R,6R,7S)-5-(furan-2-yl)-7-phenyl-4,5,6,7-tetrahydro-triazolopyrimidin-6-amino as a bridged ligand coordinating two zinc atoms has been synthesized and studied.The first coordination complex with (5 catena-poly[[[acetatochloridozinc(II)]-μ--5-(furan-2-yl)-7-phenyl-4,5,6,7-tetrahydrotriazolopyrimidin-6-amine] monohydrate], {[Zn(C2H3O2)Cl(C15H15N5O)]·H2O}n, is the first coordination complex in which the neutral tetrahydrotriazolopyrimidine derivative acts as bridging ligand between two zinc molecules. As a result, polymeric chains of the coordination complex are found. The coordination of the zinc metal atom occurs with the lone pairs of the triazolo nitrogen atom and amino group. The positive charge of the zinc atom is compensated by the chlorine anion and deprotonated acetic acid. The coordination complex exists as a monohydrate in the crystalline phase. The water molecules bind neighbouring polymeric chains by the formation of O—H⋯O, O—H⋯Cl and N—H⋯O hydrogen bonds.The title complex, systematic name  One of the key stages of the mechanism is a nucleophilic attack of the electron-rich enol carbon atom onto the electron-deficit azomethine carbon, with the formation of a C—C bond in the cyclization. If the suggested hypothesis is true, other reagents with a polar C=C bond similar to the C=C bond in enoles should possess similar reactivity. Using this analogy, we performed a multicomponent reaction between 3-amino-1,2,4-triazole, β-nitrostyrene and furfural. As expected, a derivative of tetrahydro-triazolopyrimidine 1 was obtained in high regio- and stereoselectivity. Further reduction of the nitro group in this compound unexpectedly resulted in formation of the zinc polycomplex 2. A single crystal of this compound was characterized by X-ray diffraction.Multicomponent reactions of 3-amino-1,2,4-triazole and carbonyl compounds have divergent selectivity, allowing the synthesis of alternative products from the same set of starting reagents , 161.4 (2), −78.2 (2) and 105.5 (3)° respectively].The tetrahydropyrimidine ring of the neutral organic ligand adopts an asymmetric half-chair conformation Fig. 1 with puccis-oriented. The furan ring is disordered over two positions with an occupancy ratio of 0.707 (11):0.293 (11) and twisted in relation to the N1—C3 endocyclic bond . This may be due to the strong bifurcated intramolecular N—H⋯π hydrogen bonds °].The amino group and furan ring are A⋯O3i hydrogen bonds  indicates the presence of strong hydrogen bonds in the crystal structure. The main contribution to the Hirshfeld surface is provided by H⋯H contacts (44.5%), shown in Fig. 4b. The contributions of O⋯H/H⋯O (15.3%) and C⋯H/H⋯C (14.8%) contacts associated with X—H⋯O and X—H⋯π hydrogen bonds are much smaller . The smallest contributions in the total Hirshfeld surface are provided by Cl⋯H/H⋯Cl (8.5%) and N⋯H/H⋯N (7.3%)  interactions associated with X—H⋯Cl and X—H⋯N hydrogen bonds.The pair of sharp spikes in the two-dimensional fingerprint plot Fig. 4a indicaer Fig. 4c, 4d. T%) Fig. 4e, 4f inet al., 2016et al., 1999et al., 2011et al., 1994et al., 2010et al., 2009et al., 2018et al., 2009et al., 2008et al., 2012et al., 2011A search of the Cambridge Structural Database  equipped with an outer IR temperature sensor. The reaction was performed in a sealed microwave process vial using the ‘very high’ mode, which decreased the initial power to 90 W. Reaction time under microwave conditions refers to the time that the reaction mixture was kept at the set temperature (fixed hold time).Microwave irradiation experiments were carried out using an Emrys:)-5-(Furan-2-yl)-6-nitro-7-phenyl-4,5,6,7-tetrahydro-triazolotriazolopyrimidin-6-amine with zinc(II) acetate-chloride (2): To a solution of 4.0 mmol of 1 in 5.0 mL of acetic acid was added 4.5 mL of concentrated hydrochloric acid. The mixture was cooled down in an ice–water bath and 1.0 g of zinc dust was slowly added to the mixture portionwise. After the addition, the cooling bath was removed and the mixture was stirred for 30 min and then refluxed until the reducing agent was completely dissolved. The reaction mixture was left undisturbed overnight, and the single crystal used for the X-ray diffraction study was taken directly from the reaction mixture. The isolated yield of 2 was 67%.Uiso(H) = 1.5Ueq(C) for methyl groups, O—H = 0.98 Å, Uiso(H) = 1.5Ueq(O) for the water molecule, Car—H = 0.93 Å, Csp3—H = 0.97 Å, N—H = 0.89 Å and Uiso(H) = 1.2Ueq(parent atom) for all other hydrogen atoms. The furan ring is disordered over two positions with an occupancy ratio of 0.707 (11):0.293 (11).Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989021012226/jy2010sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021012226/jy2010Isup2.hklStructure factors: contains datablock(s) I. DOI: 2122655CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


A tetranuclear complex with an open-cubane like structure was synthesized from 2-methoxy-6-(pyridin-2-yl-hydrazonomethyl)-phenol and characterized using micro-analytical and spectroscopic techniques, and single-crystal X-ray diffraction analysis. HL), namely, cyclo-tetrakis(μ-2-methoxy-6-{[2-(pyridin-2-yl)hydrazin-1-ylidene]methyl}phenolato)tetranickel(II) tetrakis(perchlorate) acetonitrile monosolvate dihydrate, [Ni4(C13H12N3O2)4](ClO4)4·C2H3N·2H2O, and characterized using micro-analytical and spectroscopic techniques. The crystal-structure determination reveals the formation of a distorted Ni4O4 cubane-like core architecture encapsulated by four hydrazone Schiff base (HL) molecules. A open-cube tetranuclear architecture is created in which nickel(II) ions of the NiN2O3 unit are connected by μ2-O anions of the phenolate moiety of HL. In this complex, each Ni centre has a slightly distorted square-pyramidal coordination environment. The supramolecular architectures are stabilized via the presence of various intermolecular hydrogen bonds and  stacking interactions.A tetranuclear complex with an open-cubane-like core structure was synthesized from 2-methoxy-6-(pyridin-2-yl-hydrazonomethyl)phenol ( The synthesis of such polynuclear metal complexes can often be promoted with the use of polydentate Schiff base ligands possessing nitrogen and oxygen donor atoms. Such Schiff bases are known to form high nuclearity complexes with interesting architectures, and the hydroxyl groups and other donor atoms are often suitable for the synthesis of polynuclear complexes  2-methoxy-6-(pyridin-2-yl-hydrazonomethyl)-phenol, which can bind one or two metal ions, simultaneously. The stoichiometric reaction of nickel(II) perchlorate hexahydrate with this ligand resulted the formation of Ni4O4 distorted cubane-like structure described herein.Polynuclear metal(II) complexes have attracted much attention owing to their structural variety and significant applications in biology, catalysis, molecular recognition and magnetism (Alcantara HL) was prepared by the reaction of 2-hydrazinopyridine and 2-hydroxy-3-methoxybenzaldehyde in a 1:1 ratio in ethanol. The reaction of nickel perchlorate hexahydrate and the HL ligand yielded a tetranuclear open-cubane-like complex with an Ni4O4 core-type architecture. The tetranuclear complex is formulated as [Ni4(L)4](ClO4)4·C2H3N·2H2O  ligand coordinates in a pentadentate mode , thus forming eight fused metal chelate rings (four five-membered and four six-membered rings). Such a coordination pattern results in a distorted square-pyramidal coordination sphere around each nickel(II) ion. The distortion in the square-pyramidal geometry is shown by the τ index –3.668 (3) Å].In the polynuclear crystal, intermolecular hydrogen-bonding interactions are detected involving C—H and N—H donors from the hydrazone Schiff base and acceptor oxygen atoms of perchlorate counter-ions and solvate water molecules Fig. 3. The impet al., 2016et al., 2011et al., 2012et al., 2003et al., 2014et al., 2013et al., 2012A search of the Cambridge Structural Database : 1648, for (>C=N) and 3480 (–OH). The tetranuclear nickel complex was synthesized by taking an equimolar methanolic solution (10 ml) of the HL ligand  and nickel perchlorate hexahydrate . The resulting solution was stirred for 3 h. The obtained green crystals, suitable for diffraction studies, were filtered off and washed with cold methanol and kept in a CaCl2 desiccator. Yield 62%. Analysis calculated for C54H55Cl4N13Ni4O26: C, 38.63; H, 3.30; N, 10.84%. Found: C, 38.28; H, 3.28; N, 10.98%. FTIR : 1626 (>C=N), 1537 (C—O), 487 (Ni—O) and 421 (Ni—N).A mixture of 2-hydrazinopyridine  and 2-hydroxy-3-methoxybenzaldehyde  in 30 mL of ethanol was refluxed for 3 h. The resulting light-yellow solution was cooled to room temperature. The obtained crystalline material was filtered off, washed with ethanol and kept in a CaClUĩso(H) = 1.5Ueq(O). Three of the four perchlorate anions are disordered over two orientations and were restrained to have tetrahedral geometries with occupancies of 0.57 (6)/0.43 (6), 0.412 (13)/0.488 (13), and 0.806 (12)/0.194 (12), respectively.Crystallographic data and refinement details are presented in Table 310.1107/S2056989021012408/yy2002sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021012408/yy2002Isup2.hklStructure factors: contains datablock(s) I. DOI: 2096266CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


