The pore size distributions indicate that the textural characteristics change after oxide modification by poly(dimethylsiloxane) (PDMS). Composites are characterized by mainly mesoporosity and macroporosity of aggregates of oxide nanoparticles or oxide@PDMS nanoparticles and their agglomerates. The FT-IR spectra show that PDMS molecules cover well the oxide surface, since the intensity of the band of free silanols at 3748 cm−1 decreases with increasing PDMS concentration and it is absent in the IR spectrum at CPDMS ≥ 20 wt% that occurs due to the hydrogen bonding of the PDMS molecules to the surface hydroxyls. SEM images reveal that the inter-particle voids are gradually filled and aggregates are re-arranged and increase from 20 to 200 nm in size with the increasing polymer concentration. The highest hydrophobicity (contact angle θ = 140° at CPDMS = 20–40 wt%) is obtained for the CeO2–ZrO2–SiO2@PDMS nanocomposites. The heat of composite immersion in water shows a tendency to decrease with increasing PDMS concentration.SiO However, PDMS conformation can be changed in an adsorption layer depending on the PDMS content and MO structure . A variation in the PDMS content can affect many properties of NC. Thus, such PDMS/zirconia/silica characteristics as the texture, hydrophobicity, and interfacial behavior strongly differ from those of PDMS/silica [In recent decades, hydrophobic hybrid metal or metalloid oxide (MO)–polymer composites are widely used in a variety of applications such as self-cleaning hydrophobic coatings –3, chemi2O3) , and mix2O3) –22 are o2O3)  having a2O3) –26. The 2O3) , 27 is c2O3) , 29, slo2O3) –33, and 2O3)  in compa2O3) , 21) but2O3) , 34. The2O3) , compati2O3) –39. The 2O3)  restrictS/silica . Therefo2 and ZrO2 in MO/PDMS increases the thermal stability [2–ZrO2–SiO2 could be a promising material. Preparation of similar complex nanooxides with a high specific surface was previously described in detail [Upon creation of NC with MO/PDMS, a particular interest should be paid to complex MO, in which each component can enhance certain properties of the whole composite. Since the presence of such oxides as CeOtability , 34, 41,n detail , 43.Since the creation of the composites is primarily intended to obtain hydrophobic materials, the most important task is complete and correct characterization of their hydrophobic properties vs. structures. The hydrophobic properties of fine materials could be determined not only by measuring the contact angles , 45 but 2@PDMS and CeO2–ZrO2–SiO2@PDMS composites analyzed using SEM, adsorption, spectral, contact angle, and calorimetry methods.Thus, in the current study, the main attention is paid to the textural, morphological, and hydrophilic/hydrophobic properties of the polymer SiO2)  and CeO2–ZrO2/SiO2 were used as substrates for adsorption modification by poly(dimethylsiloxane). Silica-supported ceria–zirconia nanocomposites were prepared using a liquid-phase method and subjected to thermal treatment at 550 °C for 1 h. The content of grafted CeO2 was 3 and 10 wt%, and the ZrO2 content was constant at 10 wt% . The synthesis and physicochemical characteristics of CeO2–ZrO2–SiO2 nanooxides were described in detail previously [Fumed silica  was adsorbed onto silica and ceria–zirconia–silica to prepare samples containing 5, 10, 20, and 40 wt% of the polymer. Before the adsorption, oxide samples were dried at 110 °C for 1 h, and then a solution of PDMS (1 wt%) in hexane was added and the suspension was stirred. The suspension was dried at room temperature for 24 h and then at 80 °C for 3 h. All samples with PDMS were in the powder form similar to the initial silica. The composition and textural characteristics of the NC are listed in Table Commercial liquid poly(dimethylsiloxane) PDMS-400  were calculated according to the standard BET method [Vp was evaluated from nitrogen adsorption at p/p0 = 0.98–99 . The nitrogen desorption data were used to compute the pore size distributions  using a self-consistent regularization (SCR) procedure under nonnegativity condition (fV ≥ 0 at any pore radius R) at a fixed regularization parameter α = 0.01 with voids (V) between spherical nonporous nanoparticles packed in random aggregates (V/SCR model) [fV~dV/dR, ∫fVdR~Vp were re-calculated to the incremental PSD (IPSD) at ΦV(Ri) = (fV(Ri+1) + fV(Ri))(Ri+1 − Ri)/2 at ∑ΦV(Ri) = Vp. The fV and fS functions were also used to calculate contributions of micropores (Vmicro and Smicro at 0.35 nm < R < 1 nm), mesopores (Vmeso and Smeso at 1 nm < R < 25 nm), and macropores (Vmacro and Smacro at 25 nm < R < 100 nm).To analyze the textural characteristics of initial oxides and oxide@PDMS nanocomposites, low-temperature (77.4 K) nitrogen adsorption–desorption isotherms were recorded using an ASAP 2405N  adsorption analyzer after outgassing the samples at 110 °C for 2 h in a vacuum chamber. The values of the specific surface area (T method . The totR model) . The dif−1 range using a ThermoNicolet FT-IR spectrometer with a diffuse reflectance mode. For quantitative analysis, some IR spectra were normalized using the intensity of the Si–O vibration overtone at 1865 cm−1 as an inner standard.FT-IR spectra of powdered samples (ground with dry KBr at the mass ratio 1:9) were recorded over the 4000–400 cmThe particulate morphology was analyzed using field emission scanning electron microscopy employing a QuantaTM 3D FEG  apparatus operating at the voltage of 30 kV.θ) measurements. Using the tilting plate method, the contact angles were measured on the pressed (180 bars for 15 min) plates of samples inclined toward the optical axis and in that position liquid gathered on one side of the droplet and retracted on the other one. A 6-μl droplet [A Digidrop GBX Contact Angle Meter (France) equipped with a video camera and firm software was used in the contact angle , θa is the advancing contact angle, and θr is the receding contact angle of water [The apparent surface free energy of given surfaces was calculated using the contact angle hysteresis (CAH):of water .n-decane) liquids, the samples were degassed at 130 °C and 0.01 Pa for 2 h and then used (100 ± 5 mg per 3 ml of wetting liquid) without contact with air. Since the total heat of immersion depends on the mass and surface area of a sample, the obtained results were normalized to both 1 g of a sample and 1 m2 of the surface area. To obtain the averaged Q values, the measurements were performed two to three times for each system and the average errors were ≤7%.Microcalorimetric investigation of oxides and composites was carried out using a DAC1.1A  differential automatic calorimeter. Before the measurements of the heat of immersion of samples in polar (water) or nonpolar  size distribution functions Fig.  show thaSBET) does not demonstrate a significant reduction after grafting of ceria–zirconia. All oxide samples  and macroporous (aggregates of modified SiO2). In general, the average pore radii  are by almost twice larger in CZS1 and CZS2, as compared to unmodified silica. The textural characteristics of the materials change due to the adsorption of PDMS  for SiO2@P40, CZS1@P40, and CZS2@P40, respectively, after the PDMS adsorption in the amount of 40 wt% (Table Vp) as well as Vmeso and Vmacro. After addition of PDMS, the average pore radii  decrease. In general, the addition of polymer can change the porosity characteristics because each long macromolecule can be able to bind several oxide nanoparticles and their aggregates in more compacted structures. This leads to the decrease in the volume of voids between particles [The specific surface area Table , SBET dles Fig. a are maiii Table , Rp,V aarticles .2–ZrO2–SiO2 nanooxides was conducted with poly (dimethylsiloxane) due to the formation of hydrogen bonds ≡ MiO–H···O(Si(CH3)2–)2  between the oxygen atoms in the PDMS polymer chain and hydrogen atoms of the surface hydroxyls (–OH) of the oxide particles.Adsorption modification of initial silica and CeO−1 for the initial oxides and oxide@PDMS composites. The broad band of high intensity at 3050–3700 cm−1 centered at 3300 cm−1 corresponds to the O–H stretching vibrations of surface hydroxyls and adsorbed water [−1 represents the bending vibrations of adsorbed water [2@P20 and CZS2@P20  was determined using the ACA/RCA hysteresis approach [Evaluation of the hydrophobic properties of the composites was carried out by measurements of the contact angle of wetting with water. The apparent surface free energy  and PDMS (hydrophobic) with water differ largely.The surface free energies are significantly higher for SiOtes Fig. . It was Conditions of interactions of water with composites in the microcalorimetry method differ from those measuring the CA by a lack of the gas phase, which affects the wetting. The Gibbs free energy of the system decreases upon immersion of a composite into a liquid. This occurs due to compensating high surface energy of a solid surface interacting with a liquid that is accompanied by the heat release .According to the molecular theory of wetting , interfaQw) and decane (Qd) calculated per 1 g of SiO2@PDMS and CeO2–ZrO2–SiO2@PDMS nanocomposites at different CPDMS are given in Table 2 with Ce- and Zr-oxides results in increasing the heat of immersion in water, while the heats of immersion in decane are almost unchanged (Table 2–ZrO2 can enhance the surface hydrophilicity due to the formation of new active centers of the hydrophilic nature such as OH groups and bridge M–O(H)–M bonds.The heats of immersion in water  reduction of specific surface of composites with the increasing PDMS content, and (ii) changes in the structure of the surface layer interacting with water.After modification of nanooxides with PDMS the surface hydrophilic active centers forming the hydrogen bonds with PDMS become less accessible or inaccessible for water molecules. For all composites SiO2 , a sharp decrease in the immersion heat in water is observed with the increasing PDMS content. However, with a further increase in the content of PDMS, some increase in the heat of immersion in water was manifested, though CA displayed high values  were used as substrates for adsorption modification by PDMS-400 in the amounts of 5, 10, 20, and 40 wt%. Effects of nanostructured oxides on the textural and morphological characteristics of polymer silica@PDMS and ceria–zirconia–silica@PDMS NC were studied employing structure  and morphology (SEM) techniques. Microcalorimetry and measurements of the contact angle of wetting with water were performed to investigate the hydrophobic properties of the materials.Nanooxides  at any content of PDMS.The nanocomposites displayed high values of the main parameters related to the hydrophobicity (ACA and RCA). The contact angles of water drops for the complex oxide with adsorbed PDMS are about 140° at The heats of immersion in water significantly decrease with increasing PDMS content in the composites due to two factors: formation of the hydrogen bonds between the hydrophilic active sites of oxides and PDMS leading to the decrease in their accessibility to the interactions with the water molecules and reduction of the specific surface area of composite with the increasing PDMS concentration. However, at a high content of polymer (40 wt%), the interactions of composites with water can increase due to higher aggregation of PDMS molecules each other resulting to the adsorbed layer structure changes.Thus, this study presents promising results for an easy and cost-effective alternative using such composites as protective coatings with appropriate hydrophobic properties.


