We report the silencing of CA1 mRNA in PC3 and MDA cells. The levels of mRNA coding CA1 protein in the knock‐down mRNA (CA1 siRNA) cells have been measured by RT‐PCR and were approximately 5% (PC3) and 20% (MDA‐MB‐231), respectively, of the level of control (Mock siRNA) used during silencing. In PC3 and MDA‐MB‐231 cells, the mRNAs for COL1A1 and COL4A4 were up‐regulated. The mRNAs for CTHRC1, LAMC2, and WNT7B were not changed when compared to the control. The morphology of the cells during the treatments remained the same. On the Western blots, the lysate from the silenced cells showed lower levels of CA I as well. The first isoform of CAs, CA I, was first discovered in vertebrate erythrocytes in vitro. We have shown that the expression of the CA1 mRNA was up‐regulated in the tumour cells during the treatment of the above‐mentioned patients’ sera. The mRNAs coding for proteins associated with basal lamina assembly, cytoskeleton, WNT7B and collagen triple helix repeat containing 1 (CTHRC1) were down‐regulated Interestingly, in our previous work we reported that some patients who relapsed after high‐dose therapy (HDT) and autologous stem cell transplantation (ASCT) were positive for autoantibodies against CA I, and the tumours in these patients spontaneously regressed ®CRL‐1435TM) and a human breast adenocarcinoma cell line MDA‐MB‐231 (ECACC 92020424) were maintained in a high‐glucose (4.5 mg/ml) DMEM  supplemented with 10% foetal calf serum (FCS)  and gentamicin  at 37°C in humidified air with 5% CO2. All chemicals were purchased from Sigma‐Aldrich  unless stated otherwise.A human prostate adenocarcinoma cell line derived from metastatic site PC3 (ATCC6 cells) in a 10‐cm Petri dish. After 24 hrs, the cells were transfected with siCA1 to silence CA I expression and with siMock as a control using DharmaFECT™  according to the manufacturer's recommendations. The next day, the medium was replaced and the cells were incubated for 48 hrs. Two days later, the cells were collected and frozen for reverse transcriptase quantitative PCR analyses or lysed in a RIPA buffer and analysed by Western blot.For the transient silencing of the CA1 gene, PC3 and MDA‐MB‐231 cells were seeded (3 × 10g), and the total protein concentrations in supernatants were determined by BCA assay. Samples of 100 μg total proteins were separated using electrophoresis in the 10% SDS‐PAGE and blotted onto the PVDF membrane . The membrane was blocked for 2 hrs in a blocking buffer containing 5% non‐fat milk in PBS with 0,1% Tween 20. Incubation with primary antibodies was performed overnight at 4°C. A CA I mouse monoclonal antibody  was diluted 1:1000 in 3% BSA in TBST. Then, the membrane was washed with 0.1% Tween 20 in PBS, incubated for 1 hr (RT) with secondary antibody (Sigma‐Aldrich), washed again and developed with the ECL detection system.The cell lysates were centrifuged  was performed as previously described versus 36.37)  and MDA‐MB‐231 (breast carcinoma). Tumour cells with silenced CA1 gene (siCA1) compared with their negative control (siMock) did not show any morphological changes (Figs in vitro and changes the gene expression profile. All of these genes for proteins of the basal lamina and cytoskeleton were down‐regulated and in agreement with the observed morphological changes Subsequently, we checked the effect of silencing of the CA1 gene on the expression of the mRNAs for all selected ECM genes: COL1A1, COL4A4, CTHRC1, LAMC2, WNT7B (Figs Interestingly, results from RT‐qPCR were opposite to the results mentioned above. PC3 cells with silenced CA1 gene, mRNAs for COL1A1, COL4A4 were up‐regulated while the mRNAs for CTHRC1, LAMC2, and WNT7B were not changed when compared with the control/Mock RNA (Figs We have performed the silencing of the CA1 mRNA in the tumour cell lines PC3 and MDA‐MB‐231. We decided to use the PC3 cells because the CA1 mRNA is abundantly present in this line and the MDA‐MB‐231 cells because the CA1 mRNA is present only marginally in vitroWhen the tumour cells were grown in the presence of the anti‐CA I positive sera of the patients with spontaneous tumour regression, the expression of the CA1 gene has been increased. At the same time, we observed the downregulation of genes coding collagen type I alpha 1, collagen type IV alpha 4, laminin subunit gamma 2, CTHRC1 and WNT7B. This effect was accompanied with morphological changes of the tumour cells grown The decrease/increase of the CA I enzyme in the red blood cells of the human patients has been observed in patients with hyperthyroidism/hypothyroidism The authors confirm that there are no conflict of interests.


The present study demonstrates an ambient solution phase capping free synthesis of superionic AgCuS nanocrystals. Nanoscale size reduction, order–disorder phase transition and band gap evolution tailor the thermoelectric properties in AgCuS. Copper and silver based chalcogenides, chalco-halides, and halides form a unique class of semiconductors, as they display mixed ionic and electronic conduction in their superionic phase. These compounds are composed of softly coupled cationic and anionic substructures, and undergo a transition to a superionic phase displaying changes in the substructure of their mobile ions with varying temperature. Here, we demonstrate a facile, ambient and capping agent free solution based synthesis of AgCuS nanocrystals and their temperature dependent (300–550 K) thermoelectric properties. AgCuS is known to show fascinating p–n–p type conduction switching in its bulk polycrystalline form. Temperature dependent synchrotron powder X-ray diffraction, heat capacity and Raman spectroscopy measurements indicate the observation of two superionic phase transitions, from a room temperature ordered orthorhombic (β) to a partially disordered hexagonal (α) phase at ∼365 K and from the hexagonal (α) to a fully disordered cubic (δ) phase at ∼439 K, in nanocrystalline AgCuS. The size reduction to the nanoscale resulted in a large variation in the thermoelectric properties compared to its bulk counterpart. Temperature dependent Seebeck coefficient measurements indicate that the nanocrystalline AgCuS does not display the p–n–p type conduction switching property like its bulk form, but remains p-type throughout the measured temperature range due to the presence of excess localized Ag vacancies. Nanocrystalline AgCuS exhibits a wider electronic band gap (∼1.2 eV) compared to that of the bulk AgCuS (∼0.9 eV), which is not sufficient to close the band gap to form a semimetallic intermediate state during the orthorhombic to hexagonal superionic phase transition, thus AgCuS nanocrystals do not show conduction type switching properties like their bulk counterpart. The present study demonstrates that ambient solution phase synthesis and size reduction to the nanoscale can tailor the order–disorder phase transition, the band gap and the electronic conduction properties in superionic compounds, which will not only enrich solid-state inorganic chemistry but also open a new avenue to design multifunctional materials. We found that, unlike its bulk counterpart, nanocrystalline AgCuS does not exhibit p–n–p type conduction switching during the first superionic phase transition. Nanocrystalline AgCuS exhibits a higher electronic band gap (1.2 eV) compared to that of its bulk counterpart (0.9 eV) and the Raman spectroscopy shows the absence of Cu–S bond vibration during the (β–α) transition, which collectively disfavor the creation of a semimetallic intermediate electronic state during the orthorhombic (β) to hexagonal (α) phase transition that was responsible for the p–n–p type conduction switching in bulk AgCuS.13Herein, we present a facile and room temperature solution based synthesis for capping free AgCuS nanocrystals and investigate their temperature dependent (300–550 K) structural phase transition and thermoelectric properties. The synthesis process is very simple and effective with a high yield (∼90%). Temperature dependent synchrotron powder X-ray diffraction, Raman spectroscopy and heat capacity measurement indicates the observation of two superionic phase transitions, from a room temperature ordered orthorhombic (β) to a partially disordered hexagonal (α) at ∼365 K and from the hexagonal phase (α) to a fully disordered cubic phase (δ) at ∼439 K, in nanocrystalline AgCuS. A sharp decrease of the Seebeck coefficient value from 783 μV K2.ii) nitrate trihydrate , silver(i) nitrate , sulfur powder  and sodium borohydride , and tetraethylene glycol  were used for synthesis without further purification.Copper was added to the reaction mixture with stirring at room temperature (300 K) under N2 atm. By variation of the reaction time, we were able to synthesize nanocrystals of different sizes. Nanocrystals of ∼40 nm and ∼120 nm were synthesized with a reaction time of 30 min and 12 hours respectively. After the reaction the black colored product was isolated by centrifugation. The AgCuS nanocrystal powder was washed several times with ethanol to remove surface adsorbed tetraethylene glycol and was finally dried under a vacuum at 60 °C for 2 h. The yield of the reaction is ∼90%.Capping agent free AgCuS nanocrystals were synthesized by a bottom up wet chemical method. At first, silver nitrate , copper nitrate , sulfur powder  and tetraethylene glycol (15 ml) were placed in a round bottom flask, then the reaction mixture was sonicated for 15 minutes to get a well dispersed solution. NaBH–1.TGA was performed using a TGA/DSC 2 STAR instrument in the temperature range of 300–730 K under nitrogen atmosphere with a ramp rate of 5 K minA Bruker IFS 66v/S spectrometer was used for recording the FTIR spectra.α (λ = 1.5406 Å) radiation on a laboratory Bruker D8 diffractometer. Temperature dependent synchrotron powder X-ray diffraction measurements were carried out under N2 flow with an X-ray beam of λ = 1.0279 Å, at BL-18B (Indian beamline), Photon Factory, KEK, Tsukuba, Japan. The energy of the beam was set by a Si(111) double crystal monochromator, which was cross checked with a Si (640b NIST) standard. All the measurements were carried out in Bragg–Brentano geometry with a divergence slit (300 μm), an anti-scattering slit (350 μm) and a receiving slit (300 μm). High temperature measurements were carried out with Anton Paar DHS 1100 heat cell.Room temperature powder X-ray diffraction of the samples was recorded using Cu KFESEM imaging has been performed using a NOVA NANO SEM 600  operated at 15 kV. Energy dispersive spectroscopy (EDAX) analysis has been performed with an EDAX Genesis instrument attached to the FESEM column.2 20 STWIN TEM operating at 200 kV. EDAX elemental mapping was performed during STEM imaging. Background was subtracted  during the data processing for EDAX elemental mapping (with 500 eV minimum region of interest width).The TEM experiment has been done using a JEOL (JEM3010) TEM fitted with a Gatan CCD camera operating at 300 kV accelerating voltage and also using a FEI TECNAI G3 : HCl = 1 : 3) followed by diluting with milipore water. Ag standard , Cu standard  and S standard  were used to determine the compositions in ICP.ICP-AES measurements were carried out using a Perkin-Elmer Optima 7000DV instrument. ICP-AES measurements were carried out by dissolving nanocrystals in aqua regia (HNOXPS measurements were performed with a Mg-Kα (1253.6 eV) X-ray source with a relative composition detection better than 0.1% on an Omicron Nanotechnology spectrometer.