CH3 CN@open-C60 : An Effective Inner-Space Modification and Isotope Effect Inside a Nano-Sized Flask.

Despite several small molecules being encapsulated inside cage-opened fullerene derivatives, such species have not considerably affected the structures and properties of the outer carbon cages. Herein, we achieved an effective inner-space modification for an open-cage C60 derivative by insertion of a neutral CH3 CN molecule into the cavity. The CH3 CN@open-C60 thus obtained showed an enhanced polarity, thus affording an easy separation from a mixture containing the empty cage by column chromatography on silica gel, without the preparative HPLC that was needed for previous cases. The less negative reduction potentials with respect to those of empty cage reflect the decreased energy level of the LUMO, which is supported by the DFT calculations. NMR spectroscopy, single-crystal X-ray analysis, and theoretical calculations revealed that both the presence of the encapsulated CH3 CN and cage deformation caused by the CH3 CN play an essential role in the change of the electronic properties. Furthermore, the favored binding affinity of deuterated acetonitrile CD3 CN with internal C60 surface is discussed.

Chiral Calix[3]pyrrole Derivatives: Synthesis, Racemization Kinetics, and Ring Expansion to Calix[9]- and Calix[12]pyrrole Analogues.

Chiral pyrrolic macrocycles continue to attract interest. However, their molecular design remains challenging. Here, we report a calixpyrrole-based chiral macrocyclic system, calix[1]furan[1]pyrrole[1]thiophene (1), synthesized from an oligoketone. Macrocycle 1 adopts a partial cone conformation in the solid state, and undergoes racemization via ring inversion. Molecular dynamics simulations revealed that inversion of the thiophene is the rate determining step. Pyrrole N-methylation suppressed racemization and permitted chiral resolution. Enantioselective N-methylation also occurred in the presence of a chiral ammonium salt, although the stereoselectivity is modest. A unique feature of 1 is that it acts as a useful synthetic precursor to yield several calix[n]furan[n]pyrrole[n]thiophene products (n=2-4), including a calix[12]pyrrole analogue that to our knowledge constitutes the largest calix[n]pyrrole-like species to be structurally characterized.

An H2 O2 Molecule Stabilized inside Open-Cage C60 Derivatives by a Hydroxy Stopper.

An H2 O2 molecule was isolated inside hydroxylated open-cage fullerene derivatives by mixing an H2 O2 solution with a precursor molecule followed by reduction of one of carbonyl groups on its orifice. Depending on the reduction site, two structural isomers for H2 O2 @open-fullerenes were obtained. A high encapsulation ratio of 81 % was attained at low temperature. The structures of the peroxosolvate complexes thus obtained were studied by 1 H NMR spectroscopy, X-ray analysis, and DFT calculations, showing strong hydrogen bonding between the encapsulated H2 O2 and the hydroxy group located at the center of the orifice. This OH group was found to act as a kinetic stopper, and the formation of the hydrogen bonding caused thermodynamic stabilization of the H2 O2 molecule, both of which prevent its escape from the cage. One of the peroxosolvates was isolated by HPLC, affording H2 O2 @open-fullerene with 100 % encapsulation ratio, likely due to the intramolecular hydrogen-bonding interaction.

Strain-Induced Ring Expansion Reactions of Calix[3]pyrrole-Related Macrocycles.

The recent discovery of calix[3]pyrrole, a porphyrinogen-like tripyrrolic macrocycle, has provided an unprecedented strain-induced ring expansion reaction into calix[6]pyrrole. Here, we synthesized calix[n]furan[3-n]pyrrole (n=1∼3) macrocycles to investigate the reaction scope and mechanism of the ring expansion. Single crystal X-ray analysis and theoretical calculations revealed that macrocyclic ring strain increases as the number of inner NH sites increases. While calix[1]furan[2]pyrrole exhibited almost quantitative conversion into calix[2]furan[4]pyrrole within 5 minutes, less-strained calix[2]furan[1]pyrrole and calix[3]furan were inert. However, N-methylation of calix[2]furan[1]pyrrole induced a ring-expansion reaction that enabled the isolation of a linear reaction intermediate. The mechanism analysis revealed that the ring expansion consists of regioselective ring cleavage and subsequent cyclodimerization. This reaction was further utilized for synthesis of calix[6]-type macrocycles.