Organic nitrogen (N) concentrations far exceed those of inorganic N in most soils and, despite much investigation, the composition and cycling of this complex pool of soil organic matter (SOM) remain poorly understood Fig. .1, 2, 3,15N‐labelled substrates as stable isotope tracers has contributed much to our understanding of N cycling in the soil system  of nucleic acids (deoxyribonucleic acid (DNA) or ribonucleic acid (RNA)).15N‐enriched nucleic acids, which, following fingerprinting, may enable the taxonomic identity of actively assimilating microorganisms to be established.15N enrichments required (>50%); the potential for cross feeding and trophic cascades under highly 15N‐labelled substrates and longer incubation times; and the complexity of matching terminal restriction fragments  with sequences from clone libraries.15N‐SIP does not afford insights into the N cycling of the soil community as a whole and the fate of applied 15N.A more recently developed technique that interrogates the active microbial community in more detail is 15N‐SIP of soil organic N using mass spectrometry. Many studies have used conventional gas chromatography/mass spectrometry (GC/MS) in compound‐specific 15N‐SIP studies ; plant N uptake ; small leaks can be detrimental due to the high abundance of N2 in air; and there is potential for interfering ionic species, such as [CO]+, at m/z 28, 29 and 30),15N‐SIP using GC/C/IRMS to investigate soil N cycling.The latter studies have begun to hint at the potential of 15N‐SIP using GC/C/IRMS to investigate the fate of N in soils and obtain a measure of the assimilation of an applied 15N‐labelled substrate by the soil microbial biomass. We demonstrate the utility of the approach for the study of any N‐containing soil amendment via the results of laboratory incubations applying inorganic  and organic  substrates. We derive quantitative estimates of newly synthesised soil protein, which is representative of the functioning of the soil microbial biomass and biomass protein production using the applied substrate. The high selectivity and sensitivity of GC/C/IRMS15N‐tracer doses that minimise perturbations to native soil conditions and ensure the highly 'diluted' metabolic products of the 15N tracer are readily detectable in the incubated soils. The power of the approach lies in the analysis of AAs as these are major 'building blocks' of all life, forming the proteins which regulate essential biochemical reactions. Proteinaceous matter  generally comprises 20–50% of total soil N and is ubiquitous in living organisms, so is a major 'organic product' of microbial activity/assimilation.15N detectable in the total hydrolysable AA pool offers a measure of (or 'proxy' for) the assimilation of applied 15N‐labelled substrate by the soil microbial biomass and an estimate of newly synthesised soil protein. We discuss the range of potential insights and highlight the wider applicability of the approach in the investigation of complex N cycling ecosystems.Herein we describe the advantages of compound‐specific AA ate; 15NO–) and oret al.15N‐labelled ammonium chloride , 15N‐labelled potassium nitrate , 15N‐labelled glutamic acid  or for the control samples, DDW (200 μL). Substrates were introduced by injection and the needle was drawn up through the soil as the plunger was depressed in order to achieve an optimal distribution. The incubation experiments were halted by immersion in liquid nitrogen (N2) after periods of 1.5, 3, 6 and 12 h and 1, 2, 4, 8, 16 and 32 days in the dark and stored at 20 °C until freeze‐drying. All incubations were carried out in triplicate so there were three tubes for each time point of each treatment  at North Wyke Research Station near Okehampton, Devon, UK; the same site used by Knowles –1 in 0.1 M HCl; Sigma‐Aldrich) was added as an internal standard. Hydrolysis with 5 mL 6 M HCl was carried out at 100 °C for 24 h under an atmosphere of N2. Acid hydrolysis extracts both free and proteinaceous AAs as well as catalysing the breakdown of living microbial biomass.2 as the presence of oxygen (O2) can induce the thermal breakdown of hydroxyl‐ and sulfur‐containing AAs .Finely ground, freeze‐dried incubation soil samples (100 mg) were weighed into culture tubes and 100 μL of norleucine .N‐acetyl, O‐isopropyl derivatives for analysis.Hydrolysates were collected by centrifugation, dried under a stream of Nδ15N analyses were carried out using a Eurovector elemental analyser  coupled to a Micromass Isoprime isotope ratio mass spectrometer  at the Lancaster node of the Natural Environment Research Council Life Sciences Mass Spectrometry Facility . Soil (ca 10 mg) was weighed into tin capsules, combusted and subsequently reduced over heated copper (Cu) wires in the elemental analyser before the resultant N2 was passed into the isotope ratio mass spectrometer for determination of % TN contents and δ15N values.Bulk soil percentage total N (% TN) and VF‐23ms column  and a flame ionisation detector  was used for quantification of individual AAs as their N‐acetyl, O‐isopropyl derivatives by comparison with the internal standard, Nle. The N‐acetyl, O‐isopropyl AAs were identified by their known elution order2; The BOC Group plc, Guildford, UK), at a flow rate of 3 mL min–1. The temperature programme utilised was: 40 °C (1 min) to 120 °C at 15 °C min–1, then to 190 °C at 3 °C min–1 and finally to 260 °C (12 min) at 5 °C min–1. Data were acquired and analysed using Clarity chromatographic station for Windows by DataApex .A model 5890 Series II gas chromatograph  fitted with a δ15N values of individual AAs as their N‐acetyl, O‐isopropyl derivatives were determined using a ThermoFinnigan Trace 2000 gas chromatograph coupled with a ThermoFinnigan DeltaPlus XP isotope ratio mass spectrometer via a ThermoFinnigan Combustion III Interface . Samples were introduced using a GC Pal autosampler  and via a programmable temperature vaporisation (PTV) inlet (Thermo Electron Corporation). The carrier gas was helium  at a flow rate of 1.4 mL min–1 and the gas chromatograph was fitted with a DB‐35 column . The temperature programme utilised was: 40 °C (5 min) to 120 °C at 15 °C min–1, to 180 °C at 3 °C min–1, then to 210 °C at 1.5 °C min–1 and finally to 270 °C at 5 °C min–1. The oxidation reactor was composed of Cu, nickel (Ni) and platinum (Pt) wires  and maintained at 980 °C and the reduction reactor was composed of Cu wires and maintained at 650 °C. AA δ15N values were determined relative to that of a monitoring gas of known (previously determined using in‐house AA standards) N isotopic composition introduced directly into the ion source via an open split in four pulses at the beginning and end of each run. The δ15N values of the in‐house AA standards were determined off‐line by elemental analysis/isotope ratio mass spectrometry (EA/IRMS) by Thermo Fisher Scientific  and by the NERC Centre for Ecology & Hydrology  using primary reference materials . In order to adhere to the identical treatment principle and ensure the GC/C/IRMS system was functioning properly, each sample was bracketed by the in‐house AA standard mixture of known δ15N values and sample AA δ15N values accepted only when at least 75% of the AAs in the standard mixture run either side of the sample were within ±1 ‰ and the others were within ±1.5 ‰, and when this was also true on average over the course of the run. Data were acquired and analysed using Isodat NT 3.0 (Thermo Electron Corporation). Figure m/z value recorded.The 15N enrichment (E) in hydrolysable AAs can be summed to represent newly synthesised soil protein in the soil at that time:If the total hydrolysable AA pool is taken to be representative of the soil protein pool, then any E of an AA may be expressed as the number of moles of 15N derived from the applied substrate that are present in that AA in the soil:nN is the number of moles of N in the AA  and AFE is the atom fraction excess of the AA after incubation compared with the control:The 15N in the AA, i.e.:AF is the atom fraction of δ15N value as in Knowles et al.:RStd is the 15N/14N ratio of AIR, the international isotopic standard for N. The data may also be expressed in terms of the percentage of the applied 15N incorporated into each AA, as in Knowles et al.:N is the number of moles of excess 15N applied . Percentage incorporations reflect both the concentration and 15N enrichment  of the AA (i.e. how much was incorporated if the AA at x concentration was 15N‐enriched by x ‰) and the percentage of applied 15N incorporated into newly synthesised soil protein is determined by summing these results for individual AAs. Note that these percentage incorporation data will be affected by the conservation of applied 15N in the system; thus, if 15N is lost from the system (e.g. over time), AA percentage incorporations may become skewed as there is less 15N available for incorporation than expected. The incubation experiment design described aims to limit any 15N losses from the system in order to obviate this issue, but it is equally possible to calculate percentage incorporations at time, t, based on the moles of applied 15N retained  in the system at time, t  almost by definition results in no notable changes in the % TN of the soil over the course of the experiment. Tables 4]+, [NO3]– or Glu , 25.9% (SE: 0.523) and 48.4% (SE: 2.71) of total soil N throughout the 15[NH4]+, 15[NO3]– or 15N‐Glu experiments, respectively  determination of the 4]+, 15NO– or 15N‐15[NH4]+  or via the glutamine synthetase (GS), glutamate synthase  pathway.