in Ophthalmology and Vision Research. Also, the permission to conduct the study was obtained from institutional review board of Cheng-Hsin General Hospital . As for the animals, six-week-old Wistar rats (BioLasco) were purchased and reared in an environment set at 40∼60% humidity and 19∼23°C temperature. Throughout the experimental days, the animals were kept on a 12 h light/dark setting along with 12∼15 air refreshment per hour and also given food and water at their own pleasure.Then, the animals were randomly placed into normal, ischemic (Vehicle + I/R), preischemic treatment groups (SAC 0.0018 mg + I/R and SAC 0.018 mg + I/R) and postischemic treatment group (SAC I/R + 0.0018 mg).An intraperitoneal injection was employed to anesthetize the rats. Specifically, 100 mg/kg ketamine (Pfizer) and 5 mg/kg xylazine (Sigma-Aldrich) are given to the rat, which causes the effect of sedation along with analgesia. Another intraperitoneal injection of 140 mg/kg sodium pentobarbital (SCI Pharmtech) was conducted to humanely kill the rat (Scientific Procedures Acts 1986).Each rat (200–250 g) was anesthetized using the method described above and set onto a stereotaxic frame. As described in Chao et al. , a rat mAfter the rats were euthanized, blood sample is slowly taken from the rat's left ventricle, via technique known as cardiac puncture . Then, t−1 was given via a strobe 2 cm before the rat's eyes. In this case, fifteen continuous recordings were done at each 2 s interval, which is set at 10 s−1. Afterwards, their amplitudes were maximized and calibrated to give a mean, via usage of an amplifier P511, regulated power supply RPS107, and stimulator PS22 (Grass-Telefactor). The b-wave amplitude ratio of I/R eye to that of the normal eye was measured, in order to compare between different experimental treatments as illustrated by Chao et al. [For the flash ERG measurements, they were carried out on day 0 (nonadministered rats) along with the day after vehicle + I/R, SAC + I/R, or I/R + SAC procedure. Then, the dark adaptation was carried out for a total of 8 hours, and anesthesia was conducted for ERG recording with pupil dilation. Then, a stimulus of 0.5 so et al. .μl of 5% fluorogold (Sigma-Aldrich) at 3.8, 4.0, and 4.2 mm below its skull. Of note, this step was carried out 3 days prior euthanasia of rat. Then, the retina was dissected and collected, as described by Chao et al. [The anesthetic rat is placed onto the stereotaxic frame, and a 2 cm deep cut is created on the rat's scalp and 2 small holes drilled into its skull . Afterwao et al. . Finallyo et al. .μg/30 μl/well, and underwent separation through 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis . Once that is completed, the samples were transferred from the gel onto polyvinylidene fluoride membrane. The membranes itself were soaked with 5% fat-free skimmed milk at 4°C and blocking buffer  for 16 hours. Then, the membranes were incubated with a series of primary antibodies  at 25°C for 1 hour, namely, mouse anti-β-actin monoclonal antibody , rabbit anti-Wnt3a monoclonal antibody , mouse anti-HIF-1α antibody , and rabbit polyclonal anti-VEGF antibody . The blots were then incubated with the relevant secondary antibodies, which were horseradish peroxidase-conjugated goat anti-rabbit IgG  and anti-mouse IgG  at 25°C for 1 hour. Lastly, the blots were developed through enhanced chemiluminescent analysis system (HyCell) and exposed to an X-ray film (Fujifilm), and then the proteins levels were analyzed through scanning densitometry.The samples of the retina were isolated and sonicated in lysis buffer  after the rat is euthanized. Then, they were divided to similar amount of 30 t test was utilized to compare between two groups, whereas one-way analysis of variance (ANOVA) was carried out for comparison of three or more than 3 independent groups. After the ANOVA tests, Dunnet's test was selected to compare the control  versus the rest of the groups . Of note, results were presented as mean ± SE, and a P value that is smaller than 0.05 was considered as significant.An unpaired To test the optimal and safe cellular concentration of fortified SAC onOGD-insulted cells, lower  and higher  concentrations of pre-OGD administration of SAC were tested and then compared against the control group .As demonstrated in P < 0.001; at 1 mM) attenuation of the OGD-induced cellular injury.In contrast, as shown in Overall, the result implies that SAC with concentrations below 1 mM or 0.0018 mg/day should have some form of protective effect against ischemic injury. Thus, this data along with ERG's results (mentioned later) lead to the selection of SAC 0.0018 mg for testing in animal studies, namely, fluorogold labelling and western blot analysis.SAC 0.0018 mg”, which is as follows: baseline (220.2 ± 18.1 vs. 217.0 ± 18.2 vs. 221.8 ± 16.9 vs. 224.0 ± 18.1), week 1 (258.2 ± 16.0 vs. 257.3 ± 14.5 vs. 262.2 ± 15.8 vs. 269.5 ± 18.2), week 2 (282.2 ± 13.8 vs. 280.0 ± 8.3 vs. 304.5 ± 14.7 vs. 292.0 ± 12.1), week 3 (324.0 ± 12.3 vs. 310.7 ± 6.7 vs. 325.7 ± 9.1 vs. 328.2 ± 8.2), and week 4 (321.0 ± 9.5 vs. 322.3 ± 8.7 vs. 323.3 ± 