The double-bond system of the acrylonitrile moiety is significantly non-planar and displays one very wide angle C—C(CN)=C. 14H14N2S3, the double-bond system of the acrylonitrile moiety is significantly non-planar, with absolute cis torsion angles of 13.9 (2) and 15.1 (2)°. The ring system and the double bond system subtend an interplanar angle of 11.16 (4)°. The wide angle C—C(CN)=C of 129.40 (12)° may be associated with a balance between planarity and avoidance of a very short S⋯S contact.In the title compound, C The high degree of structural diversity has proved to be important in the search for new effective treatments . Compound 2 was synthesized by the reaction of 2-cyanomethylbenzothiazole 1 with carbon disulfide in the presence of sodium ethoxide, followed by alkylation with ethyl iodide. The structure of 2 was originally based on its elemental analysis and spectroscopic data . In order to establish the structure of the compound unambiguously, the crystal structure was determined.As an extension of this research  is strikingly wide, at 129.40 (12)°; for comparison, the corresponding angles in the five structures mentioned below (with refcodes) range from 122–126°. One might speculate that this large angle and the deviation from planarity about the double bond represent aspects of a compromise between (i) achieving coplanarity of the heterocycle with the double-bond system and (ii) avoiding too short an S⋯S contact. The intramolecular S⋯S distances are S1⋯S3 = 3.1155 (5) and S2⋯S3 = 3.0496 (5) Å. The ethyl groups project to opposite sides of the molecule.The molecule of A⋯C7A′  3.741 (2) Å. The sulfur atom S1 lies 3.61 Å from the centroid of the six-membered ring in the facing molecule; such potential S⋯π interactions have been discussed by e.g. Ringer et al. =C gave 27 hits, but none in which any further atom at the double bond was sulfur. A search for the group C—C(CN)=C(S—C)2, with the first carbon atom three-coordinate, both sulfur atoms two-coordinate and not involving cyclicity, gave only five hits. The refcodes, references and absolute cis torsion angles NC—C=C—S were as follows: CIYDIY, Kumar et al. : υ 3056 (ArCH), 2924 (CH3), 2213 (CN), 1502 (C=N); 1H NMR : δ 1.27–1.34 , 3.16–3.23 , 7.50–7.57 , 8.04–8.15 ; analysis, calculated for C14H14N2S3 (306.47): C% 54.87; H% 4.60; N% 9.14; S% 31.39; found: C% 54.85, H% 4.58; N% 9.16; MS m/z (%): 306 , 276 (100%), 273 (57%), 248 (26%), 217 (76%), 204 (26%), 146 (20%).A mixture of sodium ethoxide (0.08 mol) and 2-cyanomethylbenzothiazole (0.04 mol) in absolute ethanol (100 ml) was refluxed for 20 min. After cooling, carbon disulfide (0.04 mol) was added gradually and then the solution was warmed for 20 min. Ethyl iodide (0.08 mol) was then added, and the reaction mixture was stirred overnight at room temperature. The solution was poured onto ice–water and the solid product thus formed was filtered off. The product was purified by dissolving it in hot petroleum ether, filtering, and allowing the solution to cool. The solid that formed was recrystallized from DMF to give pale-yellow crystals, m.p. = 366–368 K, yield 72%; IR . The U(H) values were fixed at 1.5 or 1.2 times the equivalent Uiso value of the parent carbon atoms for methyl and non-methyl hydrogen atoms, respectively.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989022002572/ex2055sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989022002572/ex2055Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022002572/ex2055Isup3.cmlSupporting information file. DOI: 2156777CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


The Hirshfeld surfaces and two-dimensional fingerprint plots for the polymorphs were calculated using CrystalExplorer17, which indicated contacts significantly shorter than the sum of the van der Waals radii in the vicinity of the piperazine N4 and triazole O1 atoms corresponding to the strong hydrogen bonds accepted by these atoms. In this study, treatment of anhydrous trazodone powder with ammonium carbamate in warm water crystallised two new polymorphs or dihydrates of trazodone after 5 h, whose structures were determined by X-ray single crystal diffraction. Each dihydrate contains infinite zigzag hydrogen-bonded chains of water molecules, which are stabilised by the N4 acceptor atom of the piperazine ring and the pendant carbonyl O1 atom of the triazole ring, as well as other water molecules. The strong dipole moment expected for the O1 atom makes it a good hydrogen bond acceptor for stabilising the chains of water molecules. Each molecule of trazodone has a similar conformation in both hydrates, except for the propyl chains, which adopt different conformations: anti-gauche in the β hydrate (triazole N-C-C-C and C-C-C-piperazine N) and anti-anti in the γ hydrate. Both piperazine rings adopt chair conformations, and the  Water, ubiquitous for the presence of life, can have important and sometimes adverse effects on the manufacture of drugs and their performance. It can be introduced through an active ingredient, excipients, or the atmosphere, and can increase interactions between a drug and its excipients, dissolve soluble components or induce phase transitions . The absWhen a compound or a drug co-crystallises with water, a new crystalline phase, termed a hydrate, is formed. Hydrates are a subset of a larger class of crystalline solids, called solvates, which have the solvent of crystallisation included in the structure. Hydrates may form by crystallisation from solution or by the uptake of water vapour in the solid state. An example is metacetamol, an isomer of paracetamol, which forms a hemihydrate . Metacet19H22ClN5O, systematic name 2-{3-[4-(3-chlorophenyl)piperazin-1-yl]propyl} triazolopyridine-3(2H)-one, is an antidepressant medication discovered in the 1960s and now widely prescribed throughout the world  channels occupied by the water molecules of crystallisation compare  with essThe α polymorph is affected by severe disorder of both the trazodone molecule and water molecules , and not19H22ClN5O·2H2O mapped over dnorm is shown in The Hirshfeld surfaces and two-dimensional fingerprint plots for the β and γ polymorphs were calculated using CrystalExplorer17  using pr19H22ClN5O·2H2O  in warm water (30 mL) was mixed with ammonium carbamate  in warm water (30 mL) and left to stand for 2 h. A white product settled to the bottom of the beaker, which was harvested. The powdery starting material was replaced by a microcrystalline phase, and visual inspection indicated that two different crystal morphologies (laths and blocks) had formed.19H22ClN5O·2H2O (colourless lath 0.24 × 0.09 × 0.02 mm) and γ-C19H22ClN5O·2H2O (colourless block 0.27 × 0.22 × 0.12 mm) were established using intensity data collected on a Rigaku AFC11 CCD diffractometer  at 100 K, and analytical absorption corrections were applied during data reduction . The structures were routinely solved by ‘dual-space’ methods using the program SHELXT [F|2 with SHELXL-2018 [Uiso(H) = 1.2 Ueq(carrier) was applied in all cases. Full details of the structures and refinements are available in the deposited cifs.The crystal structures of β-Cm SHELXT , and theLXL-2018 . The C-b19H22ClN5O·2H2O (C19H26ClN5O3): Mr = 407.90, monoclinic, space group P21/n (No. 14), a = 16.1252 (3) Å, b = 6.67160 (9) Å, c = 19.8031 (3) Å, β = 108.7862 (19)°, V = 2016.94 (6) Å3, Z = 4, T = 100 K, μ = 1.932 mm−1, ρcalc = 1.343 g cm−3, 12332 reflections measured (6.2 ≤ 2θ ≤ 136.5°), 3658 unique (Rint = 0.031), R(F) = 0.033 (3407 reflections with I > 2σ(I)), wR(F2) = 0.097 , Δρmin,max (e Å−3) = −0.30, +0.27, CCDC deposition number 2017101.Crystal data for β-C19H22ClN5O·2H2O (C19H26ClN5O3): Mr = 407.90, orthorhombic, space group Pca21 (No. 29), a = 20.08500 (16) Å, b = 7.62653 (6) Å, c = 26.2713 (2) Å, V = 4024.21 (5) Å3, Z = 8, T = 100 K, μ = 1.937 mm−1, ρcalc = 1.347 g cm−3, 16,899 reflections measured (6.7 ≤ 2θ ≤ 136.5°), 6113 unique (Rint = 0.021), R(F) = 0.030 (6073 reflections with I > 2σ(I)), wR(F2) = 0.084 , Δρmin,max (e Å−3) = −0.26, +0.23, Flack absolute structure parameter = 0.453 (12), CCDC deposition number 2017102.Crystal data for γ-CIn this study, two new dihydrates of trazodone resulted from the same crystallisation reaction, and were characterised by single crystal X-ray structure determinations. Each crystal structure has zigzag chains of hydrogen-bonded water molecules. The propyl chains have different conformations for each structure. The β polymorph has an anti-gauche conformation and the γ polymorph has an anti-anti conformation: the latter is presumed to be the more stable polymorph. The Hirshfeld surfaces show regions of strong hydrogen bonding at the O1 and N4 atoms.n-heptane [The phenomenon of the spontaneous crystallisation of two polymorphs (one of which must be metastable) from the same solution is an interesting one, and previous reported examples include the formation of α and β L-glutamic acid from aqueous solution , and two-heptane ; a detai-heptane . It is w


Talaromyces minioluteus belongs to the section Trachyspermi, has a worldwide distribution and has been found on various substrates, especially on various (stored) food commodities and indoor environments. This species is phenotypically and phylogenetically closely related to T. chongqingensis and T. minnesotensis. The phylogenetic and morphological analyses of 37 strains previously identified as T. chongqingensis, T. minnesotensis and T. minioluteus revealed that this clade incudes eight species: the accepted species T. chongqingensis, T. minnesotensis and T. minioluteus, the newly proposed species T. calidominioluteus, T. africanus and T. germanicus, and the new combinations T. gaditanus (basionym Penicillium gaditanum) and T. samsonii (basionym Penicillium samsonii). In this study, we give insight of the phylogenetic relationships and provide detailed descriptions of the species belonging to this clade. Macromorphological features, especially colony growth rates, texture and conidial colors on agar media, are important characters for phenotypic differentiation between species. Talaromyces minioluteus belongs to the section of Trachyspermi and is phylogenetically related to T. chongqingensis, T. minnesotensis and T. udagawae. It has a worldwide distribution and is previously isolated from various substrates, including soil, indoor environments  and various foods, and has been reported as a postharvest pathogen of quince, tomato and citrus fruits [s fruits . Variouss fruits ,3,4.Penicillium with four subgenera. Penicillium subgenus Biverticillium species  were characterized by its symmetrically, biverticillate conidiophores with acerose phialides and having slow growth on 25% glycerol nitrate agar. Early molecular analysis showed that these species were phylogenetically distinct from species classified in the other three subgenera [Penicillium was polyphyletic with the inclusion of subgenus Biverticillium and it was thus suggested to be excluded from Penicillium [Talaromyces and subgenus Biverticillium species cluster together, forming a monophyletic clade within the Trichocomaceae [Talaromyces was redefined to also include the majority of species previously classified in subgenus Biverticillium [Penicillium minioluteum was given the new combination Talaromyces minioluteus.In the past, based on a morphological approach, Pitt  introducubgenera ,7. Peniccomaceae ,9. Basedicillium  and for Talaromyces minioluteus was originally described by Dierckx [P. minioluteum with P. funiculosum (currently T. funiculosus) based on examination of Biourge’s P. minioluteum strain. In contrast to Raper and Thom [P. minioluteum and this was confirmed in the taxonomic study of van Reenen-Hoekstra et