δ15N values of Glu reflects the initial incorporation of 15[NH4]+, with the subsequent decline after 2 days reflecting redistribution of the 15N into newly synthesised AAs, hence their δ15N values rise.Incorporation of 15[NO3]– experiment are extremely interesting, further emphasising the importance of this approach  as any 15N enrichment in the hydrolysable AA pool must be derived from the applied substrate via microbial processing during the experiment. The assessment is more complicated however when the applied substrate is a hydrolysable AA (e.g. 15N‐Glu) as this must be accounted for in the analytical approachQuantifying the fate of N‐containing substrates (inorganic or organic) in different soils is essential to understanding the N cycle in natural or semi‐natural ecosystems but is especially important in agricultural systems where managing fertiliser applications has ecological and economic relevance. The new insights gained into N cycling through this novel approach offer potential to enhance fundamental understanding in this area. Using Eqns. ime Fig. . These cFig. 15NH+ and 15[4]+, [NO3]– and Glu here, clear differences in the assimilation of these substrates into newly synthesised hydrolysable soil AAs are revealed. Alternatively, the technique can also be used to compare the fate of particular N amendments in different soils to provide hitherto unattainable estimates of the relative 'activity' of the microbial biomass of the soils under selected incubation conditions. In both cases, a measure of newly synthesised protein can be obtained by summing the 15N enrichments of all the AAs at each time point for each treatment  are initially assimilated rapidly, with the amount assimilated increasing considerably between each time point until a transient equilibrium with slower soil N turnover/loss develops , on the other hand, the dynamics are more complex and the rate of assimilation is always closer to that of turnover (Figs. 15N substrate by the soil microbial biomass or the amount of newly synthesized soil protein, it does provide enhanced insights compared to other currently available methods. It is reassuring that the percentages of applied 15N recovered in soil microbial biomass N studies (e.g.Due to the dynamic nature of the soil system any estimates of rates (NH+ and Glurates (NH+ and Gludies e.g., 13 are 15N‐SIP approach using GC/C/IRMS described herein to investigate the fate of N amendments (e.g. via fertilisation; Fig. 15N‐labelled substrates/amendments applied at environmentally relevant concentrations and appropriately low 15N enrichments to minimise perturbations and 15N discrimination/isotopic fractionation, respectively. Bulk N isotope analysis cannot provide such insights and GC/MS 15N‐SIP studies require highly 15N‐labelled substrates for sufficient product AA 15N enrichment, making such studies more susceptible to 15N discrimination/isotopic fractionation effects and much more expensive.The method provides a sensitive and relatively selective means of assessing microbial assimilation of Valuable insights into microbial biochemical assimilation pathways can be gained and differences are readily revealed in the microbial processing of N‐containing amendments of differing chemical/biochemical natures, e.g. inorganic versus organic or different types of inorganic or organic amendment.Estimates are provided for newly synthesised soil protein, which are inaccessible based on currently available methods.Detailed quantitative insights can be gained into the dynamics of N cycling from an applied substrate through the soil protein pool.Scope exists for using this new approach to probe soil N cycling in relation to a wide range of soil biota, ecosystem variables and anthropogenic management regimes. Opportunities for further refinement of the method are exemplified by our recent paper,The method is potentially adaptable to investigate N cycling into other N‐containing biochemical pools, e.g. amino sugars.The novel compound‐specific


Candidatus Liberibacter europaeus” ASNZ1, assembled from broom psyllids (Arytainilla spartiophila) from New Zealand. The assembly comprises 15 contigs, with a total length of 1.33 Mb and a G+C content of 33.5%.Here, we report the draft genome sequence of “ Arytainilla spartiophila) was released in New Zealand as a biocontrol agent against Scotch broom (Cytisus scoparius) , in which it can also reach high titers , which was then ultracentrifuged at 100,000 × g for 2 h. DNA extracted from each 200-µl fraction was screened with a quantitative PCR (qPCR) assay for the presence of “Ca. Liberibacter europaeus” using primers targeting a region of the 16S rRNA gene. The DNA from the fraction with the highest copy number was whole-genome amplified using a REPLI-g kit (Qiagen) per the manufacturer’s recommendations. This DNA was sequenced using Illumina 2000  and PacBio RS II  technology. The Illumina sequences were assembled using SOAPdenovo2 (Ca. Liberibacter europaeus” were mapped to the draft genome sequence to confirm that the contigs are part of the “Ca. Liberibacter europaeus” genome. The resulting draft genome sequence consists of 15 contigs with a G+C content of 33.5%. The total length of the assembly is 1,331,154 bp. The low G+C content and small size of the genome are consistent with those of sequenced genomes from other Liberibacter species (Ca. Liberibacter europaeus” ASNZ1 genome is predicted to have 1,123 coding sequences, 3 rRNA operons, and 44 tRNAs. This genome sequence will enable genomic comparisons of “Ca. Liberibacter europaeus” with other Liberibacter species in order to better understand the lifestyles of this emerging group of plant pathogens and endophytes.“Pdenovo2 . An iterCa. Liberibacter europaeus” ASNZ1 has been deposited in GenBank under accession number PSQJ00000000. The version described in this paper is the first version, PSQJ01000000.The draft genome sequence of “


Light-harvesting antennae are the machinery for exciton pumping in natural photosynthesis, whereas cascade energy transfer through chlorophyll is key to long-distance, efficient energy transduction. Numerous artificial antennae have been developed. However, they are limited in their cascade energy-transfer abilities because of a lack of control over complex chromophore aggregation processes, which has impeded their advancement. Here we report a viable approach for addressing this issue by using a light-harvesting porous polymer film in which a three-dimensional π-network serves as the antenna and micropores segregate multiple dyes to prevent aggregation. Cascade energy-transfer engines are integrated into the films; the rate and efficiency of the energy-funneling engines are precisely manipulated by tailoring the dye components and contents. The nanofilms allow accurate and versatile luminescence engineering, resulting in the production of thirty emission hues, including blue, green, red and white. This advance may open new pathways for realising photosynthesis and photoenergy conversion. Light harvesting initiates photosynthetic processes, whereas antennae serve as a photon engine that pump excitons to a reaction center, where photoenergy is transformed into chemical energy121245141516171819Porous organic polymers (POPs)21222324262728293132343637384041Herein, we report light-harvesting POP films into which controllable cascade energy transfer is integrated, leading to a new generation of customisable luminescent materials that are engineered to be easily tailored. The POP films are synthesized with a controlled thickness via the previously reported electrochemical method and emit bright blue luminescence. The three-dimensional π-network drastically enhances the interface of energy transfer to relay and acceptor dyes, whereas the nanopores within the film permit segregated molecular docking of multiple laser dyes with desired properties and in specific proportions. Mechanistic studies reveal that the light-harvesting films allow the integration of a cascade energy-transfer engine that functions in a vectorial and highly efficient manner and enables total control over the speed and efficiency of the energy transfer through tuning of the distribution and types of dye introduced. We show the accuracy and versatility of luminescence engineering with the POP films in producing thirty different luminescence hues, including blue, green, red and white . By virtN-substituted carbazole groups at its periphery serve as electroactive modules for polymerisation  from the films4748434445To fabricate light-harvesting POP films, we employed orthogonal 2,2′,7,7′-tetraphenyl-9,9′-spirobifluorene as the focal core; this compound strongly absorbs in the ultraviolet region and emits highly efficient deep-blue luminescence, whereas the four risation . N-Subst2 were utilised in this study. This method allows good control over the film thickness by simply tuning the number of scan cycles. A series of thin films with controlled thickness were synthesised, where each cycle resulted in an increase in the film thickness of approximately 2 nm  measurements revealed that the content of the monomeric carbazole unit in the films is negligible . The sizely 2 nm . We condm  increased in a sigmoidal fashion and reached 94% as the coumarin 6 content approached 3.19 mol%  serves as an energy-accepting counterpart when trapped within the pores of the POP⊃C6 films. Nile red emits a deep-red fluorescence at 600 nm with an excitation band at 554 nm ; this exΦENT = 94%), whereas the energy relay from coumarin 6 to Nile red is solely dependent on the content of Nile red with an increased efficiency in a sigmoidal fashion reaching 93% as the Nile red content increased to 5.41 mol%  of two steps by reducing the energy transfer efficiency. The incomplete energy transfer would leave blue, green and red luminescence, opening a window to produce white emission. For this purpose, we decreased and optimised the coumarin 6 content at 0.63 mol%, whereas the Nile red content was tuned from 0.3 mol% to 0.75 mol% . The eneWe plotted the luminescent colour tracks of the aforementioned thirty POP films by mapping their CIE coordinates . The oriWe calculated the effective distance between coumarin 6 and Nile red in the cascade energy relay process. We assumed that four TPSC molecules circled a pore (in agreement with the simulation and BET results), leading to a pore size of 2.3 nm. For the POP⊃C6×NR film with 3.19 mol% coumarin 6 and 5.41 mol% Nile red, the maximum and minimum distances between coumarin 6 and Nile red molecules were estimated to be approximately 9 and 4 nm, respectively. The distance between coumarin 6 and Nile red is much lower than the Förster distance of energy transfer, thus enabling the cascade energy relay to occur effectively within the POP⊃C6 × NR films.