–1 between 290 and 550 K in N2 atmosphere.A Netzsch DSC 200F3 was used for the DSC measurements with a heating/cooling rate of 20 K min2 atmosphere. The excitation wavelength of the laser was 514 nm.Temperature-dependent (300–473 K) Raman spectroscopy measurements were carried out with a LABRAMHR spectrometer in N3) using an AIMIL die-press at 60 kN m–2 pressure. The rectangular shaped sample was sealed in a quartz tube under vacuum (10–5 Torr) and then sintered for 3 h at 573 K which is above the upper limit of the thermoelectric measurement temperature. We could achieve a density of ∼94%, which is really good as the present nanoparticles do not contain any capping agents. The FESEM image after sintering indicates that the sample became more compact and the adjacent nanoscale grains are closely attached to each other with a slight increase in the grain size .α/Λ) data was calculated from the reflectance data using the Kubelka–Munk equation: α/Λ = (1 – R)2/(2R), where R is the reflectance and α and Λ are the absorption and scattering coefficients, respectively. The energy band gap was derived from an α/Λ (a.u.) vs. E (eV) plot.To probe the optical energy gap, optical diffuse reflectance measurements were performed on the nanocrystalline powder sample at room temperature. The spectra were recorded at the range of 600 nm to 2400 nm using a Perkin Elmer Lambda 900, UV/Vis/NIR spectrometer. Absorption , of the present fast–fast coincidence assembly is ∼220 ps. For the present positron annihilation measurement, a 10 μCi 22Na source of positrons (enclosed between 1.5 μm thin nickel foils) has been sandwiched between two identical and plane faced pellet type AgCuS nanocrystalline samples. The measured positron annihilation lifetime spectrum has been analyzed by computer programme PATFIT-88.85Sr has been used for the detection of the 511 keV γ-ray. The DBAR spectrum has been recorded in a dual ADC based – multiparameter data acquisition system . For the temperature dependent DBPAR measurement, the source sample sandwich has been placed inside a furnace (30–600 °C with ±2 °C at the sample site).7 counts have been recorded in a dual ADC based multiparameter data acquisition system . The Doppler broadening of the annihilation 511 keV γ-ray spectrum has been analyzed by evaluating the conventional line-shape parameters (S-parameter). The S-parameter is calculated as the ratio of the counts in the central area of the 511 keV photo peak (|511 keV – Eγ| ≤ 0.85 keV) and the total area of the photo peak (|511 keV – Eγ| ≤ 4.25 keV).Positron annihilation lifetime has been measured with a fast–fast coincidence assembly consisting of two constant fraction differential discriminators (Fast ComTech Model number 7029A). The detectors are 25 mm-long × 25 mm tapered to 13 mm diameter cylindrical BaF3.Cmc21). The β-AgCuS crystal structure consists of distorted hexagonal close packing (hcp) of sulfur atoms, and in the hcp sulfur layer three coordinated Cu atoms are occupied (P63/mmc) at 361 K, and finally at 439 K to a fully cation disordered superionic cubic δ-AgCuS phase (space group Fm3m).AgCuS is a polymorphous semiconductor that shows a superionic phase transition with changing temperature .13,26,27occupied . The plaoccupied .26,27 At+ and Cu+ were used as the metal precursors, and sulfur powder was used as a sulfur source. NaBH4 was used for the reduction of the metal nitrate salts. Tetraethylene glycol (TEG) was used as the solvent. TEG is a short chain aliphatic organic compound which contains two hydroxyl groups (–OH group) on the terminal carbon. Due to the presence of two terminal –OH groups, it can adsorb on the reactive surface of the as synthesized nanocrystal. In order to remove the surface adsorbed TEG, we have repeatedly washed the sample with ethanol (TEG is soluble in ethanol) and finally dried it under vacuum at 60 °C for 2 h. FTIR and TGA measurements have been used to probe the removal of the TEG after washing with ethanol  of the as synthesized nanocrystals could be indexed based on the orthorhombic β-AgCuS phase   . In orde, Cmc21) . Tempera, Cmc21)  along wi0.97CuS0.98, which is close to the nominal composition.FESEM images of the as synthesized sample indicate that AgCuS nanoparticles are agglomerated in nature. Absence of an additional capping agent during the synthesis results in the agglomeration of the AgCuS nanoparticles. The FESEM image indicates the average particle size of the nanocrystals obtained after 30 min of the reaction is ∼40 nm . The aveThe TEM image of the AgCuS nanocrystals obtained after 30 min of reaction also indicates the agglomerated nature of the particles . The SAEi) 3d3/2 and Ag(i) 3d5/2, respectively  2p1/2 and Cu(i) 2p3/2 respectively. An additional small peak centered at 942.6 eV was evidenced, which is due to the presence of Cu(ii) , which was otherwise not detected by PXRD. A peak located at 161 eV is due to S 2p  of nanocrystalline AgCuS obtained after 30 min of reaction was measured using a differential scanning calorimetry (DSC) technique and demonstrates two phase transitions above room temperature (Cp) are generally dramatic and sharp during the (β–α) transition compared to that of an (α–δ) transition.The temperature dependent specific heat  transition, which was present in the bulk AgCuS sample  indicates a sharp anomaly during the (β–α) transition (∼361 K); while the increase in σ during the (α–δ) transition (∼433 K) is not very significant  is quite similar to that for the bulk AgCuS.σ for the nanocrystalline sample is higher compared to that of bulk AgCuS  than in its bulk counterpart (nbulk = 1.3 × 1015 cm–3 at 300 K). Higher p-type carrier concentration in the nanocrystalline AgCuS sample is due to the formation of more Ag vacancies in the nanoscale, which act as a p-type dopant and inject more positively charged holes into the nanocrystalline sample.In nificant . The treS) in the temperature 300–550 K range, indicating p-type conduction  phase transition (S during the (β–α) phase transition temperature is consistent with the sharp increases in σ at ∼361 K. We have compared the temperature dependent S data of the AgCuS nanocrystals to the previously reported bulk AgCuS in S = ∼1757 μV K–1) during the orthorhombic to hexagonal (β–α) phase transition.S during the hexagonal to cubic (α–δ) phase transition at 437 K  also show the reversibility of the transition and hence confirm the temperature stability of the thermoelectric properties above the phase transition temperature  is a first derivative of Gibbs free energy (G), , where, P is pressure and T is temperature. First order phase transitions are generally associated with a hysteresis due to the change in volume during the transition.Nanocrystalline AgCuS (obtained after 30 min of reaction) shows a positive sign Seebeck coefficient  nanocrystalline AgCuS also shows p-type conduction in the 300–550 K range, but interestingly during the phase transition (β–α) the S value decreases to a much lower value (∼133 μV K–1 at 374 K) than that for the smaller nanocrystals (∼40 nm) obtained after 30 min of reaction  the conduction properties of the system are moving towards the conduction properties of bulk AgCuS.We have also performed the temperature dependent reaction . S vs. TZT) of merit was estimated from the thermoelectric data, which has been presented in Fig. S7  during the orthorhombic (β) to hexagonal (α) phase transition. Fig. S8  has been assigned to the free annihilation of positrons,τ2) is due to the annihilation of positrons in the defect site, mainly the Ag vacancy.τ3) originates from the formation of positronium at the sample surface or in the large voids inside the sample. Since the nanocrystal size (∼40 nm) is less than the positron diffusion length, the positrons are mainly trapped in the vacancy like defects at the surfaces/grain boundaries. Here the grain surface is enriched with Ag vacancy type defects, thus τ1 is an admixture with the positron lifetime in Ag mono-vacancy type defects present at the grain boundaries. The intermediate lifetime component is due to the trapping of positrons in Ag vacancy clusters. Positron annihilation lifetime measurements on the nanocrystalline AgCuS clearly show that the nanocrystalline surface is enriched with Ag vacancy like defects and defect clusters. This finding indicates that Ag vacancies are indeed responsible for the p-type conduction in nanocrystalline AgCuS.Positron annihilation spectroscopy is a powerful technique for the detection of vacancies or defect types in solids. In i.e. due to smaller size), the Ag vacancy is localized in nature which demands creation of more Ag vacancies during the (β–α) transition. This resulted in the leaching out of the Ag from the AgCuS nanocrystals followed by the segregation at the surface or grain boundaries. Ag leaching from the AgCuS nanocrystals injects more positively charged carriers, which makes the AgCuS nanocrystal p-type throughout the measured temperature range. Temperature dependent PXRD data also shows the appearance of the Ag peak along with the hexagonal (α) and cubic (δ) AgCuS phases on heating from orthorhombic (β) AgCuS nanocrystals  transition. The Doppler broadening spectra have been analyzed by evaluating the S-parameter defined by the ratio of the counts in the central area of the annihilation photo-peak and the total area of the photo-peak. In crystals .Previous electronic structure calculations on the bulk AgCuS during the (β–α) transition indicated that the contribution from hybridized Cu–S orbitals to the overlapping valence and conduction band gave rise to the semimetallic character of the intermediate structure that was responsible for the p–n–p type conduction switching. Although the signature of Cu–S bond vibration was present in the Raman spectra of the bulk AgCuS during the (β–α) transition, the Raman spectra of nanocrystalline AgCuS does not show any Cu–S bond vibration during the (β–α) transition. This result also indicates the absence of a semimetallic intermediate electronic state during the (β–α) transition in nanocrystalline AgCuS.4.Cp) and Raman spectroscopy measurements clearly indicate that the nanocrystalline sample undergoes two distinct order–disorder phase transitions at high temperatures. A superionic order–disorder phase transition tailors the thermoelectric properties in AgCuS nanocrystals and the fundamental understanding was developed based on temperature dependent Raman spectroscopy, positron annihilation spectroscopy and optical absorption experiments. Size reduction to the nano-dimension resulted in the vanishing of p–n–p conduction switching in AgCuS due to the following combined reasons: (a) higher band gap of AgCuS nanocrystals than that of the bulk counterpart, which is not appropriate for the creation of a semimetallic electronic state and the shifting of the Fermi level in the conduction band during the first order–disorder phase transition, (b) absence of hybridized Cu–S states during the (β–α) transition, which were otherwise responsible for the creation of an intermediate semimetallic state, (c) high surface to volume ratio in nanocrystalline AgCuS, which gives rise to the formation of more Ag vacancies and increases the p-type carrier concentration and (d) Ag leaching from AgCuS nanocrystals at elevated temperature injects more positively charged carriers into the system, which makes the AgCuS nanocrystal p-type throughout the measured temperature range. Our findings demonstrated that the p–n–p type conduction switching and thermoelectric properties of the bulk noble metal based chalcogenides can be tuned by reducing the crystallite size to nanometer scale. The discovery of an order–disorder phase transition in nanoscale materials and its effect on the electronic structure can bring novel phenomena with unusual properties.In conclusion, nanocrystalline AgCuS was synthesized for the first time by a simple capping free solution phase reaction at room temperature. Temperature dependent synchrotron powder X-ray diffraction, heat capacity (Supplementary informationClick here for additional data file.


