Thus our present study unveils a multifunctional material that can be used in three different fields without further modifications.Increasing environmental pollution, shortage of efficient energy conversion and storage devices and the depletion of fossil fuels have triggered the research community to look for advanced multifunctional materials suitable for different energy-related applications. Herein, we have discussed a novel and facile synthesis mechanism of such a carbon-based nanocomposite along with its energy and environmental applications. In this present work, nitrogen-doped carbon self-assembled into ordered mesoporous structure has been synthesized via an economical and environment-friendly route and its pore generating mechanism depending on the hydrogen bonding interaction has been highlighted. Incorporation of metal oxide nanoparticles in the porous carbon network has significantly improved CO This has led to different global challenges like environmental pollution, greenhouse effect, and energy shortage. In this regard, there is an urgent need for materials that can capture this greenhouse gas and can be used in energy-related applications, all in one. Conventional amine scrubbing process for capturing CO2 has several disadvantages like high regeneration cost, extensive corrosion, and loss of amines during their generation process1. So novel porous materials having high CO2 adsorption capacity attracted considerable attention in recent times mainly because of their low cost, ease of synthesis, wide availability, good chemical stability, and high specific surface area3. As an alternative of fossil fuel and to mitigate the increasing CO2 concentration in the atmosphere, renewable alternative energy storage and conversion technologies like battery and fuel cell have also been thought of as a part of this work. Rechargeable lithium-ion batteries (LIBs) are the most efficient and convenient electrochemical storage devices for portable as well as stationary applications. Along with the lithium storage capacity, an ideal anode material for LIB should possess high electrical conductivity and low volume expansion coefficient. On the other hand, Oxygen Reduction Reaction (ORR) is the most crucial cathode reaction in fuel cell and metal-air batteries to determine the efficiency and longevity of the device. To improve the sluggish reaction kinetics, platinum (Pt) and platinum-based alloy catalysts have been reported to show the best electrocatalytic activity. But, the high cost, less availability and poor durability of Pt and Pt-based alloys have led to the search for an alternative non-platinum based catalyst4.The increasing consumption of fossil fuels has largely enhanced the accumulation of CO2 gets anchored easily by Lewis acid (CO2) – Lewis base (N) interaction. The presence of basic nitrogen groups increase the interaction between the large quadrupole moment of CO2 molecules and the polar nitrogen sites5. The N-doped sites also help in adsorption of the oxygen molecule and act as active catalytic sites towards ORR6. Porous carbon materials with high specific surface area, tunable pore size and large pore volume have played a very vital role in all the above-mentioned applications.Nitrogen doping also enhances the gas adsorption capacity by increasing the basic sites in the sample where CO2 capture at moderate temperature due to their low cost and wide availability. Many experimental studies on high CO2 capture capability of metal oxides have already been reported. These metal oxides act as heterogeneous catalysts and so the interaction between CO2 and the oxide surfaces is of great interest8. Physicochemical adsorption of CO2 on iron oxide decorated graphene nanocomposite has already been reported by A. K. Mishra et al.9. Tamilarasan et al. had worked with polyaniline-magnetite nanocapsules showing high CO2 capture capability as well as fast sorption kinetics of the nanocomposite10. In the case of LIBs, the specific capacity of any graphitic carbon is limited by the theoretical capacity of these materials. So, incorporation of any metal/metal oxides, that can take part in conversion type redox reaction, improves the specific capacity considerably12. In this regard, combining the two storage mechanisms (i.e. insertion/de-insertion and conversion) using a single material can be found as an efficient way to enhance the storage capacity of LIBs. Fe3O4 incorporated nitrogen doped porous carbon has been extensively studied as an efficient anode material for LIBs16. Iron-oxide nanoparticles were chosen to be a superior catalyst owing to their low cost, high stability and intrinsic catalytic activity towards ORR in alkaline medium.Metal oxides, as well as hydroxides, serve as a good candidate for CO2 capture, lithium-ion battery and ORR catalyst in alkaline fuel cell. It has been investigated that this nanocomposite has the potential to serve as an effective CO2 adsorbent, anode material for lithium-ion battery and efficient ORR catalyst for fuel cell operations because of its interconnected porous network, high nitrogen doping and presence of metal oxide nanoparticles (Fe3O4) on its surface. In the case of lithium-ion battery, dynamic electrochemical spectroscopy (DEIS) measurement has been carried out in order to estimate the actual potential window of the significant electrochemical reactions involved in it. In order to address the burning environmental challenges associated with increasing concentration of greenhouse gases and a shortage of renewable energy storage and conversion devices, our approach is the first of its kind where a single environment-friendly composite can be used in three above-mentioned fields.In this present work, we have synthesized metal oxide-porous carbon composite by simple self-assembly and coprecipitation techniques. The pore-foaming agent used in forming the porous carbon framework is sodium hydrogen carbonate that is nontoxic and cheap. Briefly, the pore formation mechanism based on hydrogen bonding interactions has also been explained here. Further, this particular nanocomposite has been explored for CO17. Kim et al. explained in details the formation pathway of magnetite nanoparticles by coprecipitation method. In typical coprecipitation technique, as the ammonia solution is added to the iron salt solution, interfacial contact forms in between the iron-rich solution (pH ~ 1.5) and the base (pH ~ 11). So diffusion occurs in both ways: the base tries to diffuse into the iron-rich solution while iron ions try to diffuse in the opposite direction. So precipitation occurs at the interface. The authors have mentioned that during the reaction mechanism many intermediate products are formed. As the two solutions begin to mix, two types of nucleation process will be initiated simultaneously. On one hand, the low pH iron side increases the pH by reacting with ammonia solution which basically reacts with the less stable Fe3+ ions to form the following intermediates: akaganeite → goethite → (hematite → maghemite) → magnetite. On the other hand, ammonia reacts with both Fe2+ as well as Fe3+ to form ferrous hydroxide → lepidocrocite → (maghemite) → magnetite17. This briefly explains the formation pathway of magnetite nanoparticles by coprecipitation method. This particular method explains a wide particle size distribution of the synthesized nanoparticles. The coprecipitation method is a very eco-friendly method for producing magnetite nanoparticles. It neither produces any toxic intermediates nor requires any complex precursors. However, controlling the particle size distribution is difficult in this process because the growth of the crystal depends only on kinetic factors17. The detailed synthesis procedure has been shown in Fig. The coprecipitation technique, used to synthesize iron oxide, is a simple and efficient chemical pathway where aging of a stoichiometric mixture of ferrous and ferric salts in aqueous medium takes place3−)2 anions units (highlighted by red circle) originating from the bicarbonate salt precursor via N–H∙∙∙O bond formation (Step 2 and 3) to form an anionic host lattice with the interconnected open network (Step 4)18. Sodium bicarbonate promotes self-assembly among the precursors before pyrolysis that is beneficial for the synthesis of porous carbon19. It is well reported that glucose can be transformed into 5-hydroxy-methylfurfural (5-HMF) under thermal decomposition20. Thus introducing glucose in this anionic host lattice framework results in the formation of N–H∙∙∙O and C–H∙∙∙N hydrogen bonding in between 5-HMF and the host (highlighted by the red circle) (Step 5). Finally, during pyrolysis, melamine and sodium bicarbonate decomposed and 5-HMF dehydrated to form the porous network. Evolution of water molecules from 5-HMF and carbon dioxide from the decomposition of sodium bicarbonate has led to the formation of the interconnected mesopores in the host lattice.The pore formation mechanism in NPC is explained in Fig. 3O4/NPC  which disappears after annealing at 700 °C. Figure 3O4 phase having a cubic spinel structure21. Thermogravimetric Analysis (TGA) of Fe3O4/NPC was carried out in air atmosphere from room temperature up to 800 °C at a heating rate of 20 °C min−1 , the curves show a high N2 uptake suggesting that some microporous nature is also present in the samples. The nitrogen uptake then becomes moderate at relatively intermediate pressures. As can be seen in Table 2 g−1 and a total pore volume of 1.01 cm3 g−1. Unlike N doped porous carbon (NPC), the surface area as well as the pore volume decreased but the average mesopore size slightly increased indicating a widening of the mesopores and decline of mesoporous ordering with the incorporation of Fe3O4 nanoparticles in the porous carbon framework22. The increase in pore size most likely might be due to the partial penetration of the nanoparticles leading to an expansion in the nanostructure. It might also be due to the enhanced rigidity caused by the Fe3O4 nanoparticles to decrease the pore shrinkage23. The increase in the size of the pores can be identified in the SEM and TEM images as well as shown in Fig. Gas adsorption as well as battery and fuel cell performance largely depend on the specific surface area and the pore size distribution of the nanocomposite. Fig. 3O4/NPC respectively. SEM image of the mesoporous carbon support material (NPC) shows the presence