al. [P. funiculosum, P. minioluteum produces more restricted colonies with striking yellow mycelium and does not, or slowly, grows at 37 °C. Furthermore, T. minioluteus strains can produce red soluble pigments, though not consistently [P. minioluteum with Biourge’s strain (IMI 89377ii). The ex-neotype strains of P. minioluteum FRR 1714 (=CBS 196.88) and CBS 642.68 differ morphologically causing confusion in the taxonomy of this species. The latter strain fully fits Dierckx’s original description and is therefore considered correct [T. ruber [ Dierckx  as formi Dierckx  synonymiand Thom , Pitt [5a et al. . In contistently . Pitt [5T. ruber .Penicillium gaditanum, Penicillium samsonii and Penicillium purpurogenum var. rubrisclerotium are related to T. minioluteus [Penicillium gaditanum was originally described in the P. funiculosum series [P. samsonii was described in series Islandica (sect. Simplicium) [P. minioluteum [Penicillium gaditanum was not included in the monographic treatment of Talaromyces [P. samsonii with T. minioluteus was confirmed. The taxonomic position of P. purpurogenum var. rubrisclerotium is unclear. Yilmaz et al. [P. purpurogenum var. rubrisclerotium resolved in a clade with T. amestolkiae, but CBS 270.35, the ex-type strain of this species resolved in a distinct clade closely related to T. minioluteus. Talaromyces minnesotensis and T. chongqingensis are two recently described species related to T. minioluteus [ioluteus ,4. Penicm series ,14, whilplicium) ,15. Baseioluteum . Penicilaromyces  was follz et al.  noted thioluteus ,17.BenA) sequences, Visagie et al. [T. minioluteus represents a species complex. The objective of our study is to resolve the taxonomic status of strains previously assigned to T. minioluteus using a polyphasic approach that includes macro- and micro-morphological data combined with the phylogenetic analysis of BenA, calmodulin (CaM), ITS and RNA polymerase II second largest subunit (RPB2) gene sequences. We show that the T. minioluteus-clade includes eight species  and detailed descriptions of these species and the phylogenetically related T. udagawae are provided here.Based on the analysis of internal transcribed spacer rDNA region (ITS) and β-tubulin  and the internal working collection of the Food and Indoor Mycology department (DTO), both housed at the Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands. The strain selection was based on a homology search in these collections using ITS, ® UltraClean® Microbial Kit  according to manufacturer instructions. The genetic markers BenA, CaM, ITS and RPB2 were amplified using the primer pairs Bt2a/Bt2b [w/w), in which 1 μL of template DNA was mixed with 0.98 μL dNTPs (1 mM), 0.25 μL of each primer (10 μM) and 0.05 μL of BioTaq DNA polymerase (5 U μL). The MgCl2 concentration was 1.46 mM. The targeted loci were amplified with a cycle program comprising an initial denaturation step at 95 °C for 5 min, followed by 35 cycles of 30 s at 95 °C (denaturation), 40 s at 55 °C  for benA, CaM and ITS and 48 and 52 °C for RPB2, and 2 min at 72 °C (primer extension) and ending with a final extension of 7 min at 72 °C. Automated sequencing of both strands of the PCR amplicons was performed on a 3730xl DNA Sequencer  using the BigDye Terminator chemistry. The primers in the sequencing reactions were the same as those used for PCR amplifications. Consensus sequences for each locus were assembled using SeqMan Pro v.15 . Novel sequences generated in this study were deposited in the NCBI nucleotide database (GenBank) under accession numbers depicted in Total genomic DNA was extracted from colonies grown on malt extract agar (MEA) for 4 d with the DNeasyt2a/Bt2b , CMD5/CMt2a/Bt2b , V9G/LS2t2a/Bt2b ,22 and Rt2a/Bt2b ), respecBenA, CaM, ITS and RPB2 sequence datasets as well as on the combined BenA, CaM, ITS and RPB2 dataset. The individual loci in the combined dataset were analysed as separate partitions. Phylogenetic trees were constructed based on Maximum Likelihood (ML) using RAxML-HPC2 on XSEDE 8.2.10 [Single gene alignments were generated in MAFFT v. 7  and subsE 8.2.10  through E 8.2.10  and BayeE 8.2.10 . The subBenA and ITS sequences deposited in GenBank and belonging to the T. minioluteus-clade were re-analyzed. These sequences were merged in our dataset, aligned and a phylogram was constructed according to the methods described above.In order to get insight in the distribution of the investigated species, The selected strains were three-point inoculated on the agar media Czapek yeast autolysate agar (CYA), MEA (Oxoid), yeast extract sucrose agar (YES), creatine sucrose agar (CREA), CYA supplemented with 5% NaCl (CYAS), dichloran 18% glycerol agar (DG18) and oatmeal agar (OA). Agar media formulations are as described by Samson et al. (2014) . Plates T. minioluteus-clade were inferred using single gene sequence datasets of the four studied genetic markers , including the phenotypically distinct species T. udagawae. Four lineages are represented by the accepted species T. chongqingensis, T. minioluteus, T. minnesotensis and T. udagawae. The ex-type strains of P. gaditanum (CBS 169.81) and P. samsonii (CBS 137.84) are located in two distinct lineages and are combined in Talaromyces . The new species T. africanus, T. calidominioluteus and T. germanicus are proposed for the three remaining lineages. The multi-gene phylogeny resolved the studied strains into four main clades. The segregation into these four clades is strongly supported by Bayesian Inference, while statistical support is lacking in the ML analysis. The most species rich clade includes T. minioluteus, T. gaditanus, T. chongqingensis and T. samsonii and is sister to the clade that includes the new species T. calidominioluteus and T. africanus. The sister to these is a clade that contains T. minnesotensis and T. germanicus. The phenotypically distinct species T. udagawae takes a basal position to the other three clades.The phylogenies inferred on the T. minioluteus-clade can be segregated into three groups according to their growth rate on CYA at 30 °C. Talaromyces calidominioluteus and T. africanus grow faster than the others species (18–29 mm), T. minioluteus, T. samsonii and T. gaditanus grow moderately fast (9–16 mm) whereas T. germanicus, T. chongqingensis and T. udagawae grow the slowest (5–8 mm)  . The myc texture . All spe texture , and genThree new species and two new combinations are introduced in this study. All species were compared with their morphologically and phylogenetically closest relatives. Diagnostic characters are summarized in Talaromyces africanus Houbraken, Pyrri and Visagie, sp. nov. MycoBank MB 841228. Etymology: Referring to Africa, the continent where the type was isolated.Diagnosis: Colony diameter on CYA incubated at 30 °C 18–20 mm, conidia subglobose, finely roughened.Typus: South Africa, house dust, February 2012, collected and isolated by C.M. Visagie .BenA = OK338782; CaM = OK338808; RPB2 = OK338833).ITS barcode: OK339610 : CREA no growth; CYA 21–23; CYA 30 °C 18–20; CYA 37 °C no growth; CYAS 2–3; DG18 15–17; MEA 26–28; OA 18–20; YES 20–22.Colony characters: CYA 25 °C, 7 d: Colonies low, radially sulcate with concentric rings, margin entire, plane, moderate; sporulation dense; conidia en masse greyish turquoise (24E6); mycelium white; colony texture velvety to weakly floccose; exudates present clear; soluble pigments absent; reverse reddish brown (8E6). MEA 25 °C, 7 d: Colonies with a few sulcates near the center, low, margin plane, low, wide regular; sporulation dense; conidia en masse greyish turquoise (24D5–E5); mycelium white; colony texture velvety to weakly floccose; exudates abundant, yellow; soluble pigments absent; reverse reddish orange to copper red (C8). YES 25 °C, 7 d: Colonies radially sulcate, raised at the center, margin plane, low, narrow, regular; sporulation dense; conidia en masse dull green to greyish green (25E4–5); mycelium white; exudates absent; soluble pigments absent; reverse dark brown (7F4) at the center, copper red (7C8). DG18 25 °C, 7 d: Colonies slightly radially sulcate at the center, low, margin plane, low, moderate, regular; sporulation strong; conidia en masse dark green (25F7–8); mycelium white; colony texture velvety; exudates absent; soluble pigments absent; reverse brownish orange (6C8). OA 25 °C, 7 d: Colonies plane, low, margins entire, plane, narrow; sporulation moderate to dense; conidia en masse dull green to greyish green (27E4–5); mycelium white; colony texture velvety to granulose; exudates clear droplets; soluble pigments absent; CREA 25 °C, 7 d: no growth.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate; Stipes smooth-walled, non-vesiculate, 80–200 × 2–3 µm; Metulae adpressed, 3–6 per stipe 10–13.5(–15) × 2.5–3.5 µm; Phialides acerose, 3–8 per metula, 10–14 × 2–3 µm; Conidia in long, distorted chains, finely roughened, occasionally smooth, subglobose, with distinct connectives on both sides, 2.5–3.5 × 2–2.5 µm.Notes: Talaromyces africanus is phylogenetically closely related to T. calidominioluteus 25–29 mm, conidia smooth, broadly ellipsoidal.Typus: the Netherlands (imported from Brazil), melon, isolated by J. Houbraken, Oct. 2007 .BenA = OK338786; CaM = OK338817; RPB2 = OK338837).ITS barcode: OK339612 : CREA 0–6; CYA 20–30; CYA 30 °C (20–)25–29; CYA 37 °C no growth; CYAS 2–10; DG18 15–20; MEA 22–30; OA 17–24; YES 21–29.Colony characters: CYA 25 °C, 7 d: Colonies low, slightly sunken in center, plane or with concentric rings, margin entire; sporulation moderate to strong, sometimes poor (DTO 390–E8); conidia en masse dark green (26F5–28F3); mycelium pale yellow ; colony texture, slightly lanose, sometimes velvety; exudates present as small or large, orange-brown droplets; soluble pigments present, in orange shades , red, or absent; reverse concentric rings of light brown (6D8), then brownish orange (6C6), dark brown (6F8), edge orange (6B6–7). MEA 25 °C, 7 d: Colonies low, plane, margin entire; sporulation strong, occasionally moderate; conidia en masse grey green (25D5–28E3) to dark green (30F3); mycelium white to pastel yellow (1A4–2A4); colony texture velvety to weakly floccose or cottony; exudates absent or present as orange-brown droplets; soluble pigments absent; reverse orange (6B6–8) to orange brown to brown (6C8–D8). YES 25 °C, 7 d: Colonies raised in center, concentric and radially sulcate, margin entire; sporulation strong, poor in DTO 390-E8; conidia en masse dull green (26E3–4) to dark green (28E–3F3); mycelium pale yellow (1A2–3) to yellow (2A6); exudates absent; soluble pigments absent; reverse orange (6B7) to brownish orange (6C7–7C8). DG18 25 °C, 7 d: Colonies flat, non-sulcate to radially sulcate, margin entire; sporulation strong (poor in DTO 390-E8); conidia en masse dark green (25F8–27F4); mycelium white to yellowish white to pastel yellow (2A2–4); colony texture velvety; exudates absent; soluble pigments absent; reverse yellowish orange (4B8) or greyish orange to orange (6B5–7) or brownish orange (6C8). OA 25 °C, 7 d: Colonies plane, low, margins entire; sporulation dense; conidia en masse greyish green (27E5–7); mycelium greenish yellow (1A8); colony texture velvety; exudates golden yellow droplets; soluble pigments absent. CREA 25 °C, 7 d: no growth to poor growth, acid production absent, base formation absent.