Because of the many different calcium‐binding proteins distributed throughout cells, signalling precision requires localized rises in the cytosolic Ca2+ concentration. In electrically non‐excitable cells, for example epithelial cells, this is achieved by primary release of Ca2+ from the endoplasmic reticulum via Ca2+ release channels placed close to the physiological target. Because any rise in the cytosolic Ca2+ concentration activates Ca2+ extrusion, and in order for cells not to run out of Ca2+, there is a need for compensatory Ca2+ uptake from the extracellular fluid. This Ca2+ uptake occurs through a process known as store‐operated Ca2+ entry. Ideally Ca2+ entering the cell should not diffuse to the target site through the cytosol, as this would potentially activate undesirable processes. Ca2+ tunnelling through the lumen of the endoplasmic reticulum is a mechanism for delivering Ca2+ entering via store‐operated Ca2+ channels to specific target sites, and this process has been described in considerable detail in pancreatic acinar cells and oocytes. Here we review the most important evidence and present a generalized concept.Ca AChacetylcholineANO1anoctamin‐1CaCC2+‐activated Cl− channelCaCCKcholecystokininCRAC channel2+ release‐activated Ca2+ channelCaERendoplasmic reticulum3IPinositol trisphosphate3RIP3 receptorIPMCU2+ uniportermitochondrial CaNAADPnicotinic acid adenine dinucleotide phosphateNFATnuclear factor of activated T‐cells2PIPphosphatidyl inositol bisphosphatePMplasma membranePMCA2+‐activated ATPaseplasma membrane CaRyRryanodine receptorSERCA2+ activated ATPasesarco/endoplasmic reticulum CaSOARSTIM1 Orai1‐activating regionSOCE2+ entrystore‐operated CaSTIMstromal interaction moleculeTMEM16Atransmembrane member 16ATRPtransient receptor potential2+ signalling depends on specific Ca2+ sensors and transport proteins expressed differentially on organelle and plasma membranes, as well as Ca2+ buffers with different affinities and kinetics in different cellular compartments   into the cytosol ‐evoked Ca2+ release into the nucleoplasm through the inner nuclear membrane  being tunnelled through the ER lumen directly into the nuclear envelope. In this article, we describe and review the most important evidence for Ca2+ tunnelling, primarily based on studies of pancreatic acinar cells and oocytes.Effective and precise intracellular Ca Südhof, . However Putney, . Diffusilem Fig. . The pro2+ signals is encoded in their spatial, temporal and amplitude features. These Ca2+ dynamics combine to activate a defined subset of Ca2+‐dependent downstream effectors to transduce the cellular response . These elementary events due to the opening of one or a few channels can either remain localized resulting in Ca2+ signals in the microdomain around the channel(s), or coalesce through complex mechanisms into more global Ca2+ events that often encompass the entire cell  activation in T‐cells in response to antigen stimulation  at the plasma membrane and results in the production of IP3 and diacylglycerol. IP3 diffuses and gates open IP3 receptors at the ER membrane, releasing store Ca2+ to mediate the first phase of the Ca2+ signal. Should the Ca2+ release phase result in significant store depletion, it leads to the activation of Ca2+ influx at the plasma membrane through store‐operated Ca2+ entry (SOCE). SOCE is mediated by members of the stromal interaction molecule (STIM) and Orai family  junctions that are 20 nm apart /CRAC‐activating domain, which interacts with Orai1, traps it within the STIM1‐defined ER–PM junctions, and gates it open, thus allowing Ca2+ flow into the cell. SOCE activation not only results in store refilling but also shapes Ca2+ signal dynamics. There is therefore a tight functional link between IP3‐dependent Ca2+ release in response to agonist stimulation and Ca2+ influx through SOCE.Ca2+, liberating messengers – IP3 in the case of ACh and cyclic ADP‐ribose and nicotinic acid adenine dinucleotide phosphate (NAADP) in the case of CCK stimulation – thereby releasing Ca2+ from intracellular stores  and/or the hormone cholecystokinin (CCK). Interaction with specific surface membrane receptors activates signal transduction mechanisms that generate intracellular Ca2+ concentration ([Ca2+]i) changes evoked by ACh or CCK consist of repetitive short‐lasting elevations mostly confined to the apical region /anoctamin‐1 (ANO1)  are found in the basolateral membrane  opening of Cl− channels and exocytosis  inevitably activates Ca2+ pumps in the plasma membrane  resulting in extrusion of Ca2+ , ACh stimulation was discontinued and a period of Ca2+ entry was enabled by switching the pipette potential from negative to positive. No change was observed in [Ca2+]i during this Ca2+ entry period, but after discontinuation of Ca2+ entry (switching the pipette voltage back to negative), a new period of ACh stimulation caused a local rise of [Ca2+]i in the apical pole near the apical membrane, exactly as under normal conditions. A rise in [Ca2+]i near the cell‐attached pipette during Ca2+ entry could only be observed when the ER Ca2+ pumps  were arrested by thapsigargin  and that the whole of the ER was re‐equilibrated with regard to [Ca2+]ER within a few seconds   apical Ca2+ release, occurred through the basolateral membrane after a delay of about 6–7 s  current previously discovered in immune cells  during individual apical Ca2+ spikes shows  , there is primary generation of NAADP  allowing for easy spatial resolution of Ca2+ dynamics, which becomes particularly important for studies focused on the generation and propagation of Ca2+ waves, and elementary Ca2+ release events because their large spatial footprint in the oocyte makes them more amenable to investigation. The size of the oocyte also favours biochemical analyses and importantly linking them directly to Ca2+ signalling and other cell physiological processes at the single cell level , which are critical for oocyte biology and fertilization as they contribute significantly to the maintenance and regulation of the membrane potential. The Ca2+ transient generated at fertilization is initially localized at the site of sperm entry but gradually sweeps across the entire egg in the form of a Ca2+ release wave, which activates CaCCs and depolarizes the cell to prevent polyspermy ‐ethylenediamine (TPEN) and thapsigargin . TPEN is a transition metal chelator with low Ca2+ affinity that is freely membrane permeant and chelates high lumenal ER Ca2+, thus simulating Ca2+ store depletion and inducing SOCE   . Various approaches to block the IP3R or SERCA were used to show that the high CaCC current in response to SOCE when IP3 is present requires active IP3Rs  and of the Ca2+ effector (CaCC), and the distance to be covered by Ca2+ ions in the cytoplasm from the mouth of the Orai channel to the CaCC is incompatible with a diffusion mechanism given the speed of activation of CaCCs and the measured size and distribution of SOCE puncta as compared to CaCC‐rich membrane domains  has been directly monitored. The rate of rise of [Ca2+]ER decreases, as expected for a diffusional process, with increasing distance from the uncaging site , Ca2+‐sensitive K+ channels are strongly activated, supporting the idea that SOCE ‘fuels’ the IP3 receptors when the stores are empty to provide an efficient activation of the K+ channel , the SERCA pump and the IP3R. Store depletion stabilizes the STIM1–Orai1 puncta at ER–PM junctions thus providing the source for Ca2+ entry from the extracellular space. Ca2+ flowing through SOCE channels is taken up by the SERCA pump into the ER lumen preventing its diffusion out of the SOCE microdomain. In turn ER Ca2+ is released through IP3Rs thus delivering it to the appropriate effectors  with high efficiency and specificity. The Ca2+ tunnelling machinery has been adapted to very different cell physiological needs in the oocyte as compared to the pancreatic acinar cell. In the oocyte it modulates resting membrane potential and the temporal features of Ca2+ signals, whereas in the acinar cell it drives enzyme secretion and fluid flow. Given that the molecular machinery underlying Ca2+ tunnelling is ubiquitous, it is likely that this pathway is involved in Ca2+ signalling in a plethora of other physiological functions. The remarkable functional link between SOCE, SERCA and IP3R conscripted to allow Ca2+ tunnelling results in the delivery of Ca2+ to effectors that could easily be missed experimentally and interpreted as signalling downstream of SOCE directly. It is therefore likely that Ca2+ tunnelling activates additional cell physiological events that remain to be defined.Interestingly, the molecular mechanisms underlying CaNone declared.All authors have approved the final version of the manuscript and agree to be accountable for all aspects of the work. All persons designated as authors qualify for authorship, and all those who qualify for authorship are listed.O.H.P. is a Medical Research Council Professor (G19/22/2).