12.5 vs. 330.7 ± 17.7). Of note, these BW results were represented as mean ± SE (g). From the values above, it could be implied that the animals followed a normal growth rate over time (increase of BW), which is not affected by the presence of vehicle or SAC.As shown in SAC 0.0018 mg” across weeks, which enables a comparison between different groups. The ERG a-wave amplitudes in 0.07 ± 0.02), week 1 (0.10 ± 0.03 vs. 0.09 ± 0.02 vs. 0.07 ± 0.02 vs. 0.05 ± 0.01), week 2 (0.09 ± 0.03 vs. 0.08 ± 0.02 vs. 0.10 ± 0.02 vs. 0.06 ± 0.02), week 3 (0.07 ± 0.02 vs. 0.07 ± 0.02 vs. 0.06 ± 0.02 vs. 0.06 ± 0.02), and week 4 (0.09 ± 0.03 vs. 0.07 ± 0.02 vs. 0.09 ± 0.02 vs. 0.06 ± 0.01). Throughout the 4 weeks of observation, there was no significant difference of a-wave amplitudes between each group and at each week.By means of ERG, preischemia daily doses of vehicle or SAC  have been proved to be safe, as shown in Tables 0.41 ± 0.07), week 1 (0.49 ± 0.05 vs. 0.44 ± 0.05 vs. 0.44 ± 0.06 vs. 0.45 ± 0.06), week 2 (0.43 ± 0.07 vs. 0.37 ± 0.06 vs. 0.46 ± 0.01 vs. 0.47 ± 0.08), week 3 (0.41 ± 0.07 vs. 0.35 ± 0.04 vs. 0.44 ± 0.06 vs. 0.40 ± 0.01), and week 4 (0.35 ± 0.04 vs. 0.38 ± 0.05 vs. 0.44 ± 0.03 vs. 0.41 ± 0.04). Similarly, these data are also not significantly different across different groups and various weeks.As for Of note, the ERG a-wave amplitudes are relatively smaller and sometimes harder to be measured than b-wave amplitudes. Thus, it is usually disregarded for the test of therapeutic properties of drugs; instead, it is presently for safety tests and evidence of retinal normality.https://pubmed.ncbi.nlm.nih.gov/31651183/ [SAC 0.0018 mg” for comparison.As reported in previous publication 1651183/ , blood ln = 4∼11): for glucose (136.55 ± 4.67 vs. 124.71 ± 6.83 vs. 128.76 ± 3.73 vs. 130.39 ± 8.41), blood urea nitrogen (15.89 ± 0.51 vs. 15.39 ± 0.42 vs. 15.75 ± 0.65 vs.17.14 ± 1.25), creatinine (0.41 ± 0.01 vs. 0.43 ± 0.01 vs. 0.42 ± 0.02 vs. 0.47 ± 0.02), AST (109.71 ± 5.91 vs. 104.25 ± 5.46 vs. 112.86 ± 9.12 vs. 107.71 ± 2.30), ALT (35.67 ± 1.62 vs. 31.43 ± 0.84 vs. 33.71 ± 2.16 vs. 34.86 ± 1.72), ALKP (101.40 ± 5.22 vs. 113.00 ± 4.99 vs. 113.17 ± 5.38 vs. 108.14 ± 3.94), triglyceride (39.11 ± 6.08 vs. 35.57 ± 3.47 vs. 38.00 ± 7.48 vs. 35.22 ± 3.78), cholesterol (41.89 ± 2.34 vs. 38.11 ± 3.01 vs. 36.00 ± 2.16 vs. 40.33 ± 1.35), high-density lipoprotein , low-density lipoprotein , red blood cells (8.12 ± 0.16 vs. 8.00 ± 0.11 vs. 7.79 ± 0.18 vs. 8.19 ± 0.13), hematocrit (46.03 ± 0.65 vs. 44.04 ± 0.49 vs. 44.94 ± 0.77 vs. 44.24 ± 0.60), mean corpuscular volume (MCV: 55.68 ± 0.72 vs. 55.46 ± 0.50 vs. 55.46 ± 0.85 vs. 54.51 ± 0.67), mean corpuscular hemoglobin , and platelet (1018.89 ± 33.50 vs. 1022.00 ± 36.15 vs. 1148.50 ± 46.56 vs. 1032.50 ± 69.83). In addition, there were also no significant differences among various groups (n = 6∼8) for albumin (1.51 ± 0.04 vs. 1.68 ± 0.04 vs. 1.58 ± 0.03) and WBC (6.19 ± 0.29 vs. 7.140.39 vs. 6.80 ± 0.51).After 4 weeks of treatment, there were no significant differences among various groups (P=0.005) vs. 1.12 ± 0.10 (P < 0.001) vs. 1.49 ± 0.23. As for P < 0.001) vs. 15.81 ± 0.26 vs. 15.41 ± 0.22. Finally, the mean corpuscular hemoglobin concentration found in P=0.006) vs. 34.73 ± 0.18 vs. 34.15 ± 0.35.Of note, the only significant side effects were the following parameters displayed in P < 0.001) resulted in ERG b-wave ratio reduction of 0.20 ± 0.04 (n = 8), relative to that of normal (control) retina . The ischemia-induced ERG b-wave reduction was either tended to be alleviated or significantly blunted by pre- or postischemia oral ingestion of SAC, namely, SAC 0.018 mg + I/R , SAC 0.0018 mg + I/R , and I/R + SAC 0.0018 mg .As shown in Figures n = 4), the Vehicle + I/R treatment led to significant (P < 0.001) reductions in RGC numbers per field of 185.48 ± 7.50  elevated number of RGCs cells, namely, 329.52 ± 17.25 cells/field for SAC 0.0018 mg + I/R  upregulation of target proteins, namely, Wnt3a  are target genes of Wnt pathway, which largely correlated with the development of ischemic-related ocular diseases [The results show that I/R significantly affected the retina in a number of ways, via cellular viability Figures  and 4, ediseases .α and in return increases VEGF [α  for 4 weeks was proven to be the safest and most effective, via observation of body weight changes , ERG altα/VEGF overexpression , relative to conventional treatment methods such as anti-VEGF.Preischemia administration of fortified SAC (low dose: 0.0018 mg) is proven to be experimentally safe as shown by the animal safety experiments and electrophysiological/histopathological data, in terms of the ability of SAC to alleviate retinal ischemia injury. Of medical importance and novelty to this paper, the ischemia-induced elevation of upstream protein Wnt3a and its associated downstream target HIF-1