of innumerable pores in the carbon framework. The decoration of this porous framework with Fe3O4 nanoparticles can be seen in Fig. 3O4 nanoparticles decorated porous carbon is shown in Fig. 3O4/NPC shown in the inset. It suggests almost uniform distribution of Fe3O4 nanoparticles on the surface of the porous carbon framework. The size of the pores in Fe3O4/NPC is seen to be slightly larger than those of NPC. This is in confirmation with the results obtained from BET analysis.The morphological studies of the prepared nanocomposite has been carried out using electron microscopy techniques. Fig. 2 C=C, carbon-nitrogen complex C–N (sp3) and oxygenated carbon complex (COx) respectively. N 1 s peak is deconvoluted into four components with binding energies 398.0 eV, 400.21 eV, 402.24 eV and 404.17 eV . The other peak at 710.6 eV signifies the coexistence of Fe3+ as well as Fe2+ of Fe (2p3/2)28. Other two peaks with binding energies 725.2 eV and 713.1 eV are allocated to the Fe3+ oxidation state of Fe (2p1/2) and Fe (2p3/2)29. The XPS spectra of both NPC and Fe3O4 before and after coupling are presented in Fig. The high resolution XPS spectrum of C1s peak Fig.  is decon eV Fig. . The firnts Fig.  with bintra Fig.  is decon2 adsorption studies have been carried on Fe3O4 decorated N doped porous carbon using Sievert’s apparatus using Pressure Swing adsorption technique. Fe3O4 nanoparticles are basic in nature and therefore Fe3O4/NPC consists of a large number of basic functional sites that attract acidic CO2 towards it and enhances adsorption. The nitrogen functionalities also play a significant role in the enhanced adsorptive behavior of the nanocomposite. Fe3O4/NPC has shown a high CO2 adsorption capacity of 40.5 mmol g−1 at 25 °C and at 20 bar equilibrium pressure in comparison to NPC which has shown a capacity of 17.2 mmol g−1 under the same conditions of temperature and pressure  and Fe3O4 (0.027 mmol g−1) at 25 °C and 1 bar pressure, Fe3O4/NPC (4.25 mmol g−1) has shown a comparatively high adsorption capacity that might be due to the synergistic effect of Fe3O4 as well as the porous carbon support. Fe3O4/NPC shows high isosteric heat of adsorption values of about 38 kJ mol−1. The adsorption energies have been found to be much greater than 17.2 kJ mol−1, the enthalpy of liquefaction of CO2. These results imply that Fe3O4 particles enhance CO2 affinity and interaction strength mainly in the range of physisorption  at a scan rate of 0.1 mV s−1. In Fig. +3/Fe+2 to Fe° which is in good agreement with literature39. The corresponding reactions are given as follows:In order to have insight knowledge about the redox reactions occurring during charge-discharge, cyclic voltammetry (CV) was carried out in the potential range of 0.01 V–3.0 V  arising from lithium extraction-insertions respectively. During the anodic sweep, two prominent peaks at 1.5 V and 1.9 V signify corresponding two-step oxidation process of Fe° to Fe+3/Fe+2 respectively37. Along with all the above-mentioned peaks, a pair of submerged redox peaks at 0.05 V and 0.21 V corresponds to lithium intercalation and de-intercalation in the porous carbon framework. The EIS spectra of the same anodic half-cell have been shown in Fig. The peak around 0.8 V can be assigned to the electrolyte decomposition at the electrode surface and subsequent formation of solid electrolyte interface (SEI) layer. The peak has been found to disappear in 240. Two sets of spectra have been recorded during charging and discharging process in the potential window of 0.01 V to 3.0 V at a regular potential interval of 0.1 V. During discharge,  and the associated charge transfer process. After 1.5 V the increase in the radius of semicircles describes the associated impedance due to the formation of the SEI layer, which makes the insertion of Li+ ion difficult. In the potential window of 0.01 V to 1.0 V, the diffusion tails of the spectra exhibit minimum impedance.In order to elucidate the dynamic variation of the impedance during charging and discharging, dynamical electrochemical impedance spectroscopy (DEIS) has been carried out with the assembled anodic half-cell. In the DEIS technique, the variable frequency response of an AC signal is superimposed with a DC voltage in the same potential scan of the anodic half-cell. The superimposed DC potential on the corresponding AC amplitude holds the electrochemical state of the cell to a complete stationary state, which is advantageous over EIS. In order to have an in-depth knowledge about the electrode reaction as a function of the state of charge (SOC), this technique is beneficial over EIS because it does not hold the potential at the requisite potential point leading to a non-stationary statege, Fig.  the decr+ ion de-insertion process due to the formation of the thick SEI layer. The same behavior can be interpreted from the diffusion tails of the impedance spectra above 2.0 V as well. The impedance associated with the Li+ ion diffusion in the porous network gets hindered because of the formation of the SEI layer.During charging Fig.  the semi3O4/NPC was investigated through cyclic voltammetry with a scan rate of 50 mV s−1 by saturating the 0.1 M KOH electrolyte with nitrogen and oxygen gas. Figure 3O4/NPC in nitrogen and oxygen saturated electrolyte. The cyclic voltammogram in the N2 saturated electrolyte exhibits double layer capacitance, while, in the oxygen-saturated electrolyte, a prominent oxygen reduction peak can be observed at −0.15 V.The electrocatalytic activity of Fe−1 in oxygen saturated 0.1 M KOH electrolyte at different rotation rates varying from 400 rpm to 1600 rpm as shown in Fig. 41. The onset potential and half-wave potential of Fe3O4/NPC at 1600 rpm are −0.03 V and −0.1 V respectively and the mass activity at −0.1 V is 35.8 mA mg−1. More the positive onset and half-wave potentials, higher is the catalytic activity. This is mainly due to the high specific surface area and mesoporous nature of NPC that enhances the density of active sites and mass transport of the ions, electrons, oxygen and water molecules through the catalyst42. It is well reported that pyridinic N present in the carbon network consists of a lone pair of electrons. Conjugation of pyridinic N lone pair of electrons with the graphene π system makes easy adsorption of O2 molecules on these sites. On the other hand, graphitic N in the NPC network facilitates the electron transfer from the carbon host to the oxygen anti-bonding orbitals44. Yousung Jung et al. have reported that graphitic N becomes pyridinic N by opening the cyclic C-N ring after electron and proton transfer reaction and has shown that both pyridinic N and graphitic N can act as active sites for ORR45. Hence the presence of pyridinic N and graphitic N in NPC network of Fe3O4/NPC enhances the limiting current density and improves the onset potential of ORR46. Herein, we have compared the onset and half-wave potential of Fe3O4/NPC with that of few reported Fe based nitrogen doped porous carbon catalyst  technique was carried out. The linear sweep voltammograms were obtained in the potential window of −0.5 to 0.1 V at a scan rate of 10 mV sst Table  and withst Table . Moreove3O4/NPC shown in Fig. −1 vs ω−1/2) were obtained from polarization curves at various rotation rates as shown in Fig. 47. The ORR in alkaline medium takes place in two ways: i) two-electron pathway which leads to the formation of peroxide ion (HO2−) as an intermediate  by applying suitable ring potential. The number of electrons transferred (n) per oxygen molecule reduction and the percentage of peroxide ion formation is calculated from following equations . The calculated values of n and the percentage of HO2− formed at different potentials varying from −0.5 V to −0.2 V is shown in Fig. 2− varies from 5% to 1.5%, which indicates that the intermediate product formed is very less and mostly the reaction takes place in the four-electron pathway. Feng and Mullen et al. have reported the dependence of the peroxide ion yield on different loading of Fe3O4 for Fe3O4/N-GAs. They have shown that by increasing the Fe3O4 loading from 4.1 to 46.2 wt %, the yield of HO2− decreased and with around 20 wt% of Fe3O4 they got approximately 48% of peroxide49. Whereas, in the present work, Fe3O4/NPC with 20 wt % of Fe3O4 loading showed 12 folds lesser HO2− yield. The low yield of HO2−, the improvement in the onset and half-wave potential and the four-electron pathway of ORR for Fe3O4/NPC is due to the presence of pyridinic N and graphitic N sites50. The number of electrons transferred (n) per oxygen molecule reduction and the percentage of peroxide ion formation for NPC catalyst is shown in Fig. In order to confirm the presence of peroxide intermediate, rotating ring-disk electrode (RRDE) Fig.  measuremons Eqs. :6\docume3O4/NPC was tested through chronoamperometry technique at −0.25 V  in O2 saturated 0.1 M KOH electrolyte at 1600 rpm as shown in Fig. 3O4/NPC shows good stability up to 20,000 sec with 91% of current retention. The better durability of Fe3O4/NPC is because of the active sites in Fe3O4/NPC that can promote the ORR with less overpotential and the mesopores that can facilitate the mass transport of ions. So the synergistic effect of Fe3O4 nanoparticles along with the highly porous nitrogen-doped carbon can show excellent catalytic activity towards ORR.The durability of Fe2 capture as well as for ORR catalyst and lithium-ion storage simultaneously. The as-prepared nanocomposite has shown an excellent CO2 uptake capacity (40.5 mmol g−1 at 25 °C and 20 bar pressure), a maximum reversible specific capacity of 930 mA h g−1 at 100 mA g−1 current density as an anode material in lithium-ion battery and superior catalytic activity as Pt-free catalyst in alkaline medium with 12 fold decrement in peroxide yield. Largely, we can conclude that nitrogen doped porous carbon with metal oxide nanoparticles decorated on them can be considered as a versatile nanocomposite that has promising applications in gas adsorption technology as well as in energy storage and conversion devices.In summary, we have synthesized an environment friendly nitrogen doped porous carbon-metal oxide composite with a high specific surface area. The mechanism of the pore formation along with the coprecipitation method to synthesize metal oxide nanoparticles has been discussed. To the best of our knowledge, there are no such reports available on such a multifunctional material being used for CO3 acts as a porogen for creating porosity in the carbon framework. The furnace was allowed to cool down to room temperature and the sample was collected. The sample, thus obtained, was washed profusely with water and ethanol to remove any unreacted chemicals and sodium content in the sample. Then it was allowed to dry at 60 °C overnight and the final sample is named as NPC.Nitrogen-doped mesoporous carbon was synthesized using a simple thermal decomposition technique. At first, glucose , sodium hydrogen carbonate  and melamine  were mixed thoroughly in 1:1:1 ratio and carbonized at 800 °C for 2 h in a tubular furnace with a heating rate of 5 °C/min. Melamine acts as the nitrogen source and NaHCO3O4 nanoparticles were synthesized using coprecipitation technique. 