In vitro experiments have shown that there is significant cooperation between Pdx1 and Shh with regard to the production and maintenance of insulin‐producing cells (IPCs). In this study, the combined effect of Pdx1 overexpression and Shh manipulation on the function of adipose tissue‐derived IPCs was determined. A eukaryotic expression vector (Pdx1‐pCDNA3.1(+)) was constructed and transfected into a Chinese hamster ovary (CHO) cell line. Adipose tissue‐derived mesenchymal stem cells (ADMSCs) obtained from rats were assigned to two groups [control (C) and manipulated (M)] and differentiated into IPCs. Manipulated cells were treated with a mixture of FGF‐β and cyclopamine and recombinant Shh protein at days 3 and 11, respectively, and transfected with Pdx1‐pCDNA3.1(+) at day 10. The expression of multiple genes related to function of beta cells was analyzed using real‐time PCR. The functionality of IPCs in vitro was analyzed through dithizone (DTZ) staining and ELISA. IPCs were injected into the tail vein of diabetic rats, and blood glucose and insulin concentrations were measured. CHO cells transfected with Pdx1‐pCDNA3.1(+) showed a significantly higher expression of Pdx1 compared with nontransfected cells. Manipulated IPCs exhibited a significantly higher expression of MafA, Nkx2.2, Nkx6.1, Ngn3, insulin, and Isl1 and a higher insulin secretion in response to glucose challenge in relation to control cells. Rats that received manipulated IPCs exhibited a higher ability to normalize blood glucose and insulin secretion when compared to controls. Our protocol might be used for more efficient cell therapy of patients with diabetes in the future.Pancreatic and duodenal homeobox 1 (Pdx1) and Sonic hedgehog (Shh) are the key regulators of beta‐cell function.  ADMSCsadipose tissue‐derived mesenchymal stem cellsCDsclusters of differentiationCHOChinese hamster ovaryDABdiaminobenzidineDMEMDulbecco's modified Eagle's mediumDTZdithizoneIPCsinsulin‐producing cellsLBlysogeny brothNAnicotinic acidPdx1pancreatic and duodenal homeobox 1Pen/Streppenicillin/streptomycinShhsonic hedgehogT1DMtype 1 diabetes mellitusPdx1) is a transcriptional activator of insulin promoter Pdx1, MafA, and NeuroD mRNA expression and ultimately results in the activation of insulin gene expression Pdx1 is viewed as an essential factor for pancreatic development and β‐cell maturation Pdx1 activity promotes the inhibition of insulin‐like growth factors and apoptotic β‐cell death Pdx1 gene expression and subsequent pancreatic β‐cell functionality Pdx1 overexpression and Shh manipulation on the differentiation of ADMSCs toward IPCs.Among the existing treatment methods for type 1 diabetes mellitus (T1DM), stem cell‐based therapy is a practical approach with permanent results −1 ketamine and 10 mg·kg−1 xylazine. The pancreatic tissue and adipose tissue from the splanchnic region were isolated and washed three times with sterile PBS  containing 3% penicillin/streptomycin .Normal Sprague‐Dawley male rats aged 2–3 months and weighing 180–200 g were selected for the experiment. All the animals were kept in accordance with the Guide for the Care and Use of Laboratory Animals by the National Academy of Sciences  and Ahvaz Jundishapur University of Medical Sciences (AJUMS.REC.1393.100). The rats were euthanized using a mixture of 100 mg·kg2 concentration was adjusted to 1.5 mm. The specific primers (Bioneer Corporation) used for the amplification of full‐length Pdx1 gene included 5′‐ATATAAGCTTGATATGGAAAGTGAGGAGCAGGAGC‐3′ and 5′‐ATATGGATCCTCACCGGGGTTCCTGCGG‐3′. The primers were designed using primer premier 5.0 software  with restriction sites at the 5′ (HindIII) and 3′ (EcoRI) ends. The PCR was carried out using a thermal cycler . The thermal program given consisted of 35 cycles as follows: initial denaturation at 95 °C for 5 min, denaturation at 94 °C for 1 min, annealing at 58 °C for 1 min, extension at 72 °C for 1 min, and a final extension at 72 °C for 5 min. The purification of the Pdx1 PCR product from agarose gel was performed using the Gel DNA Recovery Kit  as per the manufacturer's recommendations. The purified Pdx1 PCR product and pcDNA3.1+ vector (ThermoFisher Scientific) were double‐digested with EcoRI and HindIII restriction enzymes  at 37 °C for 2 h. The digested fragments were electrophoresed on 1% agarose gel stained by Safe stain (SinaClon BioSciences). The digested fragments were purified using the Gel DNA Recovery Kit (SinaClon BioSciences) based on the manufacturer's instructions. The purified linear vector and insert were subjected to ligation reaction using T4 DNA ligase (Fermentas). The reaction was deactivated through incubation at 65 °C for 15 min. Two microliters of the ligation product was transformed into calcium chloride‐competent Escherichia coli Top10F’ cell . The transformed cells were selected on lysogeny broth (LB) medium agar plates using ampicillin (100 μg·mL−1). Several colonies were assayed by colony PCR using the universal primers T7 and BGH. Positive recombinant clones were cultured overnight at 37 °C. The plasmid was purified using the AccuPrep Nano‐Plus Plasmid Mini Extraction Kit  and sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit in an ABI 3130 Genetic Analyzer .Total RNA was isolated from the rats’ pancreas using RNX™ reagent as per the manufacturer's recommendations . The RNA concentration was determined at the wavelength of 260 nm using a NanoDrop spectrophotometer . The unimpaired RNA was confirmed by OD 260/280 nm between 1.8 and 2. The reverse transcription reaction was performed using CycleScript RT PreMix cDNA synthesis kit  in the total amount of 20 μL as recommended by the manufacturer. PCR was carried out in the total volume of 20 μL using Taq DNA Polymerase 2× Master Mix RED . The final MgClPdx1‐pCDNA3.1(+) vector and the expression of Pdx1 was determined using western blot analysis. CHO cells were cultured in Dulbecco's modified Eagle's medium (DMEM)‐HG medium containing 10% FBS and 1% Pen/Strep. After reaching a confluence of 70%, the cells were transfected with 20 μg of purified Pdx1‐pCDNA3.1(+) recombinant vector using electroporation. A gene pulser  was used to produce a pulse of 140 V for 15 msec. The CHO cells transfected with pCDNA3.1(+) were used as the control group. The transfected cells were selected using 1.5 mg·mL−1 of neomycin. The cells were incubated at 37 °C for 2 weeks. The medium was changed every 3 days. The transfected CHO cells were centrifuged for 5 min at 448 g washed three times with sterile PBS, and mixed with 100 μL of RIPA buffer . After 30 min of incubation on ice, the samples were centrifuged for 20 min at 11 200 g. The supernatant was collected for further analysis. The total protein concentration was determined using the Bradford method and BSA (1 mg·mL−1) as the standard solution. A total of 10 μL of cell lysate was mixed with 10 μL of Laemmli sample buffer containing 7.5% β‐mercaptoethanol and was heated for 10 min at 70 °C. The samples were loaded onto 12% SDS/PAGE gel and transferred into a nitrocellulose membrane . The membranes were blocked with 5% nonfat milk in PBS. After washing with PBS/Tween, the primary anti‐Pdx1 and GAPDH antibodies  were added at the ratio of 1 : 1000. The primary antibody detection was performed using goat anti‐rabbit HRP‐conjugated antibody (Abcam) at a 1 : 1000 ratio. The membranes were incubated with diaminobenzidine (DAB) for 10 min at room temperature and dried at a 37 °C incubator for 15 min. The densitometric quantification of Pdx1 protein in relation to GAPDH as the calibrator was performed using imagej software . The western blot analysis was performed in three independent experiments for each sample.To ensure the efficient overexpression of Pdx1 protein in eukaryotic cells, Chinese hamster ovary (CHO) cells were transfected with g for 8 min, and the supernatant was discarded. The pellet of the cells was resuspended in DMEM containing high glucose and 1% Pen/Strep and 10% FBS and was transferred to 25‐mL culture flasks. The flasks were incubated at 37 °C with 5% CO2. Four days after isolation, the medium was changed.The adipose tissue was harvested from the perivesical region and preserved in cold PBS containing 3% Pen/Strep. The adipose tissue was minced under sterile conditions and digested with collagenase type 1  for 25 min at 37 °C by shaking. The cell suspension was centrifuged at 161 6 cells·mL−1, the cells were washed three times with PBS and harvested using 0.05% trypsin. The cells were washed three times with PBS. Specific antibodies against cell surface clusters of differentiation  were dissolved in 3% BSA/PBS. A total of 1 μg·mL−1 of the antibody solution was added to the cells. The samples were incubated in the dark at room temperature for 30 min. The cells were washed three times with PBS and resuspended in 1 mL of ice‐cold PBS containing 10% FBS and 1% sodium azide. The fluorescence activity of the samples was measured using a Galaxy flow cytometer . The results were analyzed with flowjo 8.8.7 software . Two negative controls, including an isotype control and a stainless control, were provided for each sample.At the third passage, after the number of cells reached 1 × 10m isobutylmethylxanthine , 1 μm dexamethasone (Sigma‐Aldrich), 10 μm insulin , 100 μm indomethacin (Sigma‐Aldrich), and 10% FBS in DMEM‐LG (Gibco) was added. The cells were kept in the adipogenic medium for 21 days. The medium was refreshed every 3 days. After completing the differentiation process, the cells were washed three times with PBS. Oil Red O was dissolved in isopropanol and added to the flasks. The flasks were incubated at room temperature for 1 h. The cells were washed with PBS, and the oil droplets were visualized by light microscopy.At the third passage, for the adipogenic differentiation of ADMSCs, an adipogenic medium containing 0.5 mm β‐glycerophosphate (Sigma‐Aldrich), 10 nm dexamethasone (Sigma‐Aldrich), and 10% FBS in DMEM‐LG. The differentiation medium was changed every 3 days. After 14 days, the cells were washed three times with cold PBS and stained with 40 mm Alizarin Red. The presence of calcium deposits was examined using light microscopy.At the third passage, ADMSCs were differentiated to osteocytes using an osteogenic medium containing 50 μg·mL ascorbic acid (Sigma‐Aldrich), 10 mPdx1 overexpression. The basic protocol consisted of a 7‐day treatment with DMEM/F12, 1% insulin/transferrin/selenium (ITS), 2% FBS, and the change of the medium with a mixture of DMEM‐LG, 1% nicotinic acid (NA), 1% ITS, and 10% FBS