τ0) of 2.12 ns (τDA) of the POP⊃C6 films monitored at 410 nm of the POP emission spectrum, was significantly decreased from 1.98 to 0.25 ns in a sigmoidal fashion (kENT)kENT = τDA−1 − τ0−1, increased from 0.03 × 109 to 3.5 × 109 s−1  monitored at 503 nm on the coumarin 6 emission spectrum was 3.25 ns (τDA), and finally reached a value of only 0.34 ns , the lifetime , the energy transfers from the POP film to coumarin 6 and relay from coumarin 6 to Nile red are both incomplete processes . The fil7 to 109 s−1 – by tuning the nature and content of the dye incorporated into the film. Remarkably, the rate-determining step of cascade energy transfer is solely dependent on the relative amounts of dyes in the films. Our results reveal that the films trigger vectorial and highly efficient cascade energy transfer through their channels at a high yet manageable speed, which enables predesignable and precise engineering of their luminescence. Because dye molecules in this system can be replaced with redox-active components, the cascade films can be useful for the development of photocatalytic and photoenergy conversion systems.The cascade energy-transfer processes are controlled over a range of efficiencies and a rather wide range of rate constants – from 109 s−1. We demonstrate the accuracy and versatility of luminescence engineering by developing films that luminesce with thirty different emission hues, including the three primary colours and pure white. Another significant finding is that the films gain the highest fluorescence quantum efficiencies of the corresponding laser dyes as a result of site segregation effect, whereas the antenna effect of the POP skeletons greatly amplifies the green and red emissions by 30- and 24-fold compared with the emissions generated by direct excitation of the laser dyes. Our strategy may open new possibilities for the development of cascade systems for luminescent and lasing materials, artificial photosynthesis and photoenergy conversion.We have integrated controlled cascade energy transfer and relay engines into light-harvesting POP films. The porous polymer network with extended π-cloud delocalisation increases the interface for exciton pumping and energy transfer, whereas the nanopores offer ‘independent rooms' for the spatially segregated molecular dockings of laser dyes. The synthetic control over the incorporated dye components and their relative compositions results in the generation of controlled cascade energy transfer systems that can be tuned in a straightforward and precise manner. The efficiency and speed of the cascade energy transfer were controlled by the dye components and their loading in the films and were exploited to achieve energy-transfer efficiencies of near 100% and energy-transfer rate on the order of 10+ nonaqueous electrode was used as the reference electrode. ITO was used as the working electrode, and titanium metal was used as the counter electrode. A mixture of TPSC (0.5 mg mL−1) and TBAPF6 (0.1 M) in CH2Cl2 (5 mL) was used as the electrolyte solution. The CV potential range was set between −0.8 and 1.0 V, and the scan rate was 50 mV s−1. The thickness of the films was controlled by the cycle number used in their formation. The resulting films were washed with acetonitrile to remove unreacted precursors and supporting electrolytes, dried under vacuum and stored under Ar in the absence of light.The electro-oxidative coupling reaction was performed using a standard one-compartment, three-electrode electrochemical cell attached to an ALS/CHi 610C electrochemical workstation. The Ag/Ag2Cl2 solutions (20 mL) of coumarin 6 at different concentrations  for 12 h at room temperature. The resulting films were rinsed several times with CH2Cl2 until the wash solution was still clear after use. The resulting films were dried under vacuum and stored under Ar in the absence of light. The loading of coumarin 6 in the POP⊃C6 films were determined by XPS mesurements are 0.11 mol%, 0.16 mol%, 0.25 mol%, 0.34 mol%, 0.41 mol%, 0.49 mol%, 0.57 mol%, 0.57 mol%, 0.63 mol%, 0.76 mol%, 1.01 mol%, 1.80 mol% and 3.19 mol% (The POP⊃C6 films were prepared by immersing the POP films into CH.19 mol% .2Cl2 solutions (20 mL) of Nile red at different concentrations  for 12 h at room temperature. The resulting POP⊃C6 films were rinsed several times with CH2Cl2 until the wash solution was still clear after use, at which point the films were dried under vacuum and stored under Ar in the absence of light. The loadings of Nile red in the POP⊃NR films were determined by XPS mesurements; the results were summarized in The POP⊃NR films were prepared by immersing the POP films into CH2Cl2 solutions (20 mL) of Nile red at different concentrations . The POP⊃C6×NR films (coumarin 6 content = 0.63 mol%) used in engineering the luminescence of white light were prepared by immersing the POP⊃C6 films into CH2Cl2 solutions (20 mL) of Nile red at different concentrations  for 12 h at room temperature. The resulting POP⊃C6×NR films were rinsed with CH2Cl2 until the wash solution was still clear after use, dried under vacuum and stored under Ar in the absence of light. For the POP⊃C6×NR films (coumarin 6 content = 3.19 mol%) used in green-to-red luminescence engineering, the Nile red content was 0.3 mol%, 0.39 mol%, 0.45 mol%, 0.62 mol%, 0.75 mol%, 0.89 mol%, 1.07 mol%, 1.18 mol%, 1.53 mol%, 2.04 mol%, 3.16 mol% and 5.41 mol%, respectively (The POP⊃C6×NR films (coumarin 6 content = 3.19 mol%) for green-to-red luminescence engineering were prepared by immersing the POP⊃C6 films into CHectively . For theectively .Supplementary Information


The use of carbonyl‐stabilized ammonium‐ and sulfonium ylides allows for the synthesis of highly‐functionalized trifluoroacetyl‐substituted cyclopropanes. It turned out that the diastereoselectivities strongly depend on the nature of the chosen ylide and the employed base. The products could be obtained in good yields under operationally simple conditions. These highly functionalized cyclopropanes have so far only sparingly been investigated,2 and 3 to the easily accessible Michael acceptors 4 would result in a powerful method to access a variety of these highly functionalized target molecules 2 and 3 as ylide‐precursor for the cyclopropanation reaction with acceptor 4a. Table 2, Me3N was chosen as the amine component because of its proven superior leaving group ability compared to other achiral tertiary amines.[2Cl2 at room temperature for 20 h. This was also carefully double‐checked with the optimized conditions and it turned out that longer reaction times and/or higher temperatures lead to a pronounced decomposition of the product 1a under the basic conditions. While the use of sodium‐ and potassium carbonate did not result in any product formation (entries 1 and 2), the use of an excess of K3PO4 lead to the slow formation of a mixture of the three trans,trans,cis‐diastereomers  of cyclopropane 1a (entry 3). Switching to solid Cs2CO3 next, the conversion could be increased significantly (entries 4–8). It turned out that 2 equiv. of this base leads to a reasonable combined 68 % isolated yield of the three diastereomers (entry 4), while a higher amount resulted in a more pronounced by‐product formation (entry 6). The three diastereomers were obtained in slightly varying ratios depending on the amount of base and also depending on the stoichiometric ratio of ammonium salt 2 and acceptor 4a (entries 7 and 8). In most of these Cs2CO3 experiments D3 turned out to be the slightly favored diastereomer and we were also able to analyze this isomer by single‐crystal X‐ray analysis, which revealed the relative trans,trans,cis‐configuration depicted in Figure all‐cis diastereomer could be detected. Unfortunately, we have not been able to crystallize the other two diastereomers of 1a. However, we realized that D1 always contains noticeable amounts of its corresponding hydrate 5a  and the same was also observed for some of the other derivatives shown in Scheme cis‐oriented carbonyl group, which would only be possible for one specific trans,trans,cis‐diastereomer as illustrated in Figure 5b) of the o‐methoxy‐containing cyclopropane 1b, which proved this cis‐configuration of the trifluoroacetyl group and the acetophenone group . Here an excess of base or the use of an aqueous base lead to a significantly reduced yield. By changing for organic bases, it was only with DBU that we observed a reasonable formation of the cyclopropane 1a . Very interestingly, while 2 equiv. of DBU resulted in a slight preference of D2, an excess of DBU lead to an increased selectivity towards this diastereomer (entry 11). In a next attempt, we used the sulfonium salt 3a for this reaction (entries 12–14). Noteworthy, a similar result as for the ammonium salt 2a was observed under the DBU‐conditions (compare entries 11 and 12). However, when changing for NaOH or Cs2CO3 as bases (entries 13 and 14), we observed a strong preference for the D1 diastereomer in this sulfonium ylide‐mediated cyclopropanation. This complementary outcome illustrates very nicely the well‐documented different reactivity trends of ammonium and sulfonium ylides, which are a consequence of the different nucleophilicities and leaving group abilities of these ylides.When we used stronger inorganic bases like solid NaOH, Summing this screening up, we have identified complementary conditions that give the diastereomers with different preferences by using ammonium or sulfonium salts in the presence of different bases. The most promising conditions , it turned out that the (electronic) nature of the reagents can have a pronounced influence on the observed diastereoselectivities under the three different sets of conditions. The use of solid Cs2CO3 with the ammonium ylides . We also tested the potential of ester‐ and amide‐stabilized ylides to access the cyclopropanes 1k and 1l. Unfortunately, those ylides were found to be less suited for these reactions. We have not been able to obtain the amide‐based 1l under any conditions and the ester‐containing 1k could only be accessed in low yields (but with a very high diastereomeric purity) when using the corresponding ammonium salt in the presence of Cs2CO3. We were however not able to increase the yield by optimizing the reaction conditions.Testing different acceptors 1a : General details as well as the analytical details and characterization data of all the novel cyclopropanes can be found in the online supporting information.General Cyclopropanation Procedure: To a stirred suspension of the base (2 equiv. Cs2CO3 or 10 equiv. DBU) in 3 mL of CH2Cl2, the ammonium salt 2 (0.1 mmol)  and the acceptor 4 (0.1 mmol) were added. The reaction mixture was stirred for 20 h at room temperature. Subsequently, the reaction was quenched by addition of 10 mL of H2Odeion and extracted three times with 10 mL of EtOAc. The combined organic phases were dried with Na2SO4 and the solvents evaporated to dryness. The crude product was purified by column chromatography over silica gel with gradients of heptane/EtOAc (100:0 – 20:1 – 10:1 – 5:1 – 2:1) to yield the cyclopropanes 1 in the reported yields.Supporting InformationClick here for additional data file.