An Efficient Method of Observing Diatom Frustules via Digital Holographic Microscopy.

Herein, we propose a convenient method to enable pretreatment of target objects using digital holographic microscopy (DHM). As a test sample, we used diatom frustules (Nitzschia sp.) as the target objects. In the generally used sample preparation method, the frustule suspension is added dropwise onto a glass substrate or into a glass chamber. While our work confirms good observation of purified frustules using the typical sample preparation method, we also demonstrate a new procedure to observe unseparated structures of frustules prepared by baking them on a mica surface. The baked frustules on the mica surface were transferred to a glass chamber with 1% sodium dodecyl sulfate solution. In this manner, the unseparated structures of the diatom frustules were clearly observed. Furthermore, metal-coated frustules prepared by sputtering onto them on a mica surface were also clearly observed using the same procedure. Our method can be applied for the observation of any target object that is pretreated on a solid surface. We expect our proposed method to be a basis for establishing DHM techniques for microscopic observations of biomaterials.

Calix[3]pyrrole: A Missing Link in Porphyrin-Related Chemistry.

A long-standing question in porphyrin chemistry is why pyrrole monomers selectively form tetrapyrrolic macrocycles, whereas the corresponding tripyrrolic macrocycles are never observed. Calix[3]pyrrole, a tripyrrolic porphyrinogen-like macrocycle bearing three sp3-carbon linkages, is a missing link molecule that might hold the key to this enigma; however, it has remained elusive. Here we report the synthesis and strain-induced transformations of calix[3]pyrrole and its furan analogue, calix[3]furan. These macrocycles are readily accessed from cyclic oligoketones. Crystallographic and theoretical analyses reveal that these three-subunit systems possess the largest strain energy among known calix[n]-type macrocycles. The ring-strain triggers transformation of calix[3]pyrrole into first calix[6]pyrrole and then calix[4]pyrrole under porphyrin cyclization conditions. The present results help explain the absence of naturally occurring three-pyrrole macrocycles and the fact that they are not observed as products or intermediate during classic porphyrin syntheses.

Coordination-Induced Spin-State Switching of an Aminyl-Radical-Bridged Nickel(II) Porphyrin Dimer between Doublet and Sextet States.

A bis(NiII -porphyrinyl)aminyl radical with meso-C6 F5 groups was prepared as a spin-delocalized stable aminyl radical with a doublet spin state. Upon addition of pyridine, both NiII centers became hexacoordinated by accepting two axial pyridines, which triggered a spin-state change of the NiII centers from diamagnetic (S=0) to paramagnetic (S=1). The resulting high-spin NiII centers interact with the aminyl radical ferromagnetically to give rise to an overall sextet state (S=5/2). Importantly, this coordination-induced spin-state switching can be conducted in a reversible manner, in that washing of the high-spin radical with aqueous hydrochloric acid regenerates the original doublet radical in good yield.

Benzonorcorrole NiII Complexes: Enhancement of Paratropic Ring Current and Singlet Diradical Character by Benzo-Fusion.

Fused benzene rings to antiaromatic compounds generally improve their stability but attenuate their antiaromaticity. The opposite case is now reported. NiII benzonorcorroles were synthesized and the effect of benzo-fusion on the antiaromaticity was elucidated. The benzo-fusion resulted in significant decrease of the HOMO-LUMO gaps and enhancement of the paratropic ring current effect. Furthermore, the introduction of the benzo groups induced singlet diradical character in the antiaromatic porphyrinoid.

Imaging and Quantitative Analysis on the Etching of Diatom Frustules via Digital Holographic Microscopy.