for the subsequent 7 days −1 FGF‐β (Sigma‐Aldrich) and 0.25 μm cyclopamine (Sigma‐Aldrich). The reactivation of Shh was performed at day 11 through the incubation of the cells in a medium containing 150 ng·mL−1 recombinant Shh (Sigma‐Aldrich). The overexpression of Pdx1 was performed at day 10 of differentiation by the transfection of the cells with Pdx1‐pCDNA3.1(+) vector as described in the ‘Determination of functionality of Pdx1‐pCDNA3.1(+) vector’ section. The differentiation medium was refreshed every 2 days.The differentiation protocol used is summarized in Fig. Pdx1, Nkx2.2. Nkx6.1, MafA, Isl1, Ngn3, and insulin) between the two groups. The analysis was performed using the Ampliqon RealQ Plus Master kit (Ampliqon) and the LightCycler® SYBR Green I Master (Ampliqon). The primer (Bioneer Corporation) characteristics are listed in Table m), 3 μL cDNA (100 ng), and 8.5 μL nuclease‐free water. The PCR protocol consisted of a 5‐min denaturation at 95 °C followed by 45 cycles at 95 °C for 15 s and at 60 °C for 30 s. Two separate reactions without cDNA or with RNA were performed in parallel as the controls. Relative quantification was performed according to the comparative 2−ΔΔCt method using lightcycler 96® software. The validation of the assay to check whether the primer had similar amplification efficiencies for the target genes and GAPDH was performed as previously described Real‐time PCR was performed to compare the expression of the genes related to the pancreatic β cells  in sterile deionized water. The samples were incubated with KRB containing 5.5 mm glucose at 37 °C for 30 min. The supernatant was then exchanged with KRB containing 25 mm glucose and 30 mm KCl, and the incubation was performed at 37 °C for 30 min. The supernatant was collected, and the insulin concentration was determined using a rat‐specific insulin ELISA kit  based on the manufacturer's recommended protocol. The insulin concentration was reported in μIU·mL−1.The IPCs’ ability for insulin secretion was assayed using ELISA. At day 14 of differentiation, the IPCs were washed three times with Krebs‐Ringer bicarbonate buffer (KRB) containing 120 m−1 of dithizone  was prepared in dimethyl sulfoxide (Sigma‐Aldrich) and filtered through a 0.2‐μm filter. At the last stage of differentiation, 3 mL of working solution was added to each 25‐cm2 flask and incubated at 37 °C for 30 min. The cells were washed three times with PBS, and crimson red‐stained clusters were observed using a phase contrast microscope .A working solution of 100 ng·mLn = 15) aged 8 weeks and weighing 180–200 g were used. A total of 50 mg·kg−1 of streptozotocin  was dissolved in citrate buffer and injected intraperitoneally to the rats to induce experimental diabetes mellitus. One week after the injection, the rats with basal blood glucose above 500 mg·mL−1 were taken to be diabetic. The rats were divided into three groups. One group (n = 5) received undifferentiated ADMSCs, another control group (n = 5) was injected with unmanipulated IPCs, and the remaining rats (n = 5) received manipulated IPCs. The differentiated IPCs (manipulated and unmanipulated) were trypsinized and washed three times with PBS at day 14 of differentiation. Then, 1 × 106 of the isolated cells were suspended in 200 μL of DMEM‐HG. A mixture of 100 mg·kg−1 ketamine and 10 mg·kg−1 xylazine was used to anesthetize the rats. The cells were injected to the tail vein of the rats. Fasting blood glucose concentrations were measured once a week using a glucometer . At the end of the sixth week, the rats received a 25 mm glucose solution. After 10 min, the rats were anesthetized with a mixture of 100 mg·kg−1 ketamine and 10 mg·kg−1 xylazine. Then, 2 mL of blood was taken from them. The rats’ blood serums were isolated by centrifugation at 2000 r.p.m. for 5 min, and their insulin concentrations were determined using ELISA.Normal male Sprague‐Dawley rats . All analyses were carried out in triplicate. One‐way ANOVA followed by Tukey's post hoc analyses was used to test differences between various means including the expression level of different genes and insulin concentration. All experimental data were presented as mean ± SEM. The level of significance for all tests was set at P < 0.05.Data analyses were carried out using the Pdx1 gene is broken from both ends by restriction enzymes and the 850‐bp band is separated from the vector, which suggests the existence of Pdx1 gene in the pCDNA3.1(+) vector. After transforming the recombinant pCDNA3.1(+) plasmid containing the Pdx1 sequence to E. coli, direct colony PCR was used to ensure an accurate transformation. As shown in lane 3 in Fig. Pdx1 gene and 50‐bp flanking regions of the plasmid. This finding confirms the accuracy of the recombinant plasmid transformation in the bacteria. The sequencing of the recombinant plasmids was also performed with universal primers to confirm the accuracy of the Pdx1 sequence after amplification and cloning. The sequence obtained was translated using ExPASy Translate tool (https://web.expasy.org) and then analyzed using the online tools nBLAST and pBLAST. Based on this finding, the cloned Pdx1 gene sequence had a 100% homology to the Pdx1 sequences submitted to the GenBank (GenBank accession number: NM_022852.3). The ability of the CHO cells transfected with the Pdx1‐pCDNA3.1(+) construct for Pdx1 expression was confirmed through western blot analysis. After applying specific Pdx1 antibody, a single band at ~ 40 KD was detected. The CHO cells transfected with Pdx1‐pCDNA3.1(+) expressed higher levels of Pdx1 compared with the pCDNA3.1(+)‐transfected cells. Pdx1 overexpression in CHO cells transfected with the Pdx1‐pCDNA3.1(+) construct was thus confirmed  within 2 weeks. At the end of the third week, the average amount of blood glucose was raised to 315 ± 1.9 mg·dL−1. From then on, blood glucose decreased gradually. At the sixth week after the transplantation, the average glucose concentration reached 147 ± 1.1 mg·mL−1. The rats that received positive control IPCs showed the same pattern of changes in blood glucose. Nonetheless, they showed a more severe hyperglycemic condition. The diabetic rats that received undifferentiated ADMSCs showed no detectable changes in their blood glucose concentration (Fig. The transplantation of the manipulated IPCs to STZ‐diabetic rats resulted in a sharp reduction in the mean blood glucose concentration (92 ± 1.2 mg·mLion Fig. B.et al. Pdx1 promoter and adjusts Pdx1 expression, while Pdx1 in turn plays an important role in the regulation of insulin gene expression. Previous studies have also shown the key role of Pdx1 in the differentiation of different stem cells into IPCs, the activation of insulin transcription, and the maintenance of IPCs Pdx1 overexpression and Shh manipulation to optimize the differentiation of ADMSCs toward IPCs. The isolated ADMSCs exhibited general fibroblast‐like phenotypes of MSCs. The adherent ADMSCs expanded rapidly and expressed stem cell markers including CD90 and CD105, while they were negative for CD34 and CD45. The multipotency of ADMSCs was confirmed by their successful differentiation into osteocytes and adipocytes. The main differentiation protocol resulted in the production of functional IPCs. The differentiated cells obtained exhibited an endocrine pancreatic cell morphology and insulin secretion capacity in response to glucose. A study by Raikwar et al. Pdx1‐engineered embryonic stem cells into IPCs similarly yielded an elevation of Ngn3, Isl1, and insulin expression both in vitro and in vivo. By contrast, the study by Kubo et al. Pdx1 and Ngn3 during the differentiation of embryonic mice cells into IPCs resulted in the production of immature IPCs. The study by Cao et al. showed the effect of Pdx1 overexpression on IPCs’ functionality. The IPCs obtained in the study by Cao et al. reversed the hyperglycemic condition but were unable to express the late‐stage genes related to the pancreatic β‐cell development. They concluded that the exact differentiation of hepatic cells into functional IPCs requires additional external factors et al. showed that Pdx1 has a hedgehog‐responsive element on its promoter. Hedgehog suppression therefore downregulates Pdx1 and ultimately decreases insulin expression Pdx1, MafA, Ngn3, Isl1, Nkx2.2, Nkx6.1, and insulin mRNA compared to the control group Pdx1 overexpression concurrent with Shh manipulation on differentiation outcomes. The IPCs obtained expressed significantly higher amounts of Pdx1, MafA, Ngn3, Isl1, Nkx2.2, Nkx6.1, and insulin compared to the control group. The differentiated cells obtained secreted remarkably higher amounts of insulin than the cells in the control group. This finding demonstrates the effectiveness of Pdx1 overexpression concurrent with Shh manipulation in the production of functional IPCs in vitro. The manipulated IPCs obtained in the present study were capable of reducing blood glucose levels to near euglycemia. The potency of the manipulated cells for the secretion of insulin showed a significant increase compared to the control IPCs in vivo. In line with the present findings, Thomas et al. Pdx1 promoter activity and Pdx1 and insulin expression at the late stages of pancreatic development. Hebrok et al. et al. on the differentiation of human neural progenitor cells into IPCs showed no detectable expression of Shh during the four stages of differentiation toward IPCs. In their study, 300 ng·mL−1 of Shh protein administered at the final stage of differentiation resulted in the elimination of Pdx1, FoxA3, and insulin expression The transplantation of artificial IPCs is a promising treatment for T1DM Pdx1 overexpression induced the generation of functional IPCs. The concurrent use of Pdx1 overexpression and Shh manipulation significantly increased the functionality of the generated artificial IPCs. Further studies are needed to clarify the molecular mechanisms underlying the concurrent use of Pdx1 and Shh in the generation of mature functional IPCs.The present study reveals successful functional IPC generation through the described differentiation protocol. Shh manipulation and DD, MHT, EMP, and MRT designed the study. DD, MHT, and MRT performed the study, researched the data, analyzed the results, wrote the manuscript, and revised the article critically for important intellectual content. EAB, AAG, MAG, and MO analyzed the data and drafted and revised the manuscript. All authors gave final approval of the version to be published. DD is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.