FCGR2A, FCGR2B, FCGR2C, FCGR3A, and FCGR3B, which are located in an 82.5‐kb segmental tandem duplication on chromosome 1q23.3, which shows extensive copy‐number variation (CNV). Deletions of FCGR3B have been suggested to increase the risk of inflammatory diseases such as systemic lupus erythematosus and rheumatoid arthritis (RA). In this study, we identify the deletion breakpoints of FCGR3B deletion alleles in the UK population and endogamous native American population, and show that some but not all alleles are likely to be identical‐by‐descent. We also localize a duplication breakpoint, confirming that the mechanism of CNV generation is nonallelic homologous recombination, and identify several alleles with gene conversion events using fosmid sequencing data. We use information on the structure of the deletion alleles to distinguish FCGR3B deletions from FCGR3A deletions in whole‐genome array comparative genomic hybridization (aCGH) data. Reanalysis of published aCGH data using this approach supports association of FCGR3B deletion with increased risk of RA in a large cohort of 1,982 cases and 3,271 controls .Fcγ receptors are a family of cell–surface receptors that are expressed by a host of different innate and adaptive immune cells, and mediate inflammatory responses by binding the Fc portion of immunoglobulin G. In humans, five low‐affinity receptors are encoded by the genes  Fcγ receptors are a family of cell–surface receptors that are expressed by a host of different innate and adaptive immune cells, and mediate inflammatory responses by binding the Fc portion of immunoglobulin G (IgG) [Nimmerjahn and Ravetch, FCGR2A (MIM# 146790), FCGR2B (MIM# 604590), FCGR2C (MIM# 612169), FCGR3A (MIM# 146740), and FCGR3B (MIM# 610665). All five genes are encoded within or surrounding an 82.5‐kb tandemly arranged segmental duplication on chromosome 1q23.3  between the two segmental duplications with the location of the breakpoint determining whether FCGR3A or FCGR3B genes are deleted, and whether fusion FCGR2A/C genes are generated [Machado et al., FCGR2B pseudogenes and FCGR2C/A fusion genes [Machado et al., In humans, there are five low‐affinity receptors FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, and FcγRIIIb encoded by the genes R2A MIM# 46790, FCFCGR3B deletion with SLE [Morris et al., FCGR3B copy number from FCGR3A copy number, although it should be noted that MLPA does not distinguish the nucleotide change that defines FCGR3A from FCGR3B. In contrast, the largest study used array comparative genomic hybridization (aCGH) to call copy number at this locus in the Wellcome Trust Case Control Consortium (WTCCC) and failed to find an association [Craddock et al., FCGR3A and FCGR3B, which would contribute to a lack of power to observe such an association.In this context, genetic association studies at this locus that involve genotyping copy number, sequence variants, or both, can often be unreliable due to a combination of technical variation due to noisy assays measuring copy‐number variation and biological variation because of the underlying complexity of the locus [Haridan et al., FCGR3A deletions from FCGR3B deletions in aCGH data. This allows us to reanalyze aCGH data from RA cases and healthy controls, and test the association of the FCGR3B deletion with RA.In this study, we develop our understanding of the variation within this region by reanalyzing the fosmid sequences previously generated in the light of published findings [Machado et al., Four‐hundred eighty British samples were obtained from the Human Random Control series . Native American samples were obtained from the Ashaninka population from five villages along the Junin River in Central Peru or from the Matsiguenga population living in the community of Shimaa, Peru. The Ashaninkas and Matsiguengas live in the Amazon Yunga tropical forest, and speak languages belonging to the Arawak family. DNA collection was collected under informed consent and approved by local ethics institutional review boards. WTCCC data were provided courtesy of the WTCCC Access Committee and Dr. Matthew Hurles (Wellcome Trust Sanger Institute). Full details of the cohorts have been published previously [Craddock et al., For each fosmid, the reads were mapped to the cloning vector sequence (cloning vector pEPIFOS‐5) using version 0.7.6 of the SMALT aligner. Reads mapping to the vector with an alignment score of less than 30 were retained, and the rest were discarded as it was assumed they were contaminated with vector sequence. Filtered reads were assembled using version 2.9 of the GS De Novo Assembler (newbler). The assembled contigs labeled as large by the assembler were retained. Filtered reads were mapped to these contigs, and the consensus algorithm of GAP5 version 2.0.0b10 [Bonfield and Whitwham, FCGR3 typing using a PRT/REDVR approach, as described previously [Hollox et al., Following Quantile‐normalized log ratio data from Agilent 2×105K aCGH experiments were used [Conrad et al., Results) was conducted using CNV tools on RA and control individuals showing heterozygous deletion, and a Gaussian mixture model run with two components to attempt to stratify FCGR3B deletions and FCGR3A deletions on the basis of values on the first PC, and case/control status included as a factor in the mixture model to minimize differential bias in calling between cohorts. In all analyses, the values from each aCGH probe were weighted equally.PC analysis on a narrower set of aCGH probes , the reciprocal B to A gene conversion of 9.1 kb (NC_000001.9:g.159888461_159897571con159806619_159815757), and a smaller B to A gene conversion of 2.4 kb (NC_000001.9:g.159888397‐159890836con159806551_159808988). All three gene conversions occur between Although these gene‐conversion alleles may complicate assessment of breakpoint position in certain circumstances, the phylogenetic network Fig.  shows thFCGR3A deletion, FCGR3B deletion, or heterozygous for FGCR3A and FCGR3B deletions. We chose the British as one population, which is an outbred population at the edge of northwest Europe, for two reasons. First, we had not previously mapped a European deletion allele, yet most genetic association studies involving the FCGR3B deletion alleles are on European populations [McKinney and Merriman, Previously, mapping deletion breakpoints in a globally diverse selection of alleles suggested that different deletion breakpoints were distinct yet clustered into two major hotspot regions [Machado et al., FCGR3B deletion. Sequence analysis showed that both were heterozygous for the NAHR breakpoint, with one allele showing a breakpoint in region B, generating CNR1, and the other in a region distal to breakpoint B, generating CNR4 .Using PRT, we identified two UK samples that were homozygous for the NR4 Fig. ; Table 2FCGR3B deletion alleles  [Zuccherato, FCGR3B deletions, and one individual who carried both a FCGR3A deletion and a FCGR3B deletion. Using a PCR amplification and Sanger sequencing approach, we located all the deletion breakpoints from the FCGR3B deletion homozygotes to a 10‐kb region immediately distal to the gene‐conversion region .Using the approach above, we called heterozygous FCGR3B deletion (CO092 and UKTS8802), the breakpoints all map to the same regions as previously identified [Machado et al., FCGR3B deletion alleles has been reported [Zuccherato, FCGR3B deletion breakpoints were identical, showing that these deletions in these populations are identical by descent. Therefore, in outbred populations, a combination of identity‐by‐descent and identity‐by‐state is the norm. However, with increased levels of endogamy in the population, recurrent sampling of the identical‐by‐descent deletion alleles is more likely.Our data show that for the two UK samples homozygous for the FCGR3B deletion breakpoints. It is likely that the longer region of gene conversion is more likely to have been generated by a double crossover event, although long gene‐conversion tracts are known to exist [Chen et al., We also identify the reciprocal duplication breakpoint generated by NAHR within this region from analysis of the fosmid sequence, as predicted by the NAHR model. We found three gene conversion alleles within the region that contains the FCGR3B deletion allele has been shown to be associated with RA in several studies. In some studies, the association is with all forms of RA [McKinney et al., FCGR3B copy number and/or small sample sizes that are likely to be underpowered. In this study, we replicate an association of the FCGR3B deletion allele with RA  in a large cohort of 1,982 cases and 3,271 controls, which is three times the size of the largest previous study. Furthermore, an odds ratio of 1.61 (95% CI 1.16–2.24) is a size of effect consistent with previous studies.The FCGR3A and FCGR3B deletions using aCGH data is not absolutely clear, so there may be some misclassification of FCGR3A and FCGR3B deletions where signals are at the boundary of the two Gaussian curves in the mixture model. Second, as we have shown, FCGR3B deletion alleles can have different breakpoints in the UK population, and it is probable that structurally different alleles will have different effects on RA risk. Indeed, a previous study has suggested that the association of inflammatory disease with FCGR3B CN may not be due to the loss of FCGR3B as such, but rather the juxtaposition of FCGR2C regulatory elements next to a full FCGR2B coding sequence, causing ectopic expression of the inhibitory receptor FCGR2B on NK cells [Mueller et al., FCGR2C/B fusion genes, including a FCGR2B null variant [Nagelkerke et al., FCGR2B and FCGR2C are affected by gene‐conversion events, and the role of gene conversion in modulating variation in gene expression in deleted and nondeleted chromosomes remains to be determined.Nevertheless, this study has limitations. First, the distinction between FCGR3B deletion alleles [Hollox et al., Recent studies have shown that some structural variation can be imputed from flanking dense SNP haplotypes [Sekar et al., FCGR2B/FCGR2CSupplementary Figure 1 Deletion breakpoints, gene conversion regions and other features involving Supplementary table 1 8‐SNP Haplotype compositionSupplementary table 2 Frequency of 8‐SNP haplotypes in Human Genome Diversity project populationsFCGR3B deletionSupplementary table 3. Haplotypes of individuals homozygous for FCGR3A deletion and RASupplementary table 4 Association analysis of Click here for additional data file.