Frustules, whose length spans from a few micrometers to more than a hundred micrometers, have been the subject of various modifications to improve their physical properties because of their complex porous silica structure. However, three-dimensional measurements of these changes can be challenging because of the complex 3D architecture and limitations of known methods. In this study, we present a new method that applies digital holographic microscopy (DHM) to analyze controlled etched frustules and observe real-time degradation of frustules at the single-cell level. Frustules obtained from Craspedostauros sp. diatoms were etched in 1 N NaOH for 5 min at 25 and 60 °C, respectively, and the frustule's valve was analyzed using DHM. DHM uses a combination of holography and tomography to reconstruct a 3D refractive index image of the frustule. Measurements of the width, volume, and surface area are achieved. Results showed that at 60 °C of etching, a significant difference with the unetched frustule was observed for all measurements but with high fluctuation values. Finally, real-time observation of the degradation of the frustule is observed when immersed in a high concentration of NaOH. This is the first time the real-time etching of the frustule is observed at the single-cell level. This research provides an easy estimation of the 3D measurements of frustules that may provide new fundamental information and applications.

Distinguishing Antioxidant Molecules with Near-Infrared Photoluminescence of DNA-Wrapped Single-Walled Carbon Nanotubes.

In this study, two biomolecule solutions were distinguished using the capacity difference in the near-infrared photoluminescence (PL) of single-walled carbon nanotubes (SWNTs). Biosensing techniques using sensitive responses of SWNTs have been intensively studied. When a small amount of an oxidant or reductant solution was injected into the SWNT suspensions, the PL intensity of the SWNTs is significantly changed. However, distinguishing between different molecules remains challenging. In this study, we comparably injected saponin and banana solutions, which are known antioxidant chemicals, into an SWNT suspension. The SWNTs were solubilized by wrapping them with DNA molecules. The results show that 69.1 and 155.2% increases of PL intensities of SWNTs were observed after injection of 20 and 59 µg/mL saponin solutions, respectively. Subsequently, the increase in PL was saturated. With the banana solution, 18.1 and 175.4% increases in PL intensities were observed with 20 and 59 µg/mL banana solutions, respectively. Based on these results, the two antioxidant molecules could be distinguished based on the different PL responses of the SWNTs. In addition, the much higher saturated PL intensities observed with the banana solution suggests that the banana solution increased the capacity of the PL increase for the same SWNT suspension. These results provide helpful information for establishing biosensing applications of SWNTs, particularly for distinguishing chemicals.

Alkali metal ion binding using cyclic polyketones.

Cyclic oligoketones composed of 3,3-dimethylpentane-2,4-diones showed crown ether-like alkali metal ion binding behavior with association constants up to 1.7 × 104 M-1 in chloroform/acetonitrile (v/v, 9/1). The binding properties have been used for catalysis in the Finkelstein reaction in a low-polarity solvent. Furthermore, novel ion-binding hosts were generated by terminal functionalization of linear polyketones.

Determination of the critical chain length for macromolecular crystallization using structurally flexible polyketones.

Critical chain length that divides small molecule crystallization from macromolecular crystallization is an important index in macro-organic chemistry to predict chain-length dependent properties of oligomers and polymers. However, extensive research on crystallization behavior of individual oligomers has been inhibited by difficulties in their synthesis and crystallization. Here, we report on the determination of critical chain length of macromolecular crystallization for structurally flexible polyketones consisting of 3,3-dimethylpentane-2,4-dione. Discrete polyketone oligomers were synthesized via stepwise elongation up to 20-mer. Powder and single crystal X-ray diffraction showed that the critical chain length for polyketones existed at an unexpectedly short chain length, 5-mer. While shorter oligomers adopted unique conformations and packing structures in the solid state, higher oligomers longer than 4-mer produced helical conformations and similar crystal packing. The critical chain length helped with understanding the inexplicable changes in melting point in the shorter chain length region resulting from chain conformations and packing styles.

Reversible Redox System of 2-Oxypyritriphyrin(1.2.1) Accompanying Interconversion between 3-Pyridone and 3-Hydroxypyridine Units.

To develop a system in which a π-conjugation circuit switched by a redox reaction between 3-pyridone and 3-hydroxypyridine, a ring-contracted analog of oxypyriporphyrin, 2-oxypyritriphyrin(1.2.1) was synthesized for the first time. 2-Oxypyritriphyrin(1.2.1) contains a 14π aromatic conjugation circuit with the keto-form of the 3-oxypyridine ring. When 2-oxypyritriphyrin was treated with NaBH4 , not only the 3-oxypyridine group, but also the meso-carbons were also reduced to give a colorless porphyrinogen analog. The reduced compound could be oxidized again to provide 2-oxypyritripyhrin in a reversible manner.