200 mg of porous carbon (NPC), thus obtained, was dispersed in deionized water for 6 h. Then two types of Fe salts were slowly added to this solution and allowed to stir overnight. FeSO4.7H2O  and FeCl3.6H2O  were taken in the stoichiometric ratio of 2:3 and added. Then the solution was heated up to 90 °C. Ammonia solution (25%) (RANKEM) was added drop-wise to the above solution until the pH became 9, which basically act as the reducing agent. The solution was stirred for another 30 min at 90 °C and then allowed to cool to room temperature. The black precipitate, thereby obtained, was washed repeatedly with DI water until it became neutral, filtered and dried. The obtained sample was again annealed at 700 °C for 1 h in an inert atmosphere to obtain pure Fe3O4 phase in the sample52  at 40 kV and 100 mA. Measurements were carried out from 10° to 80° with a step size of 0.02°. The specific surface area, total pore volume, and pore size analysis were done via nitrogen adsorption/desorption isotherms at liquid nitrogen temperature of 77 K using Micromeritics ASAP 2020 surface area and porosity analyzer using Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halanda (BJH) theories respectively. X-ray photoelectron spectroscopy was carried out using Specs X-ray photoelectron spectrometer with Mg Kα as the X-ray source and PHOIBOS 100MCD analyzer to find out the chemical composition of the samples. All the peaks are fitted using an iterated Shirley background. The line-shape used in every fit is a Doniach Sunjic form convoluted with a Gaussian/Lorentzian shape. Thermogravimetric analysis was carried on using SDT Q 600 from TA Instruments. The sample was heated from room temperature to 1000 °C in air atmosphere at a flow rate of 150 ml min−1 in order to find out the weight percent of Fe3O4 in the composite.The morphologies of the as-prepared samples were studied using Inspect F50 Scanning Electron Microscope from FEI and Technai G20 Transmission Electron Microscope. The elemental analyses were also determined using Inspect F50 instrument. The crystalline nature of the samples was characterized by Rigaku Smartlab X-Ray diffractometer with nickel-filtered Cu K−6 mbar) for 3 h to remove moisture content and any dissolved impurities from the sample surface and to regenerate the adsorptive sites. 200 mg of sample was loaded in a quartz tube for each measurement. For CO2 adsorption studies, the temperature was varied from 25 °C to 100 °C and pressure was varied from 2 bar to 20 bar. A number of cycles were repeated to check the adsorption capacity and the results were found to be consistent.The adsorption isotherm studies were carried out for carbon dioxide gas at high pressure and temperature using volumetric analysis method in Sievert’s apparatus. Before each measurement, the samples were degassed at 250 °C in presence of high vacuum  with polyvinylidene fluoride (PVDF) as binder (15%) and conductive carbon (10%) thoroughly in N-methyl-2-pyrrolidone (NMP) solvent. This slurry was uniformly coated over copper foil (with a thickness of 0.009 mm) using doctor blade method. Post-coating, the foil was dried at 120 °C for 8 hours in a vacuum oven. The electrodes with 12 mm diameter were cut from the coated foil. 2032 coin cells were assembled in an argon-filled glove box  using lithium foil as reference and counter electrode. Celgard 2400 microfiber dipped in electrolyte solution made of 1 (M) lithium hexafluorophosphate (LiPF6) dissolved in ethylene carbonate (EC) and dimethyl carbonate (DEC) (1:1 v/v) served as the separator. A Biologic SP-300 electrochemical workstation was used to estimate the electrochemical performances. The galvanostatic charge/discharge had been carried out in the potential range of 0.01 V to 3.0 V with current densities in the range of 100 mA h g−1 to 2 A h g−1. All impedance spectra were taken in the frequency range of 1 MHz to 10 mHz. All the measurements were carried out at room temperature.The electrode slurry for battery operation was prepared by mixing as synthesized FeCatalytic activity towards ORR was investigated using cyclic voltammetry data. The electrochemical measurements were taken using three-electrode system with Biologic SP-300 workstation and Rotating Electrode Speed Controller (Pine Research Instrumentation) was used to control the rotation rate of working electrode during RDE and RRDE measurements. A platinum wire was used as a counter electrode and Ag/AgCl dipped in 1(M) KCl was used as the reference electrode. A glassy carbon (GC) electrode  modified with electrocatalyst slurry served as the working electrode. The electrocatalyst slurry was prepared by taking the calculated amount of catalyst dispersed in DI water and Nafion solution (5 wt %). Then small amount slurry was drop-casted on the glassy carbon electrode (GCE). The electrolyte used for this measurement was 0.1 M KOH solution. Prior to use, the GCE surface was rubbed with alumina paste of particle size 1 μm, 0.3 μm, and 0.05 μm and the electrolyte was saturated with oxygen.supp info


In the article titled “Evaluation of Complication Rates after Breast Surgery Using Acellular Dermal Matrix: Median Follow-Up of Three Years” , there w(1) The title should be changed to “Evaluation of complication rates after breast surgery using two different kinds of Acellular Dermal Matrix and one Bovine Pericardium Collagen Membrane: median follow-up of three years.” (2) Tables 1, 2, and 4 along with their legends should be corrected as below.(3) In the “Material and Methods,” the sentence reading “Sixteen patients had a history of radiotherapy prior to ADM implementation; see Table 3” should be changed to “Sixteen patients had a history of radiotherapy prior to ADM implementation; see Table 2.”(4) In the “Discussion,” the sentence reading “Other limitations of our study include the relative brevity of follow-up with a mean time period of 36 months” should be changed to “Other limitations of our study include the relative brevity of follow-up with a median time period of 36 months.”(5) The legends of Figures 3 and 4 should be corrected where “capsular contracture and” should be added to the legend of Figure 3 and “and capsular contracture, followed by” should be added to the legend of Figure 4. The corrected figures are as below.


In this present study, the process proposed here was further improved by optimizing the relevant factors using the response surface methodology based on Box–Benkhen Design. The optimum condition that afforded the highest yield (98%) of 2‐methyl‐3‐n‐butylaminoyl‐1,4‐benzoquinone was obtained as follows: molar ratio of amines to hydroquinones 1.16:1, activity ratio of laccase to lipase 1.14:2, and reaction temperature 38.9°C. The results obtained indicate that this process may be useful as a green alternative method for higher yield production of mitomycin analogs.To reach the excellent yield as well as environmental friendliness, an efficient one‐pot process for the synthesis of 2‐methyl‐3‐ ANOVAanalysis of varianceBBDBox Behnken designMMCmitomycin CRSMresponse surface methodology1Mitomycin C (MMC) is one of the most important members of mitomycins family that has been widely used as strong antineoplastic antibiotic for treating various tumors associated with gastric, pancreatic, colorectum, mammary, lung, etc. Trametes species and Myceliophthora thermophila can catalyze the synthesis of aminoquinones by the nuclear amination of p‐hydroquinones with primary aromatic amines Trametesvillosa was successfully used to mediate the oxidation and cross‐coupling of two para‐dihydroxylated benzoic acid derivatives with 4‐aminobenzoic acid to synthesize aminobenzoquinones derivatives M. thermophila laccase (MtL) and P. cinnabarinus laccase (PcL), respectively. Whereas this method gives a lower product yield that is less than 35% after 6 h of reaction. Moreover, the product formation in above mentioned laccase‐catalyzed reactions was influenced by many factors including the source of laccase, the electrophilicity of the hydroquinones, the nucleophilicity of the amines, ratio of amines to hydroquinones, temperature, and solubility, etc. Enzymatic domino reactions have been paid considerable attention in the past decade, which offers two distinct advantages including good yield of the final product and an asymmetric fashion for easily obtaining nonracemic products both as a result of the highly chemo‐, regio‐ or stereoselectivity of enzyme catalysts n‐butylaminoyl‐1,4‐benzo‐quinone, using 2‐methyl‐1,4‐hydroquinone and n‐butylamine as substrates co‐catalyzed by Pleurotus ostreatus laccase and Trichosporon laibachii lipase in a one‐pot process. The general reaction for the laccase/lipase catalyzed synthesis of 2‐methyl‐3‐n‐butylaminoyl‐1,4‐benzoquinone is shown in Scheme n‐butylaminoyl‐1, 4‐benzoquinone) by RSM analysis were obtained.In this context, it is appropriate to mention here that the optimized conditions for enhancing the ability of laccase/lipase in catalyzing high yield production of aminoquinone derivatives by domino reaction has not been reported in literature so far. Herein, we investigated the feasibility of a cascade enzymatic synthesis of aminoquinone derivative, 2‐methyl‐3‐n‐butylaminoyl‐1, 4‐benzoquinone, a mitomycin‐like compound using laccase/lipase as co‐catalysts. Under the optimum condition, the highest yield of 98% was achieved. This process may be useful as a green alternative method for higher yield production of mitomycin analogs.Mitomycin C is one of the most important members of mitomycins family with extensive application in the