Columnar supramolecular phases with polarization along the columnar axis have potential for the development of ultrahigh‐density memories as every single column might function as a memory element. By investigating structure and disorder for four columnar benzene‐1,3,5‐trisamides by total X‐ray scattering and DFT calculations, we demonstrate that the column orientation, and thus the columnar dipole moment, is receptive to geometric frustration if the columns aggregate in a hexagonal rod packing. The frustration suppresses conventional antiferroelectric order and heightens the sensitivity towards collective intercolumnar packing effects. The latter finding allows for the building up of mesoscale domains with spontaneous polarization. Our results suggest how the complex interplay between steric and electrostatic interactions is influenced by a straightforward chemical design of the molecular synthons to create spontaneous polarization and to adjust mesoscale domain size. Organic materials featuring ferroelectric polarization−2 was obtained by columnar stacking of a phthalonitrile derivative.−2 for thin films of oriented LCsanti alignment of the columnar polarization, resulting in non‐polar phases.Recently, the first example of an intrinsically ferroelectric, axially polar LC with a remnant polarization of 1.7 μC cmanti alignment of the macrodipoles ≈39.5° (1) and 34.6° (3) compared to the N‐centered systems with Φ(CarCarCOO)≈35.1° (2) and 23.7° (4). Larger torsion angles cause a steeper inclination of the C=O bonds towards the columnar axis and will thus lead to larger macrodipoles.Based on this model, compounds 4 Figure  a were s4 Figure  b, whichp representative for the columnar dipole for the smaller molecules 1, 2, and 4 by quantum‐chemical calculations on finite clusters taken from the single‐crystal structure solutions (Section S3). In line with the structural considerations, p amounts to 12 D and 11 D for 1 and 2, respectively, and to 6.5 D for 4. For the latter, the p value is reduced further because of the anti alignment of the C−F and C=O bonds, which is induced by intramolecular NH⋅⋅⋅F hydrogen bonds , closely followed by 2  and a significant reduction for 3  owing to the larger dcc value and 4  based on the smaller p value.To derive a more quantitative picture of the macrodipole strengths, we estimated the average molecular dipole moments 1,2, and 3, each stack appears as a superposition of two columns with opposite macrodipole orientations in the conventional structure solution  and next‐nearest neighbor (n.n.n.) columns.J1and J2are subsequently varied until the best match between the simulated and experimental diffraction patterns  for 4. Furthermore, the excellent match between the observed and simulated X‐ray powder diffraction data  demonstrates that both the disorder and the mesoscale domain formation are inherent properties of the bulk materials. Although domains with spontaneous polarization have been suggested for axially polar phases before,The resulting arrangements of up‐ and down‐oriented macrodipoles Figure  c reprodarger 50–0 nm for 1 and 2), both J1and J2are positive, indicating that the electrostatic interaction, and thus an antiferroelectric alignment between n.n. and n.n.n. columns, dominates. This inevitably leads to disorder for up‐ and down‐oriented columns on local length scales and non‐polar unit cells  should be addressed to the authors.SupplementaryClick here for additional data file.


Accurately determined and isotopically distinct source values are essential for this process. The δ18OPO4 values of animal wastes have, up to now, received little attention.Phosphorus losses from agriculture pose an environmental threat to watercourses. A new approach using the stable oxygen isotope ratio of oxygen in phosphate (δ4) was extracted from cattle faeces using anion resins and the contribution of microbial PO4 was assessed. The δ18OPO4 value of the extracted PO4 was measured by precipitating silver phosphate and subsequent analysis on a thermal conversion elemental analyser at 1400°C, with the resultant carbon monoxide being mixed with a helium carrier gas passed through a gas chromatography (GC) column into a mass spectrometer. Faecal water oxygen isotope ratios  were determined on a dual‐inlet mass spectrometer through a process of headspace carbon dioxide equilibration with water samples.Phosphate  values ranged between +17.9 and +19.9‰, while using groundwater δ18OH2O values gave a range of +13.1 to +14.0‰. The faecal δ18OPO4 values ranged between +13.2 and +15.3‰.Microbiological results indicated that much of the extracted PO18OPO4 values were equivalent to those reported elsewhere for agricultural animal slurry. However, they were different from the Eδ18OPO4 value calculated from the faecal δ18OH2O value. Our results indicate that slurry PO4 is, in the main, derived from animal faeces although an explanation for the observed value range could not be determined.The fresh faecal δ It is fundamental to many biological processes because it is involved in energy transfer and is the constituent of several organic molecules.43−), hydrogen phosphate (HPO42−) and dihydrogen phosphate (H2PO4−) which can collectively be termed 'phosphate' (subsequently referred to as PO4 in the manuscript). This new approach uses the ratio between the 18O and 16O in PO4  to understand better P sources and transformations. Comprehensive reviews have been written by Davis et al4 are stable. Therefore, bonds are only broken through biological mediation, and in these cases PO4 exchanges O with the ambient water within which it is in solution.4 ions incorporating one O atom from the ambient H2O. This process is extremely fast and leads to a complete O exchange between H2O and PO4 over time because PO4 as well as pyrophosphate can bind at the active site of pyrophosphatase.18OPO4) which is predictable and initially described by Longinelli and Nuti18OPO4 is the stable O isotope ratio of PO4 at equilibrium in ‰, T is the temperature in degrees Celsius and δ18OH2O is the stable oxygen isotope ratio of water in ‰.Stable isotope ratios have been used to track elements during transfers between different pools and to understand the respective roles of abiotic and biotic processes during these transfers.4, the following criteria should be met:18OPO4 values for significant PO4 sources are well characterised The δ4 possess distinct δ18OPO4 signaturesThe individual sources of PO18OPO4 values for PO4 sources are not equal to the Eδ18OPO4 valuesThe δ18OPO4 signatures for PO4 sources are maintained and not rapidly transformed or modified by fractionation caused by metabolic processes.The δFor effective use of this approach for tracing the sources of PO18OPO4 values that most PO4 sources have and that they often overlap or they are similar to the Eδ18OPO4 value.18OPO4 value for water‐extractable PO4 despite its heterogenous composition. Furthermore, this study reported that its value was noticeably lower than the Eδ18OPO4 value in the rivers. Granger et al4 was probably animal faeces, the δ18OPO4 value probably reflected the Eδ18OPO4 value of PO4 within the animal due to high microbial turnover, and that the Eδ18OPO4 value was strongly influenced by the higher body temperature relative to the ambient water temperature in the aquatic environment receiving the slurry.One of the confounding issues surrounding this area of research is the narrow range of δ18OPO4 value, to see how consistent this value was, and whether it was similar both to the values of animal slurry already measured and to the calculated Eδ18OPO4 value for the animal. The forms of P in animal faeces can be split into three broad categories. Toor et al18OPO4 values – (iii) the microbial P. For the purposes of this study, we did not attempt to examine the δ18OPO4 values of organic forms of P, but, instead, aimed to characterise the inorganic 'free' PO4, and the 'microbial' PO4 of cattle faeces. There is no reported method for doing this in animal faeces so we attempted to apply and adapt an approach used for soils to test the following hypothesis: The δ18OPO4 value of inorganic 'free' PO4 and the 'microbial' PO4 will be the same and will reflect the Eδ18OPO4 value calculated for fresh cattle faeces.In the present study, we sought to analyse fresh cattle faeces to establish its δ22.1The details of the animals sampled are presented in Table 18OH2O value. Secondly, a 1 g faeces sub‐sample for microbial analysis was placed in a 25‐mL polystyrene screw‐capped container , diluted with 9 mL of Ringer's solution, , and stored at 4°C for analysis within 24 h. Thirdly, a 20–30 g sub‐sample was taken, placed in a pre‐weighed foil tray, weighed, and then dried to a constant weight at 105°C overnight to determine dry matter (DM) content.Animals were not preselected for the study; simply, the first animal to defecate was selected. The animal ID number was noted and about 150 g of faeces was collected from the ground using sterile containers. Samples of fresh faeces were collected directly after being voided onto the soil surface in clean aluminium containers and returned immediately to the laboratory for sub‐sampling and preparation. First, a sub‐sample of 2–3 g faeces was placed into a 12‐mL glass exetainer, sealed and frozen at −20°C, ready for determination of its δ2.24 were based on extraction methods described for soils;4 and 'microbial' PO4, respectively. Tests using faeces found that there was no difference in the amounts of PO4 recovered from faeces with, or without, hexanol (results not presented). This suggested that either there was no microbiological content within the faeces, or that hexanol did not lyse the cells. As it seemed unlikely that there would be no faecal microbial content, it was hypothesised that osmotic stress was causing the lysis of most of the microbial cells present and therefore the addition of hexanol would not further increase the amount of extractable PO4. This hypothesis was based on the standard practice of microbiologists in using a buffered solution when extracting gut microbiology for culture.The method development experiments for distinguishing inorganic and microbial PO4. A PO4 spike was added to a container of Ringer's solution into which anion resins were placed. After a 16‐h shaking period, it was found that PO4 recovery was unaffected by the Ringer's solution (results not shown) and on this basis the study was continued.Ringer's solution contains mainly anions, to prevent the osmotic stress of the microbiology, so a recovery test was undertaken to see if it would adversely affect the ability of the anion resins to collect PO2.2.1Escherichia coli, a faecal indicator organism. The sample to be tested was diluted through serial dilutions to obtain a small number of colonies on each agar plate; 0.1 mL of the diluted sample was spread on the surface of a Membrane Lactose Glucuronide Agar (MLGA) (Oxoid) plate. Samples were initially vortex mixed before appropriate serial dilutions, from which 0.1 mL was spread plated