Status epilepticus (SE) is a life-threatening neurological disorder. The hippocampus, as an important area of the brain that regulates cognitive function, is usually damaged after SE, and cognitive deficits often result from hippocampal neurons lost after SE. Fyn, a non-receptor Src family of tyrosine kinases, is potentially associated with the onset of seizure. Saracatinib, a Fyn inhibitor, suppresses epileptogenesis and reduces epileptiform spikes. However, whether saracatinib inhibits cognitive deficits after SE is still unknown.In the present study, a pilocarpine-induced SE mouse model was used to answer this question by using the Morris water maze and normal object recognition behavioral tests.We found that saracatinib inhibited the loss in cognitive function following SE. Furthermore, we found that the number of hippocampal neurons in the saracatinib treatment group was increased, when compared to the SE group.These results showed that saracatinib can improve cognitive functions by reducing the loss of hippocampal neurons after SE, suggesting that Fyn dysfunction is involved in cognitive deficits after SE, and that the inhibition of Fyn is a possible treatment to improve cognitive function in SE patients. Epilepsy is a chronic neurological disorder characterized by a persistent occurrence of seizures. Status epilepticus (SE) is characterized by prolonged seizures or intermittent seizures and unconsciousness , which pA receptors and is associated with both excitatory and inhibitory ion channels, which are potentially associated with the onset of seizure .Sarscatinib  was administered (25 mg/kg) orally starting 2 h after diazepam injection and repeated twice daily for the first 3 days followed by a single dose each day for the next 11 days during the 2 weeks after SE. Same volume of saline was gave to CON mice.Mice that received continuous video-EEG monitoring were implanted with silver wire electrodes (0.125 mm in diameter) into the hippocampal dentate gyrus (DG) region after anesthesia with pentobarbital . The electrode implantation site used the following coordinates with the bregma as the reference: bregma: − 2.3 mm, lateral: 1.8 mm and depth: 2.0 mm. The reference electrode was placed in the frontal cortex. All implanted surgery was performed at 7 days before the induced SE. EEG activity was recorded 12 h every day for up to 7 days at 14 days after induced SE using PowerLab 8/35 software  . Epileptic spikes (sharp (< 50 ms) positive or negative deflections with amplitudes exceeding twice the baseline EEG) were detected and scored by the Gotman spike using PowerLab software. Mice behavior was monitored using video and reviewed by an investigator who was blinded to the identity of the groups (n = 5 each in every group).One month after induced SE, the mice were anesthetized with pentobarbital  and transcardially perfused with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde in 0.1 M PBS to sacrifice the mice. The brain was dissected and fixed in 4% paraformaldehyde overnight at 4 °C and then cryoprotected in 30% sucrose in PBS for 72 h at 4 °C. Coronary sections (8 μm) were cut with a cryostat and mounted onto glass slides. Sections were washed in PBS and rehydrated in ethanol of decreasing concentrations. After washing with PBS, the sections were incubated with anti-NeuN antibody  overnight at 4 °C and washed with PBS. The sections were then incubated with secondary antibody and visualized using 3,3-diaminobenzidine. NeuN-positive neurons in a 100 μm line long DG of hippocampus were counted in five sections from identical levels in each mouse (n = 5/groups).p < 0.05 was considered statistically significant.Data are expressed as the mean ± SEM. One-way or two-way analysis of variance was used to assess differences between two groups and the least significant difference (LSD) or Dunnett T3 post hoc test was used to compare multiple groups after normality tests. A value of Mice often exhibited hypoactivity, curling up, tremors, head bobbing, and myoclonic movement of the limbs after pilocarpine injection. SE usually began by rearing with or without falling and jumping at the onset of SE.t(8)=2.986, p =0.007). Furthermore, video-EEG showed that saracatinib injection decreased the duration =2.763, p =0.025) and frequency =3.715, p =0.006) of spontaneous seizure after SE  = 3.68; CON vs SE: 33.59 ± 3.8 vs 50.29 ± 4.87 on 3 days, p = 0.13; F = 7.116; CON vs SE vs Sar + SE: 22.13 ± 3.4 vs 45.83 ± 5.8 vs 30.11 ± 4.18 on 4 days, p = 0.01 in CON vs SE, p = 0.025 in SE vs Sar + SE; F = 10.307; CON vs SE vs Sar + SE: 17.42 ± 2.88 vs 41.72 ± 3.94 vs 26.45 ± 4.69 on 5 days, p = 0.009 in CON vs SE; p = 0.012 in SE vs Sar + SE) and swimming path length from the starting position to the target platform  = 3.02; CON vs SE: 1589.37 ± 213.03 vs 2490.87 ± 296.99 on 3 days, p = 0.23; F = 8.54; CON vs SE: 1138.7 ± 166.62 vs 2381.65 ± 258.46 on 4 days, p = 0.15; F = 13.81; CON vs SE vs Sar + SE: 802.907 ± 103.21 vs 1786.48 ± 155.98 vs 1070.8637 ± 113.52 on 5 days, p = 0.001 CON vs SE; p = 0.16 SE vs Sar + SE) were decreased in Sar + SE mice, when compared with the SE mice. However, the swimming speed was similar in all groups mice  = 0.54; CON vs SE vs Sar + SE: 62.09 ± 2.69 vs 61.72 ± 3.02 vs 57.85 ± 3.79 on 3 days, p > 0.05; F = 0.475, CON vs SE vs Sar + SE: 57.46 ± 2.87 vs 55.94 ± 3.3 vs 59.62 ± 2.6 on 4 days, p > 0.05; F = 0.11, CON vs SE vs Sar + SE: 59.01 ± 2.36 vs 57.85 ± 2.7 vs 57.43 ± 2.46 on 5 days, p > 0.05). There results suggested that decreased escape latency and swimming path length in the Sar + SE group mice were not due to an impaired capacity of swim. When the target platform was removed in the probe trails, the number of crossing target platform areas  = 5.917; CON vs SE vs Sar + SE: 3.22 ± 0.43 vs 1.25 ± 0.25 vs 2.5 ± 0.5, p = 0.012 in CON vs SE; p = 0.04 in SE vs Sar + SE) and staying time in the quadrant  = 5.202; CON vs SE vs Sar + SE: 20.8 ± 1.97 vs 12.84 ± 2.14 vs 18.237 ± 0.922, p = 0.01 in CON vs SE; p = 0.38 in SE vs Sar + SE) where the target platform was previously located were increased in the Sar + SE group, when compared with the SE group. Furthermore, when we performed the NOR test, the PI was similar in both SE group mice and Sar + SE group mice  = 0.326; CON vs SE vs Sar + SE: 0.58 ± 0.017 vs 0.553 ± 0.02 vs 0.545 ± 0.049, p > 0.05), but the RI of the Sar + SE mice was increased  = 6.944; CON vs SE vs Sar + SE: 0.64 ± 0.029 vs 0.453 ± 0.039 vs 0.578 ± 0.036, p = 0.015 in CON vs SE; p = 0.023 in SE vs Sar + SE). Together, these results showed that saracatinib inhibited deficits in cognitive function after SE.Cognitive function deficits are a common sequelae after SE, so we examined whether saracatinib inhibited deficits in cognitive function after SE. Using the MWM test, escape latency Fig. a  = 12.32; CON vs SE vs Sar + SE: 30.32 ± 1.65 vs 20.48 ± 1.09 vs 26.44 ± 1.42, p = 0.005 in CON vs SE; p = 0.032 in SE vs Sar + SE), suggesting that saracatinib relieved the cognitive function deficits by rescuing hippocampal neuronal loss during SE.Because hippocampal neuron loss is an important reason for the deficits in cognitive function after SE, we next determined the number of hippocampal neurons. Neurons in the DG region in the hippocampus were visualized by anti-NeuN antibody using immunohistochemical staining Fig. a. The reSE is a neurological condition with high mortality, involving continuous seizure activity, which is often considered as one of the precipitating factors for temporal lobe epilepsy (TLE). The hippocampus and piriform cortex are brain areas commonly damaged by SE. In addition, the amygdala, thalamus, neocortex, and cerebellum are also brain areas that are usually damaged after SE . NeuronaThe Src kinase family is a family of non-receptor tyrosine kinases that plays important roles in regulating signal transduction. Fyn, as a member of the Src family kinases, plays critical roles in regulating cognitive function in AD and in frontotemporal dementia (FTD) patients , 26. FynLoss of hippocampal neurons is the most common reason for cognitive deficits. Neuronal loss frequency occurs in both patients with TLE and in animal models of SE , 22, 32.The Fyn inhibitor, saracatinib, decreased cognitive deficits by attenuating the loss of hippocampal neurons in a pilocarpine-induced SE mouse model, indicating that inhibition of Fyn activity can potentially improve cognitive deficits in patients with TLE. However, further investigation is necessary to identify the mechanism responsible for this neuroprotective effects.


Cyclic GMP‐AMP synthase (cGAS) is a cytosolic DNA sensor that catalyzes the synthesis of the cyclic GMP‐AMP dinucleotide 2′3′‐cGAMP. 2′3′‐cGAMP functions as inducer for the production of type I interferons. Derivatives of this important second messenger are highly valuable for pharmaceutical applications. However, the production of these analogues requires complex, multistep syntheses. Herein, human cGAS is shown to react with a series of unnatural nucleotides, thus leading to novel cyclic dinucleotides. Most substrate derivatives with modifications at the nucleobase, ribose, and the α‐thio phosphate were accepted. These results demonstrate the catalytic promiscuity of human cGAS and its utility for the biocatalytic synthesis of cyclic dinucleotide derivatives. Enzymatic shortcut: Cyclic dinucleotides, which are of great interest to study immunology and immune oncology, can be synthesized in a one‐step biotransformation significantly shortening the chemical synthesis route. The enzyme displays a surprisingly large substrate scope. Cyclic dinucleotides are second messengers that can be found in prokaryotes and also in eukaryotes.Vibrio cholerae.During recent years, the structure, regulation, mechanism, and kinetics of human cGAS and other homologues have been described.4b, 5 Herein, we show that the substrate promiscuity of human cGAS can be exploited to produce a series of 2′3′‐cGAMP analogues in a one‐step reaction superior to classical synthesis routes. The substrate derivatives were chosen with modifications at different positions in the nucleobase, ribose or α‐phosphate Scheme . Each su2‐ATP (entry 3) and ATP‐α‐S (entry 16) and only minor amounts for the substrate 8‐Cl‐ATP (entry 13). Higher specific activities between 40 and 43 mU mg−1 were determined for the derivatives 8‐Br‐dATP (entry 11), 8‐Br‐ATP (entry 10) and GTP‐α‐S (entry 17) with conversions between 56 and 93 %. The specific activities are