treatment of various tumors associated with gastric, pancreatic, colorectum, mammary, lung, etc. Chemical methods for synthesis of Mitomycin C and its analogues still fail to reach the excellent yield as well as environmental friendliness due to the poor reaction selectivity and tedious reaction steps, etc. This paper presents an efficient one‐pot process for the synthesis of 2‐methyl‐3‐22.1T. laibacchii CBS5791 was used as catalyst in combination with laccase from P. ostreatus for oxidation/amination reaction of 2‐methyl‐1,4‐hydroquinone with n‐butylamine. The immobilized T. laibacchii lipase was prepared using an in situ immobilization method as described in our previous work in which an aqueous two‐phase system with 12% PEG 4000/13% K2HPO4 was used to purify the lipase, diatomites was used as a supporter, and glutaraldehyde was used as a cross‐linker P. ostreatus was purified and immobilized on glass beads by using the methods reported by Macellaro et al. P. ostreatus laccase was prepared in 50 mM NaP buffer, pH 6.5 at room temperature for 1 h. 2‐Methyl‐1,4‐hydroquinone  and n‐butylamine  were both purchased from Shanghai Aladdin Biochemical Technology and used as received without further purification.An immobilized lipase from 2.2n‐butylaminoyl‐1,4‐benzoquinone (c) was synthesized via the oxidation reaction of 2‐methyl‐1,4‐hydroquinone (a) followed by amination of the formed intermediate 2‐methyl‐1,4‐benzoquinone with n‐butylamine (b) in a 20‐mL reaction vessel. The reactions were performed using the immobilized T. laibacchii lipase in conjugation with immobilized P. ostreatus laccase pH 7. Methanol  was used as a co‐solvent to improve the dissolution of the substrates. The reaction mixture (10 mL) contained 25–50 U/mL of each biocatalyst. The reactions were carried out at 40–50°C and shaken at 200 rpm in a water bath shaker for 20–24 h. Samples were withdrawn at specific time intervals from reaction mass and analyzed by HPLC. All reactions were carried out in duplicate.The target product 2‐methyl‐3‐4 and finally evaporated under reduced pressure. Then the concentrated organic layer was applied to a Strata™ SDB‐L column with styrene‐divinylbenzene polymer as absorbent material  for SPE. The reaction product c was eluted with methanol/0.5% aqueous acetic acid . The by‐products and residual substrate were eluted with methanol/deionized water . The pure methanol was used to elute the diaminated products d. The methanol of samples was removed by rotary evaporation at 30°C.The immobilized lipase was removed by filtration and the supernatant solution was extracted three times with ethyl acetate. The ethyl acetate layer was concentrated by being dried over MgSO2.3n‐butylaminoyl‐1,4‐benzoquinone (c) and 2‐methyl‐3,5‐n‐butylaminoyls‐1,4‐benzoquinone (d) were 4.13 and 4.74 min, respectively.The samples were withdrawn at various intervals and were analyzed with an Agilent 1200 series device equipped with a G1312BB inpump SL pump, G1367LHip‐ALS SL auto sampler, and a G1315A DAD detector . The column employed was an Agilent ZORBAX Eclipse XDB‐C18 column . The mobile phase consisted of a gradient of 60–100% methanol in water within 12 min at a flow rate of 0.5 mL/min and the sample injection volume was 5 µL. The column was maintained at 30°C and the detection wave length was 227 nm. The retention times of 2‐methyl‐3‐2.4c was chosen as response variable. Table c by lipase/laccaseco‐catalyzed were investigated based on a three‐level, three‐variable Box–Behnken design by using the Design Expert software . The experiment design and the yield results are presented in Table c.Y is the predicted response; A0 is the constant; and iA, iiA, and ijA are the regression coefficients estimated by the multiple regressions for the linear, quadratic, and cross‐product effects of the factors, respectively; iX and jX are the independent variables. The coefficient of determination (R2) and the adjusted coefficient of determination (R2Adj) were used to check the fit of regression model. A regression analysis and the plotting of response surface were performed to fit quadratic polynomial equations for all response variables and obtain the optimum conditions. Significance of data was tested using the analysis of variance.Three factors including the mole ratio of amines to hydroquinones (MR), activity ratio of laccase to lipase (AR), and reaction temperature (T) were selected as independent variables and the yield of product 33.1X1), activity ratio of laccase to lipase (X2), and reaction temperature (X3) on the yield of product c (Y). The terms found to be significant were combined into the following fitted quadratic polynomial equation Y=85n‐butylamine co‐catalyzed by lipase/laccase in producing 2‐methyl‐3‐n‐butylaminoyl‐1,4‐benzoquinone is appropriately described with Equation X1 and X2 as well as X2 andX3 carried positive coefficients. That indicated these factors have a direct positive effect on the yield of product c. In contrast, all the quadratic terms  and the interaction terms (X1X3) had a negative effect on the yield of product c.The oxidation/amination reaction of 2‐methyl‐1, 4‐hydroquinone with R2 of 0.9941 indicates that 99.41% of the total sample variation for the yield of product c can be explained by the model. The present R2 value reflects that the predicted values obtained by model fitting sufficiently correlate with the observed values. The adjusted R2 value that is higher than 0.98 also indicates a very good fit between the observed and predicted responses. Larger the F‐value is, more significant the corresponding coefficient would be, by contrast, a very small p‐value (<0.0001) would indicate the significance of the corresponding coefficient. In this study, the F‐ratio of quadratic regression model was 94.05 with a low probability value (p < 0.0001), which means the model is well accurate for predicting the yield of product c. It can be observed from analysis of variance (Table X1X3 and X2X3) had significant impact on the yield of product c (p < 0.001), while the model term of X1X2 was significant at 5% level (p < 0.05). The variables with the largest effect on the yield of product c were the linear term of X2 and the quadratic term of X2 according to the F‐value. It was that the activity ratio of laccase to lipase (X2) being the key variable that afforded the highest impact on the formation of product c. The insignificant Lack‐of‐Fit F‐value of 4.35 further proves that the data in the experimental domain are well represented by the model and thus the model equation used is sufficient for predicting the yield of product c under any combination of values of the variables. Therefore, it is reasonable to use the quadratic polynomial model generated to explain the actual relationship between the response and the variables.The results of analysis of variance and regression analysis of the fitted quadratic regression model are shown in Table ce Table  and Equa3.2X1), activity ratio of laccase to lipase (X2) and reaction temperature (X3) on oxidation/amination reaction of 2‐methyl‐1,4‐hydroquinone with n‐butylamine catalyzed by lipase/laccase were investigated based on contour plots. According to Equation X1 and X2 as well as X2 and X3 with positive coefficients were synergistic, while the mutual interaction between X1 and X3 with negative coefficients was antagonistic. Among these interaction terms, the interaction effect of X1X3 with F‐value of 103.44 in comparison with that of X1X2 and X2X3 has the highest impact on the yield of product c. Figure c at constant activity ratio of laccase to lipase of 1:2. It was evident that the reaction was sensitive to minor alterations of the test variables X1, X2, and X3. The variables with the largest effect on the yield of product c were the linear term of X2 and the quadratic term of X2 according to the F‐value. The effect of activity ratio of laccase to lipase  on the yield of product c was more impacting than the mole ratio of amines to hydroquinones  and reaction temperature  in this enzymatic catalysis reaction. Generally, an elliptical contour plot means the interactions between the corresponding variables are significant, while a circular contour plot indicates that the interactions between them are negligible. The contour plots of the response in Figure X1 and X3, exhibited a strong and perfect interaction as implied by the obtained elliptical contours with F‐value of 103.44  was statistically more significant than the interaction between reaction temperature and activity ratio of laccase to lipase . However, the mutual interaction between mole ratio of amines to hydroquinones and activity ratio of laccase to lipase was less favorable  when compared to these two interactions with F‐value of 103.44 and 28.88, respectively.The influence of activity ratio of laccase to lipase on the response at constant mole ratio of amines to hydroquinones and reaction temperature (zero level) are shown in Figures c increased with temperature up to the optimum temperature, after which the degree of production achieved had a tendency to decrease . This indicates that the model can be considered quite reliable for predicting the effect of variables including mole ratio of amines to hydroquinones, activity ratio of laccase to lipase, and reaction temperature on the yield of product.4c was observed under the optimum condition with molar ratio of amines to hydroquinones 1.16:1, activity ratio of laccase to lipase 1.14:2, and reaction temperature 38.9°C. A model based on RSM experimental results was developed that was verified as quite reliable for predicting the effects of significant factors on the response. The optimized conditions revealed by the BBD design may prove beneficial for higher yield of monoaminated mitomycin‐like products, an important aspect in the large scale production of mitomycin analogs. However, in order to have better understanding of one‐pot process for synthesis of aminonaphthoquinones catalyzed by co‐catalytic system of laccase/lipase, more researches on the synergistic catalytic mechanism of bienzyme are needed.The present study successfully evaluated the optimized conditions suggested by the response surface methodology based on BBD utilizing laccase/lipase as co‐biocatalysts for producing mitomycin analogs (aminonaphthoquinones) via a one pot process. Considerably high yield of product The authors have declared no conflict of interest.