aseptically. Once plates were dry, they were incubated at 44.0°C (±0.5°C) for between 18 and 24 h. After the total incubation period, all plates were examined and plates with between 30 and 300 colonies were counted.Determination of the number of bacteria was undertaken using the standard plate count method for 2.32.3.14; (i) Resin PO4: 25–100 g placed in a 5‐L HDPE sealable bottle, diluted with 3 L Ringer's solution, and 72 anion‐exchange resin  squares (4 cm × 4 cm) added; and (ii) Microbial PO4: 1–2 g placed in a 5‐L HDPE bottle and diluted with 3 L deionised water, and 72 anion‐exchange resin squares added. The bottles were placed on an orbital shaker set at 100 rpm, in a 4°C walk‐in refrigerator. After 16 h, the bottles were removed and the extracting solution sub‐sampled for microbial analysis by diluting 1 mL of extractant solution in 9 mL Ringer's solution and stored at 4°C before analysis within 24 h. Resins were then recovered by pouring the extraction solution from the 5‐L bottle though a 4 mm sieve ensuring that all resins were recovered from the bottle. As the sample was highly organic in nature we felt it necessary to test and, if needed, account for any potential hydrolysis of organic P during the extraction of PO4 from the resins. Resins from each extraction were divided into two sub‐sets of 36, placed in a 250‐mL polypropylene screw‐capped bottle and washed several times with their respective, fresh, matrix solutions. When clean, PO4 was liberated from the resins using 75 mL of 0.2 M nitric acid (HNO3). For each of the two sub‐sets of 36 resins collected from a single extraction matrix, δ18OH2O unlabelled (−5.7‰) and labelled (+81.6‰) 0.2 M NHO3 was used to test for hydrolysis of organic P by the acid. The corrected δ18OPO4 value is then calculated using a revised version18OPO4 is the corrected final stable oxygen isotope ratio for PO4 considering the effect of any hydrolysis of organic P, δ18OPsp is the stable oxygen isotope ratio of the PO4 collected using 18O‐spiked HNO3, δ18OPus is the stable oxygen isotope ratio of the PO4 collected using unspiked HNO3, δ18OAus is the stable oxygen isotope ratio of the water in the unspiked HNO3, and δ18OAsp is the stable oxygen isotope ratio of water in the 18O‐spiked HNO3.Two further sub‐samples were extracted for PO3PO4) using the purification protocol described by Tamburini et al.4. The PO4 is precipitated first as ammonium phosphomolybdate before it is dissolved and reprecipitated as magnesium ammonium phosphate which is dissolved again. The resultant PO4 in solution is converted into Ag3PO4 through the addition of an Ag‐ammine solution which is then placed in an oven for 1 day at 50°C. Although the Tamburini protocol uses a DAX‐8 resin early in the extraction its use is not necessary unless organic contamination is present in the subsequent Ag3PO4 .Phosphate in the extracts was converted into silver phosphate , the sample containing side of which was immersed in liquid N2.42.4.1th) to avoid any acid interference with the molybdenum chemistry.Phosphate concentrations were determined colourimetrically on an Aquachem 250 analyser  using a molybdenum blue reaction2.4.2418O/16O ratio was undertaken by weighing approximately 300 μg of Ag3PO4 into a silver capsule to which a small amount of fine glassy carbon powder was added.18OPO4 values were calculated by comparison with an internal Ag3PO4 laboratory standard, ALFA‐1 . In the absence of an international Ag3PO4 reference material, we derived this value for ALFA‐1 by comparison with the Ag3PO4 standard 'B2207' , which has been measured in an inter‐laboratory comparison study to have a δ18O value of +21.7‰ versus VSMOW. Samples were run in triplicate, with a typical precision σ ≤0.3‰. Sample purity was assessed by determining the CO yield compared with the yield of Ag3PO4 standards, and samples were rejected where this differed by 10%.Measurement of the PO18OH2O values were determined on an Isoprime Aquaprep coupled to an Isoprime 100 dual‐inlet isotope ratio mass spectrometer  through a process of headspace CO2 equilibration with water samples. The isotope ratios are reported as δ18OH2O values versus VSMOW, based on comparison with laboratory standards calibrated against IAEA standards VSMOW and SLAP, with analytical precision typically σ ≤0.05‰.Faeces water δ2.5All statistical analyses were conducted in R.33.118OH2O values ranged between −1.19 and +0.41‰ with a mean of −0.73‰ (±0.65) (Table 4 collected from faeces when using Ringer's solution ranged from 67 to 93 μg PO4‐P g−1 DM with a mean of 78 (±9.1) μg PO4‐P g−1 DM. This was found to be significantly less  than that collected using deionised water which ranged from 3885 to 8635 μg PO4‐P g−1 DM with a mean of 5713 (±1856) μg PO4‐P g−1 DM.The fresh faeces were found to have a DM ranging from 9.3 to 16.6% with a mean of 11.4% ±2.5) while the δ.5 while 3.2E. coli concentrations ranging from 6.1 to 7.85 CFU g−1 DM  or deionised water . It was concluded therefore that there was no need to correct data using the equation described by McLaughlin et al.18OPO4 values. The δ18OPO4 values for the PO4 extracted from faeces are presented in Table 18OPO4 values for PO4 extracted using Ringer's solution for the first three samples are not presented as the amount of some of them was too small for standard Ag3PO4 precipitation. Of the remaining four faecal samples the values ranged from +12.0 to +19.8‰ with mean values between +12.1 and +16.3‰. The values for the seven samples extracted in deionised water ranged from +12.9 to +15.6‰ with mean values of +13.2 and +15.3‰. The greatest variation between labelled and unlabelled acid δ18OPO4 elution values occurred in the Ringer's solution dataset with the mean difference of the labelled acid extraction being +2.1‰. This result, however, was strongly influenced by one anomalously high labelled acid δ18OPO4 value of +19.8‰, leading to a difference of +6.9‰. This sample also had a slightly higher oxygen yield indicating that it was not pure Ag3PO4 which could explain the relatively high difference between the δ18OPO4 values of labelled and unlabelled acid extraction. The differences observed in the deionised water labelled and unlabelled acid elution were far smaller and ranged between −1.8 and +1.4‰ with a mean of −0.3‰. Statistical analysis of the two sets of paired data shows that there was no difference between the δ18OPO4 values obtained following extraction using Ringer's solution and that using deionised water .To assess whether organic P had been hydrolysed by the 0.2 M HNO44.1E. coli reported here are consistent with those reported in the literature for beef cattle faeces.4 in both cases does not represent 'microbial' PO4 released through cellular breakdown during the extraction process but, instead, 'free' PO4.The concentrations of 4.24 extracted in deionised water were significantly higher than in Ringer's solution. This finding is at odds with the initial recovery test undertaken on PO4 in a pure Ringer's solution matrix. However, it would seem that the combination of organic material, faecal anions, and the anions within the solution itself significantly reduced the recovery of PO4 on the resins in a way that did not occur in just the Ringer's solution alone. This interference raises questions about the validity of the δ18OPO4 values of PO4 recovered in this solution due to potential unknown fractionations that might occur as a result of preferential adsorption/desorption of the lighter/heavier isotopologues.4 from inorganic PO4 (as defined earlier) requires further development.The amounts of PO4.318OH2O value of the groundwater is relatively stable and will represent an integrated value of the annual precipitation supplying it. At this location, the δ18OH2O value is predicted to be between −5.5 and −6.0‰.18OH2O value due to evaporative losses as important. Abeni et al18OH2O values did not differ greatly despite the increased temperatures. Water is also ingested as metabolic water in food, which is likely to be isotopically heavier than local meteoric water due to fractionation;18OH2O values of various forms of body water in cattle were from 4.2 to 7.9‰ heavier than in drinking water in the summer and that for faecal water they were from 4.8 to 7.7‰ heavier. The measured δ18OH2O value in faeces in this study was found to be up to 6.4‰ heavier than in groundwater and this was not unexpected as demonstrated by the model proposed by Bryant and Froelich.18OH2O values while water lost via pathways such as urine, faeces and sweat will be similar and thus have similar δ18OH2O values to that of the animal's body water. The increase in δ18OH2O value will also be more pronounced in the summer when temperatures are higher.The fresh faeces %DM values are consistent with those reported elsewhere for cattle grazing pasture.4.418OPO4 values is widespread within the δ18OPO4 community to benchmark measured values with values that have potentially lost their original signal through intracellular cycling, specifically through the enzyme pyrophosphatase. However, there is much uncertainty as to how relevant this theoretical equilibrium is in many situations, and we acknowledge that in terms of animal gut processes other cycling pathways may predominate.The use of Eδ18OH2O values measured in faeces and with the range expected for the ground/drinking water in the region, a Eδ18OPO4 range of values from +13.2 to +14.0‰ is expected, assuming that the body water δ18OH2O value is similar to that of ground water and +18.1 to +19.9‰ if the δ18OH2O values within faeces are used and are taken to represent the animal body water (Figure The normal temperature of cattle is 38.6°C, with anything outside a range of 38.0 to 39.2°C indicating ill health.4.54 was not derived directly from the lysis of microbial cells, it was not possible to compare 'free' PO4 with 'microbial' PO4. However, the δ18OPO4 values of the 'free' PO4 ranged between +13.2 and +15.3‰ which are very similar to those reported for slurry PO4 by Granger et al18OPO4 values and the animal variables; however, the scope of the study was too limited to investigate variables such as age, gender, breed, etc. The δ18OPO4 values reported within this study indicate that the slurry δ18OPO4 values are caused by the PO4 in animal faeces. The δ18OPO4 values of the faeces themselves, however, are at or slightly above the range of Eδ18OPO4 values based on the ground/drinking water δ18OH2O values. However, all the values are at least 2.8‰ lower that the Eδ18OPO4 value range calculated from the δ18OH2O value of faecal water, water that should be far more representative of the body water of the animal.As it was shown that the resin‐extractable PO54 from fresh cattle faeces was lower using Ringer's solution than deionised water. However, this did not appear to be because of microbial cellular lysis in the deionised water extraction. It would appear to be due to some form of interference between the Ringer's solution ions, compounds in the faeces and the anion resin sheets. Because of this it was not possible to differentiate 'microbial' PO4 and 'free' PO4, and their respective δ18OPO4 values. As it has been shown that deionised water does not lyse the microbial cells it would be worth repeating the study using the more traditional resin PO4 extraction in a water/hexanol extraction solution to extract 'microbial' PO4 and to also use the microbial assays described to establish if this occurs.The extractable PO18OPO4 values of fresh cattle faeces, under the conditions reported in this study, ranged between +13.2 and +15.3‰ which are consistent with those reported elsewhere for agricultural animal slurry.The δ18OPO4 values are similar to the Eδ18OPO4 value calculated for within the animal using the δ18OH2O value of groundwater. However, they are at least 2.8‰ lower than the Eδ18OPO4 value range calculated using faecal water as a proxy for the animals' body water.The δ18OPO4 value. However, to examine these, a more detailed study is required which should also include other animals for which few data exist in the literature.There were no apparent relationships between the animal variables and the δ