An Arabidopsis hydrophilic Ca2(+) -binding protein with a PEVK-rich domain, PCaP2, is associated with the plasma membrane and interacts with calmodulin and phosphatidylinositol phosphates.

We found a new hydrophilic protein in Arabidopsis thaliana. Real-time PCR demonstrated that the protein was expressed in roots. Histochemical analysis of promoter-beta-glucuronidase fusions demonstrated its extensive expression in root hairs. The protein is rich in proline, glutamate, valine and lysine residues (PEVK-rich domain), and bound Ca(2+) even in the presence of Mg(2+) and K(+) when examined by the (45)Ca overlay assay. Treatment of seedlings with K(+), Mn(2+), Zn(2+), Na(+), ABA and gibberellic acid, and cold and drought stresses enhanced the transcription. Expression of the protein linked to green fluorescent protein in A. thaliana showed its plasma membrane localization and cell-specific expression in the epidermal cells including root hairs and the elongating pollen tubes. Therefore, we named the protein PCaP2 (plasma membrane-associated Ca(2+)-binding protein-2). The substitution of glycine at position 2 with alanine resulted in cytoplasmic localization of PCaP2. These results and the N-terminal characteristic motif suggest that PCaP2 is N-myristoylated at Gly2. We examined the capacity for binding to phosphatidylinositol phosphates (PtdInsPs), and found that PCaP2 interacts strongly with PtdIns(3,5)P(2), PtdIns(4,5)P(2) and PtdIns(3,4,5)P(3), and weakly with PtdIns(3,4)P(2). Furthermore, calmodulin was associated with PCaP2 in a Ca(2+)-dependent manner, and its association weakened the interaction of PCaP2 with PtdInsPs. These results indicate that PCaP2 is involved in intracellular signaling through interaction with PtdInsPs and calmodulin in growing root hairs. PCaP2 was previously reported as microtubule-associated protein-18. We discuss the physiological roles of PCaP2 in relation to microtubules in cells.

Label-free imaging and analysis of subcellular parts of a living diatom cylindrotheca sp. using optical diffraction tomography.

Optical diffraction tomography is an emerging label-free microscopic technique with its capability of label-free, quantitative, and rapid imaging of biological samples. In this work, we present the imaging and analysis of a living diatom Cylindrotheca sp. in seawater without using any pretreatment such as fluorescence staining. The 3D refractive index (RI) of a living diatom cell was measured, to which quantitative image analysis was perform to investigate subcellular parts of the diatom. Each part of the cell was well distinguished as RI values and distributions. From the analysis, RI values of frustules, protoplasm, vacuole, and chloroplast were estimated to be in the range of 1.352-1.388, 1.363-1.381, 1.388-1.395, and 1.403-1.436, respectively. Our results suggest that the present method will be a powerful tool not only for observing diatom cells but also for studying various cells and mesoscopic materials.•Subcellular parts of a living diatom cell was well visualized by digital holographic microscope.•Subcellular parts could be identified as differences of refractive indexes.•The observation was achieved without any pre-treatment of the living cell.

Aliphatic polyketones as classic yet new molecular ropes for structural diversity in organic synthesis.

Polyketone compounds play an important role in organic chemistry as a prominent source of reactivity and functionality. Their chemical properties vary widely depending on the ketone sequence in carbon chains. Although vicinal and ß-polyketones have been used for more than 150 years, the recent development of new ketone sequences composed of alternating 1,3- and 1,4-diketones has demonstrated the further potential of polyketones as structurally flexible molecular ropes capable of derivatization to π-conjugated chromophores and molecular assemblies. In this Feature Article, we review the synthetic strategies and reactivities of polyketones with respect to their ketone sequence. Furthermore, recent research on polyketones with hybrid ketone sequences is also discussed, focussing on their structural diversity in chemical transformations including intramolecular cyclization and stereoselective oxidation.