A heterostructure composed of In2S3 and Sb2S3 shows a unique morphology of formicary microspheres, which provide abundant channels for fast transfer of sodium ions, large surface area for a high pseudocapacitance effect, and enough voids to relieve volume expansion. A sodium‐ion battery containing the bimetallic sulfide anode exhibits a high reversible capacity of 400 mA h g−1 and long cycle life of about 1000 cycles. Similarly, a high capacity of ≈610 mA h g−1 is achieved for a lithium‐ion battery containing the anode. During sodiation/desodiation, the synergistic effect of In2S3 and Sb2S3 enhances electronic conductivity and supports the host structure, preventing collapse. The cycling performance and rate performance of the In2S3–Sb2S3 anode are further improved by wrapping the electrode with carbon nanotubes. Even at a high current density of 3.2 A g−1, this carbon composite structure still shows a capacity of about 355 mA h g−1.Metal sulfides show promise for use in alkali‐ion batteries because of their high theoretical capacities. However, their poor cycling stability and rate performance hinder their further development. To avoid these issues, In Third, the low electronic conductivity of Sb2S3 of below 1 × 10−5 S cm−1 hinders fast electron transfer in its electrodes.Sodium‐ion batteries (SIBs) are considered as potential alternatives to lithium‐ion batteries (LIBs), and in past years, development of the cathode, anode, and electrolyte of SIBs has progressed rapidly.+ storage in Sb2S3 electrodes. One effective way to enhance the reaction kinetics of Sb2S3 electrodes is to design novel nanostructures with a short diffusion distance for Na+ and large interfaces between the electrolyte and electrode.2S3 anode self‐assembled from nanosheets showed enhanced electrochemical properties compared with those of bulk Sb2S3, which was ascribed to the effective volume buffering and Na+ transport.2S3 electrode is subject to the 1D van der Waals force.2S3, with long‐range lattice order and the nanosize effect, displayed good charge‐transfer kinetics and high specific surface area.2S3 electrodes can also be produced by combination with carbon materials. For example, in situ growth of Sb2S3 on multiwalled carbon nanotubes (MCNTs) produced materials with high capacity and good stability because of the 3D porous networks of the MCNTs.2S3 embedded in graphite as a long‐life, high‐rate anode for SIBs. Graphite not only acts as a conductive matrix, but also provides a buffer between Sb2S3 nanoparticles, relieving volume expansion. Xiong et al. showed that sulfur‐doped graphene sheets can be tightly connected to Sb2S3, providing high electronic conductivity and protecting the intermediate products of Sb and Na3Sb.Various modifications have been used to try to overcome the sluggish kinetics and large volume change associated with NaM– intermetallics, where M is an electrochemically inactive component in the system.M and active alloys. M not only acts as a buffer layer between the Na–alloy phases, but also forms a conductive network to allow rapid electron transport in the electrode. Therefore, M  can be introduced into Sb2S3 electrodes to improve their performance.2S3 and Sb2S3 exhibited high capacity retention of 79% after 200 cycles, which can be ascribed to the synergistic effect of these two components.+ storage, which may provide a new strategy to design bimetallic sulfides.2 heterostructure delivered outstanding durability at high current density because of its enhanced electronic transport properties compared with those of SnO2 alone.Based on previous reports on alloying anodes, a promising strategy to improve both structural stability and electronic conductivity is constructing 2S3 and Sb2S3 (denoted as I–S) that displays synergistically enhanced electrochemical properties by a simple synthetic route. I–S contains abundant pores and a rather high specific surface area, which not only improve the Na+ storage dynamics of the material via a strong pseudocapacitance effect, but also provide space to allow volume expansion during the storage process. The indium(III) ions (In3+) introduced to Sb2S3 influence the final structure of I–S because they induce crystal growth. Moreover, the low reversibility of the conversion reaction between In2S3 and the electrochemically inert In lead to the formation of an In buffer layer, which suppresses volume expansion. To build a 3D conductive network, we prepare a composite of MCNTs and I–S particles (denoted as I–S@MCNTs).In this study, we form a heterostructure of In2Figure+ transfer. A cross‐sectional image of the I–S microspheres  and high‐resolution transmission electron microscopy (HRTEM). Figure +. Selected‐area electron diffraction (SAED) patterns obtained from different regions of a particle  and (211) planes, as well as In2S3, with signals from the (104) and (110) planes detected, which are connected by dislocation areas. The FFT image further corroborated the hierarchical structure of I–S consisting of different crystal systems. These results show that the Sb2S3 and In2S3 phases were tightly bound by chemical bonds.The I–S sample was investigated by scanning electron microscopy (SEM). s Figure c illustre Figure f contain0.63Sb1.37S3. In addition, composite formation did not change the elemental contents of I–S (In0.66Sb1.34S3@MCNTs). Note that the spherical I–S microparticles , indicating a mesoporous structure. Further analysis of pore size revealed that the I–S particles had both microporous and mesoporous structures. Thus, the electrolyte can fully contact the surface and interspace of I–S, which shortens the propagation distance of Na+.The center dark field image of I–S in Figure The low electron transmission rate of common metal sulfides may cause sluggish reaction kinetics. To avoid this problem, we composited an I–S sample with ultrafine MCNTs by in situ growth. As shown in Figure S2a of the Supporting Information, the I–S@MCNT composites contained tightly connected I–S and MCNTs, forming a 3D conductive network that can raise electron and ion transmission rates and prevent particle agglomeration. Meanwhile, the uniform distributions of In, Sb, and S in the nanoparticles reveal that composite formation did not change the original material properties of I–S .Scheme3+ and In3+) and sulfur anions (S2−) combine to form In2S3 and Sb2S3 nuclei, respectively. As the temperature and pressure in the stainless steel autoclave increase, the crystal nuclei gradually grow in a certain direction to form nanosheets along the facets with low formation energy, which is guided by the ethanediol solvent and interaction between In2S3 and Sb2S3 particles.2S3 was obtained in a solution without In3+ , whereas flower‐like In2S3 was produced in a solution without Sb3+ under the same conditions . When the reaction was maintained at high temperature for long enough, the folded nanosheets including Sb2S3 and In2S3 phases assembled by electrostatic forces into porous spheres with large specific surface areas: these are the formicary structures. To improve the electron transfer between isolated I–S particles, we composited them with MCNTs, which did not change their porous spherical morphology.Based on complementary experiments and previous reports,Figure2S3 with a rhombohedral structure and Sb2S3 with an orthorhombic structure, indicating a composite phase of In2S3 and Sb2S3. The peaks at 2θ of 29.3°, 32.3°, and 35.5° can be well indexed to the (211), (221), and (240) crystal planes of Sb2S3 , respectively.2S3, respectively.2S3 phase is in the Pbnm space group and that the In2S3 phase is in the P3m1 space group. I–S was successfully composited with MCNTs, as verified by the diffraction peak at 25.6° in the pattern obtained for in the I–S@MCNTs that corresponds to the (002) crystal plane of the MCNTs. The coexistence of Sb2S3 and In2S3 in the I–S@MCNTs was further confirmed by Raman spectroscopy . The peaks observed at 266 and 312 cm−1 are consistent with vibrations of the homopolar bonds of Sb–Sb and In–In, respectively.The X‐ray diffraction (XRD) patterns of I–S and I–S@MCNTs Figure2a showd to the 1, (221),Thermogravimetric analysis (TGA) was performed in air to confirm the carbon content of the I–S@MCNT composite. As shown in Figure 5/2 and Sb 3d3/2, respectively, indicating the presence of Sb3+. Furthermore, the O 1s peak appears in the Sb 3d spectrum, which likely originates from oxygen‐containing functional groups on the MCNTs. These functional groups can effectively improve the sodium storage capacity of the material through surface redox reactions.5/2 and In 3d3/2 signals of In3+, respectively.3/2 and S 2p1/2, respectively. This spectrum can be further fitted into four peaks originating from S atoms in various functional groups, including single doublets from S—Sb and In—Sb bonds. Meanwhile, the chemical interaction between MCNTs and I–S was confirmed by the existence of a signal from S=C=S bonds.2S3 and Sb2S3 with a formicary structure.X‐ray photoelectron spectroscopy (XPS) was used to verify the presence of In, Sb, S, C, and O in the I–S@MCNTs, as shown in Figure Figure−1 at a current density of 200 mA g−1, with a high coulombic efficiency of above 98% after 13 cycles. The capacity contribution of the MCNTs to the total capacity of the I–S@MCNT electrode was very limited  because of their low electrochemical activity  and small content in the electrode. A reference bare Sb2S3 electrode exhibited rapid capacity fading, as shown in Figures S8a and S9a of the Supporting Information. It is speculated that a synergistic effect of Sb2S3 and In2S3 enhances the cycling stability of the I–S@MCNT electrode. The lower specific capacity of the I–S@MCNT electrode compared with that of the Sb2S3 electrode can be ascribed to the formation of inert components during the cycling process, which consumed a certain amount of active components.The electrochemical performance of an I–S@MCNT electrode was evaluated by measuring galvanostatic charge/discharge curves. As shown in 3+ and In3+ showed electrochemical activity in the sodium storage reaction, meaning that two pairs of Na+ can be captured during the insertion and extraction process. However, the sodiation and the desodiation of In2S3  occur at potentials close to those of Sb2S3. Subsequently, the In‐containing components gradually converted into inert components, resulting in the existence of only one group of redox peaks in CV curves. Furthermore, the voltage plateaus in the charge/discharge profiles after 100 cycles were perfectly maintained without increasing polarization voltage. However, the increase in polarization voltage between the charge and discharge curves of the In2S3 electrode  may be the immediate reason for its poor cycling performance.The charge/discharge profiles of the I–S@MCNT electrode during the first cycle Figure b are sim−1, as shown in Figure −1 at current densities of 200, 400, 800, and 1600 mA g−1, respectively. A specific capacity of 355 mA g−1 was obtained at an ultrahigh current density of 3.2 A g−1. When the current density was returned to 200 mA g−1, the capacity fully recovered to 454 mA h g−1 after 40 cycles. Figures S8b and S9c of the Supporting Information compare the capacities of I–S@MCNT, Sb2S3, and In2S3 electrodes at different current densities, revealing the remarkably enhanced rate performance of the I–S@MCNT electrode compared with those of the other electrodes.The I–S@MCNT electrode exhibited a rate capability of up to 3.2 A g−1, the charge and discharge profiles of the I–S@MCNT electrode . However, the starting voltages of the discharge and charge profiles of the In2S3 electrode  were quite different at 1.58 and 0.1 V, respectively, deviating considerably from the set values. After 1000 cycles , illustrating the considerable improvement of electrochemical properties achieved for the I–S@MCNT electrode. The In2S3–Sb2S3 composite plays a critical role in enhancing the long‐term cyclability and rate performance of the I–S@MCNT electrode.As the current density was increased from 200 to 3200 mA g−1 after 100 cycles. The charge/discharge profiles of the I–S@MCNT electrode in an LIB  indicate a similar lithium storage process to that of sodium storage, consisting of intercalation, conversion, and alloy reactions. After long‐term cycling, we measured galvanostatic