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate, occasionally with one subterminal branch or two branches positioned at the same level on the stipe; Stipes smooth-walled, non-vesiculate, 80–200 × 2.5–3.5(–4) µm; Metulae adpressed, 3–10 per stipe, 10–13(–16) × 2.5–3.5 µm; Phialides acerose, 3–8 per metula, 9.5–13.5 × 2–3 µm; Conidia in long, distorted chains, smooth, broadly ellipsoidal, with distinct connectives on both sides, 2.5–4 × 2–2.5 µm.Notes: Talaromyces calidominioluteus generally grows faster on CYA incubated at 30 °C (18–29 mm) than the morphologically most similar species T. minioluteus (5–11 mm) and T. gaditanus (13–20 mm). Talaromyces africanus is phylogenetically most closely related and for more details, see description of this species above.Talaromyces chongqingensis X.C. Wang and W.Y. Zhuang, Biology 10 : 10. 2021. MycoBank MB 570851. Typus: China, Gaoguan Town, Daba Mountain National Nature Reserve, soil under palm tree, isolated by Xin-Cun Wang, Huan-Di Zheng and Chang Liu, Oct. 2020 .BenA = MZ361343; CaM = MZ361350; RPB2 = MZ361357).ITS barcode: MZ358001 : CREA 7–9; CYA 17–18; CYA 30 °C 6–8; CYA 37 °C no growth; CYAS 3–4; DG18 12–14; MEA 28–29; OA 15–16; YES 20–22.Colony characters: CYA 25 °C, 7 d: Colonies plane, moderately deep, margin plane, regular; sporulation poor; conidia en masse greenish at the center; mycelium beige; colony texture densely cottony; exudates absent; soluble pigments present, red to dark red (10B–C8); reverse violet brown (10D8), edge red (10B8). MEA 25 °C, 7 d: Colonies moderately deep, plane, margin entire; sporulation moderate; conidia en masse greyish turquoise (24C5); mycelium white; colony texture densely cottony; exudates absent; soluble pigments absent; reverse golden yellow (5B7–8). YES 25 °C, 7 d: Colonies plane, moderately deep, raised at center, margin plane, high, wide regular; sporulation sparse to absent; conidia en masse greyish; reverse orange (5B8); mycelium white; exudates absent; soluble pigments absent; DG18 25 °C, 7 d: colonies plane, raised at center, plane, margin entire; sporulation poor; conidia en masse (5A2); mycelium beige in DTO 060-C9; colony texture cottony to floccose; exudates absent or orange; soluble pigments absent; reverse greyish orange (5B6). OA 25 °C, 7 d: colonies plane, moderately deep, margin plane, raised, wide, regular; sporulation moderate; conidia en masse greenish grey (27C2); mycelium white; colony texture floccose; exudates absent; soluble pigments absent; CREA 25 °C, 7 d: growth, acid production absent, base formation absent.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate and several monoverticillate; Stipes smooth walled, non-vesiculate 50–100 × 2.5–3.5 µm; Metulae adpressed, 2–5 per stipe, 11.5–15 × 2.5–3.5 µm; Phialides cylindrical to acerose, 2–5 per metula, 9–12 × 2.5–3.5 µm; Conidia in long, distorted chains, smooth, ovoidal, 2.5–3.5 × 1.5–2.5 µm.Notes: The ex-type strain was not available for examination during our study and the description above is based on CBS 270.35 (=CBS 147316). The phenotype of this old strain, isolated from Zea mays (corn) in 1912, differs significantly from the original description of T. chongqingensis [T. chongqingensis as a floccose texture on MEA, sporulates well and produces greyish green colored conidia [Talaromyces chongqingensis and T. minioluteus are phylogenetically most closely related (T. minioluteus (9–14 mm). Furthermore, CBS 270.35 produces mono- and biverticillate conidiophores and ovoidal conidia, in contrast to the biverticillate conidiophores and broadly fusiform or ellipsoidal conidia produced by T. minioluteus. Unfortunately, no data is available for ex-type strain (CGMCC 3.20482) on the growth on CYA at 30 °C, a useful character for species identification in the T. minioluteus-clade. CBS 270.35 could be degenerated and might not be a good representative of the species. However, differences in the phenotype can also occur when a strain is grown at different labs. In order to make a more reliable comparison of the macromorphology of T. chongqingensis with the current data, the freshly isolated ex-type strain (CGMCC 3.20482) should be grown under the same conditions as the other strains in this study.ingensis . We refeingensis  for the  conidia . Talarom related . CBS 270Talaromyces gaditanus (C. Ramírez and A.T. Martínez) Houbraken and Soccio, comb. nov. MycoBank MB 841226. Basionym: Penicillium gaditanum C. Ramírez and A.T. Martínez, Mycopathol. 74: 165. 1981.Typus: Spain, Madrid, air, 1978, isolated by C. Ramírez .BenA = OK338775; CaM = OK338802; RPB2 = OK338827).ITS barcode: MH861318 : CREA 2–6; CYA 19–25; CYA 30 °C 12–16; CYA 37 °C no growth; CYAS 2–6; DG18 10–18; MEA 25–28; OA 15–19; YES 18–25.Colony characters: CYA 25 °C, 7 d: Colonies low, slightly sunken at center, plane, sulcate near center, margin entire; sporulation strong; conidia en masse greyish green to dark green (28E7–F7–8); mycelium greenish yellow (1A6–7) to light yellow (2A5–6); colony texture velvety, slightly funiculose; exudates absent; soluble pigments present, orange shade; reverse brown (6D8) at the center, orange (5B8–6B8) elsewhere. MEA 25 °C, 7 d: Colonies low, plane, margin entire; sporulation moderate to dense; conidia en masse greyish green (28E7), dark green (27F7–9); mycelium pastel yellow ; colony texture floccose; exudates absent or present as red droplets; soluble pigments absent; reverse brown (6D8), reddish orange (7B8). YES 25 °C, 7 d: Colonies raised at center, having a leathery appearance, plane, lightly sulcate to radially sulcate, margin entire; sporulation absent, sparse to moderate; conidia en masse light brown (7D8) in center, orange (5B8) in middle, dark green ; mycelium white, pastel yellow (1A4); exudates absent; soluble pigments absent, brownish orange (6C8); reverse yellow (3A6) edge, brownish yellow (5C8) in center, brownish orange to reddish orange (7B8) to brownish red (8C8). DG18 25 °C, 7 d: Colonies a little raised at center, plane, margin entire; sporulation poor in DTO 228-B8, dense in other isolates; conidia en masse greyish green ; mycelium pastel yellow (2A4), yellow (2A6) in DTO 228-B8, greenish yellow (1A6) in other isolates; colony texture cottony in DTO 228-B8, velvety with floccose at center; exudates absent, yellow, red in DTO 335-A5; soluble pigments absent; reverse orange (6B7), orange to brownish orange to reddish orange (6B8–7C8). OA 25 °C, 7 d: Colonies moderately deep, plane; margin entire; sporulation poor; conidia en masse greyish green (27E6); mycelium greenish yellow (1A6); colony texture floccose; exudates absent; soluble pigments reddish. CREA 25 °C, 7 d: poor growth, acid production absent, base formation absent.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate; Stipes smooth-walled, non-vesiculate, 80–200 × 2.5–3.5 µm; Metulae adpressed, 3–8 per stipe, 10–13.5 × 2.5–3.5 µm; Phialides acerose, 3–8 per metula, 8.5–12.5 × 1.5–3 µm; Conidia in long, distorted chains, smooth, fusiform, with distinct connectives on both sides, 2.5–4 × 2–2.5(–3) µm.Notes: This species was originally described as Penicillium gaditanum. Based on morphological and chemical data, Penicillium gaditanum was treated by van Reenen-Hoekstra et al. [T. minioluteus. A molecular treatment of this species was lacking and sequence analyses presented here shows that this species is related to T. minioluteus, but distinct based on the GCPSR concept. This species produces strongly floccose colonies on MEA, with bright yellow (pastel yellow) mycelium and fusiform, smooth-walled conidia. Phenotypically, this species resembles T. samsonii. Talaromyces samsonii has a violet brown to deep red reverse on CYA and (vivid) red soluble pigments, in contrast to the predominantly orange reverse and orange soluble pigments (if present) in T. gaditanus.a et al.  and Yilma et al.  as a synTalaromyces germanicus Houbraken and Pyrri, sp. nov. MycoBank MB 841227. Etymology. Referring to Germany, the country where the type was isolated.Diagnosis: Colony diameter on CYA incubated at 30 °C 6–7 mm, colony texture on MEA velvety, conidial color en masse on MEA dull green to greyish green.Typus: Germany, drywall , July 2009 .BenA = OK338799; CaM = OK338812; RPB2 = OK338845).ITS barcode: OK339619 : CREA no growth; CYA 20–22; CYA 30 °C 6–7; CYA 37 °C no growth; CYAS no growth; DG18 16–18; MEA 23–25; OA 18–19; YES 22–24.Colony characters: CYA 25 °C, 7 d: Colonies radially sulcate with a concentric ring, margin plane, low, moderately wide, regular; sporulation dense; conidia en masse dull green to greyish green (26D4–5); mycelium pastel yellow (1A4); colony texture velvety; exudates cherry red (10B8) only small drops at the center; soluble pigments present, cherry red (10B8); reverse violet brown (10F7) centrally, brownish red (10D7) elsewhere. MEA 25 °C, 7 d: Colonies low, with a few sulcations near the center, margin plane, low, wide, regular; sporulation dense; conidia en masse dull green to greyish green (25E4–5); mycelium light yellow (1A6); colony texture velvety; exudates red; soluble pigments absent; reverse reddish orange (7B8). YES 25 °C, 7 d: Colonies radially sulcate, raised and densely sulcate at the center, margin plane, low, moderate, regular; sporulation dense; conidia en masse dull green to greyish green (25D4–5); mycelium pastel yellow (1A4); exudates golden; soluble pigments tomato red (8C8); reverse brownish red (9C8). DG18 25 °C, 7 d: colonies plane, low, margin plane, low, moderate, regular; sporulation dense; conidia en masse dark green (25F7–8); mycelium white; colony texture velvety; exudates absent; soluble pigments absent; reverse copper red (7C8) at the center, orange (5B8) elsewhere. OA 25 °C, 7 d: colonies plane, low, margin plane, low, moderate, regular; sporulation dense; conidia en masse greyish green (27E5–28E5); mycelium light yellow (1A6); colony texture velvety; exudates absent; soluble pigments absent. CREA 25 °C, 7 d: no growth.