Corrigendum to "Label-free imaging and analysis of subcellular parts of a living diatom cylindrotheca sp. using optical diffraction tomography" [MethodsX: 7 (2020) 100889].

[This corrects the article DOI: 10.1016/j.mex.2020.100889.].

Unique observation method of temperature dependence of diatom floating by direct microscope.

Diatoms are one of the earth's major oxygen producers. For that reason, studying the floating phenomena of living diatom cells in water is an important research subject. Efficiency of photosynthesis of diatom cells may be heavily affected by their floating behavior. In our previous research, we devised a 'tumbled' microscope, a device created by tilting an inverted microscope (CKX53, OLYMPUS) by 90 degrees, due to which allowed observation with a sample stage perpendicular to the ground. When we observed a Petri dish filled with diatom cell suspension, the floating behavior of diatom cells were well visualized. Cyclotella meneghiniana was isolated and subcultured in bold modified basal freshwater nutrient solution liquid medium (B5282-500ML, Sigma-Aldrich) at 18 °C. Before the microscopic observation, cell suspension was cultured for two weeks after the final subculture. Observation was performed at room temperature, 30 °C, and 40 °C with a temperature sensor in the center of the chamber (inside). Observations were started as soon as the sample was installed. In a typical image obtained using the tumbled microscope, the diatom cells were found to move from the top to the bottom. In order to analyze floating velocity and trajectory, observation was continued for 35 min at room temperature, 30 °C, and 40 °C. Tracking analysis was carried out using the two-dimensional motion image measurement software Move-tr/2D. The average speed of 100 cells was 7.0 ± 4.3 µm/s at room temperature, 85.6 ± 31.9 µm/s at 30 °C and 470.1 ± 279.8 µm/s at 40 °C. In this study, we devised the unique observation to visualize the temperature dependence of diatom cells.

Fabrication of Microscope Stage for Vertical Observation with Temperature Control Function.

Samples are usually placed onto a horizontal microscope stage for microscopic observation. However, to observe the influence of gravity on a sample or study afloat behavior, it is necessary to make the microscope stage vertical. To accomplish this, a sideways inverted microscope tilted by 90° has been devised. To observe samples with this microscope, sample containers such as Petri dishes or glass slides must be secured to the stage vertically. A device that can secure sample containers in place on a vertical microscope stage has been developed and is described here. Attachment of this device to the stage allows observation of sample dynamics in the vertical plane. The ability to regulate temperature using a silicone rubber heater also permits observation of temperature-dependent sample behaviors. Furthermore, the temperature data is transferred to an internet server. Temperature settings and log monitoring can be controlled remotely from a PC or smartphone.

Synthesis, photodynamic activities, and cytotoxicity of new water-soluble cationic gallium(III) and zinc(II) phthalocyanines.

The cationic Ga(III) and Zn(II) phthalocyanines carrying N-methyl-pyridinium groups at eight peripheral ß-positionshave been synthesized. These complexes are highly soluble in dimethyl sulfoxide (DMSO) and moderately soluble in water and phosphate buffered saline (PBS); both Ga(III)Cl and Zn(II) complexes have shown no aggregation in water up to 1.2 × 10-4 and 1.5 × 10-5 M, respectively. A higher water-solubility of Ga(III)Cl complex as compared to Zn(II) complex is ascribed to the presence of an axially coordinated chloride. The spectroscopic properties, photogeneration of singlet oxygen (1O2), and cytotoxicity of these complexes have been investigated. The absolute quantum yields (ΦΔabsolute) for the photogeneration of singlet oxygen using Ga(III)Cl and Zn(II) complexes have been determined to be 4.4 and 5.3%, respectively, in DMSO solution. The cytotoxicity and intracellular sites of localization of Ga(III)Cl and Zn(II) complexes have been evaluated in human HEp2 cells. Both complexes, localized intracellularly in multiple organelles, have shown no cytotoxicity in the dark. Upon exposure to a low light dose (1.5 J/cm2), however, Zn(II) complex has exhibited a high photocytotoxicity. The result suggests that Zn(II) complex can be considered as a potential photosensitizer for Photodynamic therapy (PDT).