intermittent titration technique (GITT) curves for the I–S@MCNT electrode , which showed that its charge/discharge profiles were close to thermodynamic equilibrium.The I–S@MCNT electrode also exhibited outstanding performance as an anode for LIBs , delivering a reversible charge capacity of 613 mA h g+ storage kinetics of the I–S@MCNT electrode. As shown in the inset of Figure+ was inserted into the host structure of I–S. The conversion reaction between Na2S, Sb, In, and I–S then occurred near 0.5 V. The obvious reduction peak at 0.2–0.4 V may originate from conversion and alloying during the formation of NaxSb.A series of electrochemical experiments was carried out to investigate the Na+ into the Sb2S3 layered structure. Accordingly, anodic peak D corresponds to the deintercalation of Na+ from Sb2S3. Peaks B and C originate from conversion and alloying reactions with similar potentials. These well‐defined redox peaks reveal that the sodiation and the desodiation reactions on the electrode are the same at different scan rates. As shown in previous work,v) and recorded current (i) from CV curves measured at different sweep ratesTo investigate the sodium storage mechanism in the I–S@MCNT electrode, we measured CV curves at different scan rates, which revealed two pairs of redox peaks Figure b. Cathodb determined by the linear relationship between log v and log i describes the ionic storage process of the electrode. For a diffusion‐controlled process, b approaches 0.5, whereas it approaches 1 for a capacitance‐dominated process. Therefore, the high b values of the I–S@MCNT electrode  demonstrate its favorable capacitive kinetics .Here, the slope The current response at a specific potential has contributions from diffusion insertion and surface capacitive effectsk1v and k2v1/2 represent the contributions to the current from the capacitance‐dominated process and intercalation‐controlled process, respectively. The values of k1 and k2 can be uniquely determined at a fixed potential. The intercalation contribution is dominant near the voltage range of the redox peak, where the redox reactions of Sb3+/Sb0 and In3+/In0 may promote sodium diffusion.−1. In addition, the capacitive contribution ratios at other scan rates were also calculated, as shown in Figure −1 to a maximum of 73.4% at 5 mV s−1. Thus, we infer that the outstanding rate performance of the I–S@MCNT electrode originates from its pseudocapacitance effect that arises from its large surface area and abundant pores, which provide many active sites and fast sodium storage kinetics.Here, 2S3 electrode .The enhanced reaction kinetics of the I–S@MCNT electrode caused by the honeycomb structure and high conductivity of the MCNTs was also verified by electrochemical impedance spectroscopy (EIS), as depicted in Figure 2S, as shown by the peaks at 25.6°, 36.3°, and 38.9°, respectively, in the ex situ XRD patterns , its faint lattice fringes indicate that the discharge products have low crystallinity. The peak at 29.7° suggests that an alloying reaction occurs between sodium and antimony to form Na3Sb.2S3 was gradually regenerated with high reversibility. In particular, the intense peak at 36.3° did not disappear during Na+ extraction, showing that the partially metallic indium transformed into irreversible components. These components played a crucial role as a buffer layer, preventing the material from being pulverized by the huge volume expansion during sodiation/desodiation. This phenomenon can explain the large difference between the first and second CV curves, which originates from the different chemical compositions of the electrode in the first and second cycles.We investigated the sodium storage mechanism of the I–S@MCNT electrode in detail by conducting various ex situ measurements. When the voltage was decreased to 0.9 V (named “D0.9 V”), the conversion reaction generated metallic Sb, metallic In, and Nans Figure5a.47 N3+/Sb0 couple. The peaks at binding energies of 452.6 and 445 eV  are related to the 3d3/2 and 3d5/2 levels of In3+, respectively. The binding energies of these peaks decreased by 0.5 eV during discharging, indicating the reduction of In3+. The In0 signal was always observed in successive sodiation and desodiation reactions, indicating that some indium formed irreversibly in the electrode. Considered together, the results indicate that the storage and extraction of Na+ in the I–S@MCNT electrode involves a three‐step process of intercalation, conversion, and alloying  gradually disappeared during discharging and then appeared again in the reverse process, suggesting a reversible valence change of the Sbd spacings of 0.312 and 0.275 nm are consistent with the (021) and (101) crystal planes of metallic antimony and indium, respectively. The InSb phase appeared between the two metallic antimony phases, which relieved the volume expansion and reciprocal extrusion of sodiated antimony. The HRTEM image of the “D0.01 V” sample reveals the presence of Na3Sb formed by full alloying of Sb, as demonstrated by the (311) fringes for Na3Sb. When the I–S@MCNT electrode was charged to 1.3 V, clear (231) fringes of Sb2S3 with high crystallinity reappeared in the HRTEM image because of the highly reversible conversion reaction. The InSb phase retained in the gaps between Sb2S3 provides an effective buffer for subsequent conversion and alloying reactions.To directly observe these products, especially for the weakly crystalline compounds, HRTEM images Figure c were co+ storage process can be verified by the reappearance of metallic Sb. Based on the low crystallinity observed in the XRD patterns, the existence of intermetallic InSb is demonstrated in the Raman spectra , which show characteristic peaks of the In—Sb bond at 149.1 and 465.8 cm−1. When the electrode was fully discharged, the Sb—Sb bonds gradually disappear as Na3Sb forms. The Raman spectra were reproducible for the same electrode in different regions, indicating a uniform passivation layer formed on the electrode surface.Figure 2S3 and Sb2S3, we used density functional theory (DFT) to investigate the energy band structures and electronic cloud distributions of In2S3, Sb2S3, I–S, and their Na‐inserted structures. The insets of Figure2S3  was slightly lower than those of In2S3 (3.9352 eV) and Sb2S3 (3.9926 eV), as confirmed by the formula Ebarrier = Esodiated electrode – (Eelectrode + ENa+). Table S3 (Supporting Information) presents detailed energy data for these compounds.The band gaps of Sb3 Figure e and I–S3 Figure f gradualn Figure d, reveal+, as shown in Figure 2S3 phase expands more than that of the In2S3 phase, meaning that the stable structure of Na‐inserted In2S3 appears to act as a support skeleton between Sb2S3 phases. In the alloying reaction, the remarkable volume change of metallic Sb and Na3Sb can be relieved by the presence of the metallic In phase, which is an inert constituent of the electrode. Therefore, the In‐containing components improved the cycling stability of the I–S@MCNT electrode.The In‐containing components act as electronic transport paths and buffer layers in the I–S@MCNT electrode during insertion/extraction of Na3−1 at a current density of 400 mA g−1 after 1000 cycles. Even at a high rate of 3200 mA g−1, the electrode maintained a current density of 355 mA h g−1. We ascribe the favorable kinetic properties of the I–S@MCNT electrode to its optimized structure and the inclusion of MCNTs. Therefore, modifying metal sulfides in this manner is promising for the following three reasons: (1) it creates interfaces and a porous morphology, (2) it introduces an electrochemically inert component (such as In2S3) to support the structural stability of the electrode during redox reactions, and (3) it lowers the Na+ migration barrier by improving the electronic structure of the electrode. In addition, the poor conductivity of the sulfide nanostructure can be enhanced by coating with a carbon material.Using various methods, we investigated the electrochemical performance and novel sodium storage process of an I–S@MCNT anode for SIBs, which represents a new electrode design. The I–S@MCNT electrode showed a reversible specific capacity of 400 mA h g4Synthesis: The I–S microspheres were synthesized by the following solvothermal method. Thiocarbamide (0.01 mol) was dissolved in ethanediol (80 mL) under violent agitation to form solution A. Meanwhile, SbCl3 (0.0016 mol) and an equimolar amount of InCl3 were dissolved in stirred ethanediol (80 mL) to form solution B. Solution A was rapidly added to solution B, and then the resulting mixture was stirred for 4 h. The solution was transferred to a 200 mL stainless steel Teflon‐lined autoclave and then reacted at 200 °C for 32 h. After the autoclave had cooled naturally to room temperature, the reddish‐brown precipitate was collected by high‐speed centrifugation, washed repeatedly with absolute ethanol and deionized water, and then dried under vacuum at 80 °C overnight.To improve the electronic conductivity of I–S, we formed an in situ composite of the active materials with MCNTs as follows. MCNTs (50 mg) were dispersed in solution B by ultrasonication for 1 h and then stirred for 2 h to produce MCNTs with attached metal ions. Subsequent mixing of solutions and processing were conducted as described above, providing I–S@MCNT.Material Characterization: Wide‐angle XRD patterns were recorded with a powder X‐ray diffractometer  over the 2θ range of 10°–70°. The specific surface area of I–S was determined by N2 adsorption–desorption measurements using a micropore analyzer (ASAP‐2460). Sample morphology and microstructure were observed by SEM . TEM and HRTEM images were obtained using a JEOL JEM‐2100F electron microscope. The element distribution was determined by EDS with an Oxford attachment. Exact elemental contents were confirmed by ICP‐AES . XPS  was conducted using monochromatic Al Kα radiation. Raman spectra and images were obtained with a Thermo Fisher DXR2xi spectrometer. The MCNT content was measured by TGA, which was performed with a simultaneous thermal analyzer  in air with a temperature ramp of 10 °C min−1. The cycled electrodes were repeatedly washed with volatile dimethyl carbonate to remove the passivation layer. The electrodes and mixed powders were studied with XRD, Raman spectroscopy, TEM, and XPS.Electrochemical Measurements: All electrochemical tests were performed using two‐electrode systems assembled as 2032‐type coin cells. A powder of as‐prepared materials (70 wt%) and active carbon  was bonded with carboxymethylcellulose sodium binder (10 wt%) and then coated on copper foil. The resulting electrode films were dried in a vacuum oven at 60 °C for 24 h, and then cut into 1 × 1 cm2 squares with a mass loading of ≈1.5 mg cm−2. Metallic Na was used as the counter electrode and a glass fiber filter (Whatman GF/C) was used as the separator. The electrolyte was composed of 1 m NaClO4 and equal volumes of ethylene carbonate and diethyl carbonate containing 5% FEC. Galvanostatic charge/discharge data were measured with a battery tester  over a voltage range of 0.01–2.5 V versus Na+/Na. CV and EIS measurements were conducted on an electrochemical analyzer  in the frequency range of 10 kHz to 0.1 Hz.Computation Methods: Theoretical investigations were performed using DFT as implemented in the plane wave code Cambridge sequential total energy package (CASTEP).2S3 with an orthorhombic structure and In2S3 with a rhombohedral structure were used to calculate properties after transforming them into 2 × 2 × 2 supercells. The I–S heterostructure was constructed by stitching together the (100) crystal plane of Sb2S3 and (121) crystal plane of In2S3 without a redundant vacuum layer, producing a heterostructure with well‐defined periodicity. A k‐point mesh density of 5 × 5 × 5 was used for integration over the Brillouin zone. To obtain accurate results, we used a maximum self‐consistent field convergence threshold of <5 × 10−6 eV atom−1. The other convergence tolerances for the maximum force, maximum stress, and maximum displacement were set to 0.01 eV Å−1, 0.02 GPa, and 5 × 10−4 Å, respectively. In particular, the calculations of the total DOS and PDOS used a bond‐energy tolerance of 1 × 10−5 eV.The authors declare no conflict of interest.SupplementaryClick here for additional data file.