In this proof‐of‐concept study, an active‐template coupling is used to demonstrate a novel kinetic self‐sorting process. This process iteratively increases the yield of the target heterocircuit [3]rotaxane product at the expense of other threaded species. However, this raises the synthetic complexity of the thread, and only limited, stereochemically trivial examples have been reported.n]rotaxanes have been developed; Sauvage and co‐workers reported the coupling of kinetically inert pseudorotaxane complexes,Conceptually, the simplest approach to the synthesis of heterocircuit rotaxane synthesis rely on passive templates to direct the assembly of a thermodynamically preferred pseudorotaxane complex before covalent bond formation kinetically traps the interlocked assembly. Leigh's active template (AT) approach to mechanical bond formation is unusual in that the mechanical bond can, in theory, be formed solely under kinetic control; the covalent bond forming reaction simply takes place faster through the cavity of the macrocycle.Self‐sorting reactions typically operate under thermodynamic control, allowing the system to correct errors, although the trajectory to achieve equilibrium can be kinetically complex.This suggests an opportunity to develop self‐sorting reactions that are governed only by the kinetic stability of the products. Herein we report the realization of this proposal: a stereoselective four‐component coupling in which kinetic self‐sorting amplifies the yield of a target heterocircuit [3]rotaxane.I and II, with different internal diameter and two half‐threads, III and IV, of different steric bulk. Focusing on the fate of the larger of the two rings , four threaded products are possible: [2]rotaxane V, homocircuit [3]rotaxane VII, and stereoisomeric heterocircuit [3]rotaxanes VIII and IX. If only one of the half‐threads is bulky enough to retain the larger macrocycle this mixture is simplified as macrocycle I can dethread in all cases except product VIII, in which its path is blocked by smaller macrocycle II, which is in turn held in place by the smaller stopper, an example of Schalley's cascade stoppering.I renders it available for further AT coupling, suggesting that this “ratcheted” self‐sorting process will amplify the yield of VIII above the natural selectivity of the reaction. Overall, [3]rotaxane VIII is the only stable interlocked product derived from I, alongside the products of dethreading (recovered I non‐interlocked thread X and [2]rotaxane VI) and the products of direct AT couplings of macrocycle II.Our proposed AT self‐sorting process Figure  requires1 and 2 , whereas macrocycles 2 b (entry 3) and 2 c (entry 4) only form the singly interlocked product. To account for the high yield of [3]rotaxane in the case of macrocycle 1, we previously proposed a mechanistic pathway involving dinuclear reactive intermediates A and B (Scheme 2 to form significant quantities of [3]rotaxane was ascribed to the lack of Cu−O interactions to stabilize doubly threaded intermediates (2 a) and the inability of smaller macrocycles to coordinate Cu(π) in a threaded manner (2 b and 2 c).Initial experiments were performed with bulky azide 1 and 2 a were employed together (entry 5), a complex mixture was produced consisting of all possible interlocked products. Careful 1H NMR analysis of the crude mixture revealed a number of key points. First, under these conditions, simple [2]rotaxanes 5 and 6 a were formed in unequal quantities, possibly indicating a difference in reactivity between 1 and 2 a. However, when the reaction was run under competition conditions (entry 6), [2]rotaxanes 5 and 6 a were formed in near‐equal amounts, suggesting that the [2]rotaxane pathway is equally favored for both macrocycles. Second, although macrocycle 2 a alone forms only trace quantities of [3]rotaxane 8 a, in the presence of 1, 2 a is competitively recruited into the [3]rotaxane pathway and the combined yield of heterocircuit rotaxanes 9 a and 10 a is of the same order of magnitude as homocircuit [3]rotaxane 7. Finally, heterocircuit rotaxanes 9 and 10 were formed in an unequal ratio of isomers, suggesting that there is a significant preference in the position macrocycles 1 and 2 a occupy in the mixed‐macrocycle equivalents of intermediates A and B. Careful chromatography allowed the major isomer to be isolated and identified by ROESY NMR as [3]rotaxane 9 a , suggesting that the intermediate in which macrocycle 1 coordinates to Cu(π) is favored.When macrocycles 2 b to coordinate Cu(π), the reaction of 1 with 2 b (entry 7) led to a simpler product mixture containing [2]rotaxanes 5 and 6 b, homocircuit product 7 and single heterocircuit [3]rotaxane isomer 9 b. Finally, when 1 and 2 c are employed (entry 8), both of which contain the key benzylic ether unit, the quantity of the sole heterocircuit stereoisomer 9 c increases significantly, further suggesting that the heterocircuit pathway is enhanced when both macrocycles contribute stabilizing Cu−O interactions. Chromatography allowed 9 c to be isolated in 20 % yield based on 1 and the stereochemistry confirmed by ROESY NMR .In keeping with the inability of 9 c observed in the reaction of macrocycles 1 and 2 c, we selected this combination for investigation under self‐sorting conditions. As the sole heterocircuit product formed in the reaction between azide 3 and alkyne 4 is that in which larger macrocycle 1 is situated on the side of the axle derived from the alkyne component, we investigated the reaction of macrocycles 1 and 2 c in the presence of alkyne 4 and smaller azides 11  and recovered 1 (81 %), alongside the corresponding non‐interlocked thread, and the [2]rotaxane of macrocycle 2 c. [3]Rotaxane 14 c was not observed. The order of the macrocyclic components on the axle of 15 c was again confirmed unambiguously by ROESY NMR analysis .Although molecular modeling1 into heterocircuit [3]rotaxanes 9 c  suggests that significant de‐threading of 1 from pseudorotaxanes 12 c and 13 c, and thus self‐sorting, does not take place competitively on the time scale of the AT‐CuAAC reaction.2 c, 4, 11 c and CuI was added and the AT‐CuAAC reaction repeated, 1H NMR analysis indicated the conversion of 1 to 15 c increased to 31 % , macrocycle 2 (0.5 equiv), azide (1.20 equiv), alkyne (1.20 equiv), and [Cu(MeCN)4]PF6 (0.96 equiv) were dissolved in CH2Cl2 and the solution was stirred at 100 °C (under microwave radiation) for 2 h. The solution was allowed to return to RT, diluted with CH2Cl2 (50 mL), and washed with NH3–EDTAaq. The aqueous layer was extracted with CH2Cl2. The organic extracts were combined, dried (MgSO4), and the volume reduced in vacuo.General procedure for the synthesis of heterocircuit [3]rotaxanes 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.


The recent theoretical prediction for superconductivity in LaNiO3 thin films has also triggered intensive research efforts. LaNiO3 seems to be the only rare earth nickelate that stays metallic and paramagnetic down to lowest temperatures. So far, centimeter-sized impurity-free single crystal growth has not been reported for the rare earth nickelates material class since elevated oxygen pressures are required for their synthesis. Here, we report on the successful growth of centimeter-sized LaNiO3 single crystals by the floating zone technique at oxygen pressures of up to 150 bar. Our crystals are essentially free from Ni2+ impurities and exhibit metallic properties together with an unexpected but clear antiferromagnetic transition.The material class of rare earth nickelates with high Ni 3 has an unanticipated magnetically ordered metallic phase.The phase transitions of rare earth nickelates have attracted intensive study as they arise from the complex interplay of charge, spin and lattice degrees of freedom. Here Guo et al. present evidence that LaNiO RNiO3  with the high Ni3+ oxidation state have continued to attract enormous interest due to the famous bandwidth controlled metal-insulator (MI) transition and associated unusual charge and spin-order phenomena occurring in this system2 with even the possibility for multiferroicity3. The description of the underlying physics of these phenomena turned out to be a true intellectual challenge and it is not a surprise that important theoretical concepts have been (and still need to be) developed along the way12. More recently, the prediction of high-Tc superconductivity in LaNiO3-based heterostructures13 has triggered a flurry of new activities on LaNiO3–LaMO3 superlattices 15.The rare earth nickelates 3, all RNiO3  compounds exhibit a MI transition at lower temperatures2 with an insulating antiferromagnetic ground state. With decreasing R-ionic radius, the octahedral tilts become larger. Thus, the Ni–O–Ni bond angles become smaller, which alters the electronic bandwidth and the magnetic exchange interactions. Whereas an enhancement of the insulating properties and an increase of the MI transition temperature TMI can be observed for decreasing R-ionic radius, the antiferromagnetic transition temperature and the strength of the exchange interactions decrease with decreasing R-ionic radius and Ni–O–Ni bond angles. With La having the largest ionic radius of the series, one might expect the strongest antiferromagnetic properties for LaNiO3. However, so far it was reported that LaNiO3 does not show any magnetic order, and thus, violates the trend. It remains a paramagnetic metal down to lowest temperatures with an enhanced effective mass17.Apart from LaNiO3 single crystals by the floating zone technique. Electrical resistivity and Hall effect measurements on our single crystals show that LaNiO3 is intrinsically not a bad metal as recently discussed for the RNiO3  compounds in their paramagnetic phase10. On the contrary, we found that LaNiO3 has a high conductivity. Moreover, we were able to observe bulk antiferromagnetism in magnetization, specific heat, and neutron scattering experiments. Thus, LaNiO3 appears to be a highly metallic and antiferromagnetic transition metal oxide—a rather rare combination in oxides. Special about LaNiO3 is that it is also very close to an insulating state, making LaNiO3 an intriguing quantum material, probably close to a quantum critical point, where strong local electronic correlations at the Ni sites are likely to interfere in an intricate manner with Fermi surface effects.Here, we report on the successful growth of large LaNiO3 single crystals with the floating zone technique, see inset of Fig. 3 crystal is confirmed by a series of Laue diffraction patterns from different directions and across the length of this crystal as well as by single crystal X-ray diffraction, see inset of Fig. PmmRc) somewhat below 1100 K2, our floating zone grown single crystals are twinned.Using high oxygen pressures of 130–150 bar, we were able to grow LaNiO2,3 X-ray absorption spectroscopy (XAS) data of the LaNiO3 single crystal taken at 300 and 90 K together with that of a NiO single crystal serving as a Ni2+ reference compound. We have removed the La-M4 white lines located at 850.6 eV from the LaNiO3 spectra using the La-M4 spectrum of LaCoO3. It is well known that XAS spectra at the L2,3 edge of transition metal oxides are highly sensitive to the valence state, in particular, an increase of the valence state of the transition metal ion by one causes a shift of the XAS L22,3 spectra by one or more eV toward higher energies18. The more than one eV higher energy shift between the spectra of NiO and LaNiO3 indicates the formal Ni3+ valence state in LaNiO319, with the note that the spectrum cannot be interpreted in terms of a Ni 3d7 configuration, but, rather by a coherent mixture of 3d8 and 3d812, where each 2+ impurities in our LaNiO3 single crystal—otherwise the sharp main peak of the Ni2+ impurity spectrum would have been visible as a sharp shoulder at the leading edge18. At the O-K edge, the pre-edge peak is shifted by about 1 eV to higher energies when going from Ni3+ to Ni2+19. Figure 3 single crystal (blue) and the NiO reference compound (red). The O-K XAS spectra demonstrate even more clearly that there is