comparable to those measured for the conversion of the natural substrates ATP and GTP (74 mU mg−1). The highest specific activity of 136 mU mg−1 was measured for the product synthesis from 8‐Br‐GTP with a yield of 85 %. Thus, the addition of a bromide at the 8‐position of the nucleobase seems to be preferred for both, ATP and GTP, substrates. Another interesting substrate derivative, the α‐thio phosphate GTP analogue (entry 17), was accepted for the cyclization reaction and 93 % was converted within 24 h. This product has relevant chemical properties and is of particular importance, because the phosphothionate diester linkages are more resistant to hydrolysis than the phosphate diesters.The specific activities and conversions within 24 h were calculated for all reactions Table . No prodThe cGAS products were additionally analyzed by electrospray ionization‐liquid chromatography‐mass spectrometry (ESI‐LC‐MS) and the expected masses were compared with the detected product masses. The synthesis of nine of the 17 products was confirmed with ESI‐LC‐MS analysis  were converted into the corresponding cyclic dinucleotide derivative using cGAS within 24 h. The conversion of 5 μmol ATP and GTP served as a reference reaction. The reactions resulted in 65 % conversion of GTP with 2′‐F‐ATP, 47 % GTP with 8‐Br‐ATP, 35 % GTP with 8‐Cl‐ATP, 55 % GTP with 8‐NH2‐ATP, and 58 % GTP with 8‐N3‐ATP. These conversions were similar, but not fully comparable with the biotransformations on analytical scale, which can only be attributed to the change in the reaction scale. HPLC Purification of 2′3′‐cGAMP yielded 1.1 mg of product. The derivatives were purified by two‐step HPLC to give 0.1 mg cyclic GMP‐2′‐F‐AMP, and 4.1 mg cyclic GMP‐8‐NH2‐AMP. The syntheses of cGAMP, cyclic GMP‐2′‐F‐AMP and cyclic GMP‐8‐NH2‐AMP were successfully validated by NMR spectroscopy (see the Supporting Information for details). The products cyclic GMP‐8‐Br‐AMP, cyclic GMP‐8‐N3‐AMP and cyclic GMP‐8‐Cl‐AMP were not purified in quantifiable amounts.Based on our findings mg‐scale biotransformations of 2′‐F‐ATP, 8‐Br‐ATP, 8‐Cl‐ATP, 8‐NHIn summary, we have reported a short and facile enzymatic synthesis of novel unnatural cyclic dinucleotides based on the promiscuous activity of human cGAS. Remarkably, most tested ATP and GTP substrate derivatives with modifications at the nucleobases, riboses and α‐phosphate were converted into cyclic dinucleotides with their natural counterpart.The relevance of these cyclic dinucleotide derivatives was recently demonstrated by investigating the biological activity towards the STING receptor based on the analysis of the structure‐activity relationship and mouse model systems.Analytical scale biotransformation: For enzyme assays, 40 mM HEPES, 10 mM MgCl2, pH 7.2 was used with 0.1 mg mL−1 Herring testis DNA and 40 μg mL−1 human cGAS. The reaction was started by adding 0.5 mM substrate derivative and 0.5 mM substrate, either ATP or GTP. The reaction volume was 1 mL. Negative controls were performed analogously without the addition of enzyme. All samples were incubated at 37 °C and 300 rpm in an orbital shaker for at least 24 h. Reactions were stopped by heating at 95 °C for 5 min. The samples were analyzed with HPLC and LC‐MS. The activities given in Table Milligram‐scale biotransformation: For milligram‐scale biotransformation, 20 mL reaction solution  was used in shaking flasks. The reaction was started by adding 40 μg mL−1 human cGAS. The reaction mixture was incubated at 37 °C and 200 rpm in an orbital shaker for 24 h. The reaction was stopped by heating at 95 °C for 5 min. The product solutions were frozen and stored at −20 °C.Fractionation of biotransformation products by HPLC: The frozen samples were lyophilized to dryness, subsequently resolved in 1 mL H2O and centrifuged at 21 000 g for 5 min. The supernatant of the product solution was separated and collected as fractions with a LaChrome Elite HPLC system  equipped with an ISAspher 100‐3C18AQ column, 150×3 mm . The column temperature was set to 30 °C. The flow rate was set to 1 mL min−1. The product solution was repetitively injected with volumes of 99.5 μL. A gradient of 50 mM triethylamine acetate (TEAA) with 3 % v/v acetonitrile (solvent A) and 100 % acetonitrile (solvent B) was used for the chromatography. The solvent gradient for all products was: 0–10 min: 0 % B, 10–20 min: 0 to 30 % B, 20–22 min: 30 % B, 22–25 min 30 to 0 % B, 25–30 min 0 % B. Fractions were taken: cGAMP: 12–13 min; cyclic GMP‐2′‐F‐AMP: 11–12 min and 17–18 min; cyclic GMP‐8‐Br‐AMP: 17–19 min; cyclic GMP‐8‐Cl‐AMP: 16–18 min; cyclic GMP‐8‐NH2‐AMP: 11–13 min; cyclic GMP‐8‐N3‐AMP: 16–17 min. Elution of compounds was monitored with a UV detector at 254 nm. The collected fractions were lyophilized and product purification was validated with NMR analysis.2‐AMP: 0–18 min: 0 % B, 18–28 min: 0 to 30 % B, 28–30 min: 30 to 90 % B, 30–38 min: 90 % B, 38–40 min: 90 to 0 % B, 40–50 min: 0 % B; Fractions were taken: 25.9–27.9 min; cyclic GMP‐8‐N3‐AMP: 0–15 min: 0 to 22.5 % B, 15–17 min: 22.5 to 90 % B, 17–25 min: 90 % B, 25–27 min: 90 to 0 % B, 27–37 min: 0 % B; Fractions were taken: 7.1–9.1 min. Elution of compounds was monitored with a UV detector at 254 nm. The collected fractions were lyophilized and product purification was validated with NMR analysis.For further purification of the product derivatives, a second fractionation using chromatography was performed. In the second fractionation, the solvent gradient was individually adapted for each compound: cyclic GMP‐2′‐F‐AMP: 0–20 min: 0 to 22.5 % B, 20–22 min: 22.5 to 90 % B, 22–30 min: 90 % B, 30–32 min 90 to 0 % B, 32–42 min 0 % B; Fractions were taken: 8.7–10.2 min; cyclic GMP‐8‐Br‐AMP: 0–20 min: 0 to 15 % B, 20–22 min: 15 to 90 % B, 22–30 min: 90 % B, 30–32 min: 90 to 0 % B, 32–42 min: 0 % B; Fractions were taken: 14.3–16.3 min; cyclic GMP‐8‐Cl‐AMP: 0–5 min: 0 % B, 5–25 min: 0 to 30 % B, 25–27 min: 30 to 90 % B, 27–35 min: 90 % B, 35–37 min: 90 to 0 % B, 37–47: 0 % B; Fractions were taken:12.7–14.1 min and 16.3–18.3 min; cyclic GMP‐8‐NHNMR analysis: NMR spectra were recorded on a Bruker AV 600 Avance III HD system  with D2O as solvent and 25 μL [D4]methanol as internal standard. The solvent signals were referenced to δH 3.31 ppm and δC 49.0 ppm.2‐AMPCyclic GMP‐8‐NH: 1H NMR : δ=8.08 , 7.94 , 6.17 , 5.99 , 5.60 , 5.06 , 4.72 , 4.62 , 4.44 , 4.38 , 4.27 , 4.37, 4.14 ppm ; 13C NMR : δ=160.2 (CCO), 154.5 (CN), 153.2 (CN), 153.1 (CN), 150.1 (CH), 149.7 (CN), 142.3 (CH), 118.1 (CN), 117.1 (CN), 91.5 (CH), 86.8 (CH), 84.6 (CH), 80.6 (CH), 75.6 (CH), 74.7 (CH), 72.4 (CH), 71.7 (CH), 66.7 (CH2), 62.9 ppm (CH2).Cyclic GMP‐8‐F’‐AMP: 1H NMR : δ=8.30 , 8.29 , 7.87 , 6.47 , 5.96 , 5.67 , 5.56 , 5.12 , 4.60 , 4.57 , 4.40 , 4.25 , 4.16 ppm ; 13C NMR : δ=n.d. (CCO), n.d. (CN), 156.0 (CN), 155.1 (CH), 152.2 (CN), 148.1 (CN), 142.9 (CH), 140.7 (CH), 119.2 (CN), 118.1 (CN), 87.8 (CH), 87.6 (CH), 87.2 (CH), 84.0 (CH), 74.6 (CH), 72.2 (CH), 71.8 (CH) 69.9 (CH), n.d. (CH), 66.8 ppm (CH). (n.d.=not detected)The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re‐organized for online delivery, but are not copy‐edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file.


Developing a universal strategy to improve the specificity and sensitivity of PEGylated nanoaparticles (PEG-NPs) for assisting in the diagnosis of tumors is important in multimodality imaging. Here, we developed the anti-methoxypolyethylene glycol (mPEG) bispecific antibody , which has dual specificity for mPEG and human epidermal growth factor receptor 2 (HER2), with a diverse array of PEG-NPs to confer nanoparticles with HER2 specificity and stronger intensity.++) but not MCF7/neo1 cells (HER2+/−). The αHER2/Lipo-DiR and αHER2/SPIO could enhance the sensitivity of untargeted PEG-NPs on MCF7/HER2 (HER2++). In in vivo imaging, αHER2/Lipo-DiR and αHER2/SPIO increased the specific targeting and enhanced PEG-NPs accumulation at 175% and 187% on 24 h, respectively, in HER2-overexpressing tumors.We used a one-step formulation to rapidly modify the nanoprobes with mPEG × HER2 and optimized the modified ratio of BsAbs on several PEG-NPs . The αHER2/PEG-NPs could specifically target MCF7/HER2 cells (HER2mPEG × HER2, therefore, provided a simple one-step formulation to confer HER2-specific targeting and enhanced sensitivity and contrast intensity on HER2 positive tumors for multimodality imaging.  Optical imaging (OI) is relatively inexpensive and robust for all kinds of molecular and cellular processes in small animals, but clinical applications are hindered by limited depth penetration /[µW/cm2].MCF7/HER2 (5 × 106 cell/well) were incubated αHER2/SPIO, αDNS/SPIO or SPIO were added to the tubes and incubated at 37 °C for 1 h. After washing with PBS three times, cells precipitated at the bottom of the Eppendorf tube and were then imaged with 7T MRI  TR/TE, 3000/65 ms; echo train length, 10; flip angle, 150o; field of view, 6 cm × 6 cm; slice thickness, 1 mm; interslice gap, 0.1 mm (10% of slice thickness); and matrix, 192 × 192. BALB/c nude mice bearing MCF7/HER2 (HER2++) and MCF7/neo1 (HER2+/−) tumor (~ 100 mm3) were injected intravenously with αHER2/SPIO and αDNS/SPIO (10 mg/kg per mouse) in their mammary fat pad regions, respectively. Isoflurane anesthetized mice were imaged with 7T MRI at 0, 24 h after injection. TR/TE, 3000/65 ms; echo train length, 10; flip angle, 150; field of view, 4 cm; slice thickness, 1.2 mm; interslice gap, 0.12 mm (10% of slice thickness); and matrix, 256 × 210. The negative enhancement was calculated by [pretreated signal intensity (SI0)-treated signal intensity (SI24)]/SI0*100.MCF7/HER2 cells  and 500 units/ml recombinant human granulocyte–macrophage colony-stimulating factor (R&D systems), and then seeded 4 × 105 cells/well in 24-well plate. On day 6, 20 ng/ml recombinant human TNF-α (Sigma) and 10 ng/ml IL-1β (R&D systems) were added to the cells to activate DCs for 24 h. On day 7, the harvested DCs were counted and then incubated 50 μg/ml mitomycin C for 30 min at 37 °C at a density of 1 × 106 cells/ml, then washed extensively. Autologous CD4+ T cells were isolated on Day 7 by negative selection using CD4+ T Cell Isolation Kit II and LS columns (Miltenyi Biotech). After