Cyclic multiredox centered systems are currently of great interest, with new compounds being reported and developments made in understanding their behavior. Efficient, elegant, and high‐yielding (for macrocyclic species) synthetic routes to two novel alkynyl‐conjugated multiple ferrocene‐ and biferrocene‐containing cyclic compounds are presented. The electronic interactions between the individual ferrocene units have been investigated through electrochemistry, spectroelectrochemistry, density functional theory (DFT), and crystallography to understand the effect of cyclization on the electronic properties and structure. Cyclic structures have enthralling beauty that has attracted chemists for generations.Communication between redox‐active centers is highly studied, with the identification of useful bridging ligands, binding groups, and suitable redox‐active components, furthermore establishing a solid understanding in redox center communication and electronic coupling in mixed valence states.tBu3)2 as the catalyst.tBu3 ligand.1 over polymeric side products were achieved through the use of a 10:1 excess of 1,1′‐diiodoferrocene to alkyne. Compound 1 was purified by column chromatography, also allowing the retrieval of the excess of 1,1′‐diiodoferrocene .Cyclic compounds can be synthesized by several different methods. We devised a route to the triferrocene cyclic system through a stepwise assembly to a covalently bonded system Scheme . 1,1′‐Di2, the linear system was cyclized using an Ullmann‐like coupling mediated with copper(I) thiophene‐2‐carboxylate (CuTC). The desired product was achieved by using high‐dilution conditions, a two‐day reaction time, and isolated in a yield of 25 % after chromatographic purification. The product has been identified by NMR, EA, mass spectrometry, and X‐ray crystallography (see the Supporting Information).To form the macrocyclic product 2, a symmetrical molecule with two sets of the biferrocene motif (compound 7) was proposed. After several attempts at different synthetic routes, the procedure shown in Scheme 2. Each intermediate and the final product was isolated through either column chromatography or recrystallization and fully characterized (see the Supporting Information) but compounds 5–7 were only sparingly soluble.Following the success with the synthesis of 1H NMR spectra of the cyclic products 2 and 7 ; the first and third are associated with the singular ferrocene while the second, fourth, fifth, and sixth are related to the biferrocene unit. Owing to the symmetry of 7, all Cp−H peaks are related to the equivalent ferrocenes. This was confirmed by a 2D NMR spectrum. Conventional 1H NMR spectra are displayed and annotated in Figure The 2 °, Fe⋅⋅⋅Fe separation 5.0752(6) Å). This linkage also has a noticeable fold deformation, the centroid of one Cp ring lying circa 0.24 Å out of the plane of the other. The Fe1⋅⋅⋅Fe2 and Fe1⋅⋅⋅Fe3 separations across the macrocycle are 10.6097(7) and 12.2662(7) Å respectively.The crystal structure of 7, electrochemical investigations did not provide conclusive results; for example, at room temperature and in a range of solvents, the voltammograms contained additional features, which are likely related to solubility issues. Running the voltammograms at 60 °C in THF indicated two distinct redox transitions  but degradation was observed at high potentials. The electrochemical properties were thoroughly examined for 2, through cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in CH2Cl2, using 0.1 m [nBu4N][PF6] as the supporting electrolyte. As shown in Figure E1/2) and peak splittings (ΔE) for each redox event are shown in Table E=59 mV and a peak current ratio of the reverse to the forward peak ipc/ipa=1 are expected.Both systems contain multiple redox‐active centers that could communicate through bond or space and may, on oxidation to a mixed valence state, exhibit electronic interactions between the different redox sites that are specific to the cyclic structure. Owing to the low solubility of −1. For both of these peaks, ip scales linearly with v1/2 . Combined with the data in Table v=0.04 V s−1) at high potential (above the 2+/3+ transition). Additionally, CVs were run in [nBu4N][BArF4] solution phenyl]borate). Owing to the weak ion pairing capabilities of BArF4 and increased electrostatic interactions between the redox sites, half‐wave potential splittings were significantly enhanced.Owing to the proximity of each of the individual redox events, we found it difficult to reliably define a baseline for some of the redox transitions. However, this is straightforward for the first cathodic (3+/2+ transition) and the first anodic wave (0/1+ transition). The peak current ratio, as defined above, then amounts to 0.97. It was also calculated that the sum of all cathodic peak currents divided by the sum of all anodic peak currents is 1.01 at 0.1 V s2 with those of 1,1′′′′‐bis(ethynylphenyl)biferrocene 8 and of the second wave to that of 1,1′′′‐bis(phenylethynyl)ferrocene 9  suggest the corresponding sequence of redox events that is, that the 2nd oxidation of 2 is linked to the singular ferrocene and the first and third correspond to the biferrocene unit.The close proximity to the first and third redox waves of 2, +2, 2+2, and 3+2 in the UV/Vis/NIR range. The mono‐ and di‐cations were generated by chemically oxidizing 2 with one or a slight excess over 2 equiv of acetylferrocenium hexafluorophosphate (E1/2 =0.27 V), respectively, while 3+2 was produced by oxidation of 2 with three equivalents of Ag+[SbF6]−. Their vis/NIR spectra are shown in Figure One indicator of such electronic communication are electronic absorption bands at low energy, typically in the near infrared (NIR), for the mixed‐valence (MV) forms that relate to the transfer of electron density from the reduced to the oxidized ferrocene subunits . Such bands were recently observed for the MV states of cyclic sexiferrocene.+2 and then to 2+2 induces a growth of a band at ca. 680 nm (14 700 cm−1), which is readily identified as the typical Cp→Fe charge transfer (CT) band of a ferrocenium ion.ν˜1/2≈4000 cm−1) and a sharper, less intense one at ca. 3175 nm (Δν˜1/2≈1250 cm−1). The observation of that band in +2 and 2+2 thus confirms our previous assignment of the first oxidation as based on the biferrocene and that of the second one as involving the remote monoferrocenyl site. More importantly, and despite of the interconnecting π‐conjugated pathway, we have no evidence for an additional IVCT transition between the remote ferrocenyl site and the FeIII of the biferrocenium cation in +2 or the reduced FeII site of the biferrocenyl and the remote ferrocenium site of 2+2. Tricationic 3+2, however, exhibits a rather intense and broad absoption envelope over the entire Vis/NIR and down to the mid‐IR range . The underlying excitations are likely of bridge/Cp‐to‐metal CT origin.Stepwise oxidation to 2 and its various oxidized forms by density functional theory (DFT) calculations. Contour plots of the calculated MOs and the respective MO contributions of every subunit as derived from natural bond order (NBO) analysis are compiled in the Supporting Information as Figures S20–S23. Our results indicate that the highest occupied molecular orbital (HOMO) and the HOMO−2 of 2 are largely centered on the biferrocene (bfc) moiety whereas the HOMO−1, LUMO, and LUMO+1 receive major contributions from the 1,3‐diethynylphenylene bridges along with minor contributions from the remote ferrocene moiety Fc3. Our calculations correctly predict that on the first oxidation the electron is removed from one ferrocenyl subunit (biFc1) of the biferrocene. This follows from the β‐LUSO and the calculated spin density map, as graphically shown in the Supporting Information, Figure S24 and listed in Table S1. Further MOs with major contributions of the bfc unit are found as the α‐LUSO and the LUSO+1 and LUSO+2 of the α‐ and β‐manifolds. Again in agreement with our experiment, the second electron is lost from the remote ferrocenyl site Fc3. Quite interestingly, the spin density at the bfc entity of 2+2 is computed to be equally distributed over both biFc sites as opposed to localization at only one Fe site (biFc1) in +2. This is not necessarily at odds with the experimentally observed near invariance of the IR/NIR spectra to the second oxidation. Biferrocenium cations as they are present in +2 and 2+2 are generally situated close to or at the borderline between fully delocalized or valence‐trapped mixed valence systems of Class III or II according to the Robin–Day classification.+ at the Cp−Cp linkage can be very subtle and do not cause detectable changes of the NIR spectrum.3+2 failed to converge.We sought for deeper insight into the electronic structures of 2 exhibits three reversible oxidations related to the bi/ferrocene components of the system. The biferrocene unit is able to electronically share charge and spin densities whereas the separated ferrocene moiety is virtually decoupled from its neighbors. Weak IVCT transitions for a biferrocenium radical cation were observed for +2 and 2+2 and confirm the ordering of redox events as proposed from the comparison of the voltammograms of the individual components of macrocyle 2. The cavity present in the crystal structure of 2 suggests that these molecules could also be accessible for small neutral and charged guests. Our bespoke synthetic methodology allows for further design of small, cyclic or in‐parallel multi‐metallic systems, to also feature surface‐ligating functionalities, and these investigations are currently underway.To conclude, two new multiferrocene macrocycles have been reported, through analogous stepwise Sonogashira and Ullmann‐like coupling methods. The tri‐ferrocene macrocycle The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re‐organized for online delivery, but are not copy‐edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file.