no spectral feature from Ni2+ impurities and thus that our as-grown LaNiO3 single crystals are highly stoichiometric. This is further confirmed by thermogravimetric and inductively coupled plasma optical emission spectroscopy (ICP-OES) measurements, see Methods and Supplementary Note Figure 3 single crystal that has been powdered and measured by means of powder X-ray diffraction. There is no clear indication for the presence of a structural anomaly which otherwise occurs readily in the other nickelates RNiO3 with smaller rare earths  when cooling through the MI transition2.In Fig. ρ of our LaNiO3 single crystal that is shown in Fig. 3 powder samples17 or LaNiO3 thin films14 or of crystals grown under 50 bar oxygen pressure20. Thus, our LaNiO3 single crystals are more metallic, see also Supplementary Note 3 (for temperatures between 2 and 300 K) yields high carrier densities of about 4.8 × 1028 m−3, corresponding to ~2.7 (close to 3) electrons per formula unit, consistent with the 3+ valence of the Ni ions in LaNiO3. Up to ~35 K, the low-temperature behavior of the resistivity is Fermi-liquid-like, ρ(T) = A = 1.62.10−3 μΩ cm K−2, and n ~2.0, see Fig. A is of the same order as reported in literature21. A fit over the entire temperature range gives an exponent n ~1.50(1) with the residual resistivity 3 is close to a quantum critical point (cf. the results for PrNiO3 under pressure21).The electrical resistivity χ of our LaNiO3 single crystal, see also Supplementary Note 17, which confirms unprecedented high quality of our single crystals. Surprisingly, χ exhibits an anomalous kink at ~157 K, which we take as an indication for a hitherto unknown antiferromagnetic transition in LaNiO3. That this anomaly is not simply caused by a signal from a tiny fraction of a magnetic impurity phase  can be excluded by our specific heat (Cp) measurements. As can be seen in Fig. 3 minority phase that becomes antiferromagnetic and insulating at low temperatures23: we do not observe an upturn or a slowing down of the decrease in the resistivity on cooling. Moreover, our samples have resistivities in the μΩ cm range and have conductivities higher than reported so far. All these support the notion that the transition is an intrinsic bulk property and not due to an impurity phase. The low-temperature behavior of the specific heat supports a Fermi-liquid type of the ground state of LaNiO3, Cp(T) = −1 K−2, consistent with the value 18 mJ mol−1 K−2 reported before17, and with β = 1.87(5) × 10−4 J mol−1 K−4, thus yielding a Debye temperature N is the number of atoms in the chemical formula and R is the ideal gas constant.Figure 3 single crystals also enabled us to study this intriguing system by means of neutron diffraction and inelastic neutron scattering. These experiments were performed at the Thales, IN12 and IN8 spectrometers at the ILL in Grenoble, France. Within elastic scans, we were able to observe quarter-integer peaks at low temperatures, see Fig. RNiO324. The propagation vector observed for the insulating antiferromagnetic regime of RNiO3 amounts to (1/4 1/4 1/4) in pseudocubic notation (or (1/2 0 1/2) in orthorhombic notation)2. Moreover, the study of the temperature dependence of these quarter-integer peaks indicates an onset temperature that coincides with the magnetic ordering temperature TN that we observed in susceptibility and specific heat measurements of LaNiO3, see Fig. The availability of sizeable LaNiO3 could be also confirmed by polarized neutrons at the IN12 spectrometer. In Fig. L-scans across two quarter-integer peaks are shown for the three spin-flip (SF) channels and for a non-spin-flip channel. Only in the SF channels, neutron scattering intensities 3. Although we cannot detect a symmetry lowering from our high-resolution powder X-ray diffraction measurements—see Fig. 3 is the same as the pseudocubic propagation vector for the orthorhombic insulating nickelates RNiO3 . Based on the magnetic symmetry analysis for the high symmetry cubic structure with space group Pmm and for the propagation vector (1/4 1/4 1/4), a helical magnetic structure with moments spiraling perpendicular to the propagation vector is consistent with our single crystal neutron data, see Supplementary Note B, which indicates that magnetism in LaNiO3 is a bulk property and does not originate from a tiny (insulating) impurity phase. This small magnetic moment might explain why paramagnetic properties have been reported for LaNiO3 in the past17.The magnetic origin of these quarter-integer peaks in LaNiO3 by means of inelastic neutron scattering, thereby also providing further support that the antiferromagnetism is a bulk property. As can be seen in Fig. B in LaNiO3.We also have been able to study and observe the magnetic excitations in LaNiO3 appears to be a rare case of an antiferromagnetic and metallic transition metal oxide with a fully three-dimensional (3D) crystal and electronic structure. Other systems like 3Mn2O7 and Ca3Ru2O7 have a lower electronic and structural dimensionality where the antiferromagnetic order is resulting from a stacking of ferromagnetic layers26. So far, the chromate system ACrO3  and very recently, RuO2 are known to be the only other intrinsically antiferromagnetic and metallic transition metal oxides with such a fully 3D crystal and electronic structure28. The systems CaCrO3 and LaNiO3 have in common that the oxidation state of the transition metal ion is very high. Thus, the oxygen 2p to transition metal 3d charge-transfer energy here is apparently negative29 resulting in extreme 2p-3d covalency where the presence of holes in the oxygen band can effectively prevent the opening of the conductivity gap and at the same time mediate strongly the magnetic exchange interactions. However, in contrast to CaCrO327, the rare earth nickelate LaNiO3 in single crystalline form is much more metallic and shows conductivities in the μΩ cm range. This is probably also true for RuO228. Unique for LaNiO3 is that it is close to the insulating phase of RNiO3  indicating the importance of strong correlation effects, making it rather exceptional among all transition metal oxides.LaNiORNiO3 with small rare earth ions R, cannot be observed in our present X-ray diffraction (XRD) and neutron-diffraction measurements—see Fig. 30. Nevertheless, with the charge order effects being so weak, one could infer that LaNiO3 is perhaps better described using Fermi surface arguments28, while the other insulating nickelates RNiO3 with the smaller R-ionic sizes and with higher charge ordering temperatures can be more intuitively understood in terms of local charge or bond disproportionations12. Note, that recent ab initio calculations31 reproduced metallic and antiferromagnetic state of LaNiO3, and also predicted charge (or rather bond) disproportionation, whose magnitude is however below our experimental detection limit. One needs special dedicated experiments to probe for the eventual symmetry lowering with the appearance of inequivalent Ni ions. However, our main conclusion—the existence of antiferromagnetic ordering in highly metallic single crystals of LaNiO3, is confirmed by these calculations. According to our findings, we now also present a tentative RNiO3 phase diagram in Fig. Although charge order, which plays a very important role in nickelates 3 single crystal was grown under high oxygen pressures of 130–150 bar with a growth speed of 6–7 mm per h in a mirror furnace from Scidre that was equipped with a 5000 W Xe lamp and with counter-rotation of feeding and seeding rods—see Supplementary Movie 2O3 and NiO into rods with roughly 10 cm length and 6 mm diameter. These rods were sintered at 800–1000 °C for several days. The temperature of the melting zone was measured in situ by means of a pyrometer and is close to 1800 °C at ~140 bar The LaNiOLaue diffraction measurements were performed on a multiwire real-time back-reflection Laue camera from Multiwire Laboratories; see also Supplementary Note α1 radiation. A closed cycle helium cryostat (Phenix of Oxford Cryosystems) was used for temperature-dependent measurements.Powder X-ray diffraction measurements were performed on a Bruker D8 Discover A25 diffractometer, which is equipped with a Johansson monochromator for Cu K3 using a Bruker D8 VENTURE single crystal X-ray diffractometer equipped with a bent graphite monochromator for Mo Kα radiation (about 3× intensity enhancement) and a Photon CMOS large area detector. A crystal with roughly 20 μm diameter has been measured and a multi-scan absorption correction has been applied to the data . A total of 12,328 (observed) reflections  have been collected with an internal R-value of 6.70%, a redundancy of 36.8, and with 98.85% coverage up to R- and weighted R-values amount to 2.29% and 5.79%, respectively. The refinement of the Ni and La occupancies yields an almost stoichiometric composition: La0.995(12)Ni1.000(14)O3. The structural parameters are listed in Supplementary Table Single crystal X-ray diffraction measurements have been performed on a twined single crystal of LaNiO3: La1.001(11)Ni0.999(4)Oδ3+. Moreover, thermogravimetric measurements confirm an almost perfectly stoichiometric oxygen content with δ = −0.002, see Supplementary Note Inductively coupled plasma optical emission spectroscopy measurements yields the following composition of LaNiOX-ray absorption spectroscopy measurements have been performed at the 08B beamline of the National Synchrotron Radiation Research Center (NSRRC), Taiwan.The magnetic properties were studied using a Quantum Design Inc. MPMS-5XL SQUID magnetometer.The measurements of electrical resistivity and specific heat were carried out using a four-probe and a standard thermal relaxation calorimetric method in a Quantum Design Inc. Physical Property Measurement System.kf = 1.8 Å−1). On the IN8 spectrometer, very first quick inelastic scans have been made using a Si monochromator and a Flatcone analyzer array. Polarized neutron measurements have been performed at the IN12 Spectrometer at the ILL in Grenoble, France. A velocity selector was used for choosing the incident neutron wavevector ki of 2.25 Å−1. A horizontally and vertically focussing PG (002) monochromator and Heusler (111) analyzer have been used. The incident neutron beam was polarized by a transmission polarizer (cavity) in the neutron guide and the measured flipping ratio amounts to 22.2.Unpolarized neutron measurements have been performed at the IN8 and Thales Spectrometers at the ILL in Grenoble, France. For the elastic and inelastic measurements on the Thales spectrometer, a pyrolytic graphite (PG) (002) monochromator and analyzer as well as a velocity selector were used (The data that support the findings of this study are available from the corresponding author upon reasonable request.Supplementary InformationDescription of Additional Supplementary FilesSupplementary Movie 1


The rate and the mechanism of the Cu(II)—Aβ complexes formation determines the aggregation pathway of the Aβ species, starting from smaller but more cytotoxic oligomers and ending up in large Aβ plaques, being the main hallmark of the AD. In our study we exploit the existing knowledge on the Cu(II)—Aβ interactions and create the theoretical model of the initial phase of the copper- driven Aβ aggregation mechanism. The model is based on the direct solution of the Chemical Master Equations, which capture the inherent stochastics of the considered system. In our work we argue that due to a strong Cu(II) affinity to Aβ and temporal accessibility of the Cu(II) ions during normal synaptic activity the aggregation driven by Cu(II) dominates the pure Aβ aggregation. We also demonstrate the dependence of the formation of different Cu(II)—Aβ complexes on the concentrations of reagents and the synaptic activity. Our findings correspond to recent experimental results and give a sound hypothesis on the AD development mechanisms.The interactions between the Aβ


A new solvothermal approach is introduced to synthesize ultrathin Sb2Se3 nanowires with diameters ranging from 10 to 20 nm and with length up to 30 μm. The Sb2Se3 nanowire‐based photodetectors are firstly fabricated on polyethylene terephthalate and printing paper substrates, which exhibit excellent response to visible light with fast response time (0.18 and 0.22 s), high flexibility, and durability. The Sb2Se3 nanowire‐based photodetectors are firstly fabricated on polyethylene terephthalate (PET) and ordinary printing paper substrates, which exhibit excellent response to visible light with fast response time (0.18 and 0.22 s), high flexibility and durability.A new solvothermal approach is introduced to synthesize ultrathin Sb1D semiconductor nanowires have been an exciting research field due to their potential application in artificial photosynthesis, optical wavelength filters, photovoltaic technology, electronic and optoelectronic devices.2Se3) is a typical narrow band semiconductor with a good chemical stability and ≈1.21 eV band gap,2Se3 nanorods with diameters ranging from 40 to 70 nm and lengths in the range of 400–1200 nm. Yang et al. obtained the Sb2Se3 nanorods with diameters between 40 and 200 nm via a vapor–liquid–solid approach. Wang et al. developed a novel gas‐induced‐reduction method to obtain the Sb2Se3 nanorods with diameters of ≈200 nm and lengths of 3–5 μm at 200 °C. Ma et al. prepared Sb2Se3 nanowires with diameters of 28–34 nm and length of several hundred nanometers via a solvothermal method.2Se3 ultrathin nanowires with diameters less than 20 nm. In the present work, an alternative organometallic synthetic route is developed for the growth of ultrathin Sb2Se3 nanowires with high quality and ultrahigh aspect ratios from the reaction of triphenylantimony with dibenzyldiselenide in ethanol at 200 °C using oleylamine and polyvinylpyrrolidone (PVP) as surfactants. The diameters of the Sb2Se3 nanowires are mainly ranging from 10 to 20 nm with the length up to 30 μm. Meanwhile, the as‐obtained ultrathin Sb2Se3 nanowires have complementary absorption spectra in ultraviolet, visible and near‐infrared spectrum, which suggests potential application in the photoelectric devices.Antimony selenide (Sb2Se3 single‐nanowire photodetector on SiO2/Si substrate with Ti/Au (10 nm/100 nm) electrodes and the device exhibited an on/off ratio of 15 with rise and decay times of 0.3/0.3 s under 615 nm illumination (1.68 mW cm−2). Choi et al. also fabricated the Sb2Se3 single‐nanowire photodetector on SiO2/Si substrate with Ti/Au (20 nm/100 nm) electrodes and the on/off ratio of the device was found to be 75 under 655 nm illumination (15 mW cm−2). Liu et al. fabricated the Sb2Se3 nanowire photodetector on SiO2/Si substrate with Au electrodes, and the device displayed an on/off ratio of 150 and response/recovery times of 0.2/1.2 s under a xenon lamp (14.4 mW cm−2).2Se3 nanowire photodetectors based on flexile substrates have not been studied up to date. Herein, we report a simple fabrication of flexible photodetectors that are successfully fabricated by depositing Sb2Se3 nanowires on flexible ordinary printing paper in addition to PET substrates. The as‐fabricated photodetector based on PET exhibits an on/off photocurrent ratio of 19 at 23.8 mW cm−2 with fast response and recover time of 0.18/0.20 s. The flexile photodetector fabricated on printing paper displays an on/off photocurrent ratio of 16 at 31.3 mW cm−2 with fast response and recover time of 0.22/0.25 s. Moreover, the two photodetectors exhibit high flexibility and mechanical stability. The measured results imply that the ultrathin Sb2Se3 nanowires are considerably potential for the application of versatile, low‐cost, flexible and portable electronic and optoelectronic devices.Flexible devices have attracted increasing attention due to their potential applications in outfitting robots, medical prostheses, future paper displays, liquid crystal displays and wearable energy harvesting devices.Figure2Se3 nanowires (top) and the corresponding standard Joint Committee on Powder Diffraction Standards (JCPDS) card No. 72‐1184 (bottom), which indicates that the products are highly pure orthorhombic phase with space group of Pbnm. Figure 2Se3 crystal structure displaying a perspective of the (001) facet. It can be observed that the structure of Sb2Se3 consists of infinitely long ribbon‐like [Sb4Se6]n units extending along c‐axis. These 1D ribbons coordinate with each other by intermolecular attraction, creating distinct layers along the [100] and [010] direction.2Se3 nanowires, from which one can see that the length of the nanowires are up to 30 μm. A high‐magnification SEM  demonstrates that the resulted samples in the absence of oleylamine and PVP are composed of many 3D hierarchical architectures assembled by many 1D Sb2Se3 subunits (with diameters of 200–500 nm) and some rod‐like Sb2Se3 nanostructures (with diameters of 500–1000 nm). As shown in Figure S3c,d (Supporting Information), the products synthesized without PVP consist of orthorhombic Sb2Se3 nanorods with diameters of 200–500 nm and Sb2Se3 microtubes with diameters up to 1.5 μm. The obtained products synthesized without oleylamine are mainly Sb2Se3 nanorods with an average diameter of 500 nm . Above results indicate that the ultrathin Sb2Se3 nanowires can be selectively prepared by rationally tuning the temperature with PVP and oleylamine as surfactants. A Fourier transformed infrared (FT‐IR) spectrum of the as‐obtained products  confirms the effects of oleylamine and PVP on the growth of the ultrathin Sb2Se3 nanowires. In addition, it is revealed that the ultrathin Sb2Se3 nanowires exhibit some different optical properties as compared to bulk Sb2Se3 due to the quantum effect .FigureI–V) curves of the device measured under illumination and dark conditions, respectively. The linear curves indicate the good ohmic contacts between the Cu/Au electrodes and the Sb2Se3 nanowires which are favorable for electron transmission. Figure −2 at a voltage of 5.0 V, giving an on/off ratio of 19 (on/off current of 14.76/0.78 nA) at 23.8 mW cm−2. It can be observed that the current increases with increasing light intensity and no typical saturation behavior exists due to the low density of the incident light, which is consistent with the fact that the charge‐carrier photogeneration efficiency is proportional to the absorbed photon flux. The corresponding light‐intensity dependence of the photocurrent is demonstrated in Figure PI ≈ θP, where IP, P, and θ represent the photocurrent, light power intensity, and the response of the photocurrent to light intensity, respectively. Here the measurement results reveal a power dependence of PI ≈ P0.54. According to previous reports, the nonunity exponent may result from the complex process of electron–hole generation, recombination, and trapping within a semiconductor, which will lead to the final saturation of the photocurrent as the light intensity increases.−2) at the voltage of 1.0, 5.0 and 10.0 V , indicating the high stability of the flexile photodetector. It is well known that response and recovery time are critical parameters to evaluate the performance of a photodetector which are defined as the time required for dark current to increase to 90% of the peak value of photocurrent or vice versa. From one period of light excitation as shown in Figure 2Se3 photodetectors and many other flexile nanowire photodetectors.2Se3 nanowires and good ohmic contacts with no interfacial barrier or traps formed between the ultrathin Sb2Se3 nanowires and the Cu/Au electrodes. Moreover, flexible optoelectronics are required to withstand varied mechanical deformations, so the electrical performance of the device at different bending conditions is measured. Figure I–V characteristics of the flexile photodetector at different bending angles or twisting positions, where the positive and negative values are defined according to the folding direction of the device as demonstrated in the inset of Figure FigureI–V characteristics of the devices in the dark and under illumination, indicating good ohmic contacts between the Sb2Se3 nanowires and Cu/Au electrodes. The inset of Figure −2 at 5.0 V. It can be observed that the current increases with increasing light intensity with an on/off ratio of 16 (on/off current of 3.53/0.22 nA) at 31.3 mW cm−2, demonstrating a power dependence of I ≈ P0.44 . As demonstrated in Figure I–V curves in Figure S9 (Supporting Information) ensures the high flexibility and durability of the device. The measurement results demonstrate significant potential of the ultrathin Sb2Se3 nanowires for the development of versatile, low‐cost, flexible, and portable electronic and optoelectronic devices.The photocurrent performances of the photodetector fabricated on an ordinary printing paper are summarized in 2Se3 nanowires with diameters ranging from 10 to 20 nm and length up to 30 μm via the reaction of triphenylantimony with dibenzyldiselenide in ethanol at 200 °C for 16 h using oleylamine and PVP as surfactants. Especially, flexible photodetectors employing ultrathin Sb2Se3 nanowires film have been firstly fabricated on PET and ordinary printing paper substrates, which demonstrate high performance. The excellent flexibility and durability of the photodetectors based on the Sb2Se3 nanowires are desirable for practical application in high‐performance flexile optoelectronic devices with low‐cost, portability and mechanical stability.In summary, an alternative facile organometallic synthetic route is developed for the growth of ultrathin SbMW ≈ 30 000) was dissolved in 10 mL ethanol, respectively. Then, the two kinds of solution were mixed to form a homogenous solution that was transferred into a 25 mL Teflon‐lined stainless steel autoclave and maintained at 200 °C for 16 h. After reaction, the autoclaves were cooled down to room temperature naturally. The resulting black precipitates were washed with ethanol and hexane several times.All reagents and solvents were of analytical purity and they were obtained from commercial sources and used without further purification. In a typical synthesis, 0.3 mmol (105.4 mg) triphenylantimony and 0.2 mmol (68.0 mg) dibenzyldiselenide were dissolved in 5.0 mL oleylamine and 0.2 g of PVP . The nanowire morphologies, structure, and composition were investigated by the field emission scanning electron microscopy (SEM) (JEOL JSM‐6700F), transmission electron microscopy (TEM), and high‐resolution TEM accompanied by energy dispersive X‐ray spectroscopy on JEOL‐2010 with an accelerating voltage of 200 kV. Raman spectra were performed on a LABRAM‐HR spectroscopy with excitation of the 514.5 nm line of an Ar+ ion. X‐ray photoelectron spectroscopic (XPS) spectra were collected using a VGESCA‐LAB MKII X‐ray photoelectron spectrometer fitted with a monochromatic Al Kα X‐ray source (1487 eV) operating on a spot size of 560 μm. UV–vis–NIR absorption spectra were recorded on a Perkin Elmer Lambda 950 UV–vis–NIR spectrophotometer. The Fourier transformed infrared (FTIR) spectrum of the nanowires was detected by a Fourier infrared spectrometer iS10.The as‐prepared products were identified by X‐ray powder diffraction (XRD) patterns on a Philips X′Pert Pro Super diffractometer with graphite‐monochromatized Cu Kα radiation  and ordinary printing paper, respectively. In detail, 30.0 mg of the as‐prepared Sb2Se3 ultrathin nanowires was dispersed in 2.0 mL ethanol to make a homogeneous solution. The solution was drop‐deposited on the top side of a well‐prepared PET and printing paper plate (with size of 2.0 × 1.5 cm) by spin‐casting treatment and dried in air naturally to form an assembly layer film of the Sb2Se3 ultrathin nanowires. Afterward, ohmic contact electrode patterns (1.5 mm × 2.5 mm), Cu (5.0 nm), and Au (40.0 nm) deposition on the film by using a high‐vacuum electron beam system under the assistance of a shadow mask with spacing of 1.0 mm. The current–voltage (I–V) characteristics and the sensitivity of the photodetectors were measured using CHI600E electrochemical workstation, equipped with a xenon lamp of PLS‐SXE300/PLS‐SXE300UV with an output wavelength range of 400–780 nm.Flexible photodetectors based on SbAs 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.