counting, 2 × 105 CD4+ T Cells were added to 2 × 104 mitomycin C-treated DCs and incubated with mPEG × HER2, mPEG × DNS at concentration of 350 nM in 96-well round bottom plates. Controls included dendritic cells plus CD4+ T cells alone and with concentration of 10 μg/ml phytohemagglutinin (PHA). Cells were cultured at 37 °C for 5 days. Proliferation was assessed by ATPlite Luminescence Assay kit (Perkin Elmer). Counts per minute (cpm) for each well were determined by multimode plate reader (Perkin Elmer).To prepare monocyte-derived dendritic cells (DCs), peripheral blood mononuclear cells (PBMCs) from healthy donor blood isolated by Ficoll-Paque and monocytes isolated using Miltenyi Pan Monocyte Isolation Kits and LS columns (Miltenyi Biotech). Monocytes were resuspended in RPMI1640 supplemented with 10% FCS, 2 mM Additional file 1: Table S1. The BsAb-conjugation rate of αHER2/PEG-NPs. Table S2. The Characterization of BsAb/mPEG-NPs. Figure S1. Immunogenicity of humanized BsAbs. We cocultured dendritic cells differentiated from human PBMCs with autologous CD4+ T cells and stimulated with control medium (represented as DC+T), PHA (as positive control), PEG×HER2, PEG×DNS, respectively, for 5 days. Then, we detected the proliferation of CD4+ T cells by ATPlite assay. Bars, SD. CPM, counts per minute; PBMC, peripheral blood mononuclear cell; PHA, phytohemagglutinin.


Cordyceps militaris Improves Chronic Kidney Disease by Affecting TLR4/NF-κB Redox Signaling Pathway” [In the article titled “Pathway” , there w2)”.“All the patients were with the CKD late stage 3 or stage 4 (estimated glomerular filtration rate (eGFR) 25 to 40 mL/min)” should read “All the patients were with the CKD late stage 3 or stage 4 (estimated glomerular filtration rate (eGFR) 25 to 40 mL/min/1.73 m“All patients met the following criteria: (1) urine protein/creatinine ratio < 5” should read “All patients met the following criteria: (1) urinal protein > 1 g/24 h.”P < 0 05)” should read “After the three-month therapy, the values of eGFR (45.6 ± 8.4 mL/min/1.73 m2) were increased significantly when compared with before therapy .”“After the three-month therapy, the values of eGFR (28.3 ± 5.2) were reduced significantly when compared with the CG group (32.8 ± 9.2,


The cyclopentadienyl rings are eclipsed in both of the title compounds and their packing is dominated by H⋯H  contacts, as determined by Hirshfeld surface analyses. N,N-dimethylamino)methyl]ferrocene, [Fe(C5H5)(C8H12N)], (1), is an interesting starting material for the synthesis of planar chiral 1,2-disubstituted ferrocenes, as demonstrated by the preparation of -bis{2-[(dimethylamino)methyl]ferrocenyl}dimethylsilane, [Fe2(C5H5)2(C18H18N2Si)], (2), from the lithiated derivative of 1. The configuration of the lithium compound is unchanged after the substitution reaction and the chirality is preserved in space group P212121. In both compounds, the Cp rings adopt eclipsed conformations. Hirshfeld surface analysis was used to investigate the intermolecular interactions, and showed that H⋯H  interactions dominate in both structures with contact percentages of 83.9 and 88.4% for 1 and 2, respectively.The title compound [( N,N-Dimethylaminomethylferrocene  was first synthesized by Hauser & Lindsay  and can be converted by a further step using an electrophile -N,N-dimethyl-1-ferrocenylethylamine, or Ugi’s amine with a chiral directing group -tetramethyl-1,2-cyclohexanediamine (TMCDA) with yields in high stereoselectivity  and the mechanistic course of the reaction can be described by quantum-chemical calculations -meso-compound of bis[dimethyl(aminomethyl)ferrocenyl]dimethylsilane was characterized by Roewer and co-workers using X-ray diffraction analysis and formed during the synthesis of dimethyldichlorosilane with two equivalents of the racemic lithiated N,N-dimethylaminomethylferrocene -bis[dimethyl(aminomethyl)ferrocenyl]dimethylsilane (2) and analyze their intermolecular interactions using Hirshfeld surfaces and two-dimensional fingerprint plots.In this paper, we report the crystal structures of 1 crystallizes from n-pentane at 243 K as orange needles with monoclinic (P21/n) symmetry. There are no noticeable irregularities in the bond lengths or bond angles found: the aminomethyl side chain is oriented above its attached cyclopentadienyl ring, and the Cp rings are eclipsed, the dihedral angle between their mean planes being 1.53 (15)°. The molecular structure of 1 is presented in Fig. 1Compound 2 is an orange–red crystalline solid and occurs in enantiomerically pure form in the orthorhombic space group P212121. The structure is illustrated in Fig. 22 can be assigned the -configuration; furthermore the cyclopentadienyl rings are also in an eclipsed conformation for both iron atoms . The Si—C bonds span the range of 1.869 (3) to 1.874 (3) Å, which is consistent with the literature ° (C14—Si1—C10) as the smallest and 112.17 (13)° (C14—Si1—C16) as the largest. This flexibility is often observed for Si—C bonds  for N2 and 3.584 (3) Å for N1 are too long to be regarded as coordinate bonds to Si from the N lone pairs.Compound 1 is shown