To increase the thermoelectric performance of this material, its dimensions and thermal transport should be decreased. Two-dimensional nanoplates with nanopores are an ideal structure because thermal transport is disrupted by nanopores. We prepared Bi2Te3 nanoplates with single nanopores by a solvothermal synthesis and investigated their structural and crystallographic properties. The nanoplates synthesized at a lower reaction temperature (190 °C) developed single nanopores (approximately 20 nm in diameter), whereas the nanoplates synthesized at a higher reaction temperature (200 °C) did not have nanopores. A crystal growth mechanism is proposed based on the experimental observations.Bismuth telluride (Bi The crystal structure of Bi2Te3 is a rhombohedral tetradymite-type with the space group R-3m, and is described by a hexagonal unit cell, as shown in Fig. c-axis in the sequence –Te(1)–Bi–Te(2)–Bi–Te(1)–Te(1)–Bi–Te(2)–Bi–Te(1)–. Here, (1) and (2) denote two different chemical states of the anions. The nature of the bonding between neighboring Te(1) layers is a weak van der Waals attraction, whereas the bonding between bismuth and tellurium atoms is mainly covalent with a small ionic contribution. Because of this crystal structure, the lattice constant of the c-axis (3.045 nm) is approximately seven times as large as that of the a, b-axis (0.438 nm), leading to remarkable anisotropic behavior8.Bismuth telluride  or aqueous (hydro).Recently, nanosized Bi2Te3 nanoplates with a hexagonal shape are generally synthesized by solvothermal methods24. Typical Bi2Te3 nanoplates take the shape of a regular hexagon and have atomically flat surfaces26. Thermoelectric performance can be increased by forming optimized nanopores in the material, which decrease the thermal conductivity while preserving the electrical properties27. Therefore, if nanopores are formed in nanoplates, the thermoelectric performance of the nanoplates might be expected to further increase. However, to the best of our knowledge, there have been no reports on the crystal growth of Bi2Te3 nanoplates containing nanopores.Bi2Te3 hexagonal nanoplates with single nanopores based on a solvothermal synthesis. For comparison, Bi2Te3 hexagonal nanoplates with no nanopores were also prepared using solvothermal synthesis by changing the reaction temperature. The structural and crystallographic characteristics of the nanoplates were investigated. On the basis of our experimental observations and understanding of the crystal growth, the formation of the nanoplates with nanopores is discussed.In this study, we prepare Bi2Te3 hexagonal nanoplates were formed in a solvothermal synthesis. The basic experimental method has been described in our previous work29. The nanoplates were prepared in a Teflon-container within a stainless-steel autoclave, involving a reaction under high pressure and high temperature. Stirring was applied with a hot plate and magnetic stirrer. Bi2O3  and TeO2  were used as Bi2Te3 sources, and C2H6O2  was used as a ligand. NaOH  and polyvinyl pyrrolidone  were contained in the solution. All chemicals were used as received without further purification. The Bi2Te3 nanoplates were synthesized according to the following procedure: 0.4 g of PVP was dissolved in ethylene glycol (18 mL), followed by the addition of Bi2O3 (0.02 mol/L), TeO2 (0.07 mol/L), and 2 mL of a NaOH solution (5.0 mol/L). The resulting precursor solution was sealed in the autoclave and heated and maintained at either 190 °C or 200 °C for 4 h with magnetic stirring at 500 rpm. After the synthesis, the products were allowed to cool down below 50 °C naturally. The residue was washed several times with distilled water and absolute ethanol and the products were collected by centrifugation. Finally, the products were dried under vacuum at 60 °C for 24 h.Single-crystalline Bi2Te3 nanoplates with a field emission scanning electron microscope . The precise structure of the nanoplates was analyzed with a high-resolution transmission electron microscope  and selected area electron diffraction (SAED) at an accelerating voltage of 200 kV. The chemical composition of the nanoplates was determined with an electron probe microanalyzer . The compositions of the samples were calibrated using the ZAF4 program installed with the EPMA-1610. X-ray diffraction  analyses were performed to determine the crystal lattice structure and the phase purity of the nanoplates. We used Cu-Kα radiation at a scanning rate of 0.02°/s over the 2θ range of 10° to 80°.We analyzed the microstructure of Bi2Te3 nanoplates formed at different reaction temperatures in the solvothermal synthesis. At a reaction temperature of 190 °C  planes except for regions near the nanopores. Figure 2Te3 nanoplate synthesized at a reaction temperature of 200 °C. The shape of the nanoplate was almost the same as that synthesized at 190 °C except for the presence of the nanopore. The HRTEM image in Fig. 2Te3. The SAED pattern shown in the inset of Fig. l] zone axis of rhombohedral Bi2Te3, indicating that this nanoplate was single-crystalline and had a preferential (00l) orientation. Notably, weak forbidden reflections were observed, owing to the broken translational symmetry of the Te/Bi antisite defects, which is consistent with previous reports31.To further investigate the precise structure of the Bi2Te3 nanoplates prepared at different reaction temperatures are listed in Table The atomic composition ratios [Te/(Bi + Te)] of the Bi2Te3 nanoplates were examined by XRD analysis, as shown in Fig. 2Te3 (JCPDS 15-0863) although both nanoplates exhibited slight deviations from the exact stoichiometry. The main peaks originated from the (006), (015), (10 10), and (00 15) planes. Therefore, the phase purity and the crystal structure depended only slightly on nanopores in the Bi2Te3 nanoplates.The phase purity and crystal structure of the Bi2Te3 hexagonal nanoplates at the lower reaction temperature. Here, we propose a growth mechanism and reaction process to account for the structures of these nanoplates. Figure 2Te3 nuclei were generated in the solution, they aggregated. The atomic composition ratios of the nanoplates synthesized at different temperatures deviated slightly from the stoichiometric proportion; however, we consider that this deviation did not affect the aggregation process of the nuclei and Ostwald ripening process described below. When the radius of the aggregated nuclei became larger than the critical nucleus, Bi2Te3 nanoparticles were generated. In the Bi2Te3 system, the formation of nanoplates is also attributed to the inherent crystal structure. Because of the high surface energy of the nuclei, the aggregated Bi2Te3 particles were not in thermodynamic equilibrium and were metastable; Bi2Te3 nanoplates with a thermodynamic preference for better crystallinity. After formation of Bi2Te3 nanoplates, the Ostwald ripening process proceeded; however, the process differed at reaction temperatures of 190 °C and 200 °C32. At a reaction temperature of 190 °C, Ostwald ripening led to the formation of nanopore structures because of lower crystallinity or less dense particles in the colloidal aggregate, which gradually dissolved, whereas larger, better crystallized or denser particles in the same aggregate continued to grow at the lower reaction temperature. However, at 200 °C, nanopores were not formed in the nanoplates because the particles in the colloidal aggregate did not dissolve owing to the dense structures initially formed at the higher reaction temperature. Both types of Bi2Te3 nanoplates grew gradually to form deep nanoplates with many crystalline planes from the top to bottom34. Rhombohedral Bi2Te3 is built up from many layers extending along the top-to-bottom crystalline planes connected by van der Waals bonds, as shown in Fig. As mentioned above, the single nanopores appeared at the centers of the single-crystalline Bi2Te3 hexagonal nanoplates. The structural and crystallographic characteristics of the nanoplates were investigated. The nanoplates synthesized at a lower reaction temperature (190 °C) developed single nanopores (approximately 20 nm in diameter), whereas the nanoplates synthesized at a higher reaction temperature (200 °C) did not have nanopores. Based on the experimental and analytical results, we propose a growth mechanism and reaction process for the nanoplates and the nanopore appearance at different reaction temperatures. We expect that the Bi2Te3 nanoplates with single nanopores will feature improved thermoelectric performance.We used solvothermal synthesis at different reaction temperatures to prepare Bi12_11_SuppleInfo_ver2


DUX4, permitting aberrant DUX4 expression which is toxic to muscles. Molecular diagnosis of FSHD1 by Southern blot hybridization or FISH combing is difficult and time consuming, requiring specialist laboratories. As an alternative, we apply a novel approach for the diagnosis of FSHD1 utilizing single‐molecule optical mapping (SMOM).Facioscapulohumeral muscular dystrophy 1 (FSHD1) is a relatively common autosomal dominant adult muscular dystrophy with variable disease penetrance. The disease is caused by shortening of a D4Z4 repeat array located near the telomere of chromosome 4 at 4q35. This causes activation of a dormant gene Long DNA molecules with BssS1 enzyme marking were subjected to SMOM on the Bionano Genomics platform to determine the number of D4Z4 repeats. Southern blot and molecular combing were used to confirm the FSHD1 haplotypes.In a study of a five‐generation FSHD1 pedigree, SMOM correctly diagnosed the disease and normal haplotypes, identifying the founder 4qA disease allele as having 4 D4Z4 repeat units. Southern blot and molecular combing analysis confirmed the SMOM results for the 4qA disease and 4qB nondisease alleles.Based on our findings, we propose that SMOM is a reliable and accurate technique suitable for the molecular diagnosis of FSHD1. It is the presence of this polyadenylation signal that allows stable DUX4 expression and subsequent disease manifestation , located at 18p11.32, is associated with the less common FSHD2 , similarly altering the heterochromatin structure as with D4Z4 repeat contraction and thus allowing chromosome 4 to aberrantly express the DUX4 transcript and subsequently the toxic protein.In the normal state, the D4Z4 repeat region is highly methylated and forms heterochromatin , even in reduced array form in affected individuals, as well as the variable size of the subtelomeric region of chromosome 4 in different individuals. In addition, chromosome 10 harbors near identical D4Z4 repeats  as an alternative approach to diagnose the FSHD repeat structure. Several important problems of existing analysis methods that are resolved include differentiation of 4q35 D4Z4 repeats from the 10q26 array, measurement of actual repeat numbers at 4q35, and differentiation of 4qA from 4qB alleles. SMOM is capable of analyzing DNA fragments from hundreds of kilobases to nearly a megabase in size, and here, we demonstrate successful application of our approach by analyzing clinical DNA samples from a well‐characterized multigeneration FSHD1 pedigree.22.1Over the last 10 years, our Prenatal Diagnosis Center in conjunction with hospital physicians and clinical geneticists has been involved in the care of a large FSHD1 pedigree  plus Puregene Proteinase K (Qiagen) overnight at 50°C. The plugs were then washed with Tris‐EDTA buffer and then either stored at 4°C or proceeded directly to digestion. For digestion, the DNA plug was melted (70°C × 2 min) and then digested at 43°C for 45 min with Agarose (Thermo Fisher Scientific). The recovered DNA was dialyzed for 45 min at room temperature on floating membrane  in Tris‐EDTA buffer. The DNA concentration was measured using a Qubit dsDNA BR assay kit (Thermo Fisher Scientific) in a Qubit 3.0 Fluorometer (Thermo Fisher Scientific). Molecular size was determined by pulsed‐field gel electrophoresis. Typically, the size of the high molecular weight DNA ranged from 0.2 to 2 Mb, and >60% of the molecules were over 1 Mb.For Southern blot, molecular combing, and SMOM analyses, we used an agarose plug extraction method to isolate high molecular weight (MW) DNA as a starting DNA template. High MW DNA was prepared from 2 ml of fresh blood collected in EDTA tubes. DNA isolation was performed following the manufacturer's guidelines . In brief, RBC lysis solution (Qiagen) was used to lyse red blood cells and white blood cells (WBCs) were then pelleted by centrifugation. The WBCs were resuspended in cell suspension buffer, embedded into agarose plugs . We used 5–10 × 102.3Southern blot hybridization was performed by PFGE of restriction enzyme digested genomic DNA. Fragments were separated by electrophoresis in 0.5× TBE buffer at 6 V/cm for 39 hr. After transfer to a solid phase membrane , blots were hybridized with biotin‐labeled probes prepared using the North2 SouthTM Biotin Random Prime labeling Kit (Thermo Fischer). Membranes were washed sequentially at RT in 1x SSC prepared in deionized water and then exposed to X‐ray film to identify the target sequences.EcoRI and HindIII and then blots hybridized with labeled probe p13E‐11, allowing visualization of 4q35 and 10q26 fragments. In parallel, a second aliquot of genomic DNA was triple digested with EcoRI, HindIII, and BlnI, and blots hybridized with the same‐labeled probe p13E‐11, which only allows visualization of 4q fragments. To identify the 4qA and 4qB alleles, a third and fourth aliquot of genomic DNA was digested with HindIII, and blots hybridized with 4qA and 4qB specific probes, respectively.For the molecular diagnosis of FSHD1, a series of different restriction enzyme digestions is needed to size the D4Z4 repeat array and discriminate between the 4qA and 4qB alleles . Labeled DNA (80 ng) was loaded on a Saphyr chip, and linear molecules were electrophoresed for 24 hr through low‐voltage nanochannels on the Saphyr instrument (Bionano Genomics). During this process, the fluorescently labeled DNA molecules were imaged sequentially across the nanochannels by the Saphyr detectors to produce thousands of high‐resolution images of individual DNA molecules. For data analysis, the Bionano Solve/Access software (Bionano Genomics) was used to align labeled molecules against the labeled reference sequence and to identify signatures of structural variation. First, the raw bins of labeled long DNA molecules were corrected by applying the “autonoise” algorithm to the reference genome to identify noisy regions requiring mapping adjustment. Next, alignment was performed between the rescaled molecules and the reference map using Bionano's custom Refaligner software program. The hg19 reference sequence with a haplotype of 4qB D4Z4(8) was used for discrimination of 4q and 10q molecules , thus enabling simple counting of the number of D4Z4 repeats  Figure  who had Prior molecular analysis of four surviving subjects with a confirmed diagnosis of FSHD1 was performed by Southern blot hybridization Figure a. All suSMOM analysis of the nine subjects generated interpretable 4q35 profiles from the aligned reads Figures c and 3 aUnexpectedly, twins 67‐V and 68‐V with suspected early‐onset FSHD1 did not harbor the founding disease 4qA allele with 4 D4Z4 repeats Table . InsteadTo examine the inheritance and origin of the 4qB dup allelic variant, parental analysis was undertaken using the maternal DNA since the father (44‐IV) was already deceased. By SMOM analysis, 4qB dup was not present in the mother (45‐IV) Table  who had 4We have demonstrated that Bionano SMOM in the hands of an experienced laboratory is an appropriate and straightforward tool to confirm FSHD1 diagnosis. The judicious use of BssS1 enzyme as the nicking enzyme gave a simplified one‐step approach to distinguish chromosome 4 D4Z4 repeats from those of chromosome 10 and at the same time enable D4Z4 repeat counts and discernment of the 4qA and 4qB alleles from each other. In this study, application of SMOM generated unique, complex maps of the 4q35 region with a significant number of single‐molecule events, allowing enumeration of D4Z4 repeats along with distinction between the 4qA and 4qB alleles. Alignments produced allelic profiles whereby the map signature pattern was virtually identical for each allele. Further, SMOM provided an accurate molecular diagnosis of FSHD, deriving haplotypes consistent with gold standard Southern blotting. Moreover, SMOM results were reproducible, with identical 4q map signatures in duplicate 17‐III and 68‐V DNA samples Table .Single‐molecule optical mapping analyses of the 5‐generation pedigree identified the founding disease mutation involving a 4qA allele with four D4Z4 repeats. All subjects with this mutation had early‐onset FSHD1 with symptoms progressively worsening with age. Interestingly, in one branch of the pedigree, we revealed a structural variant of the FSHD1 region involving an apparent de novo duplication event. Based on maternal (45‐IV) 4q35 patterns, we tentatively identified the 4qB duplication as belonging to father 44‐IV, who appears to have passed on to both his first daughter and his twin daughters, an allelic variant. So far, only the younger twins show any clinical signs of apparent FSHD1 onset, which appears to be clinically manifesting as a milder form of FSHD1. Therefore, there is a possibility that this structural aberration, in the form of a duplication event, may have generated an unusual disease‐causing 4qB allele. However, a previous molecular study of 4q35 allelic variants associated with FSHD1 only reports the participation of the 4qA allele variants in disease causation (Lemmers et al, In summary, the SMOM procedures we developed and validated are relatively simple and achievable by most operational molecular laboratories and so this form of genome mapping broadens the options for FSHD1 disease diagnosis. Also, although not performed in this study, the moderate sample requirements and short time frame of SMOM to analysis compared to Southern hybridization make it a possible option for potential prenatal application, particularly for urgent requests. Further, as demonstrated in this study, SMOM has potential to identify structural variants such as deletions, duplications, or rearrangements of the locus, which may help to better explain the phenotype in some individuals. While it is still early days in the application of this new technology and costs may be currently prohibitive for general routine application, the indications are there that SMOM represents an advance in both FSHD1 diagnosis confirmation and has future practical potential in the modern molecular diagnostic laboratory.Y.G., Y.L., T.M., D.L., and D.S.C. are employees of Berry Genomics Corporation. None of the authors holds any stocks or bonds in the company. The other authors declare no conflict of interest. Click here for additional data file. Click here for additional data file. Click here for additional data file.