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate; Stipes smooth-walled, non-vesiculate, 90–150 × 2.5–3 µm; Metulae adpressed, 4–6 per stipe, 11.5–15 × 2.5–3.5 µm; Phialides acerose, 4–6 per metula, 11–15.5 × 2–3 µm; Conidia in long, distorted chains, smooth, narrow ellipsoidal to slightly fusiform, 2.5–3.5(–4) × 1.5–2.5 µm.Notes: Talaromyces germanicus grows slower on CYA, MEA and YES than the phylogenetically related T. minnesotensis. Talaromyces germanicus, T. chongqingensis and T. udagawae have similar growth rates on CYA incubated at 30 °C. The first two species can be differentiated by their colony texture and conidial color on MEA: T. germanicus produces velvety colonies with dull green to greyish green conidia, in contrast to the floccose colonies with greyish turquoise conidia of T. chongqingensis. Talaromyces udagawae produces a sexual state, a feature not observed in the other T. minioluteus-clade species.Talaromyces minioluteus (Dierckx) Samson et al., Stud. Mycol. 70: 176. 2011. MycoBank MB 560657. Basionym: Penicillium minioluteum Dierckx, Ann. Soc. Sci. Bruxelles 25: 87. 1901. MycoBank MB 157378.Neotype: Unknown location and substrate .BenA = MN969409; CaM = KJ885273; RPB2 = JF417443).ITS barcode: JN899346 : CREA 4–8; CYA 17–19; CYA 30 °C 9–14; CYA 37 °C no growth; CYAS no growth or 3–4; DG18 11–16; MEA 21–26; OA 9–16; YES 16–22.Colony characters: CYA 25 °C, 7 d: Colonies low, raised at center, radially sulcate towards margins, margin entire; sporulation moderate to dense in a ring towards the margins; conidia en masse greyish green (27C4–5); mycelium yellowish; colony texture velvety to weakly floccose; exudates absent; soluble pigments present, light brow (6D8), brownish orange (7C6–8); reverse light brown (6D8) in center, dark brown (6F8) in a ring under dense sporulation, edge sometimes orange (6B8). MEA 25 °C, 7 d: Colonies moderately deep, plane, margin entire; sporulation strong; conidia en masse dark green (27F7–8); mycelium pastel yellow (1A4); colony texture loosely funiculose; exudates absent; soluble pigments absent; reverse brownish orange (6C8), at margin orange (6B8). YES 25 °C, 7 d: Colonies slightly radially sulcate, umbonate, moderately deep, raised in center, margin entire; sporulation strong; conidia en masse dull green (27D4–E4); mycelium beige (4A3); exudates absent; soluble pigments orange to brownish orange (6B8–C8); reverse orange to brownish orange (6B8–C8). DG18 25 °C, 7 d: colonies plane, raised in center, plane, margin entire; sporulation moderate to strong; conidia en masse deep green (27D8–E8) or dull green (27D3); mycelium yellowish (2A2); colony texture cottony to floccose; exudates absent or orange; soluble pigments absent; reverse brownish orange to light brown in center (7C8–D8), margins reddish to brownish orange (7B8–C8). OA 25 °C, 7 d: colonies plane, low, margin entire; sporulation strong; conidia en masse dull green to greyish green (27D3–E3–5); mycelium beige; colony texture cottony; exudates absent; soluble pigments absent. CREA 25 °C, 7 d: no growth.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate, occasionally with an additional branch; Stipes smooth-walled, non-vesiculate, 100–250 × 2.5–3.5 µm; Metulae adpressed, 2–8 per stipe, 11–15.5 × 2.5–3.5 µm; Phialides acerose, 3–8 per metula, 10.5–15.5 × 2–2.5 µm; Conidia in long, distorted chains, smooth, broadly ellipsoidal or ellipsoidal, sometimes with a distinct connective, 2.5–3.5 × 1.5–2.5 µm.Notes: Talaromyces minioluteus is phylogenetically related to T. gaditanus, T. chongqingensis and T. samsonii. These species have, with the exception of T. chongqingensis, a similar growth rate on CYA 30 °C, CYA and MEA. Colonies of T. samsonii are more densely sporulating and have a floccose texture on MEA while T. minioluteus sporulates moderately dense and has a strongly funiculose texture. The colony reverse and soluble pigments produced by T. minioluteus are in shades of brown , while the reverse of T. samsonii is violet-brown to deep red and the soluble pigments vivid red to red.Talaromyces minnesotensis Guevara-Suarez et al., Mycoses 60: 657. 2017. MycoBank MB 820463. Typus: USA, Minnesota, from human ear, 2010, isolated by D.A. Sutton .BenA = LT559083; CaM = LT795604; RPB2 = LT795605).ITS barcode: LT558966 : CREA 0–5; CYA 20–24; CYA 30 °C 20–24; CYA 37 °C no growth; CYAS 0–9; DG18 14–19; MEA 28–30; OA 18–22; YES 19–28.Colony characters: CYA 25 °C, 7 d: Colonies low, radially sulcate with concentric rings, crateriform, margin entire, plane low, moderate, regular; sporulation dense; conidia en masse dull green to dark green (27E6–F6); mycelium pastel yellow (1A4); colony texture velvety; exudates absent; soluble pigments absent; reverse dark brown (8F7) at the center, reddish orange (7A7) elsewhere. MEA 25 °C, 7 d: Colonies plane or lightly sulcate near the center, low to moderately deep, margin entire, plane, low, moderate to wide; sporulation strong;); conidia en masse dull green to greyish green (25D4–E4–5); mycelium white to pastel yellow (1A4); colony texture velvety; exudates absent; soluble pigments absent or (10B8−C8reverse orange (5B8) or violet brown (10F8). YES 25 °C, 7 d: Colonies raised at center, lightly sulcate at the center to radially sulcate and sulcate at the center, margin entire, plane, low, moderate, regular; sporulation dense; conidia en masse greyish green (25D4–5) to greenish grey (27E2–3); mycelium white or pale yellow (1A3); exudates absent to golden red; soluble pigments absent or (9B6); reverse dark red (11C8) at the center with a greyish red (11D5) ring near the margins or copper red (7C8) with a brown (7F4) ring near center. DG18 25 °C, 7 d: Colonies plane, low, margin entire, plane, low, moderate, regular; sporulation dense; conidia en masse greyish green to dark green (27E5–F7); mycelium white (2A2–4); colony texture velvety; exudates absent; soluble pigments clear or absent; reverse orange (5B8−6B8) or corn to amber yellow (4B5–6) with a golden yellow (5B7) ring near center. OA 25 °C, 7 d: Colonies plane, low; margin entire, plane, low, moderate to wide, regular; sporulation moderate to dense; conidia en masse dull green (27D3–E3) or greyish green (27E5–7); mycelium white to greenish yellow (1A7); colony texture velutinous to granulose; exudates absent to dark red (11C8); soluble pigments absent. CREA 25 °C, 7 d: no growth to poor growth, acid production absent, base formation absent.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate; Stipes smooth-walled, non-vesiculate, 100–250 × 2–3 µm; Metulae adpressed, 3–5 per stipe, 9.5–15.5 × 2.5–3.5 µm; Phialides acerose, 3–5 per metula, 11–15.5 × 2–3 µm; Conidia in long, distorted chains, smooth, ellipsoidal, sometimes with distinct connectives, 2.5–3.5(–4.5) × 1.5–2.5 µm.Notes: Talaromyces minnesotensis is phylogenetically related to T. germanicus. The former species grows faster on CYA incubated at 30 °C.Talaromyces samsonii (Quintan.) Houbraken and Pyrri, comb. nov. MycoBank MB 841230. Basionym: Penicillium samsonii Quintan. Mycopathol. 91: 69. 1985.Typus: Lectotype designated here: CBS H-24877, MBT 10002955. Spain, Valladolid; apple  damaged by insect, isolated by J.A. Quintanilla (culture ex-type CBS 137.84 = DTO 304-C3 = DTO 169-G6 = CECT 2772 = IMI 282404 = IMI 327872).BenA = OK338798; CaM = OK338824; RPB2 = OK338844).ITS barcode: MH861709 : CREA 3–4; CYA 15–21; CYA 30 °C 11–16; CYA 37 °C no growth; CYAS 4–5; DG18 14–17; MEA 22–26; OA 12–15; YES 15–25.Colony characters: CYA 25 °C, 7 d: Colonies slightly sulcate, slightly raised at center, margin plane, low, moderate, regular; sporulation moderate to dense; conidia en masse greyish green to dark green ; mycelium sulfur yellow (1A5); colony texture velvety to velvety to weakly floccose; exudates absent; soluble pigments present, vivid red to red (10A8−B8); reverse violet brown to deep red . MEA 25 °C, 7 d: Colonies moderately deep, slightly radially sulcate, margin entire; sporulation strong; conidia en masse dark green ; mycelium light yellow to sulfur yellow (1A4–5); colony texture floccose; exudates absent; soluble pigments absent; reverse copper red to red (7B8−C8−D8). YES 25 °C, 7 d: Colonies slightly radially sulcate, umbonate, raised in center, margin entire; sporulation strong; conidia en masse dull green to greyish green (26D4–5); mycelium pastel yellow (2A4); exudates absent; soluble pigments orange to brownish orange (6B8−C8); reverse reddish brown to cherry red  at the center, blood red (10C8−D8) elsewhere. DG18 25 °C, 7 d: colonies plane, raised in center, margin plane, flat, wide, regular; sporulation moderate to strong; conidia en masse greyish green to deep green (27E6–8); mycelium yellowish (2A2); colony texture cottony to floccose; exudates absent; soluble pigments absent; reverse brownish orange to light brown in center (7C8–D8–9), margins reddish to brownish orange (7B8–C8). OA 25 °C, 7 d: colonies plane, low, margin entire; sporulation strong; conidia en masse dull green to greyish green (27E3–5) to dark green (27F6–8) at the center, greyish green (27C5) elsewhere; mycelium light yellow (1A4); colony texture velvety to cottony; exudates absent; soluble pigments absent. CREA 25 °C, 7 d: no growth.Micromorphology: Sclerotia and ascomata not observed. Conidiophores biverticillate, occasionally with an additional branch stipes smooth-walled, non-vesiculate, 90–200 × 2.5–3.5 µm wide. Metulae adpressed, 2–8 per stipe, 10.5–20.5(–23) × 2.0–4.5 µm. Phialides acerose, 3–8 per metula, 9.5–17 × 2–3.5 µm. Conidia in long, distorted chains, smooth, ellipsoidal to fusiform, sometimes with a distinct connective, 2.5–4(–5) × 1.5–3 µm.Notes: See also T. minioluteus. No holotype material of P. samsonii was deposited in a herbarium. A strain with a personal code (1032) is mentioned in the protologue and subcultures of this strain were deposited in CBS, IMI and CECT [and CECT . A subcuTalaromyces udagawae Stolk and Samson, Stud. Mycol. 2: 36. 1972. MycoBank MB 324424. Penicillium udagawae Stolk and Samson, Stud. Mycol. 2: 36. 1972.≡Typus: Japan, Misugimura, soil, isolated by S. Udagawa, 1964 .BenA = OK338783; CaM = MH861709; RPB2 = MN969148).ITS barcode: JN899350 : CREA no growth; CYA 5–9; CYA 30 °C 5–8; CYA 37 °C no growth; CYAS no growth; DG18 3–4; MEA 15–19; OA 13–17; YES 11–14.Colony characters: CYA 25 °C, 7 d: Colonies plane, raised at the center, margin plane, low, regular; sporulation absent; conidia en masse green dull green (28D4); mycelium white; colony texture floccose; exudates absent; soluble pigments absent, vivid red to cherry red (10A8−B8); reverse brownish orange (5C5). MEA 25 °C, 7 d: Colonies plane, low, margin plane, low, entire; sporulation absent; conidia en masse greyish green (25D5); mycelium pastel yellow (1A4); colony texture floccose; exudates absent; soluble pigments absent; reverse mandarin orange (6B8). YES 25 °C, 7 d: Colonies plane with a concentric ring, low, margin plane, low, entire; sporulation absent; conidia en masse dull green (25E4); mycelium white; exudates absent; soluble pigments absent; reverse greyish yellow (2C4). DG18 25 °C, 7 d: Colonies plane, low, margin plane, low, regular; sporulation absent; conidia en masse white; mycelium white; colony texture floccose; exudates absent; soluble pigments absent; reverse yellowish. OA 25 °C, 7 d: colonies plane, low, margin entire; sporulation strong; conidia en masse greyish green to jade green (27E5–7); mycelium yellow (1A6); colony texture velvety; exudates absent; soluble pigments absent. CREA 25 °C, 7 d: no growth.Microscopy: Asexual state fide Stolk and Samson [d Samson : SclerotNotes: Talaromyces udagawae can be distinguished from other T. minioluteus-clade species by the production of ascomata and its distinctive ascospores, which are ornamented with three to five regularly transverse, nearly parallel ridges. It has restrictive growth on media examined and does not grow on CREA, DG18 or CYA incubated at 37 °C. The asexual morph was not observed during this study, but was described in Stolk and Samson [d Samson .T. minioluteus could represent a species complex, and more recently, two species were described in this complex, T. chongqingensis and T. minnesotensis [T. minioluteus (CBS 642.68), also the ex-type strains of P. samsonii (CBS 137.84) and P. purpurogenum var. rubisclerotium (CBS 270.35) belong to this clade [BenA), Yilmaz et al. [T. minioluteus. Penicillium gaditanum (CBS 169.81) was not included in their phylogenetic analyses and this species was treated as synonym of T. minioluteus based on morphological and chemical data [P. gaditanum and P. samsonii are distinct species and we provide new Talaromyces combinations for these.Visagie et al.  suggestesotensis ,17. In aeus CBS 62.68, alseus CBS 62.68, alsnii CBS 17.84 and P. purpurogenum var. rubrisclerotium was uncertain. Yilmaz et al. [P. purpurogenum var. rubrisclerotium resolved in a clade with T. amestolkiae, but that its presumed ex-type strain CBS 270.35 clustered with T. minioluteus [T. chongqingensis. Thom [P. purpurogenum var. rubrisclerotium, producing sclerotia and able to grow at 37 °C. Both CBS 270.35 and CGMCC 3.20482 (ex-type of T. chongqingensis) deviate from the original description of P. purpurogenum var. rubrisclerotium [P. purpurogenum var. rubrisclerotium. Raper and Thom [P. purpurogenum var. rubrisclerotium. This strain morphological deviates from the original description in being floccose, poorly sporulating and lacking production of dark-red sclerotia, which is a hallmark character of P. purpurogenum var. rubrisclerotium [Penicillium pinophilum. Using sequence data, Peterson and Jurjević [T. sayulitensis, a close relative of T. pinophilus. Both NRRL 1064 and CBS 138204 (ex-type of T. sayulitensis) grow well on CYA at 37 °C [P. purpurogenum var. rubrisclerotium and treat this variety as a synonym of T. sayulitensis , a species phylogenetically unrelated to T. minioluteus. Because a name has no priority outside the rank in which it is published, the name P. purpurogenum var. rubrisclerotium does not compete with T. sayulitensis . The topologies of the single gene phylogenies are similar and largely adhere to the genealogical concordance phylogenetic species recognition (GCPSR) concept [T. minnesotensis in the BenA, CaM and ITS phylogenies, but has a different position in the RPB2 phylogram, where it is a sister of T. calidominioluteus and T. africanus . The morphology of CBS 147336 differs from the examined T. minnesotensis strains and may represent a new taxon  [BenA/CaM/ITS/RPB2 sequence signature are obtained.In total, eight species are resolved in the  concept . The excew taxon . MacromoT. minioluteus and related species can be identified using BenA, CaM, RPB2 or ITS sequences. In order to get insight in the distribution of the investigated species, re-identification of isolates/strains based on BenA and ITS sequences present in GenBank showed that 22 of 26 T. minioluteus strains/isolates actually represent other taxa in the clade (T. gaditanus (n = 7) followed by T. calidominioluteus (n = 6). In our study, we also found that these two species most common in the T. minioluteus-clade. Talaromyces minioluteus and related species have a worldwide distribution, but seem to be more frequently encountered in warmer climates [T. calidominioluteus is a postharvest pathogen of quince, tomato and orange fruits [All he clade . The majclimates ,37,38. Fe fruits  and/or iT. minioluteus or T. minnesotensis showed that this clade consists of eight phenotypically similar species. Based on ITS sequences deposited in GenBank, it can be concluded that there is likely more diversity in this clade (Re-evaluation of strains previously identified as is clade  and a mo


JUN, that we previously identified in ChIP-seq as sites of IGF-1Rβ enrichment. These data confirm the cell surface origin of nuclear IGF-1R, suggest the presence of nuclear αβ complexes and reveal that both IGF-1Rα- and β-subunits contribute to pro-tumorigenic functions of nuclear IGF-1R.Mature type 1 insulin-like growth factor receptors (IGF-1Rs) are heterotetrameric structures comprising two extracellular α-subunits disulphide-bonded to two transmembrane β-subunits with tyrosine kinase activity. IGF-1R is a well-known cell surface mediator of malignant growth, with an incompletely understood role upon nuclear import as a transcriptional regulator. Previous characterisation of nuclear IGF-1R focused on IGF-1Rβ. Here, we aimed to clarify the source of nuclear IGF-1R and investigate whether α-subunits contribute to nuclear IGF-1R function. Using prostate cancer cell lines DU145 and 22Rv1 we detected nuclear α- and β-subunits, with increase in nuclear signal upon IGF-treatment and reduction in response to IGF-1R inhibitor BMS-754807. Following biotinylation of cell surface proteins, biotinylated α- and β-subunits were detected in nuclear extracts of both cell lines. Furthermore, α- and β-subunits reciprocally co-precipitated from nuclear extract. Finally, we detected recruitment of both subunits to regulatory regions of chromatin, including the promoter of the oncogene The online version contains supplementary material available at 10.1007/s12672-021-00407-8. These mature receptors undergo trafficking to the plasma membrane, where the extracellular α-subunits form a complex binding site for a single IGF molecule and the β-subunits, which contain small extracellular and transmembrane domains, transmit signals via the intracellular tyrosine kinase domain  and Normal Rabbit IgG antibody (CST-2729) for 1 h at 4 °C with rotation. Precleared nuclear extracts were immunoprecipitated overnight at 4 °C with rotation with antibody for IGF-1Rα , IGF-1Rβ (CST-3027) or Normal Rabbit IgG control (CST-2729). Immunoprecipitated proteins were collected by 2 h incubation at 4 °C with rotation with Protein A agarose beads  Cat. No. P9269) and analysed alongside N, C and WCE input controls by western blotting.Immunofluorescence staining was performed as per Cell Signalling Technology protocol. Briefly, cells were fixed with 4% paraformaldehyde  for 1 h at room temperature. Blocking Solution was removed and cells were incubated overnight at 4 °C with primary antibodies diluted in Blocking Solution: IGF-1Rα  or IGF-1Rβ . After washing, cells were incubated in secondary antibody diluted in blocking solution: goat anti-mouse IgG  for cells incubated with primary antibody for the α-subunit, or goat anti-Rabbit IgG  for cells incubated with primary antibody for the β-subunit, for 1 h at room temperature with protection from light. Cover slips were mounted using mounting medium containing 4′,6-diamidino-2-phenylindole (DAPI). Slides were stored at 4 °C until visualisation on a Zeiss LSM 780 Confocal microscope  using ×63 magnification objective. Alexa Fluor 488 was excited with laser line 488 nm and emission collected between 480 and 670 nm. Alexa Fluor 594 was excited with laser line 594 nm and emission collected between 590 and 745 nm. Nuclear IGF-1R was assessed using FIJI (ImageJ) software by quantifying IGF-1R signal within the boundaries of outlined DAPI stained nuclei.JUN promoter and an intergenic region of chromosome 17 (IntChr17) described in [Cultured cells were fixed with 1% formaldehyde before being processed using Merck Millipore ChIP kit (Cat. No. 17-295) as per the manufacturers protocol, with minor modifications as in . Chromatribed in .DU145 and 22Rv1 prostate cancer cell lines showed detectable levels of nuclear IGF-1Rα- and IGF-1Rβ-subunits in nuclear extracts prepared from cells cultured in both serum-starved and IGF-treated conditions  protein input controls in both cell lines. Furthermore, both the α- and β-subunits were detectable in nuclear extracts from cells that had undergone cell surface protein biotinylation. Neither subunit was detected in control extracts that had undergone incubation with PBS rather than biotinylation reagent Fig. a, b. As Following confirmation that nuclear IGF-1R had translocated from the cell surface, we next wanted to investigate whether the two subunits of the receptor retain their interaction in the nucleus. Following cellular fractionation of DU145 and 22Rv1 cells, α- or β-subunits were immunoprecipitated from nuclear extracts. As before, both subunits were detected in the nuclear, cytoplasmic and WCE protein input controls. Following immunoprecipitation from nuclear extract, the α-subunit of IGF-1R could be detected in both DU145 and 22Rv1 immunoprecipitates. The IGF-1Rβ-subunit could also clearly be detected following co-immunoprecipitation using α-subunit antibody. Reciprocal β-subunit immunoprecipitation also detected both IGF-1Rα- and β-subunits in DU145 and 22Rv1 nuclear extracts Below is the link to the electronic supplementary material.


Acute lung injury (ALI) is a pulmonary disorder that leads to acute respiration failure and thereby results in a high mortality worldwide. Increasing studies have indicated that toll-like receptor 4 (TLR4) is a promoter in ALI, and we aimed to explore the underlying upstream mechanism of TLR4 in ALI.We used lipopolysaccharide (LPS) to induce an acute inflammatory response in vitro model and a murine mouse model. A wide range of experiments including reverse transcription quantitative polymerase chain reaction, western blot, enzyme linked immunosorbent assay, flow cytometry, hematoxylin–eosin staining, RNA immunoprecipitation, luciferase activity and caspase-3 activity detection assays were conducted to figure out the expression status, specific role and potential upstream mechanism of TLR4 in ALI.TLR4 expression was upregulated in ALI mice and LPS-treated primary bronchial/tracheal epithelial cells. Moreover, miR-26a-5p was confirmed to target TLR4 according to results of luciferase reporter assay. In addition, miR-26a-5p overexpression decreased the contents of proinflammatory factors and inhibited cell apoptosis, while upregulation of TLR4 reversed these effects of miR-26a-5p mimics, implying that miR-26a-5p alleviated ALI by regulating TLR4. Afterwards, OPA interacting protein 5 antisense RNA 1 (OIP5-AS1) was identified to bind with miR-26a-5p. Functionally, OIP5-AS1 upregulation promoted the inflammation and miR-26a-5p overexpression counteracted the influence of OIP5-AS1 upregulation on cell inflammatory response and apoptosis.OIP5-AS1 promotes ALI by regulating the miR-26a-5p/TLR4 axis in ALI mice and LPS-treated cells, which indicates a promising insight into diagnostics and therapeutics in ALI.The online version contains supplementary material available at 10.1186/s12890-021-01589-1. Acute lung injury (ALI), a severe respiratory disorder, is characterized by heterogeneous pathologic factors , 2. WithLong noncoding RNAs (lncRNAs) are longer than 200 nucleotides and participate in many biological and physiological processes , 11. It Importantly, it is reported that this ceRNA regulatory network is broadly implicated in multiple diseases including ALI –16. For MicroRNAs (miRNAs), another group of non-coding RNAs, comprise about 22 nucleotides and can post-transcriptionally regulate gene expression , 26. ForAccumulating studies have revealed that toll like receptor 4 (TLR4) is a key regulator of inflammatory response , 32. ForIn this study, we established animal and cell models of ALI by LPS treatment to explore the role as well as the regulatory function of OIP5-AS1 in ALI. We found out that OIP5-AS1 aggravated ALI development by promoting inflammation via regulating the miR-26a-5p/TLR4 axis. Our findings might offer a promising approach for ALI treatment.Escherichia coli O111:B4-derived lipopolysaccharide  in 50 μL of phosphate buffer saline (Sigma‐Aldrich) was intratracheally instilled while the mice in the sham group were injected with an equivalent volume of PBS (Sigma‐Aldrich). After 6 h, the mice were anesthetized with 1.5% pentobarbital sodium (60 mg/kg) and then sacrificed by cervical dislocation under anesthesia, and lung tissues were collected for following biochemical assays. Macrophages and neutrophils were isolated from ALI mice and sham mice according to previous studies [https://arriveguidelines.org).Total 64 BALB/c mice were raised in a room at 25 °C in a light/dark cycle of 12 h/12 h. All the mice were divided into two groups randomly: sham group (n = 8) and ALI group (n = 54). For establishment of the ALI mice model, 10 μg of  studies , 36. AllThe examination of lung W/D weight ratio was used to evaluate pulmonary edema. After 6 h of LPS treatment, the mice were euthanized, and the fresh lung tissues were weighed and the weight was recorded, following the drying at 180 °C in an oven for 24 h to examine dry weight.The mouse lung tissue samples were immobilized with 10% formalin for one day. After embedding in the paraffin, the tissues were cut into sections at the thickness of 5 μm. Furthermore, the tissue pieces were stained with hematoxylin–eosin (H&E) solution at room temperature and observed with a light microscope . The following standards were used to score the lung injury, 0: no damage or minimal damage; 1: mild damage; 2: moderate damage; 3: severe damage; 4: diffuse injury.Mice received an intratracheal injection of an adeno-associated virus 6 (AAV6) system as described previously . AAVs ca2 atmosphere. For cell transfection, 50 nM miR-26a-5p mimics (5′-UUCAAGUAAUCCAGGAUAGGCU-3′), 50 nM anti-miR-26a-5p (5′-AGCCUAUCGAUAUACUUGAA-3′), 1 μg of pcDNA3.1/OIP5-AS1, 1 μg of pcDNA3.1/TLR4 with corresponding negative controls were transfected into WI-38, TC-1 cells and PBECs using 3 μL of Lipofectamine 2000 . After 48 h of transfection, the cells were treated with 100 ng/mL LPS for 6 h and harvested for further use. Vectors were obtained from Genepharma .The human embryonic lung fibroblast WI-38, mice lung epithelial TC-1 cells , human primary bronchial/tracheal epithelial cells  were maintained in Dulbecco's modified Eagle medium (Sigma-Aldrich) containing 5% fetal bovine serum , 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C in a 5% CO−ΔΔCt method [hOIP5-AS1R: 5′-TGGCACTGCATGAGGGATTT-3′;qPCR: F: 5′-AACAGGTGCTTAGGTGGTGG-3′,mOIP5-AS1R: 5′-CCAACCCAGTCTCACATGCT-3′;qPCR: F: 5′-AAGCACAGTTGACCGCAGTA-3′,hBaxR: 5′-GCGTCCCAAAGTAGGAGAGG-3′;qPCR: F: 5′-TCATGGGCTGGACATTGGAC-3′,mBaxR: 5′-CTTCCAGATGGTGAGCGAGG-3′;qPCR: F: 5′-CTGGATCCAAGACCAGGGTG-3′,hBcl-2R: 5′-AGAAATCAAACAGAGGCCGCA-3′;qPCR: F: 5′-TTTGAGTTCGGTGGGGTCAT-3′,mBcl-2R: 5′-TGCACCCAGAGTGATGCAG-3′;qPCR: F: 5′-AACATCGCCCTGTGGATGAC-3′,hIL-1βR: 5′-CATGGCCACAACAACTGACG-3′;qPCR: F: 5′-TGAGCTCGCCAGTGAAATGA-3′,mIL-1βR: 5′-TGATGTGCTGCTGCGAGATT-3′;qPCR: F: 5′-TGCCACCTTTTGACAGTGATG-3′,hTNF-αR: 5′-GGCCTAAGGTCCACTTGTGT-3′;qPCR: F: 5′-CTGGGGCCTACAGCTTTGAT-3′,mTNF-αR: 5′-GTTTGCTACGACGTGGGCTA-3′;qPCR: F: 5′-ACTGAACTTCGGGGTGATCG-3′,hGAPDHR: 5′-GACTCCGACCTTCACCTTCC-3′;qPCR: F: 5′-GCTCTCTGCTCCTCCTGTTC-3′,mGAPDHR: 5′-ATGAAGGGGTCGTTGATGGC-3′;qPCR: F: 5′-GGAGAGTGTTTCCTCGTCCC-3′,hTLR4R: 5′-TTAGGAACCACCTCCACGCAG-3′;qPCR: F: 5′-GACGGTGATAGCGAGCCAC-3′,mTLR4R: 5′-CTCGGCACTTAGCACTGTCA-3′;qPCR: F: 5′-CCTGTGGACAAGGTCAGCAA-3′,hsa-miR-26a-5pR: 5′-GTCGTATCCAGTGCAGGGTCC-3′;qPCR: F: 5′-GCGCGCGTAACAGTCTACAGC-3′,mmu-miR-26a-5pqPCR: F: 5′-TCGGCAGGTTCAAGTAATCC-3′,R: 5′-CTCAACTGGTGTCGTGGAGT-3′;U6 (human)R: 5′-AACGCTTCACGAATTTGCGT-3′;qPCR: F: 5′-CTCGCTTCGGCAGCACA-3′,U6 (mouse)R: 5′-GCGACAAGGGAAGGGAACAA-3′;qPCR: F: 5′-CGCACTTTACGGCTACCTCT-3′,Total RNA was extracted from cells or tissues with TRIzol reagent . For miRNA analysis, the extracted miRNA (1 µg) was reverse transcribed into complementary DNA with a TaqMan MicroRNA Reverse Transcription Kit (Invitrogen). For lncRNA and mRNA analysis, RNA (1 µg) was reverse transcribed into complementary DNAs using the Oligo dT primer (Invitrogen). PCR reactions were conducted on an ABI 7500 Real-Time PCR System  with the following PCR procedure: initial denaturation at 95 °C for 30 s, with 40 cycles of denaturation at 95 °C for 5 s, as well as annealing at 60 °C for 20–30 s. The gene levels were determined by the 2t method  and wereg for 30 min and the supernatant were collected. Ago2 antibody  or IgG  were respectively incubated with 20 μL of protein G-agarose beads for 2 h at 4 °C and then cell lysate supernatant was filled in and incubated overnight at 4 °C. RNA was isolated from magnetic beads with TRIzol reagent (Invitrogen) and RT-qPCR was used to detect OIP5-AS1 and miR-26a-5p expression levels in the precipitates.The RIP assay was conducted using a Magna RIP RNA-Binding Protein Immunoprecipitation Kit . Briefly, cell lysate was centrifuged at 12,000×Lung tissues and cells were harvested and lysed by protein lysis buffer (Bio-Rad Laboratories). Then equal amount of protein samples (50 μg/lane) was separated on 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by transferring onto polyvinylidene difluoride membranes. Blocked by 5% skimmed milk for 1 h at room temperature, the primary antibodies against TLR4 , Bax , Bcl-2  and GAPDH  were incubated at 4 °C overnight. After washing by Tris-Buffered Saline (Bio-Rad Laboratories), the membranes were then cultured with horseradish peroxidase-conjugated secondary antibodies  at room temperature for 2 h. At last, the protein bands were assessed via an ECL kit (Amersham Biosciences) and the intensity was analyzed using ImageJ software. Original western blotting gels were provided in Additional file The luciferase reporter vectors (pGLO) containing the wild type 3′ untranslated region (3′ UTR) of TLR4 complementary to miR-26a-5p or mutated 3′ UTR of TLR4 were constructed by Genepharma , and were termed as pGLO-TLR4 3′ UTR Wt or pGLO-TLR4 3′ UTR Mut. The pGLO-miR-26a-5p -Wt or pGLO-miR-26a-5p-Mut was also constructed. Afterwards, the vectors (1 µg) were cotransfected with 50 nM miR-26a-5p, 50 nM anti-miR-26a-5p into PBECs, TC-1 and WI-38 cells using Lipofectamine 2000 (Invitrogen), separately. After 48 h, the relative luciferase activities were detected with luciferase reporter assay system . Relative luciferase activity was calculated as the ratio of firefly luciferase activity to Renilla luciferase activity.5 cells/mL and subjected to staining with 5 μL of Annexin V-FITC and PI for 25 min in the dark. Afterwards, the flow cytometry  was used to analyze apoptotic cells, and the data was analyzed using BD CellQuest Pro version 1.2 software (BD Biosciences). Percentage of apoptosis rate was calculated as apoptotic cells/total cells × 100%.The apoptosis rate of PBECs, TC-1 and WI-38 cells was evaluated using an Annexin V fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) apoptosis assay kit (Invitrogen) following previous procedures . In brieThe caspase-3 activity in lung tissues and cells was measured by a caspase-3 activity kit (Beyotime) according to the manufacture’s protocols. The tissues were isolated, prepared and lysed in cell lysis buffer. After protein concentration analyzing, proteins were filled in the cell lysis buffer. Subsequently, the reaction buffer and DEVD-ρNA substrate (caspase-3) were supplemented into the lysis buffer. The reaction mixtures were cultured at 37 °C for 2 h. The absorbance at 405 nm was assessed via a microplate reader .g for 10 min at 4 °C. Absorbance at 450 nm was determined via a microplate reader according to manufacturer’s protocols.ELISA was performed using the ELISA kits for TNF-α and IL-1β  in order to detect the concentrations of TNF-α and IL-1β in culture supernatant of PBECs after centrifugation 12,000×P value less than 0.05 was considered significant. All experiments were repeated at least three times.Data were presented as means ± standard deviation, as shown by the histograms with error bars in the figures. All data were assessed for normal distribution (Shapiro–Wilk test) and homogeneity of variance (Bartlett’s test). The differences between 2 groups were evaluated by independent Student’s tests and that among 3 or more groups were evaluated by one-way analysis of variance followed by Tukey’s post hoc test. To investigate the potential role of TLR4 in lung injury, the ALI mice model was established via intratracheally instilling with LPS. The intact alveolar structure without thickening or lymphocyte infiltration was observed in the lungs of sham-operated mice , total 28 lncRNAs were identified for further exploration (condition: high stringency of CLIP Data; Additional file According to starBase website (H&E staining results showed that miR-26a-5p upregulation alleviated the OIP5-AS1 overexpression caused lung injury in ALI mice (Fig. Overexpression of miR-26a-5p reversed the effects of OIP5-AS1 upregulation on Bax and Bcl-2 protein levels in lung tissues of ALI mice Fig. A. MiR-26ALI is a severe illness that threats health and lives worldwide because of the high incidence and mortality . TLR4, aWe then further explored the upstream regulatory mechanism of TLR4. Considering TLR4 was reported to be inhibited by multiple miRNAs including miR-214 and miR-1178 , 50, we It is widely accepted that lncRNAs are able to act as miRNA “sponges” to competitively combine with miRNAs to release target mRNAs . In ALI,In summary, our results revealed that OIP5-AS1 aggravated ALI by promoting inflammation and apoptosis by regulating the miR-26a-5p/TLR4 axis, which offers new insights into the therapeutic strategy of ALI.Additional file 1. Uncropped and unprocessed western blots.Additional file 2. The role of the OIP5-AS1/miR-26a-5p/TLR4 axis in inflammatory response and apoptosis of WI-38 and TC-1 cells.


Scientific Reportshttps://doi.org/10.1038/s41598-020-64874-9, published online 18 May 2020Correction to: 2)” was incorrectly given as “Puncta Density (Number per 20 μm2)”.The original version of this Article contained an error in Figure 1D, where the y-axis label “Puncta Density (Number per 200 μmThe original Figure The original Article has been corrected.


Quantum theory supported the feasibility of stabilizing ptSiIV by structural constraint, but its isolation has not been achieved yet. Here, we present the synthesis and full characterization of the first square-planar coordinated silicon(IV). The planarity provokes an extremely low-lying unoccupied molecular orbital that induces unusual silicon redox chemistry and CH-agostic interactions. The small separation of the frontier molecular orbitals enables visible-light ligand-element charge transfer and bond-activation reactivity. Previously, such characteristics have been reserved for d-block metals or low-valent p-block elements. Planarization transfers them, for the first time, to a p-block element in the normal valence state.The structure and reactivity of silicon(IV), the second most abundant element in our Earth's crust, is determined by its invariant tetrahedral coordination geometry. Silicon(IV) with a square-planar configuration (  •The first square-planar silicon(IV) is isolated•A low-lying LUMO provokes CH-agostic interaction and visible-light absorption•Transition metals are mimicked without leaving the natural oxidation state Tetrahedral silicon(IV) compounds are the building blocks of our Earth’s crust. Here, we describe the first species of silicon(IV) with a square-planar configuration. The structural deformation has substantial consequences for the compounds’ physicochemical properties and imparts features usually associated with transition metals. Upon planarization, the frontier molecular orbital gap shrinks by more than 50% and enables ligand-element charge transfer, CH-bond agostic interactions, and spontaneous reactivity with inert bonds. Small frontier molecular orbital gaps are critical for bond-activation reactivity, catalysis, and photochemistry with transition metals. Traditional approaches to mimic these characteristics with the more abundant p-block elements rely on unusual valence or oxidation states. With the realization of square-planar silicon(IV), these peculiarities start reaching p-block elements in their natural oxidation states. Our Earth’s crust is covered with compounds of silicon(IV). In each, tetracoordinated silicon(IV) arranges its four substituents in a tetrahedral fashion, while other environments are unknown. This work describes the isolation and properties of a molecular complex with square-planar silicon(IV). The flattened structural motif provokes a range of features that primes the second most abundant element for new applications in catalysis and materials science. The occurrence of only one peak for the pyrrole ring protons in the 1H NMR spectrum, and a 2-fold set of signals for the ethyl group, illustrates a highly symmetric species in line with effective D2d symmetry on the NMR time scale. Upon storing a solution of 2 in CH2Cl2 for 7 days at −40°C, block-shaped orange crystals developed  disclosed another structural peculiarity: two of the four inward-oriented methylene groups tilt toward the silicon center, reflected by varying CH….Si distances, e.g., Si–H21b (273.3(8) pm) versus Si–H29b (240.3(8) pm)  pm) D. In theicate 1− . Such ob2 in CH2Cl2 (69.5 μM) showed strong absorption bands at 301 and 360 nm, with a pronounced shoulder reaching into the visible range 3.,2 was attempted with decamethylcobaltocene (−1.9 V versus Fc/Fc+) as a reducing agent to support the electroanalytical results. Performed in C6D6, a rapid reaction to a mixture of unidentified products indicated a redox process to happen. Performed in CD2Cl2 as the solvent, conversion to the chlorido silicate [CoCp∗2][1] was observed (2⋅−). Hence, planarization enables silicon-based radical chemistry under mildly reducing conditions.Cyclic voltammetry revealed an irreversible reduction peak for nion 2⋅− . This reobserved . This fi2 in CD2Cl2 was subjected to phenylacetylene at room temperature. A rapid and quantitative silicon-ligand cooperative 1,2-addition of 2 to the carbon-carbon triple bond was verified by the occurrence of characteristic 1H- and 29Si-NMR spectroscopic patterns and mass spectrometry (−1) and a low reaction barrier (ΔG‡ = 61 kJ mol−1)  see . It repr2, which was thus analyzed by computational methods. Structure optimization of 2 validates the square-planar silicon (N–Si–N = 177°) as the energetic minimum. The LUMO in 2 has a strongly localized pz-character at silicon with an energy lower by 2.4 eV than the tetrahedral 3 (2 (165 kJ mol−1) is substantially higher than for tetrahedral 3 (6 kJ mol−1). The HOMO of 2 is located at the ligand's π-system, and its energy is higher in comparison with 3 by 0.9 V. The reduced HOMO-LUMO gap rationalizes the light absorption in the visible range, and the time-dependent density functional theory (TDDFT) computed UV-vis spectrum is in line with the experimental one (π(C=Cpyrrole) → Si(pz) charge transfer (HOMO/LUMO) and the more intense band at 301 nm as N(LP) → Si(pz) charge transfer (HOMO-4/LUMO) . Such a ligand-element charge transfer is very unusual for light p-block elements and underscores the π-donor stabilization in the square-planar silicon state.,π-delocalization of the nitrogen atom lone pair into the Si(pz) acceptor orbital, amounting to roughly 10% of the total Si–N bond interaction energy  B and S45n energy .2 was considered computationally. Although the experimentally observed methylene group tilting was not found in the DFT-computed global minimum structure, the red-shifted IR bands for the endo-methylene C–H bonds were reproduced in the computed IR frequencies (ρ(r) = 0.02 a.u., 0-metals.CH → Si(pz) second-order perturbation energies (38 kJ mol−1) and systematically depleted NBOs of the inward σCH bond in comparison with the other methylene CH-bond -tetrahedron into a planar structure. Small HOMO-LUMO gaps are the basis for the peculiar reactivity of d-block metals, low-valent p-block compounds, or frustrated Lewis pairs.The first square-planar silicon(IV) species greb@uni-heidelberg.de).Further information and requests for resources and reagents should be directed to and will be fulfilled by the lead contact, Lutz Greb  under accession numbers CCDC: 2042938, 2042939, 2042940, and 2042941. These data can be obtained free of charge from the CCDC at 4][1]  in 4 ml CH2Cl2 at ambient temperature, and stirred for further 12 h, during which it turned yellow. The solvent was removed under reduced pressure, the residue suspended in 10 ml n-hexane, stirred for 2 h, and filtered. The solvent of the filtrate was removed, and the residue was dissolved in 10 ml n-hexane, stirred for 24 h and, filtered. The clear yellow solution was stirred for another 24 h to drive the elimination of NaCl to completeness. The yellow solution was filtered again, and the solvent was removed. After drying in vacuo, the neutral silane 2 was obtained as a yellow solid 78% yield . Orange crystals of 2 were obtained by storing a concentrated solution of 2 in CH2Cl2 for 7 days at -40°C. Slow decomposition in CD2Cl2 and C6D6 was observed over 4 days at room temperature. Remark: the first step in this reaction is the cation exchange from PPh4+ to Na+. The SCXRD of this intermediate [Na]4[1]4 was obtained by storing a concentrated solution of [PPh4][1] and NaBArF20 in DCM for 5 days at room temperature without further workup (CCDC: 2042940). 1H NMR  3JHH = 7.4 Hz, 8H, CH2 Ethyl), 1.93 , 1.09 , 0.66 . 13C{1H}NMR C1), 106.5 (C2), 45.9 (CH2 Ethyl), 42.4 (C3), 24.8 (CH2 Ethyl), 9.4 (CH3 Ethyl), 9.3 (CH3 Ethyl). 29Si HMBC NMR : −55.6 ppm.Rigerously donor free sodium tetrakis(pentafluorophenyl)borate  was added to a solution of : C36H48N4Si∗H3O+: calc. 583.383 exp. 583.397.[ESI]: C3,112 (w), 2,963 (s), 2,931 (s), 2,872 (s), 2,815 (w), 1,344 (w), 1,514 (m), 1,455 (s), 1,378 (m), 1,364 (m), 1,322 (w), 1,297 (w), 1,278 (w), 1,234 (s), 1,132 (s), 1,087 (s), 973 (s), 953 (m), 926 (w), 857 (w), 736 (s), 713 (m), 681 (w).36H48N4Si, M = 564.87, monoclinic, P21/n (no. 14), a = 10.3052(4), b = 19.3263(7), c = 14.9876(7) Å, α = 90, β = 94.396(2), γ = 90, V = 2,976.2(2) Å3, Z = 4, Dc = 1.261 g cm–3, μ(Mo-Kα) = 0.112 mm–1, 2θmax = 61°, T = 100 K, orange blocks, Bruker APEX-II CCD; 9,086 independent measured reflections (Rint = 0.0505), R1(obs) = 0.0380, wR2 = 0.1046, 406 parameters. NoSphereA2-solution: R1(obs) = 0.0203, wR2 = 0.0436, 802 parameters. CCDC: 2042941(ShelXl solution)/2042939 (HAR/NoSpherA2-solution).C