in Fig. 3dnorm in the range from −0.072 to 1.201 (arbitrary units) and the related fingerprint plots generated by CrystalExplorer  and the C⋯H/H⋯C contacts (11.6%) contribute to the packing arrangement of the crystal. Intermolecular interactions of the cyclopentadienyl rings with neighbouring molecules can also be visualized.The crystal packing of compound 1. Selected examples found in the Cambridge Structural Database -meso-bis[dimethyl(aminomethyl)ferrocenyl]dimethylsilane -meso-bis[dimethyl(aminomethyl)ferrocenyl]dichlorosilane -[2-(dimethylaminomethyl)ferrocenyl]lithium  in n-pentane (1 ml) was made up and stored at 243 K and compound 1 crystallized in the form of orange needles.2 is illustrated in Fig. 7Sp)-[2-(dimethylaminomethyl)ferrocenyl]lithium (4.00 mmol) , the product (46%) could be obtained as yellowish plates.The reaction scheme for the synthesis of compound 1H NMR : δ = 0.81 , 2.02 {s, 12H; [N(CH3)2]2}, 2.80, 3.64 , 4.08 , 4.12 , 4.19 , 4.35  ppm.1H}13C NMR : δ = 0.6 , 45.4 {4C; [CH2N(CH3)2]2}, 60.5 , 69.7 , 69.8 , 72.6 , 74.2 , 76.6 , 90.5  ppm.{1H}29Si NMR : δ = −7.07  ppm.{m/z (%): 498 (20) [(M–NMe2)+], 409 (100) [(M–NMe2–CH2NMe2–Me2)+], 299 (50) [(M–FcCH2NMe2)+], 199 (50) [(M–SiMe2FcCH2NMe2–NMe2)+].ESI-(+)-MS: Rf:  = 0.20.Uiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso(H) = 1.5Ueq(C) for CH3 hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table 110.1107/S2056989020010397/hb7933sup1.cifCrystal structure: contains datablock(s) 1, 2, global. DOI: 10.1107/S2056989020010397/hb79331sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989020010397/hb79332sup3.hklStructure factors: contains datablock(s) 2. DOI: 2019451, 2019450CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The title molecule contains a 16-membered macrocycle with the conformation of the C—O—C—C—O—C—C—O—C polyether chain being  27H24N2O5, is a product of the deamination reaction from aza-14-crown-4 ether containing the γ-piperidone subunit. The title molecule contains a 16-membered macrocycle with the conformation of the C—O—C—C—O—C—C—O—C polyether chain being t–g(-)–t–t–g(+)–t . The dihedral angle between the planes of the benzene rings fused to the aza-14-crown-4-ether moiety is 31.11 (14)°. The cavity size inside the macrocycle is 4.72 Å. The macrocycle is significantly flattened as a result of the extended conjugated system. Steric repulsion between the pyridylcarboxamide fragment and the benzene ring results in a slight deviation of macrocycle from planarity. The structure also features intramolecular hydrogen bonding, which results in a deviation of the angle between the planes of amide and pyridyl groups from planarity: this angle is 16.32 (18)°. In the crystal, the molecules are linked into infinite zigzag chains via intermolecular C—H⋯π contacts. The chains are bound into layers parallel to (100) by weak intermolecular C—H⋯O hydrogen bonds.The title compound, C The molecular structure of (3) is presented in Fig. 1t–g(-)–t–t–g(+)–t  conformation. The dihedral angle between the mean planes of the benzene rings fused to the aza-14-crown-4-ether moiety is 31.11 (14)°. The cavity size inside the macrocycle, determined as a double-mean distance between the C16, C19, O5, O8 and O11 atoms and the center of this pentagon, is 4.72 Å. The macrocycle is significantly flattened because of the extended conjugated system. The steric repulsion between the 17-pyridylcarboxamide fragment and the aromatic ring (C11A/C12–C15/C15A) results in a slight deviation of the macrocycle from planarity. The molecular structure also features intramolecular hydrogen bonds  are linked into infinite zigzag chains via intermolecular C26—H⋯π(C22) contacts  was synthesized according to the procedure described previously  and 0.44 g (4.7 mmol) α-aminopyridine (2) in 10 ml o-xylene was refluxed with stirring for 5 h (monitored by TLC until the disappearance of the starting organic compound spots). The solvent was evaporated under vacuum, then the residue was purified by column chromatography (ethyl acetate:n-hexane = 5:1) and recrystallized from ethanol to obtain 1.27 g of pure compound (3) as single crystals in 58% yield. Tmlt = 482–484 K. Rf = 0.66 . IR, ν, cm−1: 1687 (C=O), 1638 (HN—C=O), 3317 (NH). 1H NMR : 3.89 , 4.01 , 4.33 , 6.91–7.75 , 7.73 , 8.18 , 8.32 , 8.36 . Mass spectrum, m/z : 456 [M]+ (4), 428 (1), 309 (4), 283 (3), 265 (3), 238 (25), 221 (18), 210 (50), 189 (10), 173 (89), 159 (20), 147 (38), 131 (100), 118 (48), 115 (51), 103 (27), 91 (81), 89 (52), 78 (65), 45 (38). Analysis calculated for C27H24N2O5, %: C, 71.04; H, 5.30; N, 6.14. Found: C, 70.82; H, 5.34; N, 6.01.A solution of 2.0 g (4.7 mmol) aza-crown ether (Uiso(H) = 1.2Ueq(N)]. The other hydrogen atoms were placed in calculated positions with C—H = 0.95–0.99 Å and refined as riding with fixed isotropic displacement parameters [Uiso(H) = 1.2Ueq(C)].Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989020010968/yk2136sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020010968/yk2136Isup2.hklStructure factors: contains datablock(s) I. DOI: 2022314CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: